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Sample records for oligotrophic alpha-proteobacterium caulobacter

  1. A modular BAM complex in the outer membrane of the alpha-proteobacterium Caulobacter crescentus.

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    Khatira Anwari

    Full Text Available Mitochondria are organelles derived from an intracellular alpha-proteobacterium. The biogenesis of mitochondria relies on the assembly of beta-barrel proteins into the mitochondrial outer membrane, a process inherited from the bacterial ancestor. Caulobacter crescentus is an alpha-proteobacterium, and the BAM (beta-barrel assembly machinery complex was purified and characterized from this model organism. Like the mitochondrial sorting and assembly machinery complex, we find the BAM complex to be modular in nature. A approximately 150 kDa core BAM complex containing BamA, BamB, BamD, and BamE associates with additional modules in the outer membrane. One of these modules, Pal, is a lipoprotein that provides a means for anchorage to the peptidoglycan layer of the cell wall. We suggest the modular design of the BAM complex facilitates access to substrates from the protein translocase in the inner membrane.

  2. Genome analysis of DNA repair genes in the alpha proteobacterium Caulobacter crescentus

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    Menck Carlos FM

    2007-03-01

    Full Text Available Abstract Background The integrity of DNA molecules is fundamental for maintaining life. The DNA repair proteins protect organisms against genetic damage, by removal of DNA lesions or helping to tolerate them. DNA repair genes are best known from the gamma-proteobacterium Escherichia coli, which is the most understood bacterial model. However, genome sequencing raises questions regarding uniformity and ubiquity of these DNA repair genes and pathways, reinforcing the need for identifying genes and proteins, which may respond to DNA damage in other bacteria. Results In this study, we employed a bioinformatic approach, to analyse and describe the open reading frames potentially related to DNA repair from the genome of the alpha-proteobacterium Caulobacter crescentus. This was performed by comparison with known DNA repair related genes found in public databases. As expected, although C. crescentus and E. coli bacteria belong to separate phylogenetic groups, many of their DNA repair genes are very similar. However, some important DNA repair genes are absent in the C. crescentus genome and other interesting functionally related gene duplications are present, which do not occur in E. coli. These include DNA ligases, exonuclease III (xthA, endonuclease III (nth, O6-methylguanine-DNA methyltransferase (ada gene, photolyase-like genes, and uracil-DNA-glycosylases. On the other hand, the genes imuA and imuB, which are involved in DNA damage induced mutagenesis, have recently been described in C. crescentus, but are absent in E. coli. Particularly interesting are the potential atypical phylogeny of one of the photolyase genes in alpha-proteobacteria, indicating an origin by horizontal transfer, and the duplication of the Ada orthologs, which have diverse structural configurations, including one that is still unique for C. crescentus. Conclusion The absence and the presence of certain genes are discussed and predictions are made considering the particular

  3. The complex logic of stringent response regulation in Caulobacter crescentus: starvation signalling in an oligotrophic environment.

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    Boutte, Cara C; Crosson, Sean

    2011-05-01

    Bacteria rapidly adapt to nutritional changes via the stringent response, which entails starvation-induced synthesis of the small molecule, ppGpp, by RelA/SpoT homologue (Rsh) enzymes. Binding of ppGpp to RNA polymerase modulates the transcription of hundreds of genes and remodels the physiology of the cell. Studies of the stringent response have primarily focused on copiotrophic bacteria such as Escherichia coli; little is known about how stringent signalling is regulated in species that live in consistently nutrient-limited (i.e. oligotrophic) environments. Here we define the input logic and transcriptional output of the stringent response in the oligotroph, Caulobacter crescentus. The sole Rsh protein, SpoT(CC), binds to and is regulated by the ribosome, and exhibits AND-type control logic in which amino acid starvation is a necessary but insufficient signal for activation of ppGpp synthesis. While both glucose and ammonium starvation upregulate the synthesis of ppGpp, SpoT(CC) detects these starvation signals by two independent mechanisms. Although the logic of stringent response control in C. crescentus differs from E. coli, the global transcriptional effects of elevated ppGpp are similar, with the exception of 16S rRNA transcription, which is controlled independently of spoT(CC). This study highlights how the regulatory logic controlling the stringent response may be adapted to the nutritional niche of a bacterial species.

  4. A genome-wide survey of sRNAs in the symbiotic nitrogen-fixing alpha-proteobacterium Sinorhizobium meliloti

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    Jänicke Sebastian

    2010-04-01

    Full Text Available Abstract Background Small untranslated RNAs (sRNAs are widespread regulators of gene expression in bacteria. This study reports on a comprehensive screen for sRNAs in the symbiotic nitrogen-fixing alpha-proteobacterium Sinorhizobium meliloti applying deep sequencing of cDNAs and microarray hybridizations. Results A total of 1,125 sRNA candidates that were classified as trans-encoded sRNAs (173, cis-encoded antisense sRNAs (117, mRNA leader transcripts (379, and sense sRNAs overlapping coding regions (456 were identified in a size range of 50 to 348 nucleotides. Among these were transcripts corresponding to 82 previously reported sRNA candidates. Enrichment for RNAs with primary 5'-ends prior to sequencing of cDNAs suggested transcriptional start sites corresponding to 466 predicted sRNA regions. The consensus σ70 promoter motif CTTGAC-N17-CTATAT was found upstream of 101 sRNA candidates. Expression patterns derived from microarray hybridizations provided further information on conditions of expression of a number of sRNA candidates. Furthermore, GenBank, EMBL, DDBJ, PDB, and Rfam databases were searched for homologs of the sRNA candidates identified in this study. Searching Rfam family models with over 1,000 sRNA candidates, re-discovered only those sequences from S. meliloti already known and stored in Rfam, whereas BLAST searches suggested a number of homologs in related alpha-proteobacteria. Conclusions The screening data suggests that in S. meliloti about 3% of the genes encode trans-encoded sRNAs and about 2% antisense transcripts. Thus, this first comprehensive screen for sRNAs applying deep sequencing in an alpha-proteobacterium shows that sRNAs also occur in high number in this group of bacteria.

  5. Regulation of chromosomal replication in Caulobacter crescentus.

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    Collier, Justine

    2012-03-01

    The alpha-proteobacterium Caulobacter crescentus is characterized by its asymmetric cell division, which gives rise to a replicating stalked cell and a non-replicating swarmer cell. Thus, the initiation of chromosomal replication is tightly regulated, temporally and spatially, to ensure that it is coordinated with cell differentiation and cell cycle progression. Waves of DnaA and CtrA activities control when and where the initiation of DNA replication will take place in C. crescentus cells. The conserved DnaA protein initiates chromosomal replication by directly binding to sites within the chromosomal origin (Cori), ensuring that DNA replication starts once and only once per cell cycle. The CtrA response regulator represses the initiation of DNA replication in swarmer cells and in the swarmer compartment of pre-divisional cells, probably by competing with DnaA for binding to Cori. CtrA and DnaA are controlled by multiple redundant regulatory pathways that include DNA methylation-dependent transcriptional regulation, temporally regulated proteolysis and the targeting of regulators to specific locations within the cell. Besides being critical regulators of chromosomal replication, CtrA and DnaA are also master transcriptional regulators that control the expression of many genes, thus connecting DNA replication with other events of the C. crescentus cell cycle. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Replication intermediate analysis confirms that chromosomal replication origin initiates from an unusual intergenic region in Caulobacter crescentus.

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    Brassinga, A K; Marczynski, G T

    2001-11-01

    The alpha-proteobacterium Caulobacter crescentus possesses a developmental cell cycle that restricts chromosome replication to a stalked cell type. The proposed C.crescentus chromosome replication origin (Cori) lies between hemE and RP001, an unusual intergenic region not previously associated with bacterial replication origins, although a similar genomic arrangement is also present at the putative replication origin in the related bacterium Rickettsia prowazekii. The cloned Cori supports autonomous plasmid replication selectively in the stalked cell type implying that replication of the entire chromosome also initiates between hemE and RP001. To confirm this location, we applied the 2-D (N/N) agarose gel electrophoresis technique to resolve and identify chromosome replication intermediates throughout a 30 kb region spanning Cori. Replication initiation in Cori was uniquely characterized by an 'origin bubble and Y-arc' pattern and this observation was supported by simple replication fork 'Y-arc' patterns that characterized the regions flanking Cori. These replication forks originated bi-directionally from within Cori as determined by the fork direction assay. Therefore, chromosomal replication initiates from the unusual hemE/RP001 intergenic region that we propose represents a new class of replication origins.

  7. Caulobacters in the Marine Environment.

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    1987-01-01

    LECIIN> PEANUT DOLICHOS SOYBEAN CONCONAV- ULEX WHEAT GERM RICINUS STRAIN AGGLUTININ BIFLORUS AGGLUTININ ALINA EUROPEAUS AGGLUTININ COMMUNI ~i MCS 3 MCS3...sampling locations. Strains were distinguishable by a variety of morphological and biochemical criteria. All required at least some percentage of...waters near Santa Barbara, California. Caulobacters as well as the morphologically similar Hyphomonas were found in virtually all samples. This

  8. Uranium Biomineralization by Caulobacter crescentus

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    Jiao, Y.; Yung, M.; Park, D.

    2014-12-01

    It is well known that microorganisms are able to mediate removal of U(VI) from solution through reduction to insoluble U(IV) oxides under anaerobic conditions, but microbial transformation of U(VI) under aerobic conditions are less well understood. Here, we describe two processes of U(VI) transformation by the aerobic bacterium Caulobacter crescentus, known for its ubiquitous presence in aquatic systems and high U(VI) tolerance. U(VI) causes a temporary growth arrest in Caulobacter and growth recovery is not due to a decrease in U solubility, a common detoxification strategy employed by other microorganisms. Through functional reporter assays, we discovered that Caulobacter is able to reduce U(VI) bioavailability through a metabolism-dependent increase of medium pH, representing a novel U detoxification strategy. Upon recovery from growth arrest, Caulobacter proliferates with normal growth kinetics, accompanied by active U(VI) biomineralization. We found that phosphate metabolism is actively involved in the formation of U-P precipitates that are similar to autunite-group minerals. Comparisons of growth and U(VI) precipitation by wild type versus a phosphatase mutant indicates that extra-cytoplasmic phosphatase activity is not only responsible for the formation of cell-surface-bound U-P precipitates, but also plays an important role in cell survival under U stress. Our results highlight the importance of aerobic bacterial metabolism for U biogeochemistry.

  9. Novel peptidoglycans in Caulobacter and Asticcacaulis spp.

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    Poindexter, J S; Hagenzieker, J G

    1982-01-01

    Peptidoglycan sacculi free of poly-beta-hydroxybutyric acid were prepared from whole cells of four species of Caulobacter and two species of Asticcacaluis and from morphological mutants of Caulobacter crescentus and Caulobacter leidyi. Acid hydrolysates of the sacculi were analyzed quantitatively, and each of the hydrolysates was found to contain significant amounts of only five ninhydrin-reactive compounds: alanine, glutamic acid, alpha , omega-diaminopimelic acid, muramic acid, and glucosam...

  10. Global regulation of gene expression and cell differentiation in Caulobacter crescentus in response to nutrient availability.

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    England, Jennifer C; Perchuk, Barrett S; Laub, Michael T; Gober, James W

    2010-02-01

    In a developmental strategy designed to efficiently exploit and colonize sparse oligotrophic environments, Caulobacter crescentus cells divide asymmetrically, yielding a motile swarmer cell and a sessile stalked cell. After a relatively fixed time period under typical culture conditions, the swarmer cell differentiates into a replicative stalked cell. Since differentiation into the stalked cell type is irreversible, it is likely that environmental factors such as the availability of essential nutrients would influence the timing of the decision to abandon motility and adopt a sessile lifestyle. We measured two different parameters in nutrient-limited chemostat cultures, biomass concentration and the ratio of nonstalked to stalked cells, over a range of flow rates and found that nitrogen limitation significantly extended the swarmer cell life span. The transcriptional profiling experiments described here generate the first comprehensive picture of the global regulatory strategies used by an oligotroph when confronted with an environment where key macronutrients are sparse. The pattern of regulated gene expression in nitrogen- and carbon-limited cells shares some features in common with most copiotrophic organisms, but critical differences suggest that Caulobacter, and perhaps other oligotrophs, have evolved regulatory strategies to deal distinctly with their natural environments. We hypothesize that nitrogen limitation extends the swarmer cell lifetime by delaying the onset of a sequence of differentiation events, which when initiated by the correct combination of external environmental cues, sets the swarmer cell on a path to differentiate into a stalked cell within a fixed time period.

  11. Reduction of Cr(VI) and survival in Cr-contaminated sites by Caulobacter crescentus

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    Hu, P.; Chakraborty, R.; Brodie, E. L.; Andersen, G. L.; Hazen, T. C.

    2008-12-01

    The Caulobacter spp. is known to be able to live in low-nutrient environments, a characteristic of most heavy metal-contaminated sites. Recent studies have shown that Caulobacter crescentus can grow in chemically defined medium containing up to 1 mM uranium. Whole-genome transcriptional analysis and electron microscopic imaging of heavy metal stresses in Caulobacter crescentus also provided insight and evidence that the bacterium used an array of defensive mechanisms to deal with heavy metal stresses. In addition to up-regulated enzymes protecting against oxidative stress, DNA repair and down-regulated potential chromium transport, one of the major gene groups respond to chromium stress is "electron transport process and cytochrome oxidases", including cytochrome c oxidases, raising the possibility that the cells can employ the cytochromes to reduce chromium. Analysis of the microbial community at the chromium contaminated DOE site at Hanford, WA revealed the presence of Caulobacter spp. As an oligotroph, Caulobacter can play a significant role in chromium reduction in the environment where the nutrients are limited. This result was confirmed by both 16S rDNA based microarray (Phylochip) as well as by MDA-based clone library data. Based on these results we further investigated the capability of this organism to reduce Cr(VI) using the well known model strain Caulobacter crescentus CB15N. Preliminary cell suspension experiments were set up with glucose as the electron donor and Cr(VI) as the electron acceptor in phosphate based M2 salts buffer. After 22 hours almost 27% of Cr(VI) was reduced in the incubations containing active cells relative to the controls containing heat killed cells. Also, in another set of controls with no electron acceptor added, cells showed no increase in cell density during that time demonstrating that the reduction of Cr(VI) by cells of Caulobacter was due to biological activity. Future experiments will investigate the components

  12. Core-oscillator model of Caulobacter crescentus

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    Vandecan, Yves; Biondi, Emanuele; Blossey, Ralf

    2016-06-01

    The gram-negative bacterium Caulobacter crescentus is a powerful model organism for studies of bacterial cell cycle regulation. Although the major regulators and their connections in Caulobacter have been identified, it still is a challenge to properly understand the dynamics of its circuitry which accounts for both cell cycle progression and arrest. We show that the key decision module in Caulobacter is built from a limit cycle oscillator which controls the DNA replication program. The effect of an induced cell cycle arrest is demonstrated to be a key feature to classify the underlying dynamics.

  13. Extracellular gluco-oligosaccharide degradation by Caulobacter crescentus.

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    Presley, Gerald N; Payea, Matthew J; Hurst, Logan R; Egan, Annie E; Martin, Brandon S; Periyannan, Gopal R

    2014-03-01

    The oligotrophic bacterium Caulobacter crescentus has the ability to metabolize various organic molecules, including plant structural carbohydrates, as a carbon source. The nature of β-glucosidase (BGL)-mediated gluco-oligosaccharide degradation and nutrient transport across the outer membrane in C. crescentus was investigated. All gluco-oligosaccharides tested (up to celloheptose) supported growth in M2 minimal media but not cellulose or CM-cellulose. The periplasmic and outer membrane fractions showed highest BGL activity, but no significant BGL activity was observed in the cytosol or extracellular medium. Cells grown in cellobiose showed expression of specific BGLs and TonB-dependent receptors (TBDRs). Carbonyl cyanide 3-chlorophenylhydrazone lowered the rate of cell growth in cellobiose but not in glucose, indicating potential cellobiose transport into the cell by a proton motive force-dependent process, such as TBDR-dependent transport, and facilitated diffusion of glucose across the outer membrane via specific porins. These results suggest that C. crescentus acquires carbon from cellulose-derived gluco-oligosaccharides found in the environment by extracellular and periplasmic BGL activity and TBDR-mediated transport. This report on extracellular degradation of gluco-oligosaccharides and methods of nutrient acquisition by C. crescentus supports a broader suite of carbohydrate metabolic capabilities suggested by the C. crescentus genome sequence that until now have not been reported.

  14. Novel peptidoglycans in Caulobacter and Asticcacaulis spp.

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    Poindexter, J S; Hagenzieker, J G

    1982-04-01

    Peptidoglycan sacculi free of poly-beta-hydroxybutyric acid were prepared from whole cells of four species of Caulobacter and two species of Asticcacaluis and from morphological mutants of Caulobacter crescentus and Caulobacter leidyi. Acid hydrolysates of the sacculi were analyzed quantitatively, and each of the hydrolysates was found to contain significant amounts of only five ninhydrin-reactive compounds: alanine, glutamic acid, alpha , omega-diaminopimelic acid, muramic acid, and glucosamine. Four types of peptidoglycans were distinguishable on the basis of the molar ratios among these five compounds. The respective ratios were as follows: in C. leidyi, 2:1:1:1:0.8; in Asticcacaulis biprosthecum, 1.7:1.6:1.1:0.7; in the cells of the remaining species, 2:1:1:1.2:0.8; and in stalks shed by the abscission mutant 2NY66, 2:1:1:1:1.67. Thus, in addition to some species differences among these caulobacters, it was found that the peptidoglycan sacculus of the stalked C. crescentus cell is chemically differentiated; the cellular peptidoglycan is richer in muramic acid than is the peptidoglycan of typical gram-negative bacteria, and the peptidoglycan of the stalk is correspondingly rich in glucosamine. Empirical formulas for the repeating units of the peptidoglycans have been inferred on the basis of the molar ratios of their amino components.

  15. The phylogenetic relationships of Caulobacter, Asticcacaulis and Brevundimonas species and their taxonomic implications.

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    Sly, L I; Cox, T L; Beckenham, T B

    1999-04-01

    The phylogenetic relationships among the species of Caulobacter, Asticcacaulis and Brevundimonas were studied by comparison of their 16S rDNA sequences. The analysis of almost complete sequences confirmed the early evolutionary divergence of the freshwater and marine species of Caulobacter reported previously [Stahl, D. A., Key, R., Flesher, B. & Smit, J. (1992). J Bacteriol 174, 2193-2198]. The freshwater species formed two distinct clusters. One cluster contained the species Caulobacter bacteroides, Caulobacter crescentus, Caulobacter fusiformis and Caulobacter henricii. C. bacteroides and C. fusiformis are very closely related (sequence identity 99.8%). The second cluster was not exclusive and contained the specis Caulobacter intermedius, Caulobacter subvibrioides and Caulobacter variabilis, as well as Brevundimonas diminuta and Brevundimonas vesicularis. The marine species Caulobacter halobacteroides and Caulobacter maris were very closely related, with a sequence identity of 99.7%. These two species were most closely but distantly related to the marine hyphal/budding bacteria Hyphomonas jannaschiana and Hirschia baltica, which formed a deep phylogenetic line with Rhodobacter sphaeroides and Rhodobacter capsulatus. Caulobacter leidyia is unrelated to the other species of Caulobacter and belongs to the alpha-4 subclass of the Proteobacteria, forming a distinct cluster with Asticcacaulis excentricus and Asticcacaulis biprosthecium. The taxonomic implications of the polyphyletic nature of the genus Caulobacter and the absence of a type culture for the type species of the genus Caulobacter vibrioides, are discussed.

  16. Metabolism of aromatic compounds by Caulobacter crescentus

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    Chatterjee, D.K.; Bourquin, A.W.

    1987-05-01

    Cultures of Caulobacter crescentus were found to grow on a variety of aromatic compounds. Degradation of benzoate, p-hydroxybenzoate, and phenol was found to occur via ..beta..-ketoadipate. The induction of degradative enzymes such as benzoate 1,2-dioxygenase, the ring cleavage enzyme catechol 1,2-dioxygenase, and cis,cis-muconate lactonizing enzyme appeared similar to the control mechanism present in Pseudomonas spp. Both benzoate 1,2-dioxygenase and catechol 1,2-dioxygenase had stringent specificities, as revealed by their action toward substituted benzoates and substituted catechols, respectively.

  17. Prosthecobacter fusiformis nov. gen. et sp., the fusiform caulobacter.

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    Staley, J T; Bont, J A; Jonge, K

    1976-01-01

    Four strains of heterotrophic, fusiform caulobacters have been isolated from freshwater sources. A single prostheca extends from one pole of mature cells, and cells attach to various substrata by means of a holdfast located at the distal tip of the appendage. Thus, superficially these bacteria bear a strong resemblance to bacteria in the genus Caulobacter. However, unlike Caulobacter these bacteria do not exhibit a dimorphic life cycle of motile, non-stalked daughter cells and immotile, stalked mother cells. Instead both mother and daughter cells are immotile, and at the time of cell separation the daughter cells are essentially identical mirror-image replicas of the mother cell. In addition, the prosthecae of these fusiform caulobacters do not have crossbands, they are somewhat wider than the stalks of Caulobacter and the pseudostalks of Asticcacaulis, and they terminate in a bulbous tip. The deoxyribonucleic acid (DNA) base composition ranges from 54.6-60.1, well below the 62-67 range for the genus Caulobacter. Based upon these and other differences a new genus and species, Prosthecobacter fusiformis, is proposed for the fusiform caulobacters.

  18. Caulobacter and Asticcacaulis stalk bands as indicators of stalk age.

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    Poindexter, J S; Staley, J T

    1996-01-01

    The prosthecae (stalks) of dimorphic caulobacters of the genera Caulobacter and Asticcacaulis are distinguished among such appendages by the presence of disk-like components known as stalk bands. Whether bands are added to a cell's stalk(s) as a regular event coordinated with the cell's reproductive cycle has not been settled by previous studies. Analysis of the frequency of stalks with i, i + 1, i + 2, etc. bands 'among more than 7,000 stalks of Caulobacter crescentus revealed that in finite...

  19. Growth Conditions Regulate the Requirements for Caulobacter Chromosome Segregation

    DEFF Research Database (Denmark)

    Shebelut, Conrad W.; Jensen, Rasmus Bugge; Gitai, Zemer

    2009-01-01

    Growth environments are important metabolic and developmental regulators. Here we demonstrate a growth environment-dependent effect on Caulobacter chromosome segregation of a small-molecule inhibitor of the MreB bacterial actin cytoskeleton. Our results also implicate ParAB as important segregation...... determinants, suggesting that multiple distinct mechanisms can mediate Caulobacter chromosome segregation and that their relative contributions can be environmentally regulated....

  20. Growth Conditions Regulate the Requirements for Caulobacter Chromosome Segregation▿ †

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    Shebelut, Conrad W.; Jensen, Rasmus B.; Gitai, Zemer

    2008-01-01

    Growth environments are important metabolic and developmental regulators. Here we demonstrate a growth environment-dependent effect on Caulobacter chromosome segregation of a small-molecule inhibitor of the MreB bacterial actin cytoskeleton. Our results also implicate ParAB as important segregation determinants, suggesting that multiple distinct mechanisms can mediate Caulobacter chromosome segregation and that their relative contributions can be environmentally regulated.

  1. Growth conditions regulate the requirements for Caulobacter chromosome segregation.

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    Shebelut, Conrad W; Jensen, Rasmus B; Gitai, Zemer

    2009-02-01

    Growth environments are important metabolic and developmental regulators. Here we demonstrate a growth environment-dependent effect on Caulobacter chromosome segregation of a small-molecule inhibitor of the MreB bacterial actin cytoskeleton. Our results also implicate ParAB as important segregation determinants, suggesting that multiple distinct mechanisms can mediate Caulobacter chromosome segregation and that their relative contributions can be environmentally regulated.

  2. Control of chromosome replication in caulobacter crescentus.

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    Marczynski, Gregory T; Shapiro, Lucy

    2002-01-01

    Caulobacter crescentus permits detailed analysis of chromosome replication control during a developmental cell cycle. Its chromosome replication origin (Cori) may be prototypical of the large and diverse class of alpha-proteobacteria. Cori has features that both affiliate and distinguish it from the Escherichia coli chromosome replication origin. For example, requirements for DnaA protein and RNA transcription affiliate both origins. However, Cori is distinguished by several features, and especially by five binding sites for the CtrA response regulator protein. To selectively repress and limit chromosome replication, CtrA receives both protein degradation and protein phosphorylation signals. The signal mediators, proteases, response regulators, and kinases, as well as Cori DNA and the replisome, all show distinct patterns of temporal and spatial organization during cell cycle progression. Future studies should integrate our knowledge of biochemical activities at Cori with our emerging understanding of cytological dynamics in C. crescentus and other bacteria.

  3. Constriction and septation during cell division in caulobacters.

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    Poindexter, J S; Hagenzieker, J G

    1981-07-01

    Morphogenesis of the division site in caulobacters had been described as constrictive in Caulobacter spp. and septate in Asticcacaulis excentricus. However, subsequent studies of other gram-negative genera had implied that constrictive division was an artefact resulting from inadequate preservation of septa; exploration of alternatives to osmium fixation, particularly with aldehydes, was recommended. In this study, the appearance of sectioned division sites was reinvestigated in caulobacter cells prepared by 20 different procedures varying with respect to fixation agents, media, schedules, and temperatures, to dehydrating agents, and to embedding resins. Three types of division site morphogenesis were observed: constriction in C. bacteroides and C. crescentus, partial septation in C. leidyi, and complete, undivided septation in A. excentricus and A. biprosthecum. The anatomy of the division site depended on the bacterial strain, not on the method of preparation of the cells for sectioning. These studies confirm the earlier observations on osmium-fixed caulobacter cells and lead to the general conclusion that gram-negative bacteria with tapered poles probably divide by constriction, whereas septation results in blunt cell poles. A pattern of spiral, rather than circular, insertion of new envelope subunits at the cell equator is proposed as a basic developmental difference between constrictive and septate fission in gram-negative bacteria. Since caulobacter prosthecae can develop as extensions of tapered poles formed by constriction, whereas subpolar or lateral prosthecae occur in species with blunt poles resulting from septation, the site of formation of a thick septum appears unsuitable as a site of subsequent envelope outgrowth.

  4. Screen for localized proteins in Caulobacter crescentus.

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    Jay H Russell

    Full Text Available Precise localization of individual proteins is required for processes such as motility, chemotaxis, cell-cycle progression, and cell division in bacteria, but the number of proteins that are localized in bacterial species is not known. A screen based on transposon mutagenesis and fluorescence activated cell sorting was devised to identify large numbers of localized proteins, and employed in Caulobacter crescentus. From a sample of the clones isolated in the screen, eleven proteins with no previously characterized localization in C. crescentus were identified, including six hypothetical proteins. The localized hypothetical proteins included one protein that was localized in a helix-like structure, and two proteins for which the localization changed as a function of the cell cycle, suggesting that complex three-dimensional patterns and cell cycle-dependent localization are likely to be common in bacteria. Other mutants produced localized fusion proteins even though the transposon has inserted near the 5' end of a gene, demonstrating that short peptides can contain sufficient information to localize bacterial proteins. The screen described here could be used in most bacterial species.

  5. Getting in the loop: regulation of development in Caulobacter crescentus.

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    Curtis, Patrick D; Brun, Yves V

    2010-03-01

    Caulobacter crescentus is an aquatic Gram-negative alphaproteobacterium that undergoes multiple changes in cell shape, organelle production, subcellular distribution of proteins, and intracellular signaling throughout its life cycle. Over 40 years of research has been dedicated to this organism and its developmental life cycles. Here we review a portion of many developmental processes, with particular emphasis on how multiple processes are integrated and coordinated both spatially and temporally. While much has been discovered about Caulobacter crescentus development, areas of potential future research are also highlighted.

  6. CauloBrowser: A systems biology resource for Caulobacter crescentus.

    Science.gov (United States)

    Lasker, Keren; Schrader, Jared M; Men, Yifei; Marshik, Tyler; Dill, David L; McAdams, Harley H; Shapiro, Lucy

    2016-01-04

    Caulobacter crescentus is a premier model organism for studying the molecular basis of cellular asymmetry. The Caulobacter community has generated a wealth of high-throughput spatiotemporal databases including data from gene expression profiling experiments (microarrays, RNA-seq, ChIP-seq, ribosome profiling, LC-ms proteomics), gene essentiality studies (Tn-seq), genome wide protein localization studies, and global chromosome methylation analyses (SMRT sequencing). A major challenge involves the integration of these diverse data sets into one comprehensive community resource. To address this need, we have generated CauloBrowser (www.caulobrowser.org), an online resource for Caulobacter studies. This site provides a user-friendly interface for quickly searching genes of interest and downloading genome-wide results. Search results about individual genes are displayed as tables, graphs of time resolved expression profiles, and schematics of protein localization throughout the cell cycle. In addition, the site provides a genome viewer that enables customizable visualization of all published high-throughput genomic data. The depth and diversity of data sets collected by the Caulobacter community makes CauloBrowser a unique and valuable systems biology resource.

  7. Caulobacter and Asticcacaulis stalk bands as indicators of stalk age.

    Science.gov (United States)

    Poindexter, J S; Staley, J T

    1996-07-01

    The prosthecae (stalks) of dimorphic caulobacters of the genera Caulobacter and Asticcacaulis are distinguished among such appendages by the presence of disk-like components known as stalk bands. Whether bands are added to a cell's stalk(s) as a regular event coordinated with the cell's reproductive cycle has not been settled by previous studies. Analysis of the frequency of stalks with i, i + 1, i + 2, etc. bands 'among more than 7,000 stalks of Caulobacter crescentus revealed that in finite (batch) cultures (in which all offspring accumulate), the proportion of stalks with i + 1 hands was regularly 50% of the proportion of stalks with i bands. This implied that the number of bands correlated with the number of reproductive cycles completed by a stalked cell. In chemostat-maintained perpetual cultures, the proportion was greater than 50% because stalked cells, with their shorter reproductive cycle times, contributed a larger proportion of offspring to the steady-state population than did their swarmer siblings. In Asticcacaulis biprosthecum cells, which bear twin prosthecae, the twins on a typical cell possessed the same number of bands. For both genera, stalk bands provide a unique morphological feature that could be employed in an assessment of age distribution and reproductive dynamics within natural populations of these caulobacters.

  8. CauloBrowser: A systems biology resource for Caulobacter crescentus

    Science.gov (United States)

    Lasker, Keren; Schrader, Jared M.; Men, Yifei; Marshik, Tyler; Dill, David L.; McAdams, Harley H.; Shapiro, Lucy

    2016-01-01

    Caulobacter crescentus is a premier model organism for studying the molecular basis of cellular asymmetry. The Caulobacter community has generated a wealth of high-throughput spatiotemporal databases including data from gene expression profiling experiments (microarrays, RNA-seq, ChIP-seq, ribosome profiling, LC-ms proteomics), gene essentiality studies (Tn-seq), genome wide protein localization studies, and global chromosome methylation analyses (SMRT sequencing). A major challenge involves the integration of these diverse data sets into one comprehensive community resource. To address this need, we have generated CauloBrowser (www.caulobrowser.org), an online resource for Caulobacter studies. This site provides a user-friendly interface for quickly searching genes of interest and downloading genome-wide results. Search results about individual genes are displayed as tables, graphs of time resolved expression profiles, and schematics of protein localization throughout the cell cycle. In addition, the site provides a genome viewer that enables customizable visualization of all published high-throughput genomic data. The depth and diversity of data sets collected by the Caulobacter community makes CauloBrowser a unique and valuable systems biology resource. PMID:26476443

  9. Phosphate starvation triggers production and secretion of an extracellular lipoprotein in Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Sophie Le Blastier

    Full Text Available Life in oligotrophic environments necessitates quick adaptive responses to a sudden lack of nutrients. Secretion of specific degradative enzymes into the extracellular medium is a means to mobilize the required nutrient from nearby sources. The aquatic bacterium Caulobacter crescentus must often face changes in its environment such as phosphate limitation. Evidence reported in this paper indicates that under phosphate starvation, C. crescentus produces a membrane surface-anchored lipoprotein named ElpS subsequently released into the extracellular medium. A complete set of 12 genes encoding a type II secretion system (T2SS is located adjacent to the elpS locus in the C. crescentus genome. Deletion of this T2SS impairs release of ElpS in the environment, which surprisingly remains present at the cell surface, indicating that the T2SS is not involved in the translocation of ElpS to the outer membrane but rather in its release. Accordingly, treatment with protease inhibitors prevents release of ElpS in the extracellular medium suggesting that ElpS secretion relies on a T2SS-secreted protease. Finally, secretion of ElpS is associated with an increase in alkaline phosphatase activity in culture supernatants, suggesting a role of the secreted protein in inorganic phosphate mobilization. In conclusion, we have shown that upon phosphate starvation, C. crescentus produces an outer membrane bound lipoprotein, ElpS, which is further cleaved and released in the extracellular medium in a T2SS-dependent manner. Our data suggest that ElpS is associated with an alkaline phosphatase activity, thereby allowing the bacterium to gather inorganic phosphates from a poor environment.

  10. Effects of (p)ppGpp on the progression of the cell cycle of Caulobacter crescentus.

    Science.gov (United States)

    Gonzalez, Diego; Collier, Justine

    2014-07-01

    Bacteria must control the progression of their cell cycle in response to nutrient availability. This regulation can be mediated by guanosine tetra- or pentaphosphate [(p)ppGpp], which are synthesized by enzymes of the RelA/SpoT homologue (Rsh) family, particularly under starvation conditions. Here, we study the effects of (p)ppGpp on the cell cycle of Caulobacter crescentus, an oligotrophic bacterium with a dimorphic life cycle. C. crescentus divides asymmetrically, producing a motile swarmer cell that cannot replicate its chromosome and a sessile stalked cell that is replication competent. The swarmer cell rapidly differentiates into a stalked cell in appropriate conditions. An artificial increase in the levels of (p)ppGpp in nonstarved C. crescentus cells was achieved by expressing a truncated relA gene from Escherichia coli, encoding a constitutively active (p)ppGpp synthetase. By combining single-cell microscopy, flow cytometry approaches, and swarming assays, we show that an increase in the intracellular concentration of (p)ppGpp is sufficient to slow down the swarmer-to-stalked cell differentiation process and to delay the initiation of chromosome replication. We also present evidence that the intracellular levels of two master regulators of the cell cycle of C. crescentus, DnaA and CtrA, are modulated in response to (p)ppGpp accumulation, even in the absence of actual starvation. CtrA proteolysis and DnaA synthesis seem indirectly inhibited by (p)ppGpp accumulation. By extending the life span of the motile nonreproductive swarmer cell and thus promoting dispersal and foraging functions over multiplication under starvation conditions, (p)ppGpp may play a central role in the ecological adaptation of C. crescentus to nutritional stresses.

  11. Analysis of the terminus region of the Caulobacter crescentus chromosome and identification of the dif site

    DEFF Research Database (Denmark)

    Jensen, Rasmus Bugge

    2006-01-01

    The terminus region of the Caulobacter crescentus chromosome and the dif chromosome dimer resolution site were characterized. The Caulobacter genome contains skewed sequences that abruptly switch strands at dif and may have roles in chromosome maintenance and segregation. Absence of dif or the Xer...

  12. Marine Caulobacters. Isolation, Characterization and Assessing the Potential for Genetic Experimentation.

    Science.gov (United States)

    1987-01-01

    drain runoff and sites well removed from commercial development. Other samples were taken at sites along the California coast, ranging from Bodega Bay...h? lrolns to enable transfer of mob+ plasm ids. II Table 2. SALT REQUIREMENTS FOR MARINE CAULOBACTERS Marine Caulobacter Growth in i/OX Growth in

  13. Computational Model of the Division Cycle of Caulobacter crescentus

    Science.gov (United States)

    Brazhnik, Paul; Li, Shenghua; Sobral, Bruno; Tyson, John J.

    2007-11-01

    Based on published experimental evidence, we propose a molecular mechanism and a quantitative computational model for cell cycle control in Caulobacter crescentus. Our model predicts the detailed temporal dynamics of regulatory gene expression during the cell cycle and differentiation process of wild-type cells as well as several mutant strains. Since many of the proteins involved in regulating the cell cycle of C.crescentus are conserved among other genera of α-proteobacteria, the proposed mechanism may be applicable to these species.

  14. The xylan utilization system of the plant pathogen Xanthomonas campestris pv campestris controls epiphytic life and reveals common features with oligotrophic bacteria and animal gut symbionts.

    Science.gov (United States)

    Déjean, Guillaume; Blanvillain-Baufumé, Servane; Boulanger, Alice; Darrasse, Armelle; Dugé de Bernonville, Thomas; Girard, Anne-Laure; Carrére, Sébastien; Jamet, Stevie; Zischek, Claudine; Lautier, Martine; Solé, Magali; Büttner, Daniela; Jacques, Marie-Agnès; Lauber, Emmanuelle; Arlat, Matthieu

    2013-05-01

    Xylan is a major structural component of plant cell wall and the second most abundant plant polysaccharide in nature. Here, by combining genomic and functional analyses, we provide a comprehensive picture of xylan utilization by Xanthomonas campestris pv campestris (Xcc) and highlight its role in the adaptation of this epiphytic phytopathogen to the phyllosphere. The xylanolytic activity of Xcc depends on xylan-deconstruction enzymes but also on transporters, including two TonB-dependent outer membrane transporters (TBDTs) which belong to operons necessary for efficient growth in the presence of xylo-oligosaccharides and for optimal survival on plant leaves. Genes of this xylan utilization system are specifically induced by xylo-oligosaccharides and repressed by a LacI-family regulator named XylR. Part of the xylanolytic machinery of Xcc, including TBDT genes, displays a high degree of conservation with the xylose-regulon of the oligotrophic aquatic bacterium Caulobacter crescentus. Moreover, it shares common features, including the presence of TBDTs, with the xylan utilization systems of Bacteroides ovatus and Prevotella bryantii, two gut symbionts. These similarities and our results support an important role for TBDTs and xylan utilization systems for bacterial adaptation in the phyllosphere, oligotrophic environments and animal guts.

  15. OmpW of Caulobacter crescentus Functions as an Outer Membrane Channel for Cations.

    Science.gov (United States)

    Benz, Roland; Jones, Michael D; Younas, Farhan; Maier, Elke; Modi, Niraj; Mentele, Reinhard; Lottspeich, Friedrich; Kleinekathöfer, Ulrich; Smit, John

    2015-01-01

    Caulobacter crescentus is an oligotrophic bacterium that lives in dilute organic environments such as soil and freshwater. This bacterium represents an interesting model for cellular differentiation and regulation because daughter cells after division have different forms: one is motile while the other is non-motile and can adhere to surfaces. Interestingly, the known genome of C. crescentus does not contain genes predicted to code for outer membrane porins of the OmpF/C general diffusion type present in enteric bacteria or those coding for specific porins selective for classes of substrates. Instead, genes coding for 67 TonB-dependent outer membrane receptors have been identified, suggesting that active transport of specific nutrients may be the norm. Here, we report that high channel-forming activity was observed with crude outer membrane extracts of C. crescentus in lipid bilayer experiments, indicating that the outer membrane of C. crescentus contained an ion-permeable channel with a single-channel conductance of about 120 pS in 1M KCl. The channel-forming protein with an apparent molecular mass of about 20 kDa was purified to homogeneity. Partial protein sequencing of the protein indicated it was a member of the OmpW family of outer membrane proteins from Gram-negative bacteria. This channel was not observed in reconstitution experiments with crude outer membrane extracts of an OmpW deficient C. crescentus mutant. Biophysical analysis of the C. crescentus OmpW suggested that it has features that are special for general diffusion porins of Gram-negative outer membranes because it was not a wide aqueous channel. Furthermore, OmpW of C. crescentus seems to be different to known OmpW porins and has a preference for ions, in particular cations. A putative model for OmpW of C. crescentus was built on the basis of the known 3D-structures of OmpW of Escherichia coli and OprG of Pseudomonas aeruginosa using homology modeling. A comparison of the two known structures

  16. OmpW of Caulobacter crescentus Functions as an Outer Membrane Channel for Cations.

    Directory of Open Access Journals (Sweden)

    Roland Benz

    Full Text Available Caulobacter crescentus is an oligotrophic bacterium that lives in dilute organic environments such as soil and freshwater. This bacterium represents an interesting model for cellular differentiation and regulation because daughter cells after division have different forms: one is motile while the other is non-motile and can adhere to surfaces. Interestingly, the known genome of C. crescentus does not contain genes predicted to code for outer membrane porins of the OmpF/C general diffusion type present in enteric bacteria or those coding for specific porins selective for classes of substrates. Instead, genes coding for 67 TonB-dependent outer membrane receptors have been identified, suggesting that active transport of specific nutrients may be the norm. Here, we report that high channel-forming activity was observed with crude outer membrane extracts of C. crescentus in lipid bilayer experiments, indicating that the outer membrane of C. crescentus contained an ion-permeable channel with a single-channel conductance of about 120 pS in 1M KCl. The channel-forming protein with an apparent molecular mass of about 20 kDa was purified to homogeneity. Partial protein sequencing of the protein indicated it was a member of the OmpW family of outer membrane proteins from Gram-negative bacteria. This channel was not observed in reconstitution experiments with crude outer membrane extracts of an OmpW deficient C. crescentus mutant. Biophysical analysis of the C. crescentus OmpW suggested that it has features that are special for general diffusion porins of Gram-negative outer membranes because it was not a wide aqueous channel. Furthermore, OmpW of C. crescentus seems to be different to known OmpW porins and has a preference for ions, in particular cations. A putative model for OmpW of C. crescentus was built on the basis of the known 3D-structures of OmpW of Escherichia coli and OprG of Pseudomonas aeruginosa using homology modeling. A comparison of the two

  17. Recombination within and between species of the alpha proteobacterium Bartonella infecting rodents.

    Science.gov (United States)

    Paziewska, Anna; Harris, Philip D; Zwolińska, Lucyna; Bajer, Anna; Siński, Edward

    2011-01-01

    Bartonella infections from wild mice and voles (Apodemus flavicollis, Mi. oeconomus, Microtus arvalis and Myodes glareolus) were sampled from a forest and old-field habitats of eastern Poland; a complex network of Bartonella isolates, referrable to B. taylorii, B. grahamii, B. birtlesii and B. doshiae, was identified by the sequencing of a gltA fragment, comparable to previous studies of Bartonella diversity in rodents. Nested clade analysis showed that isolates could be assigned to zero- and one-step clades which correlated with host identity and were probably the result of clonal expansion; however, sequencing of other housekeeping genes (rpoB, ribC, ftsZ, groEl) and the 16S RNA gene revealed a more complex situation with clear evidence of numerous recombinant events in which one or both Bartonella parents could be identified. Recombination within gltA was found to have generated two distinct variant clades, one a hybrid between B. taylorii and B. doshiae, the other between B. taylorii and B. grahamii. These recombinant events characterised the differences between the two-step and higher clades within the total nested cladogram, involved all four species of Bartonella identified in this work and appear to have played a dominant role in the evolution of Bartonella diversity. It is clear, therefore, that housekeeping gene phylogenies are not robust indicators of Bartonella diversity, especially when only a single gene (gltA or 16S RNA) is used. Bartonella clades infecting Microtus were most frequently involved in recombination and were most frequently tip clades within the cladogram. The role of Microtus in influencing the frequency of Bartonella recombination remains unknown.

  18. Compaction and transport properties of newly replicated Caulobacter crescentus DNA.

    Science.gov (United States)

    Hong, Sun-Hae; McAdams, Harley H

    2011-12-01

    Upon initiating replication of the Caulobacter chromosome, one copy of the parS centromere remains at the stalked pole; the other moves to the distal pole. We identified the segregation dynamics and compaction characteristics of newly replicated Caulobacter DNA during transport (highly variable from cell to cell) using time-lapse fluorescence microscopy. The parS centromere and a length (also highly variable) of parS proximal DNA on each arm of the chromosome are segregated with the same relatively slow transport pattern as the parS locus. Newly replicated DNA further than about 100 kb from parS segregates with a different and faster pattern, while loci at 48 kb from parS segregate with the slow pattern in some cells and the fast pattern in others. The observed parS-proximal DNA compaction characteristics have scaling properties that suggest the DNA is branched. HU2-deletion strains exhibited a reduced compaction phenotype except near the parS site where only the ΔHU1ΔHU2 double mutant had a compaction phenotype. The chromosome shows speed-dependent extension during translocation suggesting the DNA polymer is under tension. While DNA segregation is highly reliable and succeeds in virtually all wild-type cells, the high degree of cell to cell variation in the segregation process is noteworthy.

  19. Caulobacter chromosome segregation is an ordered multistep process.

    Science.gov (United States)

    Shebelut, Conrad W; Guberman, Jonathan M; van Teeffelen, Sven; Yakhnina, Anastasiya A; Gitai, Zemer

    2010-08-10

    Despite its fundamental nature, bacterial chromosome segregation remains poorly understood. Viewing segregation as a single process caused multiple proposed mechanisms to appear in conflict and failed to explain how asymmetrically dividing bacteria break symmetry to move only one of their chromosomes. Here, we demonstrate that the ParA ATPase extends from one cell pole and pulls the chromosome by retracting upon association with the ParB DNA-binding protein. Surprisingly, ParA disruption has a specific effect on chromosome segregation that only perturbs the latter stages of this process. Using quantitative high-resolution imaging, we demonstrate that this specificity results from the multistep nature of chromosome translocation. We propose that Caulobacter chromosome segregation follows an ordered pathway of events with distinct functions and mechanisms. Initiation releases polar tethering of the origin of replication, distinction spatially differentiates the two chromosomes, and commitment irreversibly translocates the distal centromeric locus. Thus, much as eukaryotic mitosis involves a sequence of distinct subprocesses, Caulobacter cells also segregate their chromosomes through an orchestrated series of steps. We discuss how the multistep view of bacterial chromosome segregation can help to explain and reconcile outstanding puzzles and frame future investigation.

  20. RK2 plasmid dynamics in Caulobacter crescentus cells--two modes of DNA replication initiation.

    Science.gov (United States)

    Wegrzyn, Katarzyna; Witosinska, Monika; Schweiger, Pawel; Bury, Katarzyna; Jenal, Urs; Konieczny, Igor

    2013-06-01

    Undisturbed plasmid dynamics is required for the stable maintenance of plasmid DNA in bacterial cells. In this work, we analysed subcellular localization, DNA synthesis and nucleoprotein complex formation of plasmid RK2 during the cell cycle of Caulobacter crescentus. Our microscopic observations showed asymmetrical distribution of plasmid RK2 foci between the two compartments of Caulobacter predivisional cells, resulting in asymmetrical allocation of plasmids to progeny cells. Moreover, using a quantitative PCR (qPCR) method, we estimated that multiple plasmid particles form a single fluorescent focus and that the number of plasmids per focus is approximately equal in both swarmer and predivisional Caulobacter cells. Analysis of the dynamics of TrfA-oriV complex formation during the Caulobacter cell cycle revealed that TrfA binds oriV primarily during the G1 phase, however, plasmid DNA synthesis occurs during the S and G2 phases of the Caulobacter cell cycle. Both in vitro and in vivo analysis of RK2 replication initiation in C. crescentus cells demonstrated that it is independent of the Caulobacter DnaA protein in the presence of the longer version of TrfA protein, TrfA-44. However, in vivo stability tests of plasmid RK2 derivatives suggested that a DnaA-dependent mode of plasmid replication initiation is also possible.

  1. Modulation of medium pH by Caulobacter crescentus facilitates recovery from uranium-induced growth arrest.

    Science.gov (United States)

    Park, Dan M; Jiao, Yongqin

    2014-09-01

    The oxidized form of uranium [U(VI)] predominates in oxic environments and poses a major threat to ecosystems. Due to its ability to mineralize U(VI), the oligotroph Caulobacter crescentus is an attractive candidate for U(VI) bioremediation. However, the physiological basis for U(VI) tolerance is unclear. Here we demonstrated that U(VI) caused a temporary growth arrest in C. crescentus and three other bacterial species, although the duration of growth arrest was significantly shorter for C. crescentus. During the majority of the growth arrest period, cell morphology was unaltered and DNA replication initiation was inhibited. However, during the transition from growth arrest to exponential phase, cells with shorter stalks were observed, suggesting a decoupling between stalk development and the cell cycle. Upon recovery from growth arrest, C. crescentus proliferated with a growth rate comparable to that of a control without U(VI), although a fraction of these cells appeared filamentous with multiple replication start sites. Normal cell morphology was restored by the end of exponential phase. Cells did not accumulate U(VI) resistance mutations during the prolonged growth arrest, but rather, a reduction in U(VI) toxicity occurred concomitantly with an increase in medium pH. Together, these data suggest that C. crescentus recovers from U(VI)-induced growth arrest by reducing U(VI) toxicity through pH modulation. Our finding represents a unique U(VI) detoxification strategy and provides insight into how microbes cope with U(VI) under nongrowing conditions, a metabolic state that is prevalent in natural environments.

  2. The genetic basis of laboratory adaptation in Caulobacter crescentus.

    Science.gov (United States)

    Marks, Melissa E; Castro-Rojas, Cyd Marie; Teiling, Clotilde; Du, Lei; Kapatral, Vinayak; Walunas, Theresa L; Crosson, Sean

    2010-07-01

    The dimorphic bacterium Caulobacter crescentus has evolved marked phenotypic changes during its 50-year history of culture in the laboratory environment, providing an excellent system for the study of natural selection and phenotypic microevolution in prokaryotes. Combining whole-genome sequencing with classical molecular genetic tools, we have comprehensively mapped a set of polymorphisms underlying multiple derived phenotypes, several of which arose independently in separate strain lineages. The genetic basis of phenotypic differences in growth rate, mucoidy, adhesion, sedimentation, phage susceptibility, and stationary-phase survival between C. crescentus strain CB15 and its derivative NA1000 is determined by coding, regulatory, and insertion/deletion polymorphisms at five chromosomal loci. This study evidences multiple genetic mechanisms of bacterial evolution as driven by selection for growth and survival in a new selective environment and identifies a common polymorphic locus, zwf, between lab-adapted C. crescentus and clinical isolates of Pseudomonas aeruginosa that have adapted to a human host during chronic infection.

  3. Physiochemical properties of Caulobacter crescentus holdfast: a localized bacterial adhesive.

    Science.gov (United States)

    Berne, Cécile; Ma, Xiang; Licata, Nicholas A; Neves, Bernardo R A; Setayeshgar, Sima; Brun, Yves V; Dragnea, Bogdan

    2013-09-12

    To colonize surfaces, the bacterium Caulobacter crescentus employs a polar polysaccharide, the holdfast, located at the end of a thin, long stalk protruding from the cell body. Unlike many other bacteria which adhere through an extended extracellular polymeric network, the holdfast footprint area is tens of thousands times smaller than that of the total bacterium cross-sectional surface, making for some very demanding adhesion requirements. At present, the mechanism of holdfast adhesion remains poorly understood. We explore it here along three lines of investigation: (a) the impact of environmental conditions on holdfast binding affinity, (b) adhesion kinetics by dynamic force spectroscopy, and (c) kinetic modeling of the attachment process to interpret the observed time-dependence of the adhesion force at short and long time scales. A picture emerged in which discrete molecular units called adhesins are responsible for initial holdfast adhesion, by acting in a cooperative manner.

  4. Caulobacter crescentus exploits its helical cell body to swim efficiently

    Science.gov (United States)

    Liu, Bin; Mendoza, Marcos; Valenzuela, Joanna

    2015-11-01

    How an organism gets its shape remains an open question of fundamental science. In this study, we measure the 3D shape of a bacterium, Caulobacter crescentus, using a computational graphic technique for free-swimming microorganisms to analyze thousands of image frames of the same individual bacterium. Rather than having a crescent shape, the cell body of the organism is found to be twisted with a helical pitch angle around 45 degrees. Moreover, the detailed size and geometry of the cell body, matches the optimized cell body obtained by the slender body theory for swimming at fixed power. This result sheds new light on the shape evolution of microorganisms, and suggests that C. crescentus has adapted to its natural habitat of fresh-water lakes and streams, lacking nutrients.

  5. Bacteria rolling: motilities of rosette colonies in Caulobacter crescentus

    Science.gov (United States)

    Zeng, Yu; Liu, Bin

    2016-11-01

    The aquatic bacterium Caulobacter crescentus has two life cycle stages with distinct motilities: freely swimming swarmer cells and immotile stalked cells. Here, we show a new type of movement performed by freely suspended rosettes, spontaneous aggregates of stalked cells aligned radially relative to each other. Reproductive rosette members generate predivisional daughter cells with flagella, inducing rotations of the rosette as a whole. Such rotations exhibit dynamic angular velocities and lead to intermittent linear movements along liquid-solid interfaces, resembling rolling movements. We reconstructed the translational and rotational dynamics of the rosette movements from high-speed filming and long-term tracking. A mechanical model was developed to explain the hydrodynamic mechanism underlying such motilities. Our study illustrated a nontrivial mechanism for clustered bacteria to achieve motilities and sheds light on the adaptive significance of the collective behaviors of microorganisms in complex fluid environments.

  6. TonB-dependent maltose transport by Caulobacter crescentus.

    Science.gov (United States)

    Lohmiller, S; Hantke, K; Patzer, S I; Braun, V

    2008-06-01

    We have shown previously that Caulobacter crescentus grows on maltodextrins which are actively transported across the outer membrane by the MalA protein. Evidence for energy-coupled transport was obtained by deletion of the exbB exbD genes which abolished transport. However, removal of the TonB protein, which together with the ExbB ExbD proteins is predicted to form an energy-coupling device between the cytoplasmic membrane and the outer membrane, left transport unaffected. Here we identify an additional tonB gene encoded by the cc2334a ORF, which when deleted abolished maltose transport. MalA contains a TonB box that reads EEVVIT and is predicted to interact with TonB. Replacement of valine number 15 in the TonB box by proline abolished maltose transport. Maltose was transported across the cytoplasmic membrane by the MalY protein (CC2283). Maltose transport was induced by maltose and repressed by the MalI protein (CC2284). In addition to MalA, MalY and MalI, the mal locus encodes two predicted cytoplasmic alpha-amylases (CC2285 and CC2286) and a periplasmic glucoamylase (CC2282). The TonB dependence together with the previously described ExbB ExbD dependence demonstrates energy-coupled maltose transport across the outer membrane. MalY is involved in maltose transport across the cytoplasmic membrane by a presumably ion-coupled mechanism.

  7. Correction of the Caulobacter crescentus NA1000 genome annotation.

    Directory of Open Access Journals (Sweden)

    Bert Ely

    Full Text Available Bacterial genome annotations are accumulating rapidly in the GenBank database and the use of automated annotation technologies to create these annotations has become the norm. However, these automated methods commonly result in a small, but significant percentage of genome annotation errors. To improve accuracy and reliability, we analyzed the Caulobacter crescentus NA1000 genome utilizing computer programs Artemis and MICheck to manually examine the third codon position GC content, alignment to a third codon position GC frame plot peak, and matches in the GenBank database. We identified 11 new genes, modified the start site of 113 genes, and changed the reading frame of 38 genes that had been incorrectly annotated. Furthermore, our manual method of identifying protein-coding genes allowed us to remove 112 non-coding regions that had been designated as coding regions. The improved NA1000 genome annotation resulted in a reduction in the use of rare codons since noncoding regions with atypical codon usage were removed from the annotation and 49 new coding regions were added to the annotation. Thus, a more accurate codon usage table was generated as well. These results demonstrate that a comparison of the location of peaks third codon position GC content to the location of protein coding regions could be used to verify the annotation of any genome that has a GC content that is greater than 60%.

  8. Correction of the Caulobacter crescentus NA1000 genome annotation.

    Science.gov (United States)

    Ely, Bert; Scott, LaTia Etheredge

    2014-01-01

    Bacterial genome annotations are accumulating rapidly in the GenBank database and the use of automated annotation technologies to create these annotations has become the norm. However, these automated methods commonly result in a small, but significant percentage of genome annotation errors. To improve accuracy and reliability, we analyzed the Caulobacter crescentus NA1000 genome utilizing computer programs Artemis and MICheck to manually examine the third codon position GC content, alignment to a third codon position GC frame plot peak, and matches in the GenBank database. We identified 11 new genes, modified the start site of 113 genes, and changed the reading frame of 38 genes that had been incorrectly annotated. Furthermore, our manual method of identifying protein-coding genes allowed us to remove 112 non-coding regions that had been designated as coding regions. The improved NA1000 genome annotation resulted in a reduction in the use of rare codons since noncoding regions with atypical codon usage were removed from the annotation and 49 new coding regions were added to the annotation. Thus, a more accurate codon usage table was generated as well. These results demonstrate that a comparison of the location of peaks third codon position GC content to the location of protein coding regions could be used to verify the annotation of any genome that has a GC content that is greater than 60%.

  9. Proposed Physical Mechanism of Chromosome Segregation in Caulobacter crescentus

    Science.gov (United States)

    Banigan, Edward; Gelbart, Michael; Gitai, Zemer; Liu, Andrea; Wingreen, Ned

    2010-03-01

    Chromosome segregation is a fundamental process for all cells, but the force-generating mechanisms that drive chromosome movements in bacteria are especially unclear. In Caulobacter crescentus, recent work has demonstrated that a structure made up of the ParA protein elongates from one cell pole and interacts with ParB, a protein binding to the chromosome near the origin of replication (ori). ParB disassembles ParA, causing ParA to pull ParB, and thus, the ori to the opposite end of the cell. We performed Brownian dynamics simulations of this system in order to uncover the physical mechanism of this motion. We find that motion of the ori is robust to several variations of the model as long as a steady-state concentration gradient of ParA is established in the moving frame of the ParB-decorated chromosome. We suggest that the mechanism is ``self-diffusiophoretic'': by disassembling ParA, ParB creates a concentration gradient of ParA so that the ParA concentration is higher in front of the chromosome than behind it. Since the chromosome is attracted to ParA via ParB, it moves up the gradient in the desired direction.

  10. Ultraviolet mutagenesis and inducible DNA repair in Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Bender, R.A.

    1984-11-19

    The ability to reactivate ultraviolet (UV) damaged phage phiCbK (W-reactivation) is induced by UV irradiation of Caulobacter crescentus cells. Induction of W-reactivation potential is specific for phage phiCbK, requires protein synthesis, and is greatly reduced in the presence of the rec-526 mutation. The induction signal generated by UV irradiation is transient, lasting about 1 1/2 - 2 h at 30/sup 0/C; if chloramphenicol is present during early times after UV irradiation, induction of W-reactivation does not occur. Induction is maximal when cells are exposed to 5-10 J/m/sup 2/ of UV, a dose that also results in considerable mutagenesis of the cells. Taken together, these observations demonstrate the existence of a UV inducible, protein synthesis requiring, transiently signalled, rec-requiring DNA repair system analogous to W-reactivation in Escherichia coli. In addition, C. crescentus also has an efficient photoreactivation system that reverses UV damage in the presence of strong visible light.

  11. Helical motion of the cell body enhances Caulobacter crescentus motility.

    Science.gov (United States)

    Liu, Bin; Gulino, Marco; Morse, Michael; Tang, Jay X; Powers, Thomas R; Breuer, Kenneth S

    2014-08-01

    We resolve the 3D trajectory and the orientation of individual cells for extended times, using a digital tracking technique combined with 3D reconstructions. We have used this technique to study the motility of the uniflagellated bacterium Caulobacter crescentus and have found that each cell displays two distinct modes of motility, depending on the sense of rotation of the flagellar motor. In the forward mode, when the flagellum pushes the cell, the cell body is tilted with respect to the direction of motion, and it precesses, tracing out a helical trajectory. In the reverse mode, when the flagellum pulls the cell, the precession is smaller and the cell has a lower translation distance per rotation period and thus a lower motility. Using resistive force theory, we show how the helical motion of the cell body generates thrust and can explain the direction-dependent changes in swimming motility. The source of the cell body precession is believed to be associated with the flexibility of the hook that connects the flagellum to the cell body.

  12. Intergenerational continuity of cell shape dynamics in Caulobacter crescentus

    Science.gov (United States)

    Wright, Charles S.; Banerjee, Shiladitya; Iyer-Biswas, Srividya; Crosson, Sean; Dinner, Aaron R.; Scherer, Norbert F.

    2015-03-01

    We investigate the intergenerational shape dynamics of single Caulobacter crescentus cells using a novel combination of imaging techniques and theoretical modeling. We determine the dynamics of cell pole-to-pole lengths, cross-sectional widths, and medial curvatures from high accuracy measurements of cell contours. Moreover, these shape parameters are determined for over 250 cells across approximately 10000 total generations, which affords high statistical precision. Our data and model show that constriction is initiated early in the cell cycle and that its dynamics are controlled by the time scale of exponential longitudinal growth. Based on our extensive and detailed growth and contour data, we develop a minimal mechanical model that quantitatively accounts for the cell shape dynamics and suggests that the asymmetric location of the division plane reflects the distinct mechanical properties of the stalked and swarmer poles. Furthermore, we find that the asymmetry in the division plane location is inherited from the previous generation. We interpret these results in terms of the current molecular understanding of shape, growth, and division of C. crescentus.

  13. The coding and noncoding architecture of the Caulobacter crescentus genome.

    Science.gov (United States)

    Schrader, Jared M; Zhou, Bo; Li, Gene-Wei; Lasker, Keren; Childers, W Seth; Williams, Brandon; Long, Tao; Crosson, Sean; McAdams, Harley H; Weissman, Jonathan S; Shapiro, Lucy

    2014-07-01

    Caulobacter crescentus undergoes an asymmetric cell division controlled by a genetic circuit that cycles in space and time. We provide a universal strategy for defining the coding potential of bacterial genomes by applying ribosome profiling, RNA-seq, global 5'-RACE, and liquid chromatography coupled with tandem mass spectrometry (LC-MS) data to the 4-megabase C. crescentus genome. We mapped transcript units at single base-pair resolution using RNA-seq together with global 5'-RACE. Additionally, using ribosome profiling and LC-MS, we mapped translation start sites and coding regions with near complete coverage. We found most start codons lacked corresponding Shine-Dalgarno sites although ribosomes were observed to pause at internal Shine-Dalgarno sites within the coding DNA sequence (CDS). These data suggest a more prevalent use of the Shine-Dalgarno sequence for ribosome pausing rather than translation initiation in C. crescentus. Overall 19% of the transcribed and translated genomic elements were newly identified or significantly improved by this approach, providing a valuable genomic resource to elucidate the complete C. crescentus genetic circuitry that controls asymmetric cell division.

  14. An SMC ATPase mutant disrupts chromosome segregation in Caulobacter.

    Science.gov (United States)

    Schwartz, Monica A; Shapiro, Lucy

    2011-12-01

    Accurate replication and segregation of the bacterial genome are essential for cell cycle progression. We have identified a single amino acid substitution in the Caulobacter structural maintenance of chromosomes (SMC) protein that disrupts chromosome segregation and cell division. The E1076Q point mutation in the SMC ATPase domain caused a dominant-negative phenotype in which DNA replication was able to proceed, but duplicated parS centromeres, normally found at opposite cell poles, remained at one pole. The cellular positions of other chromosomal loci were in the wild-type order relative to the parS centromere, but chromosomes remained unsegregated and appeared to be stacked upon one another. Purified SMC-E1076Q was deficient in ATP hydrolysis and exhibited abnormally stable binding to DNA. We propose that SMC spuriously links the duplicated chromosome immediately after passage of the replication fork. In wild-type cells, ATP hydrolysis opens the SMC dimer, freeing one chromosome to segregate to the opposite pole. The loss of ATP hydrolysis causes the SMC-E1076Q dimer to remain bound to both chromosomes, inhibiting segregation.

  15. The coding and noncoding architecture of the Caulobacter crescentus genome.

    Directory of Open Access Journals (Sweden)

    Jared M Schrader

    2014-07-01

    Full Text Available Caulobacter crescentus undergoes an asymmetric cell division controlled by a genetic circuit that cycles in space and time. We provide a universal strategy for defining the coding potential of bacterial genomes by applying ribosome profiling, RNA-seq, global 5'-RACE, and liquid chromatography coupled with tandem mass spectrometry (LC-MS data to the 4-megabase C. crescentus genome. We mapped transcript units at single base-pair resolution using RNA-seq together with global 5'-RACE. Additionally, using ribosome profiling and LC-MS, we mapped translation start sites and coding regions with near complete coverage. We found most start codons lacked corresponding Shine-Dalgarno sites although ribosomes were observed to pause at internal Shine-Dalgarno sites within the coding DNA sequence (CDS. These data suggest a more prevalent use of the Shine-Dalgarno sequence for ribosome pausing rather than translation initiation in C. crescentus. Overall 19% of the transcribed and translated genomic elements were newly identified or significantly improved by this approach, providing a valuable genomic resource to elucidate the complete C. crescentus genetic circuitry that controls asymmetric cell division.

  16. Bioremediation of soluble heavy metals with recombinant Caulobacter crescentus.

    Science.gov (United States)

    Xu, Zhaohui; Lei, Yu; Patel, Jigar

    2010-01-01

    To achieve one-step separation of heavy metal ions from contaminated water, we have developed a novel bioremediation technology based on self-immobilization of the Caulobacter crescentus recombinant strain JS4022/p723-6H, which overexpresses hexahistidine peptide on the surface of the bacterial cells and serves as a whole-cell adsorbent for dissolved heavy metals. Biofilms formed by JS4022/p723-6H are effective at retaining cadmium from bacterial growth media or environmental water samples. Here we provide additional experiment data discussing the application potential of this new technology. Supplementation of calcium to the growth media produced robust JS4022/p723-6H cells by alleviating their sensitivity to chelators. After growth in the presence of 0.3% CaCl(2)·2H(2)O, double the amount of JS4022/p723-6H cells survived the treatment with 2 mM EDTA. Free cells of JS4022/p723-6H effectively sequestered 51% of the total cadmium from a Lake Erie water sample at pH 5.4, compared to 37% retrieved by the control strain. Similar levels of adsorption were observed at pH 4.2 as well. Cells of JS4022/p723-6H were tolerant of acid treatment for 90 min at pH ≥1.1 or 120 min at pH ≥2.5, which provides an avenue for the convenient regeneration of the bacterial cells metal-binding capacity with acidic solutions. Designs of possible bioreactors and an operation system are also presented.

  17. Structure and function of Caulobacter crescentus aldose-aldose oxidoreductase.

    Science.gov (United States)

    Taberman, Helena; Andberg, Martina; Koivula, Anu; Hakulinen, Nina; Penttilä, Merja; Rouvinen, Juha; Parkkinen, Tarja

    2015-12-15

    Aldose-aldose oxidoreductase (Cc AAOR) is a recently characterized enzyme from the bacterial strain Caulobacter crescentus CB15 belonging to the glucose-fructose oxidoreductase/inositol dehydrogenase/rhizopine catabolism protein (Gfo/Idh/MocA) family. Cc AAOR catalyses the oxidation and reduction of a panel of aldose monosaccharides using a tightly bound NADP(H) cofactor that is regenerated in the catalytic cycle. Furthermore, Cc AAOR can also oxidize 1,4-linked oligosaccharides. In the present study, we present novel crystal structures of the dimeric Cc AAOR in complex with the cofactor and glycerol, D-xylose, D-glucose, maltotriose and D-sorbitol determined to resolutions of 2.0, 1.8, 1.7, 1.9 and 1.8 Å (1 Å=0.1 nm), respectively. These complex structures allowed for a detailed analysis of the ligand-binding interactions. The structures showed that the C1 carbon of a substrate, which is either reduced or oxidized, is close to the reactive C4 carbon of the nicotinamide ring of NADP(H). In addition, the O1 hydroxy group of the substrate, which is either protonated or deprotonated, is unexpectedly close to both Lys(104) and Tyr(189), which may both act as a proton donor or acceptor. This led us to hypothesize that this intriguing feature could be beneficial for Cc AAOR to catalyse the reduction of a linear form of a monosaccharide substrate and the oxidation of a pyranose form of the same substrate in a reaction cycle, during which the bound cofactor is regenerated.

  18. Multiple large filament bundles observed in Caulobacter crescentus by electron cryotomography

    DEFF Research Database (Denmark)

    Briegel, A; Dias, DP; Li, Z;

    2006-01-01

    , molecular mechanisms have remained obscure in part for lack of electron microscopy-resolution images where these filaments can be seen acting within their cellular context. Here, electron cryotomography was used to image the widely studied model prokaryote Caulobacter crescentus in an intact, near...

  19. A physical approach to segregation and folding of the Caulobacter crescentus genome

    NARCIS (Netherlands)

    Dame, R.T.; Tark-Dame, M.; Schiessel, H

    2011-01-01

    Bacterial genomes are functionally organized. This organization is dynamic and globally changing throughout the cell cycle. Upon initiation of replication of the chromosome, the two origins segregate and move towards their new location taking along the newly replicated genome. Caulobacter crescentus

  20. Genome Sequence of Selenium-Solubilizing Bacterium Caulobacter vibrioides T5M6

    DEFF Research Database (Denmark)

    Wang, Yihua; Qin, Yanan; Kot, Witold

    2016-01-01

    Caulobacter vibrioides T5M6 is a Gram-negative strain that strongly solubilizes selenium (Se) mineral into Se(IV) and was isolated from a selenium mining area in Enshi, southwest China. This strain produces the phytohormone IAA and promotes plant growth. Here we present the genome of this strain ...

  1. Plankton networks driving carbon export in the oligotrophic ocean.

    Science.gov (United States)

    Guidi, Lionel; Chaffron, Samuel; Bittner, Lucie; Eveillard, Damien; Larhlimi, Abdelhalim; Roux, Simon; Darzi, Youssef; Audic, Stephane; Berline, Léo; Brum, Jennifer R; Coelho, Luis Pedro; Espinoza, Julio Cesar Ignacio; Malviya, Shruti; Sunagawa, Shinichi; Dimier, Céline; Kandels-Lewis, Stefanie; Picheral, Marc; Poulain, Julie; Searson, Sarah; Stemmann, Lars; Not, Fabrice; Hingamp, Pascal; Speich, Sabrina; Follows, Mick; Karp-Boss, Lee; Boss, Emmanuel; Ogata, Hiroyuki; Pesant, Stephane; Weissenbach, Jean; Wincker, Patrick; Acinas, Silvia G; Bork, Peer; de Vargas, Colomban; Iudicone, Daniele; Sullivan, Matthew B; Raes, Jeroen; Karsenti, Eric; Bowler, Chris; Gorsky, Gabriel

    2016-04-28

    The biological carbon pump is the process by which CO2 is transformed to organic carbon via photosynthesis, exported through sinking particles, and finally sequestered in the deep ocean. While the intensity of the pump correlates with plankton community composition, the underlying ecosystem structure driving the process remains largely uncharacterized. Here we use environmental and metagenomic data gathered during the Tara Oceans expedition to improve our understanding of carbon export in the oligotrophic ocean. We show that specific plankton communities, from the surface and deep chlorophyll maximum, correlate with carbon export at 150 m and highlight unexpected taxa such as Radiolaria and alveolate parasites, as well as Synechococcus and their phages, as lineages most strongly associated with carbon export in the subtropical, nutrient-depleted, oligotrophic ocean. Additionally, we show that the relative abundance of a few bacterial and viral genes can predict a significant fraction of the variability in carbon export in these regions.

  2. Picocyanobacteria success in oligotrophic lakes: fact or fiction?

    Directory of Open Access Journals (Sweden)

    Cristiana CALLIERI

    2000-02-01

    Full Text Available Two approaches may be utilized to explain the predominance of picocyanobacteria (Pcy in oligotrophic lakes: the analysis of their interannual evolution in one single lake and their relative importance in different lakes along a trophic gradient. Here we discuss results from field data on picocyanobacteria over several seasons from a deep oligotrophic subalpine lake - Lago Maggiore, and variables influencing their abundance. Comparing data from lakes along a trophic gradient, no simple relationship emerges between lake’s trophic state and picocyanobacteria abundance and contribution to total phytoplanktonic biomass. That is, trophic state alone cannot explain the success/absence of picocyanobacteria that appear to be favored under P limitation, but seem more sensitive to grazing pressure and light. In some oligotrophic lakes, if light climate, grazing, and competition are favorable, picocyanobacteria can grow rapidly, out-compete competitors and become very abundant, but there are a host of factors that can influence the outcome of this competition, and ultimately influence Pcy success in lakes of all trophic types.

  3. Reclassification and emended description of Caulobacter leidyi as Sphingomonas leidyi comb. nov., and emendation of the genus Sphingomonas

    National Research Council Canada - National Science Library

    Chen, Hong; Jogler, Mareike; Rohde, Manfred; Klenk, Hans-Peter; Busse, Hans-Jürgen; Tindall, Brian J; Spröer, Cathrin; Overmann, Jörg

    2012-01-01

    'Caulobacter leidyi' DSM 4733(T) has been shown to be affiliated with the family Sphingomonadaceae instead of the Caulobacteraceae, and due to its poor characterization has been omitted from the current edition of Bergey's Manual...

  4. Characterization of Uranium Tolerance and Biomineralization Potential of Caulobacter crescentus

    Science.gov (United States)

    Park, D.

    2015-12-01

    Due to its high toxicity and mobility, U(VI) poses a major environmental threat to ecosystems. The ubiquitous aerobic bacterium Caulobacter cresecentus is an attractive candidate for U(VI) bioremediation because of its ability to survive in low-nutrient environments (5, 6), tolerate high U concentrations and mineralize U(VI) aerobically through the formation of uranyl phosphate (U-Pi) precipitates. Despite these attractive environmental properties, both a systems level understanding of the adaptive response pathways involved in U tolerance and the environmental conditions affecting the biomineralization process and stability of biogenic U-Pi minerals remain limited. By measuring changes in both mRNA and protein expression during exposure to high U levels, we have identified the core stress response pathways involved in U tolerance. Pathways associated with heat shock, lipospolysaccharide biosynthesis and transport, outer membrane lipoprotein transport and outermembrane assembly were highly induced at both the RNA and protein levels. Correspondingly, removal of integral components of proteolysis pathways including clpA, clpS and degP significantly reduced U tolerance under biomineralization conditions. Surprisingly, in contrast to many other heavy metals, U did not cause oxidative stress or DNA damage. Together, these analyses indicate that U predominately targets the outermembrane and causes mis-folding of both cytoplasmic and extracytoplasmic proteins. Efforts are currently underway to characterize the morphological and structural properties of biogenic U-Pi minerals and the environmental factors that influence their production and stability. Preliminary AFM studies suggest that U-Pi minerals formed under biomineralization conditions appear morphologically distinct from those formed abiotically between U(VI) and inorganic phosphate. Additionally, we observed that biomineralization tolerates a wide pH range (pH 6-9). Our long-range goal is the development of a

  5. Factors controlling in vitro recrystallization of the Caulobacter crescentus paracrystalline S-layer.

    Science.gov (United States)

    Nomellini, J F; Kupcu, S; Sleytr, U B; Smit, J

    1997-10-01

    The S-layer of Caulobacter is a two-dimensional paracrystalline array on the cell surface composed of a single protein, RsaA. We have established conditions for preparation of stable, soluble protein and then efficient in vitro recrystallization of the purified protein. Efficient recrystallization and long range order could not be obtained with pure protein only, though it was apparent that calcium was required for crystallization. Recrystallization was obtained when lipid vesicles were provided, but only when the vesicles contained the specific species of Caulobacter smooth lipopolysaccharide (SLPS) that previous studies implicated as a requirement for attaching the S-layer to the cell surface. The specific type of phospholipids did not appear critical; phospholipids rather different from those present in Caulobacter membranes or archaebacterial tetraether lipids worked equally well. The source of LPS was critical; rough and smooth variants of Salmonella typhimurium LPS as well as the rough form of Caulobacter LPS were ineffective. The requirement for calcium ions for recrystallization was further evaluated; strontium ions could substitute for calcium, and to a lesser extent, cobalt, barium, manganese and magnesium ions also stimulated crystallization. On the other hand, nickel and cadmium provided only weak crystallization stimulation, and zinc, copper, iron, aluminum ions, and the monovalent potassium, sodium, and lithium ions were ineffective. The recrystallization could also be reproduced with Langmuir-Blodgett lipid monolayers at an air-water interface. As with the vesicle experiments, this was only successful when SLPS was incorporated into the lipid mix. The best method for RsaA preparation, leading to apparently monomeric protein that was stable for many months, was an extraction with a low pH aqueous solution. We also achieved recrystallization, albeit at lower efficiency, using RsaA protein solubilized by 8 M urea, a method which allows retrieval of

  6. A stochastic spatiotemporal model of a response-regulator network in the Caulobacter crescentus cell cycle

    Science.gov (United States)

    Li, Fei; Subramanian, Kartik; Chen, Minghan; Tyson, John J.; Cao, Yang

    2016-06-01

    The asymmetric cell division cycle in Caulobacter crescentus is controlled by an elaborate molecular mechanism governing the production, activation and spatial localization of a host of interacting proteins. In previous work, we proposed a deterministic mathematical model for the spatiotemporal dynamics of six major regulatory proteins. In this paper, we study a stochastic version of the model, which takes into account molecular fluctuations of these regulatory proteins in space and time during early stages of the cell cycle of wild-type Caulobacter cells. We test the stochastic model with regard to experimental observations of increased variability of cycle time in cells depleted of the divJ gene product. The deterministic model predicts that overexpression of the divK gene blocks cell cycle progression in the stalked stage; however, stochastic simulations suggest that a small fraction of the mutants cells do complete the cell cycle normally.

  7. Complete genome sequence of Caulobacter crescentus bacteriophage φCbK.

    Science.gov (United States)

    Panis, Gaël; Lambert, Christophe; Viollier, Patrick H

    2012-09-01

    φCbK is a B3 morphotype bacteriophage of the Siphoviridae family that infects Caulobacter crescentus, the preeminent model system for bacterial cell cycle studies. The last 4 decades of research with φCbK as a genetic and cytological tool to study the biology of the host warrant an investigation of the phage genome composition. Herein, we report the complete genome sequence of φCbK and highlight unusual features that emerged from its annotation. The complete genome analysis of the φCbK phage provides new insight into its characteristics and potential interactions with its Caulobacter crescentus host, setting the stage for future functional studies with φCbK.

  8. Temporal controls of the asymmetric cell division cycle in Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Shenghua Li

    2009-08-01

    Full Text Available The asymmetric cell division cycle of Caulobacter crescentus is orchestrated by an elaborate gene-protein regulatory network, centered on three major control proteins, DnaA, GcrA and CtrA. The regulatory network is cast into a quantitative computational model to investigate in a systematic fashion how these three proteins control the relevant genetic, biochemical and physiological properties of proliferating bacteria. Different controls for both swarmer and stalked cell cycles are represented in the mathematical scheme. The model is validated against observed phenotypes of wild-type cells and relevant mutants, and it predicts the phenotypes of novel mutants and of known mutants under novel experimental conditions. Because the cell cycle control proteins of Caulobacter are conserved across many species of alpha-proteobacteria, the model we are proposing here may be applicable to other genera of importance to agriculture and medicine (e.g., Rhizobium, Brucella.

  9. Enhanced neutralization of HIV by antibodies displayed on the S-layer of Caulobacter crescentus.

    Science.gov (United States)

    Duval, Mark; Lewis, Christopher J; Nomellini, John F; Horwitz, Marc S; Smit, John; Cavacini, Lisa A

    2011-12-01

    Innovative methods of prevention are needed to stop the more than two million new HIV-1 infections annually, particularly in women. Local application of anti-HIV antibodies has been shown to be effective at preventing infection in nonhuman primates; however, the concentrations needed are cost prohibitive. Display of antibodies on a particulate platform will likely prolong effectiveness of these anti-HIV agents and lower the cost of goods. Here, we demonstrate that the bacterium Caulobacter crescentus and its highly expressed surface-layer (S-layer) protein can provide this antibody display platform. Caulobacters displaying protein G, alone or with CD4 codisplay, successfully captured HIV-1-specific antibodies and demonstrated functional neutralization. Compared to soluble antibodies, a neutralizing anti-HIV antibody displayed on Caulobacter was as effective or more effective at neutralizing diverse HIV-1 isolates. Moreover, when an antibody reactive with an epitope induced by CD4 binding (CD4i) was codisplayed with CD4, there was significant enhancement in HIV-1 neutralization. These results suggest that caulobacters displaying anti-HIV antibodies offer a distinct improvement in the use of antibodies as microbicides. Furthermore, these reagents can specifically evaluate anti-HIV antibodies in concert with other HIV-1 blocking agents to assess the most suitable tools for conversion to scFvs, allowing for direct display within the S-layer protein and further reducing cost of goods. In summary, C. crescentus, which can be easily produced and chemically stabilized at low cost, is well suited for engineering as an effective platform, offering an inexpensive way to produce and deliver HIV-1-specific microbicides.

  10. Factors controlling in vitro recrystallization of the Caulobacter crescentus paracrystalline S-layer.

    OpenAIRE

    Nomellini, J F; Kupcu, S; Sleytr, U B; Smit, J.

    1997-01-01

    The S-layer of Caulobacter is a two-dimensional paracrystalline array on the cell surface composed of a single protein, RsaA. We have established conditions for preparation of stable, soluble protein and then efficient in vitro recrystallization of the purified protein. Efficient recrystallization and long range order could not be obtained with pure protein only, though it was apparent that calcium was required for crystallization. Recrystallization was obtained when lipid vesicles were provi...

  11. Diverse functions for six glycosyltransferases in Caulobacter crescentus cell wall assembly.

    Science.gov (United States)

    Yakhnina, Anastasiya A; Gitai, Zemer

    2013-10-01

    The essential process of peptidoglycan synthesis requires two enzymatic activities, transpeptidation and transglycosylation. While the PBP2 and PBP3 transpeptidases perform highly specialized functions that are widely conserved, the specific roles of different glycosyltransferases are poorly understood. For example, Caulobacter crescentus encodes six glycosyltransferase paralogs of largely unknown function. Using genetic analyses, we found that Caulobacter glycosyltransferases are primarily redundant but that PbpX is responsible for most of the essential glycosyltransferase activity. Cells containing PbpX as their sole glycosyltransferase are viable, and the loss of pbpX leads to a general defect in the integrity of the cell wall structure even in the presence of the other five glycosyltransferases. However, neither PbpX nor any of its paralogs is required for the specific processes of cell elongation or division, while the cell wall synthesis required for stalk biogenesis is only partially disrupted in several of the glycosyltransferase mutants. Despite their genetic redundancy, Caulobacter glycosyltransferases exhibit different subcellular localizations. We suggest that these enzymes have specialized roles and normally function in distinct subcomplexes but retain the ability to substitute for one another so as to ensure the robustness of the peptidoglycan synthesis process.

  12. Synchronization of Caulobacter crescentus for investigation of the bacterial cell cycle.

    Science.gov (United States)

    Schrader, Jared M; Shapiro, Lucy

    2015-04-08

    The cell cycle is important for growth, genome replication, and development in all cells. In bacteria, studies of the cell cycle have focused largely on unsynchronized cells making it difficult to order the temporal events required for cell cycle progression, genome replication, and division. Caulobacter crescentus provides an excellent model system for the bacterial cell cycle whereby cells can be rapidly synchronized in a G0 state by density centrifugation. Cell cycle synchronization experiments have been used to establish the molecular events governing chromosome replication and segregation, to map a genetic regulatory network controlling cell cycle progression, and to identify the establishment of polar signaling complexes required for asymmetric cell division. Here we provide a detailed protocol for the rapid synchronization of Caulobacter NA1000 cells. Synchronization can be performed in a large-scale format for gene expression profiling and western blot assays, as well as a small-scale format for microscopy or FACS assays. The rapid synchronizability and high cell yields of Caulobacter make this organism a powerful model system for studies of the bacterial cell cycle.

  13. Whole-genome transcriptional analysis of heavy metal stresses inCaulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Ping; Brodie, Eoin L.; Suzuki, Yohey; McAdams, Harley H.; Andersen, Gary L.

    2005-09-21

    The bacterium Caulobacter crescentus and related stalkbacterial species are known for their distinctive ability to live in lownutrient environments, a characteristic of most heavy metal contaminatedsites. Caulobacter crescentus is a model organism for studying cell cycleregulation with well developed genetics. We have identified the pathwaysresponding to heavy metal toxicity in C. crescentus to provide insightsfor possible application of Caulobacter to environmental restoration. Weexposed C. crescentus cells to four heavy metals (chromium, cadmium,selenium and uranium) and analyzed genome wide transcriptional activitiespost exposure using a Affymetrix GeneChip microarray. C. crescentusshowed surprisingly high tolerance to uranium, a possible mechanism forwhich may be formation of extracellular calcium-uranium-phosphateprecipitates. The principal response to these metals was protectionagainst oxidative stress (up-regulation of manganese-dependent superoxidedismutase, sodA). Glutathione S-transferase, thioredoxin, glutaredoxinsand DNA repair enzymes responded most strongly to cadmium and chromate.The cadmium and chromium stress response also focused on reducing theintracellular metal concentration, with multiple efflux pumps employed toremove cadmium while a sulfate transporter was down-regulated to reducenon-specific uptake of chromium. Membrane proteins were also up-regulatedin response to most of the metals tested. A two-component signaltransduction system involved in the uranium response was identified.Several differentially regulated transcripts from regions previously notknown to encode proteins were identified, demonstrating the advantage ofevaluating the transcriptome using whole genome microarrays.

  14. Lake level fluctuations boost toxic cyanobacterial "oligotrophic blooms".

    Directory of Open Access Journals (Sweden)

    Cristiana Callieri

    Full Text Available Global warming has been shown to strongly influence inland water systems, producing noticeable increases in water temperatures. Rising temperatures, especially when combined with widespread nutrient pollution, directly favour the growth of toxic cyanobacteria. Climate changes have also altered natural water level fluctuations increasing the probability of extreme events as dry periods followed by heavy rains. The massive appearance of Dolichospermum lemmermannii ( = planktonic Anabaena, a toxic species absent from the pelagic zone of the subalpine oligotrophic Lake Maggiore before 2005, could be a consequence of the unusual fluctuations of lake level in recent years. We hypothesized that these fluctuations may favour the cyanobacterium as result of nutrient pulses from the biofilms formed in the littoral zone when the lake level is high. To help verify this, we exposed artificial substrates in the lake, and evaluated their nutrient enrichment and release after desiccation, together with measurements of fluctuations in lake level, precipitation and D. lemmermannii population. The highest percentage of P release and the lowest C:P molar ratio of released nutrients coincided with the summer appearance of the D. lemmermannii bloom. The P pulse indicates that fluctuations in level counteract nutrient limitation in this lake and it is suggested that this may apply more widely to other oligotrophic lakes. In view of the predicted increase in water level fluctuations due to climate change, it is important to try to minimize such fluctuations in order to mitigate the occurrence of cyanobacterial blooms.

  15. Complete genome sequence of the facultatively chemolithoautotrophic and methylotrophic alpha Proteobacterium Starkeya novella type strain (ATCC 8093T)

    Energy Technology Data Exchange (ETDEWEB)

    Kappler, Ulrike [University of Queensland, The, Brisbane, Queensland, Australia; Davenport, Karen W. [Los Alamos National Laboratory (LANL); Beatson, Scott [University of Queensland, The, Brisbane, Queensland, Australia; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Berry, Kerrie W. [U.S. Department of Energy, Joint Genome Institute; Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Hammon, Nancy [U.S. Department of Energy, Joint Genome Institute; Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Richardson, P M [U.S. Department of Energy, Joint Genome Institute; Bruce, David [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Tapia, Roxanne [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Chang, Yun-Juan [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Goker, Markus [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Klenk, Hans-Peter [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute

    2012-01-01

    Starkeya novella (Starkey 1934) Kelly et al. 2000 is a member of the family Xanthobacteraceae in the order Rhizobiales , which is thus far poorly characterized at the genome level. Cultures from this spe- cies are most interesting due to their facultatively chemolithoautotrophic lifestyle, which allows them to both consume carbon dioxide and to produce it. This feature makes S. novella an interesting model or- ganism for studying the genomic basis of regulatory networks required for the switch between con- sumption and production of carbon dioxide, a key component of the global carbon cycle. In addition, S. novella is of interest for its ability to grow on various inorganic sulfur compounds and several C1- compounds such as methanol. Besides Azorhizobium caulinodans, S. novella is only the second spe- cies in the family Xanthobacteraceae with a completely sequenced genome of a type strain. The cur- rent taxonomic classification of this group is in significant conflict with the 16S rRNA data. The ge- nomic data indicate that the physiological capabilities of the organism might have been underestimat- ed. The 4,765,023 bp long chromosome with its 4,511 protein-coding and 52 RNA genes was se- quenced as part of the DOE Joint Genome Institute Community Sequencing Program (CSP) 2008.

  16. Exploring Microbial Life in Oxic Sediments Underlying Oligotrophic Ocean Gyres

    Science.gov (United States)

    Ziebis, W.; Orcutt, B.; Wankel, S. D.; D'Hondt, S.; Szubin, R.; Kim, J. N.; Zengler, K.

    2015-12-01

    Oxygen, carbon and nutrient availability are defining parameters for microbial life. In contrast to organic-rich sediments of the continental margins, where high respiration rates lead to a depletion of O2 within a thin layer at the sediment surface, it was discovered that O2 penetrates several tens of meters into organic-poor sediments underlying oligotrophic ocean gyres. In addition, nitrate, another important oxidant, which usually disappears rapidly with depth in anoxic sediments, tends to accumulate above seawater concentrations in the oxic subsurface, reflecting the importance of nitrogen cycling processes, including both nitrification and denitrification. Two IODP drilling expeditions were vital for exploring the nature of the deep subsurface beneath oligotrophic ocean gyres, expedition 329 to the South Pacific Gyre (SPG) and expedition 336 to North Pond, located on the western flank of the Mid-Atlantic ridge beneath the North Atlantic Gyre. Within the ultra-oligotrophic SPG O2 penetrates the entire sediment column from the sediment-water interface to the underlying basement to depths of > 75 m. At North Pond, a topographic depression filled with sediment and surrounded by steep basaltic outcrops, O2 penetrates deeply into the sediment (~ 30 m) until it eventually becomes depleted. O2 also diffuses upward into the sediment from seawater circulating within the young crust underlying the sediment, resulting in a deep oxic layer several meters above the basalt. Despite low organic carbon contents microbial cells persist throughout the entire sediment column within the SPG (> 75 m) and at North Pond, albeit at low abundances. We explored the nature of the subsurface microbial communities by extracting intact cells from large volumes of sediment obtained from drill cores of the two expeditions. By using CARD-FiSH, amplicon (16s rRNA) and metagenome sequencing we shed light on the phylogenetic and functional diversity of the elusive communities residing in the

  17. Molecular diversity of diazotrophs in oligotrophic tropical seagrass bed communities.

    Science.gov (United States)

    Bagwell, Christopher E; Rocque, Jeannine R; Smith, Garriett W; Polson, Shawn W; Friez, Michael J; Longshore, John W; Lovell, Charles R

    2002-02-01

    Denaturing gradient gel electrophoresis (DGGE) was employed to resolve PCR-amplified nifH sequences from vegetated and unvegetated sediments from two oligotrophic seagrass bed sites on San Salvador Island, Bahamas, in order to assess diazotroph species composition. All DGGE profiles from these sites showed the same prominent bands. These bands were sequenced, yielding 67 different nifH sequences, which were used in phylogenetic reconstructions. Most sequences were from anaerobes, but some were affiliated with the alpha- and (gamma-+beta-) Proteobacteria. Several NifH sequences were nearly identical to those from Azospirillum brasilense and Vibrio diazotrophicus. These seagrass bed sediments support a diverse diazotroph assemblage that is, at least superficially, similar to that associated with an intertidal grass (Spartina alterniflora).

  18. A transducing bacteriophage for Caulobacter crescentus uses the paracrystalline surface layer protein as a receptor.

    OpenAIRE

    P. Edwards; Smit, J

    1991-01-01

    The bacteriophage phi Cr30, a transducing phage for Caulobacter crescentus strains, required the paracrystalline surface (S) layer for infectivity. Wild-type strains were phage resistant when rsaA, the gene for the 130K S-layer protein, was interrupted with an antibiotic resistance cassette. Strains that had lost the S layer by mutation were phage resistant, as were mutants that produce an S layer but which do not attach the structure to the cell surface. Phage sensitivity was restored to 130...

  19. Coordination between chromosome replication, segregation, and cell division in Caulobacter crescentus

    DEFF Research Database (Denmark)

    Jensen, Rasmus Bugge

    2006-01-01

    Progression through the Caulobacter crescentus cell cycle is coupled to a cellular differentiation program. The swarmer cell is replicationally quiescent, and DNA replication initiates at the swarmer-to-stalked cell transition. There is a very short delay between initiation of DNA replication......, and the completely replicated terminus regions stay associated with each other after chromosome replication is completed, disassociating very late in the cell cycle shortly before the final cell division event. Invagination of the cytoplasmic membrane occurs earlier than separation of the replicated terminus regions...

  20. ALGAL RESPONSE TO NUTRIENT ENRICHMENT IN FORESTED OLIGOTROPHIC STREAM(1).

    Science.gov (United States)

    Veraart, Annelies J; Romaní, Anna M; Tornés, Elisabet; Sabater, Sergi

    2008-06-01

    Nutrient input in streams alters the density and species composition of attached algal communities in open systems. However, in forested streams, the light reaching the streambed (rather than the local nutrient levels) may limit the growth of these communities. A nutrient-enrichment experiment in a forested oligotrophic stream was performed to test the hypothesis that nutrient addition has only minor effects on the community composition of attached algae and cyanobacteria under light limitation. Moderate nutrient addition consisted of increasing basal phosphorus (P) concentrations 3-fold and basal nitrogen (N) concentrations 2-fold. Two upstream control reaches were compared to a downstream reach before and after nutrient addition. Nutrients were added continuously to the downstream reach for 1 year. Algal biofilms growing on ceramic tiles were sampled and identified for more than a year before nutrient addition to 12 months after. Diatoms were the most abundant taxonomic group in the three stream reaches. Nutrient enrichment caused significant variations in the composition of the diatom community. While some taxa showed significant decreases (e.g., Achnanthes minutissima, Gomphonema angustum), increases for other taxa (such as Rhoicosphenia abbreviata and Amphora ovalis) were detected in the enriched reach (for taxonomic authors, see Table 2). Epiphytic and adnate taxa of large size were enhanced, particularly during periods of favorable growth conditions (spring). Nutrients also caused a change in the algal chl a, which increased from 0.5-5.8 to 2.1-10.7 μg chl · cm(-2) . Our results indicate that in oligotrophic forested streams, long-term nutrient addition has significant effects on the algal biomass and community composition, which are detectable despite the low light availability caused by the tree canopy. Low light availability moderates but does not detain the long-term tendency toward a nutrient-tolerant community. Furthermore, the effects

  1. Deep silicon maxima in the stratified oligotrophic Mediterranean Sea

    Directory of Open Access Journals (Sweden)

    Y. Crombet

    2011-02-01

    Full Text Available The silicon biogeochemical cycle has been studied in the Mediterranean Sea during late summer/early autumn 1999 and summer 2008. The distribution of nutrients, particulate carbon and silicon, fucoxanthin (Fuco, and total chlorophyll-a (TChl-a were investigated along an eastward gradient of oligotrophy during two cruises (PROSOPE and BOUM encompassing the entire Mediterranean Sea during the stratified period. At both seasons, surface waters were depleted in nutrients and the nutriclines gradually deepened towards the East, the phosphacline being the deepest in the easternmost Levantine basin. Following the nutriclines, parallel deep maxima of biogenic silica (DSM, fucoxanthin (DFM and TChl-a (DCM were evidenced during both seasons with maximal concentrations of 0.45 μmol L−1 for BSi, 0.26 μg L−1 for Fuco, and 1.70 μg L−1 for TChl-a, all measured during summer. Contrary to the DCM which was a persistent feature in the Mediterranean Sea, the DSM and DFMs were observed in discrete areas of the Alboran Sea, the Algero-Provencal basin, the Ionian sea and the Levantine basin, indicating that diatoms were able to grow at depth and dominate the DCM under specific conditions. Diatom assemblages were dominated by Chaetoceros spp., Leptocylindrus spp., Pseudonitzschia spp. and the association between large centric diatoms (Hemiaulus hauckii and Rhizosolenia styliformis and the cyanobacterium Richelia intracellularis was observed at nearly all sites. The diatom's ability to grow at depth is commonly observed in other oligotrophic regions and could play a major role in ecosystem productivity and carbon export to depth. Contrary to the common view that Si and siliceous phytoplankton are not major components of the Mediterranean biogeochemistry, we suggest here that diatoms, by persisting at depth during the stratified period, could contribute to a

  2. A new factor stimulating peptidoglycan hydrolysis to separate daughter cells in Caulobacter crescentus.

    Science.gov (United States)

    Collier, Justine

    2010-07-01

    Cell division in Gram-negative bacteria involves the co-ordinated invagination of the three cell envelope layers to form two new daughter cell poles. This complex process starts with the polymerization of the tubulin-like protein FtsZ into a Z-ring at mid-cell, which drives cytokinesis and recruits numerous other proteins to the division site. These proteins are involved in Z-ring constriction, inner- and outer-membrane invagination, peptidoglycan remodelling and daughter cell separation. Three papers in this issue of Molecular Microbiology, from the teams of Lucy Shapiro, Martin Thanbichler and Christine Jacobs-Wagner, describe a novel protein, called DipM for Division Involved Protein with LysM domains, that is required for cell division in Caulobacter crescentus. DipM localizes to the mid-cell during cell division, where it is necessary for the hydrolysis of the septal peptidoglycan to remodel the cell wall. Loss of DipM results in severe defects in cell envelope constriction, which is deleterious under fast-growth conditions. State-of-the-art microscopy experiments reveal that the peptidoglycan is thicker and that the cell wall is incorrectly organized in DipM-depleted cells compared with wild-type cells, demonstrating that DipM is essential for reorganizing the cell wall at the division site, for envelope invagination and cell separation in Caulobacter.

  3. Regulated proteolysis of a transcription factor complex is critical to cell cycle progression in Caulobacter crescentus.

    Science.gov (United States)

    Gora, Kasia G; Cantin, Amber; Wohlever, Matthew; Joshi, Kamal K; Perchuk, Barrett S; Chien, Peter; Laub, Michael T

    2013-03-01

    Cell cycle transitions are often triggered by the proteolysis of key regulatory proteins. In Caulobacter crescentus, the G1-S transition involves the degradation of an essential DNA-binding response regulator, CtrA, by the ClpXP protease. Here, we show that another critical cell cycle regulator, SciP, is also degraded during the G1-S transition, but by the Lon protease. SciP is a small protein that binds directly to CtrA and prevents it from activating target genes during G1. We demonstrate that SciP must be degraded during the G1-S transition so that cells can properly activate CtrA-dependent genes following DNA replication initiation and the reaccumulation of CtrA. These results indicate that like CtrA, SciP levels are tightly regulated during the Caulobacter cell cycle. In addition, we show that formation of a complex between CtrA and SciP at target promoters protects both proteins from their respective proteases. Degradation of either protein thus helps trigger the destruction of the other, facilitating a cooperative disassembly of the complex. Collectively, our results indicate that ClpXP and Lon each degrade an important cell cycle regulator, helping to trigger the onset of S phase and prepare cells for the subsequent programmes of gene expression critical to polar morphogenesis and cell division.

  4. The role of spatial asymmetries in the development of the bacterium Caulobacter crescentus

    Science.gov (United States)

    Tropini, Carolina; Chen, Erin; Sciochetti, Stephen; Newton, Austin; Laub, Michael; Huang, Kerwyn Casey

    2010-03-01

    Caulobacter is a model organism for cell cycle regulation and development. Upon division it differentiates into a sessile stalked cell and a motile swarmer cell. Throughout the cell cycle, the localization of several key proteins is highly regulated. We address the importance of spatial localization in signal transduction and development. Flagellar pole development is controlled by the response regulator DivK, whose phosphorylation state is controlled by the kinase DivJ and the phosphatase PleC. PleC localizes to the swarmer pole, while DivJ localizes at the stalked pole. We have constructed strains with a variety of PleC and DivJ localization patterns. Our results indicate that localization is not absolutely necessary in this system, rather localized proteins enhance the robustness to fluctuations. We further investigate the importance of spatial asymmetries in the regulation of the master cell-cycle-regulator CtrA. In its phosphorylated form, CtrA binds to the replication origin in Caulobacter in a highly cooperative fashion, and prevents DNA replication. The CtrA distribution is tightly controlled not only by localized phosphorylation and dephosphorylation but also synthesis and degradation. We find that physiological degradation rates exert only a small perturbation on the distribution generated by asymmetric phosphorylation.

  5. Critical clamp loader processing by an essential AAA+ protease in Caulobacter crescentus.

    Science.gov (United States)

    Vass, Robert H; Chien, Peter

    2013-11-01

    Chromosome replication relies on sliding clamps that are loaded by energy-dependent complexes. In Escherichia coli, the ATP-binding clamp loader subunit DnaX exists as both long (τ) and short (γ) forms generated through programmed translational frameshifting, but the need for both forms is unclear. Here, we show that in Caulobacter crescentus, DnaX isoforms are unexpectedly generated through partial proteolysis by the AAA+ protease casein lytic proteinase (Clp) XP. We find that the normally processive ClpXP protease partially degrades DnaX to produce stable fragments upon encountering a glycine-rich region adjacent to a structured domain. Increasing the sequence complexity of this region prevents partial proteolysis and generates a τ-only form of DnaX in vivo that is unable to support viability on its own. Growth is restored when γ is provided in trans, but these strains are more sensitive to DNA damage compared with strains that can generate γ through proteolysis. Our work reveals an unexpected mode of partial processing by the ClpXP protease to generate DnaX isoforms, demonstrates that both τ and γ forms of DnaX are required for Caulobacter viability, and identifies a role for clamp loader diversity in responding to DNA damage. The conservation of distinct DnaX isoforms throughout bacteria despite fundamentally different mechanisms for producing them suggests there may be a conserved need for alternate clamp loader complexes during DNA damaging conditions.

  6. Motion of single MreB bacterial actin proteins in Caulobacter show treadmilling in vivo

    Science.gov (United States)

    Moerner, W. E.; Kim, Soyeon; Gitai, Zemer; Kinkhabwala, Anika; McAdams, Harley; Shapiro, Lucy

    2006-03-01

    Ensemble imaging of a bacterial actin homologue, the MreB protein, suggests that the MreB proteins form a dynamic filamentous spiral along the long axis of the cell in Caulobacter crescentus. MreB contracts and expands along the cell axis and plays an important role in cell shape and polarity maintenance, as well as chromosome segregation and translocation of the origin of replication during cell division. In this study we investigated the real-time polymerization of MreB in Caulobacter crescentus using single-molecule fluorescence imaging. With time-lapse imaging, polymerized MreB could be distinguished from cytoplasmic MreB monomers, because single monomeric MreB showed fast motion characteristic of Brownian diffusion, while single polymerized MreB displayed slow, directed motion. This directional movement of labeled MreB in the growing polymer implies that treadmilling is the predominant mechanism in MreB filament formation. These single-molecule imaging experiments provide the first available information on the velocity of bacterial actin polymerization in a living cell.

  7. Unusual bacterioplankton community structure in ultra-oligotrophic Crater Lake

    Science.gov (United States)

    Urbach, Ena; Vergin, Kevin L.; Morse, Ariel

    2001-01-01

    The bacterioplankton assemblage in Crater Lake, Oregon (U.S.A.), is different from communities found in other oxygenated lakes, as demonstrated by four small subunit ribosomal ribonucleic acid (SSU rRNA) gene clone libraries and oligonucleotide probe hybridization to RNA from lake water. Populations in the euphotic zone of this deep (589 m), oligotrophic caldera lake are dominated by two phylogenetic clusters of currently uncultivated bacteria: CL120-10, a newly identified cluster in the verrucomicrobiales, and ACK4 actinomycetes, known as a minor constituent of bacterioplankton in other lakes. Deep-water populations at 300 and 500 m are dominated by a different pair of uncultivated taxa: CL500-11, a novel cluster in the green nonsulfur bacteria, and group I marine crenarchaeota. b-Proteobacteria, dominant in most other freshwater environments, are relatively rare in Crater Lake (marine crenarchaeota and green nonsulfur bacteria. Comparison of Crater Lake to other lakes studied by rRNA methods suggests that selective factors structuring Crater Lake bacterioplankton populations may include low concentrations of available trace metals and dissolved organic matter, chemistry of infiltrating hydrothermal waters, and irradiation by high levels of ultraviolet light.

  8. Flagellar Motor Switching in Caulobacter Crescentus Obeys First Passage Time Statistics

    Science.gov (United States)

    Morse, Michael; Bell, Jordan; Li, Guanglai; Tang, Jay X.

    2015-11-01

    A Caulobacter crescentus swarmer cell is propelled by a helical flagellum, which is rotated by a motor at its base. The motor alternates between rotating in clockwise and counterclockwise directions and spends variable intervals of time in each state. We measure the distributions of these intervals for cells either free swimming or tethered to a glass slide. A peak time of around one second is observed in the distributions for both motor directions with counterclockwise intervals more sharply peaked and clockwise intervals displaying a larger tail at long times. We show that distributions of rotation intervals fit first passage time statistics for a biased random walker and the dynamic binding of CheY-P to FliM motor subunits accounts for this behavior. Our results also suggest that the presence of multiple CheY proteins in C. crescentus may be responsible for differences between its switching behavior and that of the extensively studied E. coli.

  9. A NAD-dependent glutamate dehydrogenase coordinates metabolism with cell division in Caulobacter crescentus.

    Science.gov (United States)

    Beaufay, François; Coppine, Jérôme; Mayard, Aurélie; Laloux, Géraldine; De Bolle, Xavier; Hallez, Régis

    2015-07-01

    Coupling cell cycle with nutrient availability is a crucial process for all living cells. But how bacteria control cell division according to metabolic supplies remains poorly understood. Here, we describe a molecular mechanism that coordinates central metabolism with cell division in the α-proteobacterium Caulobacter crescentus. This mechanism involves the NAD-dependent glutamate dehydrogenase GdhZ and the oxidoreductase-like KidO. While enzymatically active GdhZ directly interferes with FtsZ polymerization by stimulating its GTPase activity, KidO bound to NADH destabilizes lateral interactions between FtsZ protofilaments. Both GdhZ and KidO share the same regulatory network to concomitantly stimulate the rapid disassembly of the Z-ring, necessary for the subsequent release of progeny cells. Thus, this mechanism illustrates how proteins initially dedicated to metabolism coordinate cell cycle progression with nutrient availability.

  10. Alternative mechanism for bacteriophage adsorption to the motile bacterium Caulobacter crescentus.

    Science.gov (United States)

    Guerrero-Ferreira, Ricardo C; Viollier, Patrick H; Ely, Bert; Poindexter, Jeanne S; Georgieva, Maria; Jensen, Grant J; Wright, Elizabeth R

    2011-06-14

    2D and 3D cryo-electron microscopy, together with adsorption kinetics assays of Cb13 and CbK phage-infected Caulobacter crescentus, provides insight into the mechanisms of infection. Cb13 and CbK actively interact with the flagellum and subsequently attach to receptors on the cell pole. We present evidence that the first interaction of the phage with the bacterial flagellum takes place through a filament on the phage head. This contact with the flagellum facilitates concentration of phage particles around the receptor (i.e., the pilus portals) on the bacterial cell surface, thereby increasing the likelihood of infection. Phage head filaments have not been well characterized and their function is described here. Phage head filaments may systematically underlie the initial interactions of phages with their hosts in other systems and possibly represent a widespread mechanism of efficient phage propagation.

  11. The curved shape of Caulobacter crescentus enhances surface colonization in flow

    Science.gov (United States)

    Persat, Alexandre; Stone, Howard A.; Gitai, Zemer

    2014-05-01

    Each bacterial species has a characteristic shape, but the benefits of specific morphologies remain largely unknown. To understand potential functions for cell shape, we focused on the curved bacterium Caulobacter crescentus. Paradoxically, C. crescentus curvature is robustly maintained in the wild but straight mutants have no known disadvantage in standard laboratory conditions. Here we demonstrate that cell curvature enhances C. crescentus surface colonization in flow. Imaging the formation of microcolonies at high spatial and temporal resolution indicates that flow causes curved cells to orient such that they arc over the surface, thereby decreasing the distance between the surface and polar adhesive pili, and orienting pili to face the surface. C. crescentus thus repurposes pilus retraction, typically used for surface motility, for surface attachment. The benefit provided by curvature is eliminated at high flow intensity, raising the possibility that diversity in curvature adapts related species for life in different flow environments.

  12. ppGpp and polyphosphate modulate cell cycle progression in Caulobacter crescentus.

    Science.gov (United States)

    Boutte, Cara C; Henry, Jonathan T; Crosson, Sean

    2012-01-01

    Caulobacter crescentus differentiates from a motile, foraging swarmer cell into a sessile, replication-competent stalked cell during its cell cycle. This developmental transition is inhibited by nutrient deprivation to favor the motile swarmer state. We identify two cell cycle regulatory signals, ppGpp and polyphosphate (polyP), that inhibit the swarmer-to-stalked transition in both complex and glucose-exhausted media, thereby increasing the proportion of swarmer cells in mixed culture. Upon depletion of available carbon, swarmer cells lacking the ability to synthesize ppGpp or polyP improperly initiate chromosome replication, proteolyze the replication inhibitor CtrA, localize the cell fate determinant DivJ, and develop polar stalks. Furthermore, we show that swarmer cells produce more ppGpp than stalked cells upon starvation. These results provide evidence that ppGpp and polyP are cell-type-specific developmental regulators.

  13. Estudo da regulação do gene cspD de Caulobacter crescentus.

    OpenAIRE

    Carolina Antunes do Prado Tavares Silva

    2011-01-01

    CspD é uma das quatro proteínas de choque frio de Caulobacter crescentus, sendo maior que as outras CSPs por possuir dois domínios de choque frio, e tem seu papel na célula ainda desconhecido. O objetivo deste trabalho foi identificar e caracterizar os fatores in cis e in trans envolvidos na regulação da expressão do gene cspD em C. crescentus. Neste trabalho foi visto que a expressão de cspD é induzida pela carência de glicose no meio, mas não pela carência de nitrogênio. Esta indução é depe...

  14. The core and O-polysaccharide structure of the Caulobacter crescentus lipopolysaccharide.

    Science.gov (United States)

    Jones, Michael D; Vinogradov, Evgeny; Nomellini, John F; Smit, John

    2015-01-30

    Here we describe the analysis of the structure of the lipopolysaccharide (LPS) from Caulobacter crescentus strain JS1025, a derivative of C. crescentus CB15 NA1000 with an engineered amber mutation in rsaA, leading to the loss of the protein S-layer and gene CCNA_00471 encoding a putative GDP-L-fucose synthase. LPS was isolated using an aqueous membrane disruption method. Polysaccharide and core oligosaccharide were produced by mild acid hydrolysis and analyzed by nuclear magnetic resonance spectroscopy and chemical methods. Spectra revealed the presence of two polysaccharides, one of them, a rhamnan, could be removed using periodate oxidation. Another polymer, built from 4-amino-4-deoxy-D-rhamnose (perosamine), mannose, and 3-O-methyl-glucose, should be the O-chain of the LPS according to genetic data. The attribution of the rhamnan as a part of LPS or a separate polymer was not possible.

  15. Quantitative Selection Analysis of Bacteriophage φCbK Susceptibility in Caulobacter crescentus.

    Science.gov (United States)

    Christen, Matthias; Beusch, Christian; Bösch, Yvonne; Cerletti, Dario; Flores-Tinoco, Carlos Eduardo; Del Medico, Luca; Tschan, Flavia; Christen, Beat

    2016-01-29

    Classical molecular genetics uses stringent selective conditions to identify mutants with distinct phenotypic responses. Mutations giving rise to less pronounced phenotypes are often missed. However, to gain systems-level insights into complex genetic interaction networks requires genome-wide assignment of quantitative phenotypic traits. In this paper, we present a quantitative selection approach coupled with transposon sequencing (QS-TnSeq) to globally identify the cellular components that orchestrate susceptibility of the cell cycle model bacterium Caulobacter crescentus toward bacteriophage φCbK infection. We found that 135 genes representing 3.30% of the Caulobacter genome exhibit significant accumulation of transposon insertions upon φCbK selection. More than 85% thereof consist of new factors not previously associated with phage φCbK susceptibility. Using hierarchical clustering of dose-dependent TnSeq datasets, we grouped these genes into functional modules that correlate with different stages of the φCbK infection process. We assign φCbK susceptibility to eight new genes that represent novel components of the pilus secretion machinery. Further, we demonstrate that, from 86 motility genes, only seven genes encoding structural and regulatory components of the flagellar hook increase phage resistance when disrupted by transposons, suggesting a link between flagellar hook assembly and pili biogenesis. In addition, we observe high recovery of Tn5 insertions within regulatory sequences of the genes encoding the essential NADH:ubiquinone oxidoreductase complex indicating that intact proton motive force is crucial for effective phage propagation. In sum, QS-TnSeq is broadly applicable to perform quantitative and genome-wide systems-genetics analysis of complex phenotypic traits.

  16. Identification of ClpP substrates in Caulobacter crescentus reveals a role for regulated proteolysis in bacterial development.

    Science.gov (United States)

    Bhat, Nowsheen H; Vass, Robert H; Stoddard, Patrick R; Shin, Dong K; Chien, Peter

    2013-06-01

    Energy-dependent proteases ensure the timely removal of unwanted proteins in a highly selective fashion. In Caulobacter crescentus, protein degradation by the ClpXP protease is critical for cell cycle progression; however, only a handful of substrates are currently known. Here, we use a trapping approach to identify putative substrates of the ClpP associated proteases in C. crescentus. Biochemical validation of several of these targets reveals specific protease recognition motifs and suggests a need for ClpXP-specific degradation beyond degradation of known cell cycle regulators. We focus on a particular instance of regulated proteolysis in Caulobacter by exploring the role of ClpXP in degrading the stalk synthesis transcription factor TacA. We show that TacA degradation is controlled during the cell cycle dependent on the ClpXP regulator CpdR and that stabilization of TacA increases degradation of another ClpXP substrate, CtrA, while restoring deficiencies associated with prolific CpdR activity. Together, our work reveals a number of new validated ClpXP substrates, clarifies rules of protease substrate selection, and demonstrates how regulated protein degradation is critical for Caulobacter development and cell cycle progression.

  17. Causes of change in nitrophytic and oligotrophic lichen species in a Mediterranean climate: Impact of land cover and atmospheric pollutants

    Energy Technology Data Exchange (ETDEWEB)

    Pinho, P.; Augusto, S.; Martins-Loucao, M.A. [Faculdade de Ciencias, Centro de Ecologia e Biologia Vegetal, Universidade de Lisboa, edificio C4, 1749-016 Lisbon (Portugal); Pereira, M.J.; Soares, A. [Instituto Superior Tecnico, Universidade Tecnica de Lisboa, Cerena, Av. Rovisco Pais, 1049-001 Lisbon (Portugal); Maguas, C. [Faculdade de Ciencias, Centro de Ecologia e Biologia Vegetal, Universidade de Lisboa, edificio C4, 1749-016 Lisbon (Portugal); Branquinho, C. [Faculdade de Ciencias, Centro de Ecologia e Biologia Vegetal, Universidade de Lisboa, edificio C4, 1749-016 Lisbon (Portugal); Antiga Fabrica da Polvora de Barcarena, Universidade Atlantica, 2745-615 Barcarena (Portugal)], E-mail: cmbranquinho@fc.ul.pt

    2008-08-15

    With the aim of determining the main drivers of changes in nitrophytic and oligotrophic macro-lichen communities in an industrial region with a Mediterranean climate, we considered both land-cover types and atmospheric pollutants. We determined the relation between the abundance of nitrophytic and oligotrophic species with environmental factors considering the distance of influence of land-cover types. The results showed that oligotrophic species decreased in the proximity of artificial areas, barren land and agricultural areas, associated with higher concentrations of NO{sub 2} and Zn, and Ti, probably dust of industrial and agricultural origin. Nitrophytic species were positively related to all the mentioned land-cover types, and with higher concentrations of Fe and N. Magnesium, probably from ocean aerosols, was negatively related to oligotrophic species and positively to nitrophytic. - Causes of change in nitrophytic and oligotrophic lichen species.

  18. Unprecedented slow growth and mortality of the rare colonial cyanobacterium, Nostoc zetterstedtii, in oligotrophic lakes

    DEFF Research Database (Denmark)

    Jensen, Kaj Sand; Møller, Claus Lindskov

    2011-01-01

    Centimeter-large colonies of Nostoc zetterstedtii from a Swedish oligotrophic lake had the lowest growth and mortality rates of any studied temperate macrophyte. Annual growth rates at two shallow sites averaged 0.57– 0.73 3 1023 d21, corresponding to doubling times of colony dry weight in 2...

  19. Importance of groundwater and macrophytes for the nutrient balance at oligotrophic Lake Hampen, Denmark

    DEFF Research Database (Denmark)

    Ommen, Daniela Oliveira; Kidmose, Jacob Baarstrøm; Karan, Sachin

    2012-01-01

    An ecohydrological study was carried out at an oligotrophic seepage lake, Lake Hampen, Denmark, to determine the seepage in and out of the lake and determine the mass budgets for nutrients. The lake is primarily surrounded by forest, although there are agricultural fields bordering a small portio...

  20. On some ecological and faunistic pecularities of fish parasites in large oligotrophic lakes of Karelia

    Directory of Open Access Journals (Sweden)

    Rumjantsev Yevgeny

    2014-11-01

    Full Text Available The ecological and faunistic pecularities of parasite fauna of fishes in large oligotrophic lakes of Karelia were studied. These lakes belong typologically to different classes. The presence of glacial species of parasites in Onega Lake and sea relict representatives in Ladoga lake is shown.

  1. Control of cell division and the spatial localization of assembled gene products in Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Nathan, P.D.

    1988-01-01

    Experiments are described that examine the role of penicillin-binding proteins (PBPs) in the regulation of cell division in Caulobacter crescentus; and the spatial localization of methyl-accepting chemotaxis proteins (MCPs) in C. crescentus swarmer and predivisional cells. In the analysis of PBP function, in vivo and in vitro assays are used to directly label C. crescentus PBPs with (/sup 3/H) penicillin G in wild type strain CB15, in a series of conditional cell division mutants and in new temperature sensitive cephalosporin C resistant mutants PC8002 and PC8003. 14 PBPs are characterized and a high molecular weight PBP (PBP 1B) that is required for cell division is identified. PBP 1B competes for ..beta..-lactams that induce filament formation and may be a high affinity binding protein. A second high molecular weight PBP (PBP 1C) is also associated with defective cell division. The examination of PBP patterns in synchronous swarmer cells reveals that the in vivo activity of PBP 1B and PBP 1C increases at the time that the cell division pathway is initiated. None of the PBPs, however, appear to be differentially localized in the C. crescentus cell. In the analysis of MCP localization, in vivo and in vitro assays are used to directly label C. crescentus MCPs with methyl-/sup 3/H. MCPs are examined in flagellated and non-flagellated vesicles prepared from cells by immunoaffinity chromatography.

  2. Physiological consequences of blocked Caulobacter crescentus dnaA expression, an essential DNA replication gene.

    Science.gov (United States)

    Gorbatyuk, B; Marczynski, G T

    2001-04-01

    Caulobacter crescentus chromosome replication is precisely coupled to a developmental cell cycle. Like most eubacteria, C. crescentus has a DnaA homologue that is presumed to initiate chromosome replication. However, the C. crescentus replication origin (Cori) lacks perfect consensus Escherichia coli DnaA boxes. Instead, the Cori strong transcription promoter (Ps) may regulate chromosome replication through the CtrA cell cycle response regulator. We therefore created a conditional dnaA C. crescentus strain. Blocking dnaA expression immediately decreased DNA synthesis, which stopped after approximately one doubling period. Fluorescent flow cytometry confirmed that DNA synthesis is blocked at the initiation stage. Cell division also stopped, but not swarmer to stalked cell differentiation. All cells became stalked cells that grew as long filaments. Therefore, general transcription and protein synthesis continued, whereas DNA synthesis stopped. However, transcription was selectively blocked from the flagellar fliQ and fliL and methyltransferase ccrM promoters, which require CtrA and are blocked by different DNA synthesis inhibitors. Interestingly, transcription from Cori Ps continued unaltered. Therefore, Ps transcription is not sufficient for chromosome replication. Approximately 6-8 h after blocked dnaA expression, cells lost viability exponentially. Coincidentally, beta-galactosidase was induced from one transcription reporter, suggesting an altered physiology. We conclude that C. crescentus DnaA is essential for chromosome replication initiation, and perhaps also has a wider role in cell homeostasis.

  3. Biomineralization of Uranium by PhoY Phosphatase Activity Aids Cell Survival in Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Yung, M C [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Jiao, Y [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2014-07-22

    Caulobacter crescentus is known to tolerate high levels of uranium [U(VI)], but its detoxification mechanism is poorly understood. Here we show that C. crescentus is able to facilitate U(VI) biomineralization through the formation of U-Pi precipitates via its native alkaline phosphatase activity. The U-Pi precipitates, deposited on the cell surface in the form of meta-autunite structures, have a lower U/Pi ratio than do chemically produced precipitates. The enzyme that is responsible for the phosphatase activity and thus the biomineralization process is identified as PhoY, a periplasmic alkaline phosphatase with broad substrate specificity. Furthermore, PhoY is shown to confer a survival advantage on C. crescentus toward U(VI) under both growth and nongrowth conditions. Results obtained in this study thus highlight U(VI) biomineralization as a resistance mechanism in microbes, which not only improves our understanding of bacterium-mineral interactions but also aids in defining potential ecological niches for metal-resistant bacteria.

  4. A physical approach to segregation and folding of the Caulobacter crescentus genome.

    Science.gov (United States)

    Dame, Remus T; Tark-Dame, Mariliis; Schiessel, Helmut

    2011-12-01

    Bacterial genomes are functionally organized. This organization is dynamic and globally changing throughout the cell cycle. Upon initiation of replication of the chromosome, the two origins segregate and move towards their new location taking along the newly replicated genome. Caulobacter crescentus employs a dedicated active partitioning (Par) system to move one copy of the parS centromere to the distal pole, while the other stays at the stalked pole. In this issue of Molecular Microbiology, Hong and McAdams describe studies on the speed of segregation of parS and regions up to 150 kb away. They show clear differences in segregation rates between parS and 50 kb flanking regions versus regions further away. To assess segregation rates the authors track fluorescent markers during movement using time-lapse microscopy. The relation between genomic and physical distance of pairs of markers reflects how the genome is folded. This relation permits testing experimental data against models from polymer physics. Such models are helpful in understanding principles of genome folding. Although long used in studies on eukaryotes, this approach has rarely been applied to bacteria. Finally, the authors give the first direct evidence for a role of the bacterial chromatin protein HU in folding the genome in vivo.

  5. The flagellar motor of Caulobacter crescentus generates more torque when a cell swims backwards

    Science.gov (United States)

    Lele, Pushkar P.; Roland, Thibault; Shrivastava, Abhishek; Chen, Yihao; Berg, Howard C.

    2016-02-01

    The bacterium Caulobacter crescentus swims by rotating a single right-handed helical filament. These cells have two swimming modes: a pusher mode, in which clockwise (CW) rotation of the filament thrusts the cell body forwards, and a puller mode, in which counterclockwise (CCW) rotation pulls it backwards. The situation is reversed in Escherichia coli, a bacterium that rotates several left-handed filaments CCW to drive the cell body forwards. The flagellar motor in E. coli generates more torque in the CCW direction than the CW direction in swimming cells. However, C. crescentus and other bacteria with single filaments swim forwards and backwards at similar speeds, prompting the assumption that motor torques in the two modes are the same. Here, we present evidence that motors in C. crescentus develop higher torques in the puller mode than in the pusher mode, and suggest that the anisotropy in torque generation is similar in the two species, despite the differences in filament handedness and motor bias.

  6. Potential role of a bistable histidine kinase switch in the asymmetric division cycle of Caulobacter crescentus.

    Science.gov (United States)

    Subramanian, Kartik; Paul, Mark R; Tyson, John J

    2013-01-01

    The free-living aquatic bacterium, Caulobacter crescentus, exhibits two different morphologies during its life cycle. The morphological change from swarmer cell to stalked cell is a result of changes of function of two bi-functional histidine kinases, PleC and CckA. Here, we describe a detailed molecular mechanism by which the function of PleC changes between phosphatase and kinase state. By mathematical modeling of our proposed molecular interactions, we derive conditions under which PleC, CckA and its response regulators exhibit bistable behavior, thus providing a scenario for robust switching between swarmer and stalked states. Our simulations are in reasonable agreement with in vitro and in vivo experimental observations of wild type and mutant phenotypes. According to our model, the kinase form of PleC is essential for the swarmer-to-stalked transition and to prevent premature development of the swarmer pole. Based on our results, we reconcile some published experimental observations and suggest novel mutants to test our predictions.

  7. Dynamical modeling of the cell cycle and cell fate emergence in Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    César Quiñones-Valles

    Full Text Available The division of Caulobacter crescentus, a model organism for studying cell cycle and differentiation in bacteria, generates two cell types: swarmer and stalked. To complete its cycle, C. crescentus must first differentiate from the swarmer to the stalked phenotype. An important regulator involved in this process is CtrA, which operates in a gene regulatory network and coordinates many of the interactions associated to the generation of cellular asymmetry. Gaining insight into how such a differentiation phenomenon arises and how network components interact to bring about cellular behavior and function demands mathematical models and simulations. In this work, we present a dynamical model based on a generalization of the Boolean abstraction of gene expression for a minimal network controlling the cell cycle and asymmetric cell division in C. crescentus. This network was constructed from data obtained from an exhaustive search in the literature. The results of the simulations based on our model show a cyclic attractor whose configurations can be made to correspond with the current knowledge of the activity of the regulators participating in the gene network during the cell cycle. Additionally, we found two point attractors that can be interpreted in terms of the network configurations directing the two cell types. The entire network is shown to be operating close to the critical regime, which means that it is robust enough to perturbations on dynamics of the network, but adaptable to environmental changes.

  8. Probing flagellar promoter occupancy in wild-type and mutant Caulobacter crescentus by chromatin immunoprecipitation.

    Science.gov (United States)

    Davis, Nicole J; Viollier, Patrick H

    2011-06-01

    In the asymmetric predivisional cell of Caulobacter crescentus, TipF and TipN mark the cellular pole for future flagellar development. TipF is essential for motility and contains a cyclic-di-GMP phosphodiesterase-like (EAL) domain that is necessary for proper function. TipN is localized to the flagellar pole before TipF and is essential for the proper placement of the flagellum in C. crescentus. Using β-galactosidase promoter-probe assays and quantitative chromatin immunoprecipitation, we investigated the influence of the C. crescentus flagellar assembly regulator TipF on flagellar gene transcription. We compared the transcriptional activity of class II-fliF-lacZ, class III-flgE-lacZ, and class IV-fljL-lacZ fusions in a ΔtipF mutant with that of other flagellar mutants and the wild-type strain. We subsequently verified the in vivo occupancy of the fliF, flgE, and fljL flagellar promoters by the flagellar regulators CtrA, FlbD, and FliX in addition to RNA polymerase. We deduce that TipF contributes to proper expression of flagellar genes in C. crescentus by acting both within and outside of the canonical flagellar gene expression hierarchy.

  9. Biomineralization of uranium by PhoY phosphatase activity aids cell survival in Caulobacter crescentus.

    Science.gov (United States)

    Yung, Mimi C; Jiao, Yongqin

    2014-08-01

    Caulobacter crescentus is known to tolerate high levels of uranium [U(VI)], but its detoxification mechanism is poorly understood. Here we show that C. crescentus is able to facilitate U(VI) biomineralization through the formation of U-Pi precipitates via its native alkaline phosphatase activity. The U-Pi precipitates, deposited on the cell surface in the form of meta-autunite structures, have a lower U/Pi ratio than do chemically produced precipitates. The enzyme that is responsible for the phosphatase activity and thus the biomineralization process is identified as PhoY, a periplasmic alkaline phosphatase with broad substrate specificity. Furthermore, PhoY is shown to confer a survival advantage on C. crescentus toward U(VI) under both growth and nongrowth conditions. Results obtained in this study thus highlight U(VI) biomineralization as a resistance mechanism in microbes, which not only improves our understanding of bacterium-mineral interactions but also aids in defining potential ecological niches for metal-resistant bacteria.

  10. The functions of DNA methylation by CcrM in Caulobacter crescentus: a global approach.

    Science.gov (United States)

    Gonzalez, Diego; Kozdon, Jennifer B; McAdams, Harley H; Shapiro, Lucy; Collier, Justine

    2014-04-01

    DNA methylation is involved in a diversity of processes in bacteria, including maintenance of genome integrity and regulation of gene expression. Here, using Caulobacter crescentus as a model, we exploit genome-wide experimental methods to uncover the functions of CcrM, a DNA methyltransferase conserved in most Alphaproteobacteria. Using single molecule sequencing, we provide evidence that most CcrM target motifs (GANTC) switch from a fully methylated to a hemi-methylated state when they are replicated, and back to a fully methylated state at the onset of cell division. We show that DNA methylation by CcrM is not required for the control of the initiation of chromosome replication or for DNA mismatch repair. By contrast, our transcriptome analysis shows that >10% of the genes are misexpressed in cells lacking or constitutively over-expressing CcrM. Strikingly, GANTC methylation is needed for the efficient transcription of dozens of genes that are essential for cell cycle progression, in particular for DNA metabolism and cell division. Many of them are controlled by promoters methylated by CcrM and co-regulated by other global cell cycle regulators, demonstrating an extensive cross talk between DNA methylation and the complex regulatory network that controls the cell cycle of C. crescentus and, presumably, of many other Alphaproteobacteria.

  11. A DNA damage-induced, SOS-independent checkpoint regulates cell division in Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Joshua W Modell

    2014-10-01

    Full Text Available Cells must coordinate DNA replication with cell division, especially during episodes of DNA damage. The paradigm for cell division control following DNA damage in bacteria involves the SOS response where cleavage of the transcriptional repressor LexA induces a division inhibitor. However, in Caulobacter crescentus, cells lacking the primary SOS-regulated inhibitor, sidA, can often still delay division post-damage. Here we identify didA, a second cell division inhibitor that is induced by DNA damage, but in an SOS-independent manner. Together, DidA and SidA inhibit division, such that cells lacking both inhibitors divide prematurely following DNA damage, with lethal consequences. We show that DidA does not disrupt assembly of the division machinery and instead binds the essential division protein FtsN to block cytokinesis. Intriguingly, mutations in FtsW and FtsI, which drive the synthesis of septal cell wall material, can suppress the activity of both SidA and DidA, likely by causing the FtsW/I/N complex to hyperactively initiate cell division. Finally, we identify a transcription factor, DriD, that drives the SOS-independent transcription of didA following DNA damage.

  12. Potential role of a bistable histidine kinase switch in the asymmetric division cycle of Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Kartik Subramanian

    Full Text Available The free-living aquatic bacterium, Caulobacter crescentus, exhibits two different morphologies during its life cycle. The morphological change from swarmer cell to stalked cell is a result of changes of function of two bi-functional histidine kinases, PleC and CckA. Here, we describe a detailed molecular mechanism by which the function of PleC changes between phosphatase and kinase state. By mathematical modeling of our proposed molecular interactions, we derive conditions under which PleC, CckA and its response regulators exhibit bistable behavior, thus providing a scenario for robust switching between swarmer and stalked states. Our simulations are in reasonable agreement with in vitro and in vivo experimental observations of wild type and mutant phenotypes. According to our model, the kinase form of PleC is essential for the swarmer-to-stalked transition and to prevent premature development of the swarmer pole. Based on our results, we reconcile some published experimental observations and suggest novel mutants to test our predictions.

  13. Dynamical modeling of the cell cycle and cell fate emergence in Caulobacter crescentus.

    Science.gov (United States)

    Quiñones-Valles, César; Sánchez-Osorio, Ismael; Martínez-Antonio, Agustino

    2014-01-01

    The division of Caulobacter crescentus, a model organism for studying cell cycle and differentiation in bacteria, generates two cell types: swarmer and stalked. To complete its cycle, C. crescentus must first differentiate from the swarmer to the stalked phenotype. An important regulator involved in this process is CtrA, which operates in a gene regulatory network and coordinates many of the interactions associated to the generation of cellular asymmetry. Gaining insight into how such a differentiation phenomenon arises and how network components interact to bring about cellular behavior and function demands mathematical models and simulations. In this work, we present a dynamical model based on a generalization of the Boolean abstraction of gene expression for a minimal network controlling the cell cycle and asymmetric cell division in C. crescentus. This network was constructed from data obtained from an exhaustive search in the literature. The results of the simulations based on our model show a cyclic attractor whose configurations can be made to correspond with the current knowledge of the activity of the regulators participating in the gene network during the cell cycle. Additionally, we found two point attractors that can be interpreted in terms of the network configurations directing the two cell types. The entire network is shown to be operating close to the critical regime, which means that it is robust enough to perturbations on dynamics of the network, but adaptable to environmental changes.

  14. The curved shape of the bacterium Caulobacter crescentus enhances colonization of surfaces in flow

    Science.gov (United States)

    Persat, Alexandre; Gitai, Zemer; Stone, Howard

    2014-11-01

    Bacteria thrive in all types of fluid environments; flow is thus a ubiquitous aspect of their lives. Bacteria have evolved a variety of cellular components contributing to their growth in specific environments. However, cellular features that help them survive and develop in flow have been rarely characterized. Here, we show that Caulobacter crescentus may have evolved its curved shape to enhance the colonization of surfaces in flow. C. crescentus curvature is preserved in the wild but straight mutants have no known growth disadvantage in standard laboratory conditions. Leveraging microfluidics and single-cell imaging, we demonstrate that curvature enhances surface colonization in flow, promoting the formation of larger microcolonies. Cells attach to a surface from a single pole, so that flow affects their orientation. In flow, viscous forces generate a torque on the curved cell body, which reorients the cell in the direction of the flow. The curved cell appears to arc above the surface, optimally orienting its unattached pole towards the surface. This reduces the distance between the surface and the pole, thereby enhancing attachment of its progeny. Additionally, we show that curved shape enhances colony spreading across the direction of the flow, generating more robust biofilm compared to straight mutants.

  15. Behavior of Caulobacter Crescentus Diagnosed Using a 3-Channel Microfluidic Device

    Science.gov (United States)

    Tang, Jay; Morse, Michael; Colin, Remy; Wilson, Laurence

    2015-03-01

    Many motile microorganisms are able to detect chemical gradients in their surroundings in order to bias their motion towards more favorable conditions. We study the biased motility of Caulobacter crescentus, a singly flagellated bacteria, which alternate between forward and backward swimming, driven by its flagella motor, which switches in rotation direction. We observe the swimming patterns of C. crescents in an oxygen gradient, which is established by flowing atmospheric air and pure nitrogen through a 3 parallel channel microfluidic device. In this setup, oxygen diffuses through the PDMS device and the bacterial medium, creating a linear gradient. Using low magnification, dark field microscopy, individual cells are tracked over a large field of view, with particular interest in the cells' motion relative to the oxygen gradient. Utilizing observable differences between backward and forward swimming motion, motor switching events can be identified. By analyzing these run time intervals between motor switches as a function of a cell's local oxygen level, we demonstrate that C. crescentus displays aerotacitc behavior by extending forward swimming run times while moving up an oxygen gradient, resulting in directed motility towards oxygen sources. Additionally, motor switching response is sensitive to both the steepness of the gradient experienced and background oxygen levels with cells exhibiting a logarithmic response to oxygen levels. Work funded by the United States National Science Foundation and by the Rowland Institute at Harvard University.

  16. Characterization of the Proteins Associated with Caulobacter crescentus Bacteriophage CbK Particles.

    Science.gov (United States)

    Callahan, Courtney T; Wilson, Kiesha M; Ely, Bert

    2016-01-01

    Bacteriophage genomes contain an abundance of genes that code for hypothetical proteins with either a conserved domain or no predicted function. The Caulobacter phage CbK has an unusual shape, designated morphotype B3 that consists of an elongated cylindrical head and a long flexible tail. To identify CbK proteins associated with the phage particle, intact phage particles were subjected to SDS-PAGE, and the resulting protein bands were digested with trypsin and analyzed using MALDI mass spectroscopy to provide peptide molecular weights. These peptide molecular weights were then compared with the peptides that would be generated from the predicted amino acid sequences that are coded by the CbK genome, and the comparison of the actual and predicted peptide masses resulted in the identification of single genes that could code for the set of peptides derived from each of the 20 phage proteins. We also found that CsCl density gradient centrifugation resulted in the separation of empty phage heads, phage heads containing material organized in a spiral, isolated phage tails, and other particulate material from the intact phage particles. This additional material proved to be a good source of additional phage proteins, and preliminary results suggest that it may include a CbK DNA replication complex.

  17. A DNA damage-induced, SOS-independent checkpoint regulates cell division in Caulobacter crescentus.

    Science.gov (United States)

    Modell, Joshua W; Kambara, Tracy K; Perchuk, Barrett S; Laub, Michael T

    2014-10-01

    Cells must coordinate DNA replication with cell division, especially during episodes of DNA damage. The paradigm for cell division control following DNA damage in bacteria involves the SOS response where cleavage of the transcriptional repressor LexA induces a division inhibitor. However, in Caulobacter crescentus, cells lacking the primary SOS-regulated inhibitor, sidA, can often still delay division post-damage. Here we identify didA, a second cell division inhibitor that is induced by DNA damage, but in an SOS-independent manner. Together, DidA and SidA inhibit division, such that cells lacking both inhibitors divide prematurely following DNA damage, with lethal consequences. We show that DidA does not disrupt assembly of the division machinery and instead binds the essential division protein FtsN to block cytokinesis. Intriguingly, mutations in FtsW and FtsI, which drive the synthesis of septal cell wall material, can suppress the activity of both SidA and DidA, likely by causing the FtsW/I/N complex to hyperactively initiate cell division. Finally, we identify a transcription factor, DriD, that drives the SOS-independent transcription of didA following DNA damage.

  18. DipM, a new factor required for peptidoglycan remodelling during cell division in Caulobacter crescentus.

    Science.gov (United States)

    Möll, Andrea; Schlimpert, Susan; Briegel, Ariane; Jensen, Grant J; Thanbichler, Martin

    2010-07-01

    In bacteria, cytokinesis is dependent on lytic enzymes that facilitate remodelling of the cell wall during constriction. In this work, we identify a thus far uncharacterized periplasmic protein, DipM, that is required for cell division and polarity in Caulobacter crescentus. DipM is composed of four peptidoglycan binding (LysM) domains and a C-terminal lysostaphin-like (LytM) peptidase domain. It binds to isolated murein sacculi in vitro, and is recruited to the site of constriction through interaction with the cell division protein FtsN. Mutational analyses showed that the LysM domains are necessary and sufficient for localization of DipM, while its peptidase domain is essential for function. Consistent with a role in cell wall hydrolysis, DipM was found to interact with purified murein sacculi in vitro and to induce cell lysis upon overproduction. Its inactivation causes severe defects in outer membrane invagination, resulting in a significant delay between cytoplasmic compartmentalization and final separation of the daughter cells. Overall, these findings indicate that DipM is a periplasmic component of the C. crescentus divisome that facilitates remodelling of the peptidoglycan layer and, thus, coordinated constriction of the cell envelope during the division process.

  19. Growth medium-dependent glycine incorporation into the peptidoglycan of Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Constantin N Takacs

    Full Text Available The peptidoglycan (PG is a macromolecular component of the bacterial cell wall that maintains the shape and integrity of the cell. The PG of Caulobacter crescentus, unlike that of many other Gram-negative bacteria, has repeatedly been shown to contain significant amounts of glycine. This compositional peculiarity has been deemed an intrinsic characteristic of this species. By performing a comprehensive qualitative and quantitative analysis of the C. crescentus PG by high-performance liquid chromatography (HPLC and mass spectrometry (MS, we show here that glycine incorporation into the C. crescentus PG depends on the presence of exogenous glycine in the growth medium. High levels of glycine were detected at the fifth position of the peptide side chains of PG isolated from C. crescentus cells grown in the complex laboratory medium PYE or in defined medium (M2G supplemented with casamino acids or glycine alone. In contrast, glycine incorporation was undetectable when cells were grown in M2G medium lacking glycine. Remarkably, glycine incorporation into C. crescentus peptidoglycan occurred even in the presence of low millimolar to sub-millimolar concentrations of free glycine. High glycine content in the PG had no obvious effects on growth rates, mode of PG incorporation or cell morphology. Hence, the C. crescentus PG is able to retain its physiological functions in cell growth and morphogenesis despite significant alterations in its composition, in what we deem to be unprecedented plasticity.

  20. Characterization of Caulobacter crescentus response to low temperature and identification of genes involved in freezing resistance.

    Science.gov (United States)

    Mazzon, Ricardo R; Lang, Elza A S; Braz, Vânia S; Marques, Marilis V

    2008-11-01

    Free-living bacteria must respond to a wide range of temperature changes, and have developed specific mechanisms to survive in extreme environments. In this work we describe a remarkable resistance of mesophilic bacterium Caulobacter crescentus to several cycles of freezing at -80 degrees C, which was able to grow at low temperatures. Exponentially growing cells and late stationary-phase cells presented higher freezing resistance at both -20 and -80 degrees C than early stationary-phase cells. Cryotolerance was observed when log-phase cultures grown at 30 degrees C were preincubated at 5, 15 or 20 degrees C before freezing at -20 degrees C. A transposon library was screened to identify mutants sensitive to freezing at -80 degrees C and three strains presenting <10% survival were isolated. Identification of genes disrupted in each mutant showed that they encoded an AddA family DNA helicase, a DEAD/DEAH box RNA helicase and a putative RND (resistance, nodulation, cell division) efflux system component. These strains showed longer generation times than wild-type cells when growing at 15 degrees C, with the RNA helicase mutant presenting a severe growth defect. These analyses suggest that the singular intrinsic resistance to freezing of C. crescentus is in fact a consequence of several independent traits, especially the maintenance of a proper degree of supercoiling of nucleic acids.

  1. A quantitative study of the division cycle of Caulobacter crescentus stalked cells.

    Directory of Open Access Journals (Sweden)

    Shenghua Li

    2008-01-01

    Full Text Available Progression of a cell through the division cycle is tightly controlled at different steps to ensure the integrity of genome replication and partitioning to daughter cells. From published experimental evidence, we propose a molecular mechanism for control of the cell division cycle in Caulobacter crescentus. The mechanism, which is based on the synthesis and degradation of three "master regulator" proteins (CtrA, GcrA, and DnaA, is converted into a quantitative model, in order to study the temporal dynamics of these and other cell cycle proteins. The model accounts for important details of the physiology, biochemistry, and genetics of cell cycle control in stalked C. crescentus cell. It reproduces protein time courses in wild-type cells, mimics correctly the phenotypes of many mutant strains, and predicts the phenotypes of currently uncharacterized mutants. Since many of the proteins involved in regulating the cell cycle of C. crescentus are conserved among many genera of alpha-proteobacteria, the proposed mechanism may be applicable to other species of importance in agriculture and medicine.

  2. Motor Switching Rates in Caulobacter Crescentus Follow First Passage Time Distribution

    Science.gov (United States)

    Tang, Jay; Morse, Michael; Bell, Jordan; Li, Guanglai

    2015-03-01

    The flagellar motor of uni-flagellated bacterium Caulobacter crescentus switches stochastically between clockwise (CW) and counterclockwise (CCW) rotation. We performed measurements of the time intervals between switches in order to gain insight on motor dynamics and regulation. Our measurements were performed both on free swimming cells and tethered cells with their flagella attached to a glass slide. A peak time of approximately one second was observed in both motor directions with counterclockwise intervals more sharply peaked. The distributions of switching times can be fitted using biased first passage time statistics. We present a model of motor switching dynamics, which is controlled by the binding of CheY-P to motor subunits FliM. A lower threshold number of FliM with CheY-P bound triggers a switch in motor rotation from CW to CCW, whereas a higher threshold triggers an opposing switch from CCW to CW. The time intervals between alternating switches may be increased or decreased by regulating CheY-P concentration, resulting in biased directional motion in the cells swimming trajectory over many motor cycles under external spatial or temporal gradients. Work funded by the United States National Science Foundation.

  3. Growth medium-dependent glycine incorporation into the peptidoglycan of Caulobacter crescentus.

    Science.gov (United States)

    Takacs, Constantin N; Hocking, Jason; Cabeen, Matthew T; Bui, Nhat Khai; Poggio, Sebastian; Vollmer, Waldemar; Jacobs-Wagner, Christine

    2013-01-01

    The peptidoglycan (PG) is a macromolecular component of the bacterial cell wall that maintains the shape and integrity of the cell. The PG of Caulobacter crescentus, unlike that of many other Gram-negative bacteria, has repeatedly been shown to contain significant amounts of glycine. This compositional peculiarity has been deemed an intrinsic characteristic of this species. By performing a comprehensive qualitative and quantitative analysis of the C. crescentus PG by high-performance liquid chromatography (HPLC) and mass spectrometry (MS), we show here that glycine incorporation into the C. crescentus PG depends on the presence of exogenous glycine in the growth medium. High levels of glycine were detected at the fifth position of the peptide side chains of PG isolated from C. crescentus cells grown in the complex laboratory medium PYE or in defined medium (M2G) supplemented with casamino acids or glycine alone. In contrast, glycine incorporation was undetectable when cells were grown in M2G medium lacking glycine. Remarkably, glycine incorporation into C. crescentus peptidoglycan occurred even in the presence of low millimolar to sub-millimolar concentrations of free glycine. High glycine content in the PG had no obvious effects on growth rates, mode of PG incorporation or cell morphology. Hence, the C. crescentus PG is able to retain its physiological functions in cell growth and morphogenesis despite significant alterations in its composition, in what we deem to be unprecedented plasticity.

  4. Molecular recognition of RhlB and RNase D in the Caulobacter crescentus RNA degradosome.

    Science.gov (United States)

    Voss, Jarrod E; Luisi, Ben F; Hardwick, Steven W

    2014-12-01

    The endoribonuclease RNase E is a key enzyme in RNA metabolism for many bacterial species. In Escherichia coli, RNase E contributes to the majority of RNA turnover and processing events, and the enzyme has been extensively characterized as the central component of the RNA degradosome assembly. A similar RNA degradosome assembly has been described in the α-proteobacterium Caulobacter crescentus, with the interacting partners of RNase E identified as the Kreb's cycle enzyme aconitase, a DEAD-box RNA helicase RhlB and the exoribonuclease polynucleotide phosphorylase. Here we report that an additional degradosome component is the essential exoribonuclease RNase D, and its recognition site within RNase E is identified. We show that, unlike its E. coli counterpart, C. crescentus RhlB interacts directly with a segment of the N-terminal catalytic domain of RNase E. The crystal structure of a portion of C. crescentus RNase E encompassing the helicase-binding region is reported. This structure reveals that an inserted segment in the S1 domain adopts an α-helical conformation, despite being predicted to be natively unstructured. We discuss the implications of these findings for the organization and mechanisms of the RNA degradosome.

  5. CspC and CspD are essential for Caulobacter crescentus stationary phase survival.

    Science.gov (United States)

    Balhesteros, Heloise; Mazzon, Ricardo R; da Silva, Carolina A P T; Lang, Elza A S; Marques, Marilis V

    2010-09-01

    The cold shock response in bacteria involves the expression of low-molecular weight cold shock proteins (CSPs) containing a nucleic acid-binding cold shock domain (CSD), which are known to destabilize secondary structures on mRNAs, facilitating translation at low temperatures. Caulobacter crescentus cspA and cspB are induced upon cold shock, while cspC and cspD are induced during stationary phase. In this work, we determined a new coding sequence for the cspC gene, revealing that it encodes a protein containing two CSDs. The phenotypes of C. crescentus csp mutants were analyzed, and we found that cspC is important for cells to maintain viability during extended periods in stationary phase. Also, cspC and cspCD strains presented altered morphology, with frequent non-viable filamentous cells, and cspCD also showed a pronounced cell death at late stationary phase. In contrast, the cspAB mutant presented increased viability in this phase, which is accompanied by an altered expression of both cspC and cspD, but the triple cspABD mutant loses this characteristic. Taken together, our results suggest that there is a hierarchy of importance among the csp genes regarding stationary phase viability, which is probably achieved by a fine tune balance of the levels of these proteins.

  6. Mutations in the Lipopolysaccharide biosynthesis pathway interfere with crescentin-mediated cell curvature in Caulobacter crescentus.

    Science.gov (United States)

    Cabeen, Matthew T; Murolo, Michelle A; Briegel, Ariane; Bui, N Khai; Vollmer, Waldemar; Ausmees, Nora; Jensen, Grant J; Jacobs-Wagner, Christine

    2010-07-01

    Bacterial cell morphogenesis requires coordination among multiple cellular systems, including the bacterial cytoskeleton and the cell wall. In the vibrioid bacterium Caulobacter crescentus, the intermediate filament-like protein crescentin forms a cell envelope-associated cytoskeletal structure that controls cell wall growth to generate cell curvature. We undertook a genetic screen to find other cellular components important for cell curvature. Here we report that deletion of a gene (wbqL) involved in the lipopolysaccharide (LPS) biosynthesis pathway abolishes cell curvature. Loss of WbqL function leads to the accumulation of an aberrant O-polysaccharide species and to the release of the S layer in the culture medium. Epistasis and microscopy experiments show that neither S-layer nor O-polysaccharide production is required for curved cell morphology per se but that production of the altered O-polysaccharide species abolishes cell curvature by apparently interfering with the ability of the crescentin structure to associate with the cell envelope. Our data suggest that perturbations in a cellular pathway that is itself fully dispensable for cell curvature can cause a disruption of cell morphogenesis, highlighting the delicate harmony among unrelated cellular systems. Using the wbqL mutant, we also show that the normal assembly and growth properties of the crescentin structure are independent of its association with the cell envelope. However, this envelope association is important for facilitating the local disruption of the stable crescentin structure at the division site during cytokinesis.

  7. A DNA Damage-Induced, SOS-Independent Checkpoint Regulates Cell Division in Caulobacter crescentus

    Science.gov (United States)

    Modell, Joshua W.; Kambara, Tracy K.; Perchuk, Barrett S.; Laub, Michael T.

    2014-01-01

    Cells must coordinate DNA replication with cell division, especially during episodes of DNA damage. The paradigm for cell division control following DNA damage in bacteria involves the SOS response where cleavage of the transcriptional repressor LexA induces a division inhibitor. However, in Caulobacter crescentus, cells lacking the primary SOS-regulated inhibitor, sidA, can often still delay division post-damage. Here we identify didA, a second cell division inhibitor that is induced by DNA damage, but in an SOS-independent manner. Together, DidA and SidA inhibit division, such that cells lacking both inhibitors divide prematurely following DNA damage, with lethal consequences. We show that DidA does not disrupt assembly of the division machinery and instead binds the essential division protein FtsN to block cytokinesis. Intriguingly, mutations in FtsW and FtsI, which drive the synthesis of septal cell wall material, can suppress the activity of both SidA and DidA, likely by causing the FtsW/I/N complex to hyperactively initiate cell division. Finally, we identify a transcription factor, DriD, that drives the SOS-independent transcription of didA following DNA damage. PMID:25350732

  8. Functional characterization of two SOS-regulated genes involved in mitomycin C resistance in Caulobacter crescentus.

    Science.gov (United States)

    Lopes-Kulishev, Carina O; Alves, Ingrid R; Valencia, Estela Y; Pidhirnyj, María I; Fernández-Silva, Frank S; Rodrigues, Ticiane R; Guzzo, Cristiane R; Galhardo, Rodrigo S

    2015-09-01

    The SOS response is a universal bacterial regulon involved in the cellular response to DNA damage and other forms of stress. In Caulobacter crescentus, previous work has identified a plethora of genes that are part of the SOS regulon, but the biological roles of several of them remain to be determined. In this study, we report that two genes, hereafter named mmcA and mmcB, are involved in the defense against DNA damage caused by mitomycin C (MMC), but not against lesions induced by other common DNA damaging agents, such as UVC light, methyl methanesulfonate (MMS) and hydrogen peroxide. mmcA is a conserved gene that encodes a member of the glyoxalases/dioxygenases protein family, and acts independently of known DNA repair pathways. On the other hand, epistasis analysis showed that mmcB acts in the same pathway as imuC (dnaE2), and is required specifically for MMC-induced mutagenesis, but not for that induced by UV light, suggesting a role for MmcB in translesion synthesis-dependent repair of MMC damage. We show that the lack of MMC-induced mutability in the mmcB strain is not caused by lack of proper SOS induction of the imuABC operon, involved in translesion synthesis (TLS) in C. crescentus. Based on this data and on structural analysis of a close homolog, we propose that MmcB is an endonuclease which creates substrates for ImuABC-mediated TLS patches.

  9. The Global Regulatory Architecture of Transcription during the Caulobacter Cell Cycle

    Science.gov (United States)

    Zhou, Bo; Schrader, Jared M.; Kalogeraki, Virginia S.; Abeliuk, Eduardo; Dinh, Cong B.; Pham, James Q.; Cui, Zhongying Z.; Dill, David L.; McAdams, Harley H.; Shapiro, Lucy

    2015-01-01

    Each Caulobacter cell cycle involves differentiation and an asymmetric cell division driven by a cyclical regulatory circuit comprised of four transcription factors (TFs) and a DNA methyltransferase. Using a modified global 5′ RACE protocol, we globally mapped transcription start sites (TSSs) at base-pair resolution, measured their transcription levels at multiple times in the cell cycle, and identified their transcription factor binding sites. Out of 2726 TSSs, 586 were shown to be cell cycle-regulated and we identified 529 binding sites for the cell cycle master regulators. Twenty-three percent of the cell cycle-regulated promoters were found to be under the combinatorial control of two or more of the global regulators. Previously unknown features of the core cell cycle circuit were identified, including 107 antisense TSSs which exhibit cell cycle-control, and 241 genes with multiple TSSs whose transcription levels often exhibited different cell cycle timing. Cumulatively, this study uncovered novel new layers of transcriptional regulation mediating the bacterial cell cycle. PMID:25569173

  10. The global regulatory architecture of transcription during the Caulobacter cell cycle.

    Directory of Open Access Journals (Sweden)

    Bo Zhou

    2015-01-01

    Full Text Available Each Caulobacter cell cycle involves differentiation and an asymmetric cell division driven by a cyclical regulatory circuit comprised of four transcription factors (TFs and a DNA methyltransferase. Using a modified global 5' RACE protocol, we globally mapped transcription start sites (TSSs at base-pair resolution, measured their transcription levels at multiple times in the cell cycle, and identified their transcription factor binding sites. Out of 2726 TSSs, 586 were shown to be cell cycle-regulated and we identified 529 binding sites for the cell cycle master regulators. Twenty-three percent of the cell cycle-regulated promoters were found to be under the combinatorial control of two or more of the global regulators. Previously unknown features of the core cell cycle circuit were identified, including 107 antisense TSSs which exhibit cell cycle-control, and 241 genes with multiple TSSs whose transcription levels often exhibited different cell cycle timing. Cumulatively, this study uncovered novel new layers of transcriptional regulation mediating the bacterial cell cycle.

  11. Distinct constrictive processes, separated in time and space,divide Caulobacter inner and outer membranes

    Energy Technology Data Exchange (ETDEWEB)

    Judd, Ellen M.; Comolli, Luis R.; Chen, Joseph C.; Downing,Kenneth H.; Moerner, W.E.; McAdams, Harley H.

    2005-05-01

    Cryo-electron microscope tomography (cryoEM) and a fluorescence loss in photobleaching (FLIP) assay were used to characterize progression of the terminal stages of Caulobacter crescentus cell division. Tomographic cryoEM images of the cell division site show separate constrictive processes closing first the inner, and then the outer, membrane in a manner distinctly different from septum-forming bacteria. The smallest observed pre-fission constrictions were 60 nm for both the inner and outer membrane. FLIP experiments had previously shown cytoplasmic compartmentalization, when cytoplasmic proteins can no longer diffuse between the two nascent progeny cell compartments, occurring 18 min before daughter cell separation in a 135 min cell cycle. Here, we used FLIP experiments with membrane-bound and periplasmic fluorescent proteins to show that (1) periplasmic compartmentalization occurs after cytoplasmic compartmentalization, consistent with the cryoEM observations, and (2) inner membrane and periplasmic proteins can diffuse past the FtsZ constriction site, indicating that the cell division machinery does not block membrane diffusion.

  12. Factors of ecologic succession in oligotrophic fish communities of the Laurentian Great Lakes

    Science.gov (United States)

    Smith, Stanford H.

    1972-01-01

    Oligotrophic fish communities of the Great Lakes have undergone successive disruptions since the mid-1800s. Major contributing factors have been intensive selective fisheries, extreme modification of the drainage, invasion of marine species, and progressive physical–chemical changes of the lake environments. Lake Ontario was the first to be affected as its basin was settled and industrialized earliest, and it was the first to be connected by canals to the mid-Atlantic where the alewife (Alosa pseudoharengus) and sea lamprey (Petromyzon marinus) which ultimately became established in the Great Lakes were abundant. Oligotrophic fish communities were successively disrupted in Lakes Erie, Huron, Michigan, and Superior as the affects of population growth, industrialization, and marine invaders spread upward in the Laurentian drainage.The degree and sequence of response of families offish and species within families differed for each factor, but the sequence of change among families and species has been the same in response to each factor as it affected various lakes at different times. The ultimate result of the disruption of fish communities has been a reduction of productivity of oligotrophic species that ranges from extreme in Lake Ontario to moderate in Lake Superior, and which has reached a state of instability and rapid change in the upper three Great Lakes by the rnid-1900s similar to the situation in Lake Ontario in the mid-1800s. Since oligotrophic species (primarily salmonines, coregonines, and deepwater cottids) are the only kinds of fish that fully occupied the entire volume of the deepwater Great Lakes (Ontario, Huron, Michigan, and Superior), the fish biomass of these lakes has been reduced as various species declined or disappeared. In Lake Erie, which is shallow, and in the shallow bays of the deep lakes, oligotrophic species were replaced by mesotrophic species, primarily percids, which have successively increased and declined. All oligotrophic

  13. Direct interaction of FliX and FlbD is required for their regulatory activity in Caulobacter crescentus

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    Dutton Rachel J

    2011-05-01

    Full Text Available Abstract Background The temporal and spatial expression of late flagellar genes in Caulobacter crescentus is activated by the transcription factor FlbD and its partner trans-acting factor FliX. The physical interaction of these two proteins represents an alternative mechanism for regulating the activity of σ54 transcription factors. This study is to characterize the interaction of the two proteins and the consequences of the interaction on their regulatory activity. Results FliX and FlbD form stable complexes, which can stand the interference of 2.65 M NaCl. The stability of FliX and FlbD was affected by the co-existence of each other. Five FliX mutants (R71A, L85K, Δ117-118, T130L, and L136K were created by site-directed mutagenesis in conserved regions of the protein. All mutants were successfully expressed in both wild-type and ΔfliX Caulobacter strains. All but FliXL85K could rescue the motility and cell division defects of a ΔfliX mutant strain. The ability of FliX to regulate the transcription of class II and class III/IV flagellar promoters was fully diminished due to the L85K mutation. Co-immunoprecipitation experiment revealed that FliXL85K was unable to physically interact with FlbD. Conclusions FliX interacts with FlbD and thereby directly regulates the activity of FlbD in response to flagellar assembly. Mutations in highly conserved regions of FliX could severely affect the recognition between FliX and FlbD and hence interrupt the normal progression of flagellar synthesis and other developmental events in Caulobacter.

  14. Cloning and cell cycle-dependent expression of DNA replication gene dnaC from Caulobacter crescentus.

    OpenAIRE

    1990-01-01

    Chromosome replication in the asymmetrically dividing bacteria Caulobacter crescentus is discontinuous with the new, motile swarmer cell undergoing an obligatory presynthetic gap period (G1 period) of 60 min before the initiation of DNA synthesis and stalk formation. To examine the regulation of the cell division cycle at the molecular level, we have cloned the DNA chain elongation gene dnaC from a genomic DNA library constructed in cosmid vector pLAFR1-7. To ensure that the cloned sequence c...

  15. Water level influences on body condition of Geophagus brasiliensis (Perciformes: Cichlidae in a Brazilian oligotrophic reservoir

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    Alejandra Filippo Gonzalez Neves dos Santos

    Full Text Available Effects of water level fluctuations on body condition of Geophagus brasiliensis were studied in a 30 km² Brazilian oligotrophic reservoir. Physiological condition (K and gonadosomatic index (GSI were compared according to water level (low and high. Females' best conditions were associated to higher resources availability during high water, since gonad development did not change between low and high water. Males' condition did not change between water levels, while the highest gonad development occurred in low water. Females presented higher reproductive investment than males, which allocated most of energy for somatic development. This strategy could be a mechanism to undergo the stress caused by oligotrophic characteristics of the reservoir enhanced during low water level.

  16. Parallel Evolution and Horizontal Gene Transfer of the pst Operon in Firmicutes from Oligotrophic Environments

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    Alejandra Moreno-Letelier

    2011-01-01

    Full Text Available The high affinity phosphate transport system (pst is crucial for phosphate uptake in oligotrophic environments. Cuatro Cienegas Basin (CCB has extremely low P levels and its endemic Bacillus are closely related to oligotrophic marine Firmicutes. Thus, we expected the pst operon of CCB to share the same evolutionary history and protein similarity to marine Firmicutes. Orthologs of the pst operon were searched in 55 genomes of Firmicutes and 13 outgroups. Phylogenetic reconstructions were performed for the pst operon and 14 concatenated housekeeping genes using maximum likelihood methods. Conserved domains and 3D structures of the phosphate-binding protein (PstS were also analyzed. The pst operon of Firmicutes shows two highly divergent clades with no correlation to the type of habitat nor a phylogenetic congruence, suggesting horizontal gene transfer. Despite sequence divergence, the PstS protein had a similar 3D structure, which could be due to parallel evolution after horizontal gene transfer events.

  17. Eddy-entrained Pearl River plume into the oligotrophic basin of the South China Sea

    Science.gov (United States)

    He, Xianqiang; Xu, Dongfeng; Bai, Yan; Pan, Delu; Chen, Chen-Tung Arthur; Chen, Xiaoyan; Gong, Fang

    2016-08-01

    The South China Sea (SCS) is the world's largest tropical marginal sea with an oligotrophic basin. In June 2015, a rare large phytoplankton bloom, which is ~500 km long, 100 km wide and lasting more than 19 days, was captured in the northern SCS basin by satellite daily chlorophyll images. Water within the bloom area had a feature of low salinity and high temperature measured by an accidental-passing cruise. Meanwhile, satellite sea level anomaly images and drifter trajectory proved there was a cyclonic eddy nearby. No typhoon and heavy rain happened in this period, so we believed the bloom was triggered by the injection of nutrient-rich Pearl River plume driven by eddy. This is the first report on eddy-entrained Pearl River plume into the SCS, which would raise a new view on irregular transportation of nutrient and carbon and its related biogeochemical influence on the oligotrophic ocean.

  18. The Caulobacter crescentus chromosome replication origin evolved two classes of weak DnaA binding sites.

    Science.gov (United States)

    Taylor, James A; Ouimet, Marie-Claude; Wargachuk, Richard; Marczynski, Gregory T

    2011-10-01

    The Caulobacter crescentus replication initiator DnaA and essential response regulator CtrA compete to control chromosome replication. The C. crescentus replication origin (Cori) contains five strong CtrA binding sites but only two apparent DnaA boxes, termed G-boxes (with a conserved second position G, TGATCCACA). Since clusters of DnaA boxes typify bacterial replication origins, this discrepancy suggested that C. crescentus DnaA recognizes different DNA sequences or compensates with novel DNA-binding proteins. We searched for novel DNA sites by scanning mutagenesis of the most conserved Cori DNA. Autonomous replication assays showed that G-boxes and novel W-boxes (TCCCCA) are essential for replication. Further analyses showed that C. crescentus DnaA binds G-boxes with moderate and W-boxes with very weak affinities significantly below DnaA's capacity for high-affinity Escherichia coli-boxes (TTATCCACA). Cori has five conserved W-boxes. Increasing W-box affinities increases or decreases autonomous replication depending on their strategic positions between the G-boxes. In vitro, CtrA binding displaces DnaA from proximal G-boxes and from distal W-boxes implying CtrA-DnaA competition and DnaA-DnaA cooperation between G-boxes and W-boxes. Similarly, during cell cycle progression, CtrA proteolysis coincides with DnaA binding to Cori. We also observe highly conserved W-boxes in other replication origins lacking E. coli-boxes. Therefore, strategically weak DnaA binding can be a general means of replication control. © 2011 Blackwell Publishing Ltd.

  19. Characterization of a unique Caulobacter crescentus aldose-aldose oxidoreductase having dual activities.

    Science.gov (United States)

    Andberg, Martina; Maaheimo, Hannu; Kumpula, Esa-Pekka; Boer, Harry; Toivari, Mervi; Penttilä, Merja; Koivula, Anu

    2016-01-01

    We describe here the characterization of a novel enzyme called aldose-aldose oxidoreductase (Cc AAOR; EC 1.1.99) from Caulobacter crescentus. The Cc AAOR exists in solution as a dimer, belongs to the Gfo/Idh/MocA family and shows homology with the glucose-fructose oxidoreductase from Zymomonas mobilis. However, unlike other known members of this protein family, Cc AAOR is specific for aldose sugars and can be in the same catalytic cycle both oxidise and reduce a panel of monosaccharides at the C1 position, producing in each case the corresponding aldonolactone and alditol, respectively. Cc AAOR contains a tightly-bound nicotinamide cofactor, which is regenerated in this oxidation-reduction cycle. The highest oxidation activity was detected on D-glucose but significant activity was also observed on D-xylose, L-arabinose and D-galactose, revealing that both hexose and pentose sugars are accepted as substrates by Cc AAOR. The configuration at the C2 and C3 positions of the saccharides was shown to be especially important for the substrate binding. Interestingly, besides monosaccharides, Cc AAOR can also oxidise a range of 1,4-linked oligosaccharides having aldose unit at the reducing end, such as lactose, malto- and cello-oligosaccharides as well as xylotetraose. (1)H NMR used to monitor the oxidation and reduction reaction simultaneously, demonstrated that although D-glucose has the highest affinity and is also oxidised most efficiently by Cc AAOR, the reduction of D-glucose is clearly not as efficient. For the overall reaction catalysed by Cc AAOR, the L-arabinose, D-xylose and D-galactose were the most potent substrates.

  20. Expression and characterization of a GH39 β-xylosidase II from Caulobacter crescentus.

    Science.gov (United States)

    Corrêa, Juliana Moço; Graciano, Luciana; Abrahão, Josielle; Loth, Eduardo Alexandre; Gandra, Rinaldo Ferreira; Kadowaki, Marina Kimiko; Henn, Caroline; Simão, Rita de Cássia Garcia

    2012-12-01

    In the present work, the gene xynB2, encoding a β-xylosidase II of the Glycoside Hydrolase 39 (GH39) family, of Caulobacter crescentus was cloned and successfully overexpressed in Escherichia coli DH10B. The recombinant protein (CcXynB2) was purified using nickel-Sepharose affinity chromatography, with a recovery yield of 75.5 %. CcXynB2 appeared as a single band of 60 kDa on a sodium dodecyl sulfate polyacrylamide gel and was recognized by a specific polyclonal antiserum. The predicted CcXynB2 protein showed a high homology with GH39 β-xylosidases of the genus Xanthomonas. CcXynB2 exhibited an optimal activity at 55 °C and a pH of 6. CcXynB2 displayed stability at pH values of 4.5-7.5 for 24 h and thermotolerance up to 50 °C. The K (M) and V (Max) values were 9.3 ± 0.45 mM and 402 ± 19 μmol min(-1) for ρ-nitrophenyl-β-D-xylopyranoside, respectively. The purified recombinant enzyme efficiently produced reducing sugars from birchwood xylan and sugarcane bagasse fibers pre-treated with a purified xylanase. As few bacterial GH39 family β-xylosidases have been characterized, this work provides a good contribution to this group of enzymes.

  1. Function and localization dynamics of bifunctional penicillin-binding proteins in Caulobacter crescentus.

    Science.gov (United States)

    Strobel, Wolfgang; Möll, Andrea; Kiekebusch, Daniela; Klein, Kathrin E; Thanbichler, Martin

    2014-04-01

    The peptidoglycan cell wall of bacteria is a complex macromolecule composed of glycan strands that are cross-linked by short peptide bridges. Its biosynthesis involves a conserved group of enzymes, the bifunctional penicillin-binding proteins (bPBPs), which contain both a transglycosylase and a transpeptidase domain, thus being able to elongate the glycan strands and, at the same time, generate the peptide cross-links. The stalked model bacterium Caulobacter crescentus possesses five bPBP paralogs, named Pbp1A, PbpC, PbpX, PbpY, and PbpZ, whose function is still incompletely understood. In this study, we show that any of these proteins except for PbpZ is sufficient for growth and normal morphogenesis when expressed at native or elevated levels, whereas inactivation of all five paralogs is lethal. Growth analyses indicate a central role of PbpX in the resistance of C. crescentus against the noncanonical amino acid d-alanine. Moreover, we show that PbpX and PbpY localize to the cell division site. Their recruitment to the divisome is dependent on the essential cell division protein FtsN and likely involves interactions with FtsL and the putative peptidoglycan hydrolase DipM. The same interaction pattern is observed for Pbp1A and PbpC, although these proteins do not accumulate at midcell. Our findings demonstrate that the bPBPs of C. crescentus are, to a large extent, redundant and have retained the ability to interact with the peptidoglycan biosynthetic machineries responsible for cell elongation, cytokinesis, and stalk growth. Nevertheless, they may preferentially act in specific peptidoglycan biosynthetic complexes, thereby facilitating the independent regulation of distinct growth processes.

  2. Superresolution imaging in live Caulobacter crescentus cells using photoswitchable enhanced yellow fluorescent protein

    Science.gov (United States)

    Biteen, Julie S.; Thompson, Michael A.; Tselentis, Nicole K.; Shapiro, Lucy; Moerner, W. E.

    2009-02-01

    Recently, photoactivation and photoswitching were used to control single-molecule fluorescent labels and produce images of cellular structures beyond the optical diffraction limit (e.g., PALM, FPALM, and STORM). While previous live-cell studies relied on sophisticated photoactivatable fluorescent proteins, we show in the present work that superresolution imaging can be performed with fusions to the commonly used fluorescent protein EYFP. Rather than being photoactivated, however, EYFP can be reactivated with violet light after apparent photobleaching. In each cycle after initial imaging, only a sparse subset fluorophores is reactivated and localized, and the final image is then generated from the measured single-molecule positions. Because these methods are based on the imaging nanometer-sized single-molecule emitters and on the use of an active control mechanism to produce sparse sub-ensembles, we suggest the phrase "Single-Molecule Active-Control Microscopy" (SMACM) as an inclusive term for this general imaging strategy. In this paper, we address limitations arising from physiologically imposed upper boundaries on the fluorophore concentration by employing dark time-lapse periods to allow single-molecule motions to fill in filamentous structures, increasing the effective labeling concentration while localizing each emitter at most once per resolution-limited spot. We image cell-cycle-dependent superstructures of the bacterial actin protein MreB in live Caulobacter crescentus cells with sub-40-nm resolution for the first time. Furthermore, we quantify the reactivation quantum yield of EYFP, and find this to be 1.6 x 10-6, on par with conventional photoswitchable fluorescent proteins like Dronpa. These studies show that EYFP is a useful emitter for in vivo superresolution imaging of intracellular structures in bacterial cells.

  3. Diversity and bioprospection of fungal community present in oligotrophic soil of continental Antarctica.

    Science.gov (United States)

    Godinho, Valéria M; Gonçalves, Vívian N; Santiago, Iara F; Figueredo, Hebert M; Vitoreli, Gislaine A; Schaefer, Carlos E G R; Barbosa, Emerson C; Oliveira, Jaquelline G; Alves, Tânia M A; Zani, Carlos L; Junior, Policarpo A S; Murta, Silvane M F; Romanha, Alvaro J; Kroon, Erna Geessien; Cantrell, Charles L; Wedge, David E; Duke, Stephen O; Ali, Abbas; Rosa, Carlos A; Rosa, Luiz H

    2015-05-01

    We surveyed the diversity and capability of producing bioactive compounds from a cultivable fungal community isolated from oligotrophic soil of continental Antarctica. A total of 115 fungal isolates were obtained and identified in 11 taxa of Aspergillus, Debaryomyces, Cladosporium, Pseudogymnoascus, Penicillium and Hypocreales. The fungal community showed low diversity and richness, and high dominance indices. The extracts of Aspergillus sydowii, Penicillium allii-sativi, Penicillium brevicompactum, Penicillium chrysogenum and Penicillium rubens possess antiviral, antibacterial, antifungal, antitumoral, herbicidal and antiprotozoal activities. Bioactive extracts were examined using (1)H NMR spectroscopy and detected the presence of secondary metabolites with chemical shifts. Our results show that the fungi present in cold-oligotrophic soil from Antarctica included few dominant species, which may have important implications for understanding eukaryotic survival in cold-arid oligotrophic soils. We hypothesize that detailed further investigations may provide a greater understanding of the evolution of Antarctic fungi and their relationships with other organisms described in that region. Additionally, different wild pristine bioactive fungal isolates found in continental Antarctic soil may represent a unique source to discover prototype molecules for use in drug and biopesticide discovery studies.

  4. Saharan dust inputs and high UVR levels jointly alter the metabolic balance of marine oligotrophic ecosystems

    Science.gov (United States)

    Cabrerizo, Marco J.; Medina-Sánchez, Juan Manuel; González-Olalla, Juan Manuel; Villar-Argaiz, Manuel; Carrillo, Presentación

    2016-10-01

    The metabolic balance of the most extensive bioma on the Earth is a controversial topic of the global-change research. High ultraviolet radiation (UVR) levels by the shoaling of upper mixed layers and increasing atmospheric dust deposition from arid regions may unpredictably alter the metabolic state of marine oligotrophic ecosystems. We performed an observational study across the south-western (SW) Mediterranean Sea to assess the planktonic metabolic balance and a microcosm experiment in two contrasting areas, heterotrophic nearshore and autotrophic open sea, to test whether a combined UVR × dust impact could alter their metabolic balance at mid-term scales. We show that the metabolic state of oligotrophic areas geographically varies and that the joint impact of UVR and dust inputs prompted a strong change towards autotrophic metabolism. We propose that this metabolic response could be accentuated with the global change as remote-sensing evidence shows increasing intensities, frequencies and number of dust events together with variations in the surface UVR fluxes on SW Mediterranean Sea. Overall, these findings suggest that the enhancement of the net carbon budget under a combined UVR and dust inputs impact could contribute to boost the biological pump, reinforcing the role of the oligotrophic marine ecosystems as CO2 sinks.

  5. Effect of arsenate As (V) on the biomarkers of Myriophyllum alterniflorum in oligotrophic and eutrophic conditions.

    Science.gov (United States)

    Krayem, M; Deluchat, V; Rabiet, M; Cleries, K; Lenain, J F; Saad, Z; Kazpard, V; Labrousse, P

    2016-03-01

    Alternate watermilfoil, Myriophyllum alterniflorum is an aquatic macrophyte found in the Limousin rivers (France) whose potential for biomonitoring of metal pollution has been demonstrated. The objective of the present study carried out in vitro was to identify biomarkers for an early detection of the pollution by a metalloid As (V) in eutrophic and oligotrophic conditions. A synthetic medium of similar composition to the waters of the River Vienne was prepared. The morphological development of watermilfoil was monitored for 30 days, with or without contamination by 100 μg L(-1) As (V). In addition, the mineralization of plants and the analysis of biomarkers (chlorophylls, photosynthetic and respiratory intensities …) were investigated after 21 days. Our results indicated that eutrophic medium, induced a decrease in chlorophyll pigments, in growth and an increase in H2O2 compared to the oligotrophic medium. While, the presence of As (V), led to a decrease in the osmotic potential, pigment content, photosynthesis and respiration rates and an inhibition of shoot branching of plants in both conditions. However, a significant increase in H2O2 content was noted in the eutrophic medium. Finally, As (V) was found to be more accumulated in roots than shoots in both conditions but was more accumulated in oligotrophic one. Therefore, we can conclude that the water trophic level modifies the response of M. alterniflorum in presence of arsenate. Thus, M. alterniflorum shows a great promise in water-quality biomonitoring.

  6. Toxicity of triclosan, penconazole and metalaxyl on Caulobacter crescentus and a freshwater microbial community as assessed by flow cytometry.

    Science.gov (United States)

    Johnson, David R; Czechowska, Kamila; Chèvre, Nathalie; van der Meer, Jan Roelof

    2009-07-01

    Biocides are widely used for domestic hygiene, agricultural and industrial applications. Their widespread use has resulted in their introduction into the environment and raised concerns about potential deleterious effects on aquatic ecosystems. In this study, the toxicity of the biocides triclosan, penconazole and metalaxyl were evaluated with the freshwater bacterium Caulobacter crescentus and with a freshwater microbial community using a combination of single- and double-stain flow cytometric assays. Growth of C.  crescentus and the freshwater community were repressed by triclosan but not by penconazole or metalaxyl at concentrations up to 250 μM. The repressive effect of triclosan was dependent on culture conditions. Caulobacter crescentus was more sensitive to triclosan when grown with high glucose at high cell density than when grown directly in sterilized lake water at low cell density. This suggests that the use of conventional growth conditions may overestimate biocide toxicity. Additional experiments showed that the freshwater community was more sensitive to triclosan than C.  crescentus, with 10 nM of triclosan being sufficient to repress growth and change the phylogenetic composition of the community. These results demonstrate that isolate-based assays may underestimate biocide toxicity and highlight the importance of assessing toxicity directly on natural microbial communities. Because 10 nM of triclosan is within the range of concentrations observed in freshwater systems, these results also raise concerns about the risk of introducing triclosan into the environment.

  7. A prevalent alpha-proteobacterium Paracoccus sp. in a population of the Cayenne ticks (Amblyomma cajennense from Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Erik Machado-Ferreira

    2012-01-01

    Full Text Available As Rocky Mountain Spotted Fever is the most common tick-borne disease in South America, the presence of Rickettsia sp. in Amblyomma ticks is a possible indication of its endemicity in certain geographic regions. In the present work, bacterial DNA sequences related to Rickettsia amblyommii genes in A. dubitatum ticks, collected in the Brazilian state of Mato Grosso, were discovered. Simultaneously, Paracoccus sp. was detected in aproximately 77% of A. cajennense specimens collected in Rio de Janeiro, Brazil. This is the first report of Paracoccus sp. infection in a specific tick population, and raises the possibility of these bacteria being maintained and/or transmitted by ticks. Whether Paracoccus sp. represents another group of pathogenic Rhodobacteraceae or simply plays a role in A. cajennense physiology, is unknown. The data also demonstrate that the rickettsial 16S rRNA specific primers used forRickettsia spp. screening can also detect Paracoccus alpha-proteobacteria infection in biological samples. Hence, a PCRRFLP strategy is presented to distinguish between these two groups of bacteria.

  8. A prevalent alpha-proteobacterium Paracoccus sp. in a population of the Cayenne ticks (Amblyomma cajennense) from Rio de Janeiro, Brazil.

    Science.gov (United States)

    Machado-Ferreira, Erik; Piesman, Joseph; Zeidner, Nordin S; Soares, Carlos A G

    2012-12-01

    As Rocky Mountain Spotted Fever is the most common tick-borne disease in South America, the presence of Rickettsia sp. in Amblyomma ticks is a possible indication of its endemicity in certain geographic regions. In the present work, bacterial DNA sequences related to Rickettsia amblyommii genes in A. dubitatum ticks, collected in the Brazilian state of Mato Grosso, were discovered. Simultaneously, Paracoccus sp. was detected in aproximately 77% of A. cajennense specimens collected in Rio de Janeiro, Brazil. This is the first report of Paracoccus sp. infection in a specific tick population, and raises the possibility of these bacteria being maintained and/or transmitted by ticks. Whether Paracoccus sp. represents another group of pathogenic Rhodobacteraceae or simply plays a role in A. cajennense physiology, is unknown. The data also demonstrate that the rickettsial 16S rRNA specific primers used forRickettsia spp. screening can also detect Paracoccus alpha-proteobacteria infection in biological samples. Hence, a PCR-RFLP strategy is presented to distinguish between these two groups of bacteria.

  9. A prevalent alpha-proteobacterium Paracoccus sp. in a population of the Cayenne ticks (Amblyomma cajennense) from Rio de Janeiro, Brazil

    OpenAIRE

    Erik Machado-Ferreira; Joseph Piesman; Zeidner, Nordin S; Soares, Carlos A.G.

    2012-01-01

    As Rocky Mountain Spotted Fever is the most common tick-borne disease in South America, the presence of Rickettsia sp. in Amblyomma ticks is a possible indication of its endemicity in certain geographic regions. In the present work, bacterial DNA sequences related to Rickettsia amblyommii genes in A. dubitatum ticks, collected in the Brazilian state of Mato Grosso, were discovered. Simultaneously, Paracoccus sp. was detected in aproximately 77% of A. cajennense specimens collected in Rio de J...

  10. TonB-Dependent Heme/Hemoglobin Utilization by Caulobacter crescentus HutA.

    Science.gov (United States)

    Balhesteros, Heloise; Shipelskiy, Yan; Long, Noah J; Majumdar, Aritri; Katz, Benjamin B; Santos, Naara M; Leaden, Laura; Newton, Salete M; Marques, Marilis V; Klebba, Phillip E

    2017-03-15

    Siderophore nutrition tests with Caulobacter crescentus strain NA1000 revealed that it utilized a variety of ferric hydroxamate siderophores, including asperchromes, ferrichromes, ferrichrome A, malonichrome, and ferric aerobactin, as well as hemin and hemoglobin. C. crescentus did not transport ferrioxamine B or ferric catecholates. Because it did not use ferric enterobactin, the catecholate aposiderophore was an effective agent for iron deprivation. We determined the kinetics and thermodynamics of [(59)Fe]apoferrichrome and (59)Fe-citrate binding and transport by NA1000. Its affinity and uptake rate for ferrichrome (equilibrium dissociation constant [Kd ], 1 nM; Michaelis-Menten constant [KM ], 0.1 nM; Vmax, 19 pMol/10(9) cells/min) were similar to those of Escherichia coli FhuA. Transport properties for (59)Fe-citrate were similar to those of E. coli FecA (KM , 5.3 nM; Vmax, 29 pMol/10(9) cells/min). Bioinformatic analyses implicated Fur-regulated loci 00028, 00138, 02277, and 03023 as TonB-dependent transporters (TBDT) that participate in iron acquisition. We resolved TBDT with elevated expression under high- or low-iron conditions by SDS-PAGE of sodium sarcosinate cell envelope extracts, excised bands of interest, and analyzed them by mass spectrometry. These data identified five TBDT: three were overexpressed during iron deficiency (00028, 02277, and 03023), and 2 were overexpressed during iron repletion (00210 and 01196). CLUSTALW analyses revealed homology of putative TBDT 02277 to Escherichia coli FepA and BtuB. A Δ02277 mutant did not transport hemin or hemoglobin in nutrition tests, leading us to designate the 02277 structural gene as hutA (for heme/hemoglobin utilization).IMPORTANCE The physiological roles of the 62 putative TBDT of C. crescentus are mostly unknown, as are their evolutionary relationships to TBDT of other bacteria. We biochemically studied the iron uptake systems of C. crescentus, identified potential iron transporters, and clarified

  11. The Caulobacter crescentus phage phiCbK: genomics of a canonical phage

    Directory of Open Access Journals (Sweden)

    Gill Jason J

    2012-10-01

    Full Text Available Abstract Background The bacterium Caulobacter crescentus is a popular model for the study of cell cycle regulation and senescence. The large prolate siphophage phiCbK has been an important tool in C. crescentus biology, and has been studied in its own right as a model for viral morphogenesis. Although a system of some interest, to date little genomic information is available on phiCbK or its relatives. Results Five novel phiCbK-like C. crescentus bacteriophages, CcrMagneto, CcrSwift, CcrKarma, CcrRogue and CcrColossus, were isolated from the environment. The genomes of phage phiCbK and these five environmental phage isolates were obtained by 454 pyrosequencing. The phiCbK-like phage genomes range in size from 205 kb encoding 318 proteins (phiCbK to 280 kb encoding 448 proteins (CcrColossus, and were found to contain nonpermuted terminal redundancies of 10 to 17 kb. A novel method of terminal ligation was developed to map genomic termini, which confirmed termini predicted by coverage analysis. This suggests that sequence coverage discontinuities may be useable as predictors of genomic termini in phage genomes. Genomic modules encoding virion morphogenesis, lysis and DNA replication proteins were identified. The phiCbK-like phages were also found to encode a number of intriguing proteins; all contain a clearly T7-like DNA polymerase, and five of the six encode a possible homolog of the C. crescentus cell cycle regulator GcrA, which may allow the phage to alter the host cell’s replicative state. The structural proteome of phage phiCbK was determined, identifying the portal, major and minor capsid proteins, the tail tape measure and possible tail fiber proteins. All six phage genomes are clearly related; phiCbK, CcrMagneto, CcrSwift, CcrKarma and CcrRogue form a group related at the DNA level, while CcrColossus is more diverged but retains significant similarity at the protein level. Conclusions Due to their lack of any apparent relationship to

  12. Regulation of the activity of the dual-function DnaA protein in Caulobacter crescentus.

    Science.gov (United States)

    Fernandez-Fernandez, Carmen; Gonzalez, Diego; Collier, Justine

    2011-01-01

    DnaA is a conserved essential bacterial protein that acts as the initiator of chromosomal replication as well as a master transcriptional regulator in Caulobacter crescentus. Thus, the intracellular levels of active DnaA need to be tightly regulated during the cell cycle. Our previous work suggested that DnaA may be regulated at the level of its activity by the replisome-associated protein HdaA. Here, we describe the construction of a mutant DnaA protein [DnaA(R357A)]. The R357 residue in the AAA+ domain of the C. crescentus DnaA protein is equivalent to the R334 residue of the E. coli DnaA protein, which is required for the Regulatory Inactivation of DnaA (RIDA). We found that the expression of the DnaA(R357A) mutant protein in C. crescentus, but not the expression of the wild-type DnaA protein at similar levels, causes a severe phenotype of over-initiation of chromosomal replication and that it blocks cell division. Thus, the mutant DnaA(R357A) protein is hyper-active to promote the initiation of DNA replication, compared to the wild-type DnaA protein. DnaA(R357A) could not replace DnaA in vivo, indicating that the switch in DnaA activity once chromosomal replication has started may be an essential process in C. crescentus. We propose that the inactivation of DnaA is the main mechanism ensuring that chromosomal replication starts only once per cell cycle. We further observed that the R357A substitution in DnaA does not promote the activity of DnaA as a direct transcriptional activator of four important genes, encoding HdaA, the GcrA master cell cycle regulator, the FtsZ cell division protein and the MipZ spatial regulator of cell division. Thus, the AAA+ domain of DnaA may play a role in temporally regulating the bifunctionality of DnaA by reallocating DnaA molecules from initiating DNA replication to transcribing genes within the unique DnaA regulon of C. crescentus.

  13. Regulation of the activity of the dual-function DnaA protein in Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Carmen Fernandez-Fernandez

    Full Text Available DnaA is a conserved essential bacterial protein that acts as the initiator of chromosomal replication as well as a master transcriptional regulator in Caulobacter crescentus. Thus, the intracellular levels of active DnaA need to be tightly regulated during the cell cycle. Our previous work suggested that DnaA may be regulated at the level of its activity by the replisome-associated protein HdaA. Here, we describe the construction of a mutant DnaA protein [DnaA(R357A]. The R357 residue in the AAA+ domain of the C. crescentus DnaA protein is equivalent to the R334 residue of the E. coli DnaA protein, which is required for the Regulatory Inactivation of DnaA (RIDA. We found that the expression of the DnaA(R357A mutant protein in C. crescentus, but not the expression of the wild-type DnaA protein at similar levels, causes a severe phenotype of over-initiation of chromosomal replication and that it blocks cell division. Thus, the mutant DnaA(R357A protein is hyper-active to promote the initiation of DNA replication, compared to the wild-type DnaA protein. DnaA(R357A could not replace DnaA in vivo, indicating that the switch in DnaA activity once chromosomal replication has started may be an essential process in C. crescentus. We propose that the inactivation of DnaA is the main mechanism ensuring that chromosomal replication starts only once per cell cycle. We further observed that the R357A substitution in DnaA does not promote the activity of DnaA as a direct transcriptional activator of four important genes, encoding HdaA, the GcrA master cell cycle regulator, the FtsZ cell division protein and the MipZ spatial regulator of cell division. Thus, the AAA+ domain of DnaA may play a role in temporally regulating the bifunctionality of DnaA by reallocating DnaA molecules from initiating DNA replication to transcribing genes within the unique DnaA regulon of C. crescentus.

  14. The cloning, expression, purification, characterization and modeled structure of Caulobacter crescentus β-Xylosidase I.

    Science.gov (United States)

    Graciano, Luciana; Corrêa, Juliana Moço; Gandra, Rinaldo Ferreira; Seixas, Flavio Augusto Vicente; Kadowaki, Marina Kimiko; Sampaio, Silvio César; Silva, José Luis da Conceição; Osaku, Clarice Aoki; Simão, Rita de Cássia Garcia

    2012-09-01

    The xynB1 gene (CCNA 01040) of Caulobacter crescentus that encodes a bifunctional enzyme containing the conserved β-Xylosidase and α-L-Arabinofuranosidase (β-Xyl I-α-L-Ara) domains was amplified by PCR and cloned into the vector pJet1.2Blunt. The xynB1 gene was subcloned into the vector pPROEX-hta that produces a histidine-fused translation product. The overexpression of recombinant β-Xyl I-α-L-Ara was induced with IPTG in BL21 (DE3) and the resulting intracellular protein was purified with pre-packaged nickel-Sepharose columns. The recombinant β-Xyl I-α-L-Ara exhibited a specific β-Xylosidase I activity of 1.25 U mg(-1) to oNPX and a specific α-L-Arabinofuranosidase activity of 0.47 U mg(-1) to pNPA. The predominant activity of the recombinant enzyme was its β-Xylosidase I activity, and the enzymatic characterization was focused on it. The β-Xylosidase I activity was high over the pH range 3-10, with maximal activity at pH 6. The enzyme activity was optimal at 45 °C, and a high degree of stability was verified over 240 min at this temperature. Moreover, β-Xylosidase activity was inhibited in the presence of the metals Zn(2+) and Cu(2+), and the enzyme exhibited K(M) and V(Max) values of 2.89 ± 0.13 mM and 1.4 ± 0.04 μM min(-1) to oNPX, respectively. The modeled structure of β-xylosidase I showed that its active site is highly conserved compared with other structures of the GH43 family. The increase in the number of contact residues responsible for maintaining the dimeric structure indicates that this dimer is more stable than the tetramer form.

  15. Reclassification and emended description of Caulobacter leidyi as Sphingomonas leidyi comb. nov., and emendation of the genus Sphingomonas.

    Science.gov (United States)

    Chen, Hong; Jogler, Mareike; Rohde, Manfred; Klenk, Hans-Peter; Busse, Hans-Jürgen; Tindall, Brian J; Spröer, Cathrin; Overmann, Jörg

    2012-12-01

    'Caulobacter leidyi' DSM 4733(T) has been shown to be affiliated with the family Sphingomonadaceae instead of the Caulobacteraceae, and due to its poor characterization has been omitted from the current edition of Bergey's Manual of Systematic Bacteriology and removed to limbo. We isolated a novel sphingoglycolipid-containing dimorphic prosthecate bacterium, designated strain 247, from a pre-alpine freshwater lake. Strain 247 and 'Caulobacter leidyi' DSM 4733(T) were characterized in detail. The rod-shaped cells were Gram-stain-negative, aerobic, catalase- and oxidase-positive, and formed a stalk or polar flagellum. Both strains grew optimally at 28-30 °C, and pH 6.0-8.0. The major fatty acids were C(18 : 1)ω7c, C(16 : 0) and 11-methyl C(18 : 1)ω7c. C(14 : 0) 2-OH represents the major 2-hydroxy fatty acid. Q-10 was the major respiratory quinone and the major polar lipids were diphosphatidylglycerol, phosphatidyldimethylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylcholine, three glycolipids, two phosphoaminolipids and two unidentified sphingoglycolipids. The major polyamine was sym-homospermidine. The G+C content of genomic DNA of strains 247 and DSM 4733(T) was 67.6 mol% and 67.0 mol%, respectively. According to 16S rRNA gene sequence analysis and DNA-DNA hybridization, strains DSM 4733(T) and 247 were phylogenetically closely related (99.6 % 16S rRNA gene sequence similarity, 82.9 % DNA-DNA hybridization value) and affiliated to the genus Sphingomonas. The closest recognized species was Sphingomonas aquatilis DSM 15581(T) (98.1 % sequence similarity). In addition, the presence of cystine arylamidase, absence of β-galactosidase, and the inability to utilize l-arabinose, galactose and sucrose distinguished strains DSM 4733(T) and 247 from most other members of the family Sphingomonadaceae. So far, the dimorphic life cycle that involves a prosthecate and a flagellated stage is unique for strains DSM

  16. A novel cohabitation between two diazotrophic cyanobacteria in the oligotrophic ocean.

    Science.gov (United States)

    Momper, Lily M; Reese, Brandi Kiel; Carvalho, Gustavo; Lee, Patrick; Webb, Eric A

    2015-04-01

    The cyanobacterial genus Trichodesmium is biogeochemically significant because of its dual role in nitrogen and carbon fixation in the oligotrophic ocean. Trichodesmium species form colonies that can be easily enriched from the water column and used for shipboard rate measurements to estimate their contribution to oceanic carbon and nitrogen budgets. During a July 2010 cruise near the Hawaiian Islands in the oligotrophic North Pacific Subtropical Gyre, a specific morphology of Trichodesmium puff-form colonies were examined under epifluorescent microscopy and found to harbor a colonial endobiont, morphologically identified as the heterocystous diazotrophic cyanobacterium Calothrix. Using unialgal enrichments obtained from this cruise, we show that these Calothrix-like heterocystous cyanobionts (hetDA for 'Trichodesmium-associated heterocystous diazotroph') fix nitrogen on a diurnal cycle (maximally in the middle of the light cycle with a detectable minimum in the dark). Gene sequencing of nifH from the enrichments revealed that this genus was likely not quantified using currently described quantitative PCR (qPCR) primers. Guided by the sequence from the isolate, new hetDA-specific primers were designed and subsequent qPCR of environmental samples detected this diazotroph from surface water to a depth of 150 m, reaching densities up to ∼ 9 × 10(3) l(-1). Based on phylogenetic relatedness of nifH and 16S rRNA gene sequences, it is predicted that the distribution of this cyanobiont is not limited to subtropical North Pacific but likely reaches to the South Pacific and Atlantic Oceans. Therefore, this previously unrecognized cohabitation, if it reaches beyond the oligotrophic North Pacific, could potentially influence Trichodesmium-derived nitrogen fixation budgets in the world ocean.

  17. Substrate use of Pseudovibrio sp. growing in ultra-oligotrophic seawater.

    Directory of Open Access Journals (Sweden)

    Anne Schwedt

    Full Text Available Marine planktonic bacteria often live in habitats with extremely low concentrations of dissolved organic matter (DOM. To study the use of trace amounts of DOM by the facultatively oligotrophic Pseudovibrio sp. FO-BEG1, we investigated the composition of artificial and natural seawater before and after growth. We determined the concentrations of dissolved organic carbon (DOC, total dissolved nitrogen (TDN, free and hydrolysable amino acids, and the molecular composition of DOM by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR-MS. The DOC concentration of the artificial seawater we used for cultivation was 4.4 μmol C L(-1, which was eight times lower compared to the natural oligotrophic seawater we used for parallel experiments (36 μmol C L(-1. During the three-week duration of the experiment, cell numbers increased from 40 cells mL(-1 to 2x10(4 cells mL(-1 in artificial and to 3x10(5 cells mL(-1 in natural seawater. No nitrogen fixation and minor CO2 fixation (< 1% of cellular carbon was observed. Our data show that in both media, amino acids were not the main substrate for growth. Instead, FT-ICR-MS analysis revealed usage of a variety of different dissolved organic molecules, belonging to a wide range of chemical compound groups, also containing nitrogen. The present study shows that marine heterotrophic bacteria are able to proliferate with even lower DOC concentrations than available in natural ultra-oligotrophic seawater, using unexpected organic compounds to fuel their energy, carbon and nitrogen requirements.

  18. Dust deposition in an oligotrophic marine environment: impact on the carbon budget

    Directory of Open Access Journals (Sweden)

    C. Guieu

    2014-01-01

    Full Text Available By bringing new nutrients and particles to the surface ocean, atmospheric deposition impacts biogeochemical cycles. The extent to which those changes are modifying the carbon balance in oligotrophic environments such as the Mediterranean Sea that receives important Saharan dust fluxes is unknown. DUNE project provides the first attempt to evaluate the changes induced in the carbon budget of an oligotrophic system after simulated Saharan dust wet and dry deposition events. Here we report the results for the 3 distinct artificial dust seeding experiments in large mesocosms that were conducted in the oligotrophic waters of the Mediterranean Sea in summer 2008 and 2010. Simultaneous measurements of the metabolic rates (C fixation, C respiration in the water column have shown that the dust deposition did not change drastically the metabolic balance as the tested waters remained net heterotroph (i.e. net primary production to bacteria respiration ratio < 1 and in some cases the net heterotrophy was even enhanced by the dust deposition. Considering the different terms of the carbon budget, we estimate that it was balanced with a dissolved organic carbon (DOC consumption of at least 10% of the initial stock. This corresponds to a fraction of the DOC stock of the surface mixed layer that consequently will not be exported during the winter mixing. Although heterotrophic bacteria were found to be the key players in the response to dust deposition, net primary production increased about twice in case of simulated wet deposition (that includes anthropogenic nitrogen and a small fraction of particulate organic carbon was still exported. Our estimated carbon budgets are an important step forward in the way we understand dust deposition and associated impacts on the oceanic cycles. They are providing knowledge about the key processes (i.e. bacteria respiration, aggregation that need to be considered for an integration of atmospheric deposition in marine

  19. Phylogeny by a polyphasic approach of the order Caulobacterales, proposal of Caulobacter mirabilis sp. nov., Phenylobacterium haematophilum sp. nov. and Phenylobacterium conjunctum sp. nov., and emendation of the genus Phenylobacterium.

    Science.gov (United States)

    Abraham, Wolf-Rainer; Macedo, Alexandre J; Lünsdorf, Heinrich; Fischer, Roman; Pawelczyk, Sonja; Smit, John; Vancanneyt, Marc

    2008-08-01

    Three strains of Gram-negative, rod-shaped, non-spore-forming bacteria were isolated from fresh water and human blood. As determined by analyses of 16S rRNA gene sequences, the prosthecate strain FWC 38T was affiliated to the alphaproteobacterial genus Caulobacter, with Caulobacter henricii (96.8 %) and Caulobacter fusiformis (96.8 %) as its closest relatives. The non-prosthecate strain LMG 11050T and the prosthecate strain FWC 21T both belonged to the genus Phenylobacterium with Phenylobacterium koreense (96.9 %) and Phenylobacterium immobile (96.3 %) as the closest relatives. This affiliation was supported by chemotaxonomic data (polar lipids and cellular fatty acids). Physiological and biochemical tests allowed genotypic and phenotypic differentiation of the novel strains from all hitherto recognized species of the genera Caulobacter and Phenylobacterium. The strains therefore represent novel species, for which the names Caulobacter mirabilis sp. nov. (type strain FWC 38T=LMG 24261T=CCUG 55073T), Phenylobacterium conjunctum (type strain FWC 21T=LMG 24262T=CCUG 55074T), the first described prosthecate Phenylobacterium species, and Phenylobacterium haematophilum sp. nov. (type strain LMG 11050T=CCUG 26751T) are proposed. Marker nucleotides within the 16S rRNA genes were determined for the genera Asticcacaulis, Brevundimonas, Caulobacter and Phenylobacterium and the description of the genus Phenylobacterium is emended.

  20. Diversity of picoeukaryotes at an oligotrophic site off the Northeastern Red Sea Coast.

    KAUST Repository

    Acosta, Francisco

    2013-08-20

    Picoeukaryotes are protists ≤ 3 μm composed of a wide diversity of taxonomic groups. They are an important constituent of the ocean\\'s microbiota and perform essential ecological roles in marine nutrient and carbon cycles. Despite their importance, the true extent of their diversity has only recently been uncovered by molecular surveys that resulted in the discovery of a substantial number of previously unknown groups. No study on picoeukaryote diversity has been conducted so far in the main Red Sea basin-a unique marine environment characterized by oligotrophic conditions, high levels of irradiance, high salinity and increased water temperature.

  1. A preliminary carbon budget for two oligotrophic hardwater lakes in the Forsmark area, Sweden

    Energy Technology Data Exchange (ETDEWEB)

    Nilsson, Eva [Uppsala Univ. (Sweden). Dept. of Limnology

    2001-06-01

    The Swedish Nuclear Fuel and Waste Management Co (SKB) is responsible for management and disposal of Swedish radioactive waste. The company is planning to construct repositories that will keep radioactive waste away from humans for hundreds of thousands of years. In a safety assessment of the repositories hypothetical releases are used to evaluate the robustness of the repositories. It is important to know how the radioactive nuclides would react if they were released and by which way they could enter the living biota. SFR are responsible for the disposal of low radioactive waste and close to the nuclear plant in Forsmark there is a storage for low radioactive waste. At the moment this storage is located in the bedrock far below the sea level but due to land-rise in the area it will in the future be located above sea level. Hence, it is of importance to know how the surface ecosystems in the area are functioning. A carbon budget for the aquatic ecosystem above SFR in Oeresundsgrepen exist, but it is also important to have a carbon budget for the surface systems in the Forsmark area since SFR in the future will be situated above sea level. Carbon budgets can be used to get a picture of how an ecosystem functions. The carbon flow shows how carbon is transported through a food web from lower trophic levels, e.g. plants and bacteria to higher trophic levels such as fish. Oligotrophic hardwater lakes are the most important lakes in the Forsmark area. This report aims to give a picture of a potential flow of carbon through the ecosystem in two oligotrophic hard-water lakes, Lake Haellefjaerd and Lake Eckarfjaerden. Macrophyte, mainly Chara, were calculated to make up the largest part of the biomass and production in both lakes. Benthic bacteria and microphytobenthos (benthic photosynthesising microorganisms) were other large contributors to the production. Benthic bacteria were found responsible for a major part of respiration and, hence, consumption of carbon in the

  2. Structural insights into ChpT, an essential dimeric histidine phosphotransferase regulating the cell cycle in Caulobacter crescentus.

    Science.gov (United States)

    Fioravanti, Antonella; Clantin, Bernard; Dewitte, Frédérique; Lens, Zoé; Verger, Alexis; Biondi, Emanuele G; Villeret, Vincent

    2012-09-01

    Two-component and phosphorelay signal-transduction proteins are crucial for bacterial cell-cycle regulation in Caulobacter crescentus. ChpT is an essential histidine phosphotransferase that controls the activity of the master cell-cycle regulator CtrA by phosphorylation. Here, the 2.2 Å resolution crystal structure of ChpT is reported. ChpT is a homodimer and adopts the domain architecture of the intracellular part of class I histidine kinases. Each subunit consists of two distinct domains: an N-terminal helical hairpin domain and a C-terminal α/β domain. The two N-terminal domains are adjacent within the dimer, forming a four-helix bundle. The ChpT C-terminal domain adopts an atypical Bergerat ATP-binding fold.

  3. High throughput 3D super-resolution microscopy reveals Caulobacter crescentus in vivo Z-ring organization.

    Science.gov (United States)

    Holden, Seamus J; Pengo, Thomas; Meibom, Karin L; Fernandez Fernandez, Carmen; Collier, Justine; Manley, Suliana

    2014-03-25

    We created a high-throughput modality of photoactivated localization microscopy (PALM) that enables automated 3D PALM imaging of hundreds of synchronized bacteria during all stages of the cell cycle. We used high-throughput PALM to investigate the nanoscale organization of the bacterial cell division protein FtsZ in live Caulobacter crescentus. We observed that FtsZ predominantly localizes as a patchy midcell band, and only rarely as a continuous ring, supporting a model of "Z-ring" organization whereby FtsZ protofilaments are randomly distributed within the band and interact only weakly. We found evidence for a previously unidentified period of rapid ring contraction in the final stages of the cell cycle. We also found that DNA damage resulted in production of high-density continuous Z-rings, which may obstruct cytokinesis. Our results provide a detailed quantitative picture of in vivo Z-ring organization.

  4. The Caulobacter crescentus transducing phage Cr30 is a unique member of the T4-like family of myophages.

    Science.gov (United States)

    Ely, Bert; Gibbs, Whitney; Diez, Simon; Ash, Kurt

    2015-06-01

    Bacteriophage Cr30 has proven useful for the transduction of Caulobacter crescentus. Nucleotide sequencing of Cr30 DNA revealed that the Cr30 genome consists of 155,997 bp of DNA that codes for 287 proteins and five tRNAs. In contrast to the 67 % GC content of the host genome, the GC content of the Cr30 genome is only 38 %. This lower GC content causes both the codon usage pattern and the amino acid composition of the Cr30 proteins to be quite different from those of the host bacteria. As a consequence, the Cr30 mRNAs probably are translated at a rate that is slower than the normal rate for host mRNAs. A phylogenetic comparison of the genome indicates that Cr30 is a member of the T4-like family that is most closely related to a new group of T-like phages exemplified by фM12.

  5. Caulobacter chromosome in vivo configuration matches model predictions for a supercoiled polymer in a cell-like confinement

    DEFF Research Database (Denmark)

    Hong, Sun-Hae; Toro, Esteban; Mortensen, Kim;

    2013-01-01

    is the contour length, and cell-to-cell distribution of the interloci distance r is a universal function of r/n0.22 with broad cell-to-cell variability. For DNA segments greater than about 300 kb, the mean interloci distances scale as n, in agreement with previous observations. The 0.22 value of the scaling......We measured the distance between fluorescent-labeled DNA loci of various interloci contour lengths in Caulobacter crescentus swarmer cells to determine the in vivo configuration of the chromosome. For DNA segments less than about 300 kb, the mean interloci distances, 〈r〉, scale as n0.22, where n...... exponent for short DNA segments is consistent with theoretical predictions for a branched DNA polymer structure. Predictions from Brownian dynamics simulations of the packing of supercoiled DNA polymers in an elongated cell-like confinement are also consistent with a branched DNA structure, and simulated...

  6. Identification of ω-aminotransferase from Caulobacter crescentus and site-directed mutagenesis to broaden substrate specificity.

    Science.gov (United States)

    Hwang, Bum-Yeol; Ko, Seung-Hyun; Park, Hyung-Yeon; Seo, Joo-Hyun; Lee, Bon-Su; Kim, Byung-Gee

    2008-01-01

    A putative aminotransferase gene, cc3143 (aptA), from Caulobacter crescentus was screened by bioinformatical tools and overexpressed in E. coli, and the substrate specificity of the aminotransferase was investigated. AptA showed high activity for short-chain beta-amino acids. It showed the highest activity for 3-amino-n-butyric acid. It showed higher activity toward aromatic amines than aliphatic amines. The 3D model of the aminotransferase was constructed by homology modeling using a dialkylglycine decarboxylase PDB ID: 1DGE) as a template. Then, the aminotransferase was rationally redesigned to increase the activity for 3-amino- 3-phenylpropionic acid. The mutants N285A and V227G increased the relative activity for 3-amino-3-phenylpropionic acid to 3-amino-n-butyric acid by 11-fold and 3-fold, respectively, over that of wild type.

  7. Comparing effects of oligotrophication and upstream hydropower dams on plankton and productivity in perialpine lakes

    Science.gov (United States)

    Finger, David; Schmid, Martin; Wüest, Alfred

    2007-12-01

    In recent decades, many perialpine lakes have been affected by oligotrophication due to efficient sewage treatment and by altered water turbidity due to upstream hydropower operations. Such simultaneous environmental changes often lead to public debate on the actual causes of observed productivity reductions. We evaluate the effects of those two changes by a combined approach of modeling and data interpretation for a case study on Lake Brienz (Switzerland), a typical oligotrophic perialpine lake, located downstream of several hydropower reservoirs. A physical k-ɛ scheme and a biogeochemical advection-diffusion-reaction model were implemented and applied for several hypothetical scenarios with different nutrient loads and different particle input dynamics. The simulation results are compared to long-term biotic data collected from 1999 to 2004. The analysis shows that enhanced nutrient supply increases the nutritious value of algae, stimulating zooplankton growth, while phytoplankton growth is limited by stronger top-down control. Annually integrated productivity is only slightly influenced by altered turbidity, as phosphorous limitation prevails. Simulations indicate that the spring production peak is delayed because of increased turbidity in winter caused by upstream hydropower operation. As a consequence, the entire nutrient cycle is seasonally delayed, creating an additional stress for zooplankton and fish in the downstream lake.

  8. Phytoplankton biovolume is independent from the slope of the size spectrum in the oligotrophic atlantic ocean

    KAUST Repository

    Moreno-Ostos, Enrique

    2015-08-06

    Modelling the size-abundance spectrum of phytoplankton has proven to be a very useful tool for the analysis of physical-biological coupling and the vertical flux of carbon in oceanic ecosystems at different scales. A frequent observation relates high phytoplankton biovolume in productive regions with flatter spectrum slope and the opposite in oligotrophic ecosystems. Rather than this, the relationship between high biovolume phytoplankton assemblages and flatter size-abundance spectra does not correspond with measurements of the phytoplankton community in the Atlantic Ocean open waters. As part of the Malaspina Circunnavegation Expedition, sixty seven sampling stations within the Atlantic Ocean covering six oceanographic provinces, at different seasons, produced a complete set of phytoplankton size-spectra whose slope and biovolume did not show any obvious interrelation. In these oligotrophic sites, small (procaryotes) and medium-size (nanoplankton) cells are responsible for the most part of biovolume, and their response to environmental conditions does not apply to changes in the size-abundance spectrum slope as expected in richer, large-cell dominated ecosystems.

  9. Fungal Diversity in a Dark Oligotrophic Volcanic Ecosystem (DOVE on Mount Erebus, Antarctica

    Directory of Open Access Journals (Sweden)

    Hubert Staudigel

    2013-05-01

    Full Text Available Fumarolic Ice caves on Antarctica’s Mt. Erebus contain a dark oligotrophic volcanic ecosystem (DOVE and represent a deep biosphere habitat that can provide insight into microbial communities that utilize energy sources other than photosynthesis. The community assembly and role of fungi in these environments remains largely unknown. However, these habitats could be relatively easily contaminated during human visits. Sixty-one species of fungi were identified from soil clone libraries originating from Warren Cave, a DOVE on Mt. Erebus. The species diversity was greater than has been found in the nearby McMurdo Dry Valleys oligotrophic soil. A relatively large proportion of the clones represented Malassezia species (37% of Basidomycota identified. These fungi are associated with skin surfaces of animals and require high lipid content for growth, indicating that contamination may have occurred through the few and episodic human visits in this particular cave. These findings highlight the importance of fungi to DOVE environments as well as their potential use for identifying contamination by humans. The latter offers compelling evidence suggesting more strict management of these valuable research areas.

  10. Assessing benthic oxygen fluxes in oligotrophic deep sea sediments (HAUSGARTEN observatory)

    Science.gov (United States)

    Donis, Daphne; McGinnis, Daniel F.; Holtappels, Moritz; Felden, Janine; Wenzhoefer, Frank

    2016-05-01

    Benthic oxygen fluxes, an established proxy for total organic carbon mineralization, were investigated in oligotrophic deep sea sediments. We used three different in situ technologies to estimate the benthic oxygen fluxes at an Arctic deep sea site (2500 m depth, HAUSGARTEN observatory) with limiting conditions of low oxygen gradients and fluxes, low turbulence and low particle content in the benthic boundary layer. The resolved eddy covariance turbulent oxygen flux (-0.9±0.2 (SD) mmol O2 m-2 d-1) compared well with simultaneous dissolved oxygen flux measurements carried out with a microprofiler (-1.02±0.3 (SD) mmol O2 m-2 d-1) and total oxygen uptake obtained by benthic chamber incubations (-1.1±0.1 (SD) mmol O2 m-2 d-1). The agreement between these different techniques revealed that microbial-mediated oxygen consumption was dominant at this site. The average benthic flux equals a carbon mineralization rate of 4.3 g C m-2 yr-1, which exceeds the annual sedimentation of particulate organic matter measured by sediment traps. The present study represents a detailed comparison of different in situ technologies for benthic flux measurements at different spatial scales in oligotrophic deep sea sediments. The use of eddy covariance, so far rarely used for deep sea investigations, is presented in detail.

  11. Sediment enzyme activities and microbial community diversity in an oligotrophic drinking water reservoir, eastern China.

    Science.gov (United States)

    Zhang, Haihan; Huang, Tinglin; Liu, Tingting

    2013-01-01

    Drinking water reservoir plays a vital role in the security of urban water supply, yet little is known about microbial community diversity harbored in the sediment of this oligotrophic freshwater environmental ecosystem. In the present study, integrating community level physiological profiles (CLPPs), nested polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and clone sequence technologies, we examined the sediment urease and protease activities, bacterial community functional diversity, genetic diversity of bacterial and fungal communities in sediments from six sampling sites of Zhou cun drinking water reservoir, eastern China. The results showed that sediment urease activity was markedly distinct along the sites, ranged from 2.48 to 11.81 mg NH₃-N/(g·24 h). The highest average well color development (AWCD) was found in site C, indicating the highest metabolic activity of heterotrophic bacterial community. Principal component analysis (PCA) revealed tremendous differences in the functional (metabolic) diversity patterns of the sediment bacterial communities from different sites. Meanwhile, DGGE fingerprints also indicated spatial changes of genetic diversity of sediment bacterial and fungal communities. The sequence BLAST analysis of all the sediment samples found that Comamonas sp. was the dominant bacterial species harbored in site A. Alternaria alternate, Allomyces macrogynus and Rhizophydium sp. were most commonly detected fungal species in sediments of the Zhou cun drinking water reservoir. The results from this work provide new insights about the heterogeneity of sediment microbial community metabolic activity and genetic diversity in the oligotrophic drinking water reservoir.

  12. Novel analysis of oceanic surface water metagenomes suggests importance of polyphosphate metabolism in oligotrophic environments.

    Directory of Open Access Journals (Sweden)

    Ben Temperton

    Full Text Available Polyphosphate is a ubiquitous linear homopolymer of phosphate residues linked by high-energy bonds similar to those found in ATP. It has been associated with many processes including pathogenicity, DNA uptake and multiple stress responses across all domains. Bacteria have also been shown to use polyphosphate as a way to store phosphate when transferred from phosphate-limited to phosphate-rich media--a process exploited in wastewater treatment and other environmental contaminant remediation. Despite this, there has, to date, been little research into the role of polyphosphate in the survival of marine bacterioplankton in oligotrophic environments. The three main proteins involved in polyphosphate metabolism, Ppk1, Ppk2 and Ppx are multi-domain and have differential inter-domain and inter-gene conservation, making unbiased analysis of relative abundance in metagenomic datasets difficult. This paper describes the development of a novel Isofunctional Homolog Annotation Tool (IHAT to detect homologs of genes with a broad range of conservation without bias of traditional expect-value cutoffs. IHAT analysis of the Global Ocean Sampling (GOS dataset revealed that genes associated with polyphosphate metabolism are more abundant in environments where available phosphate is limited, suggesting an important role for polyphosphate metabolism in marine oligotrophs.

  13. Characteristics and ontogeny of oligotrophic hardwater lakes in the Forsmark area, central Sweden

    Energy Technology Data Exchange (ETDEWEB)

    Brunberg, A.K.; Blomqvist, P. [Uppsala Univ. (Sweden). Dept. of Limnology

    1999-12-01

    This is the first part of a report characterising the lakes of Uppsala county, with special emphasis on the coastal lakes in the Forsmark area.The aim of the study is to characterise different main types of lakes within the Forsmark area and to create a basis for prediction of their ontogeny, that can be used also for new lakes which due to shoreline displacement will be formed during the next 10 000 years. Areas where future research is needed to fully understand the functioning of the lake ecosystems and their ontogeny should also be identified. This first part of the study identifies and describes one of the most common lake types in the area, the oligotrophic hardwater lake. The geology in the catchments of the Forsmark area includes a bedrock dominated by granites and gneisses, covered by calcareous glacial till and postglacial clay. The catchments are dominated by forest, and the oligotrophic hardwater lakes are to a large extent surrounded by mires. Inflow as well as outflow of water is often diffuse, via the surrounding mire. The lakes are small and shallow, with nutrient poor and highly alkaline water. Three key habitats have been identified within the lakes; the pelagic zone, characterised by low production of biota;, the presumably moderately productive emergent macrophyte zone, dominated by Sphagnum and Phragmites;, and the light exposed soft-bottom zone with Chara meadows and an unusually rich and presumably highly productive microbial sediment community. The oligotrophic hardwater lakes have their origin as depressions in the bottom of the Baltic Sea, which are successively transported upwards due to the land-rise process in the area. As the basins are isolated from the sea , a gradual change from a brackish to freshwater conditions occur. When the lakes have become completely isolated, the oligotrophic hardwater stage follows, due to inflow of carbonate-rich and well buffered groundwater. In the next successional stage, Sphagnum mosses start to

  14. Influence of ocean acidification and deep water upwelling on oligotrophic plankton communities in the subtropical North Atlantic

    DEFF Research Database (Denmark)

    Taucher, Jan; Bach, Lennart T.; Boxhammer, Tim

    2017-01-01

    eutrophic environments, while less attention has been paid to oligotrophic systems such as the subtropical ocean gyres. Here we report from a recent in situ mesocosm experiment off the coast of Gran Canaria in the eastern subtropical North Atlantic, where we investigated the influence of OA on the ecology...... insights into the ecological responses of plankton communities to simulated OA over the course of the 62-day experiment. One particular scientific objective of our mesocosm experiment was to investigate how OA impacts might differ between oligotrophic conditions and phases of high biological productivity...

  15. An unusual promoter controls cell-cycle regulation and dependence on DNA replication of the Caulobacter fliLM early flagellar operon.

    Science.gov (United States)

    Stephens, C M; Shapiro, L

    1993-09-01

    Transcription of flagellar genes in Caulobacter crecentus is programmed to occur during the predivisional stage of the cell cycle. The mechanism of activation of Class II flagellar genes, the highest identified genes in the Caulobacter flagellar hierarchy, is unknown. As a step toward understanding this process, we have defined cis-acting sequences necessary for expression of a Class II flagellar operon, fliLM. Deletion analysis indicated that a 55 bp DNA fragment was sufficient for normal, temporally regulated promoter activity. Transcription from this promoter-containing fragment was severely reduced when chromosomal DNA replication was inhibited. Extensive mutational analysis of the promoter region from -42 to -5 identified functionally important nucleotides at -36 and -35, between -29 and -22, and at -12, which correlates well with sequences conserved between fliLM and the analogous regions of two other Class II flagellar operons. The promoter sequence does not resemble that recognized by any known bacterial sigma factor. Models for regulation of Caulobacter early flagellar promoters are discussed in which RNA polymerase containing a novel sigma subunit interacts with an activation factor bound to the central region of the promoter.

  16. High diversity and suggested endemicity of culturable Actinobacteria in an extremely oligotrophic desert oasis

    Directory of Open Access Journals (Sweden)

    Hector Fernando Arocha-Garza

    2017-05-01

    Full Text Available The phylum Actinobacteria constitutes one of the largest and anciently divergent phyla within the Bacteria domain. Actinobacterial diversity has been thoroughly researched in various environments due to its unique biotechnological potential. Such studies have focused mostly on soil communities, but more recently marine and extreme environments have also been explored, finding rare taxa and demonstrating dispersal limitation and biogeographic patterns for Streptomyces. To test the distribution of Actinobacteria populations on a small scale, we chose the extremely oligotrophic and biodiverse Cuatro Cienegas Basin (CCB, an endangered oasis in the Chihuahuan desert to assess the diversity and uniqueness of Actinobacteria in the Churince System with a culture-dependent approach over a period of three years, using nine selective media. The 16S rDNA of putative Actinobacteria were sequenced using both bacteria universal and phylum-specific primer pairs. Phylogenetic reconstructions were performed to analyze OTUs clustering and taxonomic identification of the isolates in an evolutionary context, using validated type species of Streptomyces from previously phylogenies as a reference. Rarefaction analysis for total Actinobacteria and for Streptomyces isolates were performed to estimate species’ richness in the intermediate lagoon (IL in the oligotrophic Churince system. A total of 350 morphologically and nutritionally diverse isolates were successfully cultured and characterized as members of the Phylum Actinobacteria. A total of 105 from the total isolates were successfully subcultured, processed for DNA extraction and 16S-rDNA sequenced. All strains belong to the order Actinomycetales, encompassing 11 genera of Actinobacteria; the genus Streptomyces was found to be the most abundant taxa in all the media tested throughout the 3-year sampling period. Phylogenetic analysis of our isolates and another 667 reference strains of the family Streptomycetaceae

  17. High diversity and suggested endemicity of culturable Actinobacteria in an extremely oligotrophic desert oasis.

    Science.gov (United States)

    Arocha-Garza, Hector Fernando; Canales-Del Castillo, Ricardo; Eguiarte, Luis E; Souza, Valeria; De la Torre-Zavala, Susana

    2017-01-01

    The phylum Actinobacteria constitutes one of the largest and anciently divergent phyla within the Bacteria domain. Actinobacterial diversity has been thoroughly researched in various environments due to its unique biotechnological potential. Such studies have focused mostly on soil communities, but more recently marine and extreme environments have also been explored, finding rare taxa and demonstrating dispersal limitation and biogeographic patterns for Streptomyces. To test the distribution of Actinobacteria populations on a small scale, we chose the extremely oligotrophic and biodiverse Cuatro Cienegas Basin (CCB), an endangered oasis in the Chihuahuan desert to assess the diversity and uniqueness of Actinobacteria in the Churince System with a culture-dependent approach over a period of three years, using nine selective media. The 16S rDNA of putative Actinobacteria were sequenced using both bacteria universal and phylum-specific primer pairs. Phylogenetic reconstructions were performed to analyze OTUs clustering and taxonomic identification of the isolates in an evolutionary context, using validated type species of Streptomyces from previously phylogenies as a reference. Rarefaction analysis for total Actinobacteria and for Streptomyces isolates were performed to estimate species' richness in the intermediate lagoon (IL) in the oligotrophic Churince system. A total of 350 morphologically and nutritionally diverse isolates were successfully cultured and characterized as members of the Phylum Actinobacteria. A total of 105 from the total isolates were successfully subcultured, processed for DNA extraction and 16S-rDNA sequenced. All strains belong to the order Actinomycetales, encompassing 11 genera of Actinobacteria; the genus Streptomyces was found to be the most abundant taxa in all the media tested throughout the 3-year sampling period. Phylogenetic analysis of our isolates and another 667 reference strains of the family Streptomycetaceae shows that our

  18. Accumulation, release and turnover of nutrients (C-N-P-Si) by the blue mussel Mytilus edulis under oligotrophic conditions

    NARCIS (Netherlands)

    Jansen, H.M.; Strand, O.; Verdegem, M.C.J.; Smaal, A.C.

    2012-01-01

    To evaluate the potential role of mussels in nutrient cycling in oligotrophic fjord ecosystems, we applied a multiple-element (C-N-P-Si) approach considering several physiological processes (excretion, tissue composition, biodeposition) simultaneously. The study covered one annual cycle, reflecting

  19. Long-term monitoring of fish farms: application of Nematode/Copepod index to oligotrophic conditions.

    Science.gov (United States)

    Riera, Rodrigo; Sanchez-Jerez, Pablo; Rodríguez, Myriam; Monterroso, Oscar; Ramos, Eva

    2012-04-01

    Interannual variability (2003-2008) of meiofaunal assemblages were analyzed in sediments beneath fish cages (Impact group) and in areas not affected by aquaculture activities (Control group). Organisms responded with spatial and seasonal variation in meiofauna assemblages, with an abrupt increase of abundances in locations beneath fish cages throughout the study period. This increase was greater during the last sampling year (2008) and mainly due to high abundances of nematodes. Univariate analyses showed differences between control and impacted sites at both sites, however, only significant variations were found in Los Gigantes, which are consistent with seasonal meiofauna variations throughout the study period. These results are partially explained by differences in current velocity between both sampling areas. The Ne/Co index showed the same trend and it seems to be a reliable index in sediment slightly affected by aquaculture wastes. This index is especially recommended in oligotrophic areas (e.g. Canary Islands) where meiofaunal assemblages are poorly represented in terms of abundances.

  20. Faster growth of the major prokaryotic versus eukaryotic CO2 fixers in the oligotrophic ocean.

    Science.gov (United States)

    Zubkov, Mikhail V

    2014-04-29

    Because maintenance of non-scalable cellular components--membranes and chromosomes--requires an increasing fraction of energy as cell size decreases, miniaturization comes at a considerable energetic cost for a phytoplanktonic cell. Consequently, if eukaryotes can use their superior energetic resources to acquire nutrients with more or even similar efficiency compared with prokaryotes, larger unicellular eukaryotes should be able to achieve higher growth rates than smaller cyanobacteria. Here, to test this hypothesis, we directly compare the intrinsic growth rates of phototrophic prokaryotes and eukaryotes from the equatorial to temperate South Atlantic using an original flow cytometric (14)CO2-tracer approach. At the ocean basin scale, cyanobacteria double their biomass twice as frequently as the picoeukaryotes indicating that the prokaryotes are faster growing CO2 fixers, better adapted to phototrophic living in the oligotrophic open ocean-the most extensive biome on Earth.

  1. Water level influences on body condition of Geophagus brasiliensis (Perciformes: Cichlidae in a Brazilian oligotrophic reservoir

    Directory of Open Access Journals (Sweden)

    Alejandra Filippo Gonzalez Neves dos Santos

    2004-09-01

    Full Text Available Effects of water level fluctuations on body condition of Geophagus brasiliensis were studied in a 30 km² Brazilian oligotrophic reservoir. Physiological condition (K and gonadosomatic index (GSI were compared according to water level (low and high. Females' best conditions were associated to higher resources availability during high water, since gonad development did not change between low and high water. Males' condition did not change between water levels, while the highest gonad development occurred in low water. Females presented higher reproductive investment than males, which allocated most of energy for somatic development. This strategy could be a mechanism to undergo the stress caused by oligotrophic characteristics of the reservoir enhanced during low water level.Efeitos do nível da água na condição de Geophagus brasiliensis foram analisados em um reservatório oligotrófico. A condição fisiológica (K e o índice gonadossomático (IGS foram comparados entre os níveis da água (baixo e alto. Melhores condições de fêmeas foram associadas a maiores disponibilidades de recursos no nível alto, já que o desenvolvimento gonadal não variou. Não foram registradas diferenças na condição de machos, contudo maiores valores de IGS ocorreram no nível baixo. Fêmeas apresentaram elevado investimento reprodutivo, enquanto machos investiram mais no desenvolvimento somático. Tal estratégia pode ser um mecanismo para suportar o estresse causado pelas características oligotróficas do reservatório, intensificadas durante o período de níveis baixos da água.

  2. Natural events of anoxia and low respiration index in oligotrophic lakes of the Atlantic Tropical Forest

    Science.gov (United States)

    Marotta, H.; Fontes, M. L. S.; Petrucio, M. M.

    2012-08-01

    Hypoxia is a well-recognized condition reducing biodiversity and increasing greenhouse gas emissions in aquatic ecosystems, especially under warmer temperatures of tropical waters. Anoxia is a natural event commonly intensified by human-induced organic inputs in inland waters. Here, we assessed the partial pressure of O2 (pO2) and CO2 (pCO2), and the ratio between them (represented by the respiration index, RI) in two oligotrophic lakes of the Atlantic Tropical Forest, encompassing dry and rainy seasons over 19 months. We formulated the hypothesis that thermal stratification events could be coupled to natural hypoxia in deep waters of both lakes. Our results indicated a persistence of CO2 emissions from these tropical lakes to the atmosphere, on average ± standard error (SE) of 17.4 mg C m-2 h-1 probably subsided by terrestrial C inputs from the forest. Additionally, the thermal stratification during the end of the dry season and the rainy summer was coupled to anoxic events and very low RI in deep waters, and to significantly higher pO2 and RI at the surface (about 20 000 μatm and 1.0, respectively). In contrast, the water mixing during dry seasons at the beginning of the winter was related to a strong destratification in pO2, pCO2 and RI in surface and deep waters, without reaching any anoxic conditions throughout the water column. These findings confirm our hypothesis, suggesting that lakes of the Atlantic Tropical Forest could be dynamic, but especially sensitive to organic inputs. Natural anoxic events indicate that tropical oligotrophic lakes might be highly influenced by human land uses, which increase organic discharges into the watershed.

  3. Laboratory experiment to determine phosphate release rates from sediments of a formerly oligotrophic lake (Silbersee, Cuxhaven)

    Science.gov (United States)

    Elmarami, Hatem; Greskowiak, Janek; Hamann, Enrico; Massmann, Gudrun

    2016-04-01

    The Silbersee is a small, formerly oligotrophic lake in northwestern Germany which still contains rare character species for oligotrophic lakes (Isoëtes lacustris, Littorella uniflora) threatened by eutrophication. It was suspected that the lake sediments and the redox conditions in the hypolimnon play an important role with regard to eutrophication, potentially releasing phosphorus (P) into the water column. This was the motivation to conduct experiments to estimate the release rate of phosphorus into the lake. It had been noted that the P concentrations in the bottom water were higher during summer in the stagnation phase, when conditions turned sulfidic. Eight sediment cores were taken with a Mondsee-corer (manufactured by UWITEC) at different sites of the lake. The thickness of the sediment within the cores ranged from 15cm to 35 cm and were overlying by approximately 40cm of lake water water. The headspace was approximately 10cm. The cores were stored in a fridge first under oxic, then under anoxic conditions as observed in the lake bottom water in the different seasons. Redox conditions were maintained by bubbling with oxygen and nitrogen gas during the respective time periods. During the experiment, the temperature was held constant to match the water temperature measured at the bottom of the lake (~ 7±1°C). Concentrations of total phosphorus (TP) and dissolved phosphorus (DP), iron (Fe) and dissolved oxygen (DO) as well as pH were measured under oxic and anoxic conditions in the water column. The results showed that TP, DP and Fe concentrations were higher under anoxic conditions than under oxic conditions. The observed increase of phosporous in the water column during the anoxic phase was presumably a result of (i) reductive Fe-oxides dissolution and the corresponding loss of sorption sites and (ii) desorption of phosphorous via surface complexation reactions due to pH changes during the experiment.

  4. Impact of water-level changes to aquatic vegetation in small oligotrophic lakes

    Directory of Open Access Journals (Sweden)

    Egert VANDEL

    2016-06-01

    Full Text Available This study demonstrates the effect of drastic water-level changes to the aquatic vegetation in three small oligotrophic lakes situated in Kurtna Kame Field in north-eastern Estonia. The area holds around 40 lakes in 30 km2 of which 18 lakes are under protection as Natura Habitat lakes (Natura 2000 network. The area is under a strong human impact as it is surrounded by oil shale mines, sand quarry, peat harvesting field etc. The most severe impact comes from the groundwater intake established in 1972 in the vicinity of three studied lakes. The exploitation of groundwater led to drastic water-level drops. In 1980s the water-level drops were measured to be up to 3 to 4 meters compared to the levels of 1946. Lake Martiska and Lake Kuradijärv were severely affected and only 29% and 45% of lake area respectively and 21% of initial volume remained. Both lakes were described as oligotrophic lakes before severe human impact and held characteristic macrophytes such as Isoëtes lacustris L., Sparganium angustifolium Michx and Lobelia dortmanna L. As the water level declined the lakes lost their rare characteristic species and can now be described more as a meso- or even eutrophic lakes. When the volume of groundwater abstraction decreased in the 1990s the water levels started to recover but did not reach the natural levels of pre-industrialized era. Also the vegetation did not show any signs of recovery. In 2012 the pumping rates increased again causing a new rapid decline in water levels which almost exceed the previous minimum levels. The water-level monitoring alongside with the macrophyte monitoring data gives us a good case study on how the long term abrupt water-level changes can affect the aquatic vegetation

  5. Organic nutrient enrichment in the oligotrophic ocean: Impacts on remineralization, carbon sequestration, and community structure

    Science.gov (United States)

    Mackey, K. R.; Paytan, A.; Post, A. F.

    2007-12-01

    In oligotrophic seas where inorganic nitrogen (N) and phosphorus (P) are below the limits of detection, organic forms of these nutrients may constitute greater than 90% of the total N and P in the euphotic zone. The combined enzymatic activity of phytoplankton and heterotrophic bacteria determines the rate of nutrient remineralization, thereby influencing phytoplankton growth rates and carbon sequestration in these regions. In this study we investigated the effects of fertilization with ammonium (NH4), nitrate (NO3), nitrite (NO2), and phosphate (PO4) as well as various forms of organic N (urea, glycine) and P (deoxyribonucleic acid, 2- aminoethyl phosphonic acid, phytic acid) on the growth and taxonomic composition of the phytoplankton community in the Gulf of Aqaba, Red Sea. The impacts of these changes on nutrient cycling and biological assimilation were also assessed. Organic N additions led to phytoplankton growth when given together with PO4, yielding 2-3 fold increases in chlorophyll a (Chl a) and cell density relative to initial levels. Moreover, our results show that addition of NH4 or NO3 led to accumulation of extra-cellular NO2, suggesting that incomplete assimilatory reduction of NO3 by phytoplankton as well as chemoautotrophic oxidation of NH4 by ammonium oxidizing microbes contributed to NO2 formation. These findings conflict with earlier studies in the Gulf that attributed NO2 formation solely to the phytoplankton community. Organic P additions also led to 2-3 fold increases in Chl a and cell density relative to initial levels when given together with NH4 and NO3. Compared to other P additions, DNA led to the rapid accumulation of extra-cellular PO4, indicating substantial nucleotidase activity in excess of the amount needed to meet phytoplankton growth requirements. These results show the importance and interconnectivity of phytoplankton and heterotrophic bacteria communities in contributing to nutrient cycling and carbon sequestration in

  6. Microbial communities in dark oligotrophic volcanic ice cave ecosystems of Mt. Erebus, Antarctica

    Directory of Open Access Journals (Sweden)

    Bradley M. Tebo

    2015-03-01

    Full Text Available The Earth’s crust hosts a subsurface, dark, and oligotrophic biosphere that is poorly understood in terms of the energy supporting its biomass production and impact on food webs at the Earth’s surface. Dark oligotrophic volcanic ecosystems (DOVEs are good environments for investigations of life in the absence of sunlight as they are poor in organics, rich in chemical reactants and well known for chemical exchange with Earth’s surface systems. Ice caves near the summit of Mt. Erebus (Antarctica offer DOVEs in a polar alpine environment that is starved in organics and with oxygenated hydrothermal circulation in highly reducing host rock. We surveyed the microbial communities using PCR, cloning, sequencing and analysis of the small subunit (16S ribosomal and Ribulose-1,5-bisphosphate Carboxylase/Oxygenase (RubisCO genes in sediment samples from three different caves, two that are completely dark and one that receives snow-filtered sunlight seasonally. The microbial communities in all three caves are composed primarily of Bacteria and fungi; Archaea were not detected. The bacterial communities from these ice caves display low phylogenetic diversity, but with a remarkable diversity of RubisCO genes including new deeply branching Form I clades, implicating the Calvin-Benson-Bassham cycle as a pathway of CO2 fixation. The microbial communities in one of the dark caves, Warren Cave, which has a remarkably low phylogenetic diversity, were analyzed in more detail to gain a possible perspective on the energetic basis of the microbial ecosystem in the cave. Atmospheric carbon (CO2 and CO, including from volcanic emissions, likely supplies carbon and/or some of the energy requirements of chemoautotrophic microbial communities in Warren Cave and probably other Mt. Erebus ice caves. Our work casts a first glimpse at Mt. Erebus ice caves as natural laboratories for exploring carbon, energy and nutrient sources in the subsurface biosphere and the

  7. Interplay between the localization and kinetics of phosphorylation in flagellar pole development of the bacterium Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Carolina Tropini

    Full Text Available Bacterial cells maintain sophisticated levels of intracellular organization that allow for signal amplification, response to stimuli, cell division, and many other critical processes. The mechanisms underlying localization and their contribution to fitness have been difficult to uncover, due to the often challenging task of creating mutants with systematically perturbed localization but normal enzymatic activity, and the lack of quantitative models through which to interpret subtle phenotypic changes. Focusing on the model bacterium Caulobacter crescentus, which generates two different types of daughter cells from an underlying asymmetric distribution of protein phosphorylation, we use mathematical modeling to investigate the contribution of the localization of histidine kinases to the establishment of cellular asymmetry and subsequent developmental outcomes. We use existing mutant phenotypes and fluorescence data to parameterize a reaction-diffusion model of the kinases PleC and DivJ and their cognate response regulator DivK. We then present a systematic computational analysis of the effects of changes in protein localization and abundance to determine whether PleC localization is required for correct developmental timing in Caulobacter. Our model predicts the developmental phenotypes of several localization mutants, and suggests that a novel strain with co-localization of PleC and DivJ could provide quantitative insight into the signaling threshold required for flagellar pole development. Our analysis indicates that normal development can be maintained through a wide range of localization phenotypes, and that developmental defects due to changes in PleC localization can be rescued by increased PleC expression. We also show that the system is remarkably robust to perturbation of the kinetic parameters, and while the localization of either PleC or DivJ is required for asymmetric development, the delocalization of one of these two components does

  8. Interplay between the localization and kinetics of phosphorylation in flagellar pole development of the bacterium Caulobacter crescentus.

    Science.gov (United States)

    Tropini, Carolina; Huang, Kerwyn Casey

    2012-01-01

    Bacterial cells maintain sophisticated levels of intracellular organization that allow for signal amplification, response to stimuli, cell division, and many other critical processes. The mechanisms underlying localization and their contribution to fitness have been difficult to uncover, due to the often challenging task of creating mutants with systematically perturbed localization but normal enzymatic activity, and the lack of quantitative models through which to interpret subtle phenotypic changes. Focusing on the model bacterium Caulobacter crescentus, which generates two different types of daughter cells from an underlying asymmetric distribution of protein phosphorylation, we use mathematical modeling to investigate the contribution of the localization of histidine kinases to the establishment of cellular asymmetry and subsequent developmental outcomes. We use existing mutant phenotypes and fluorescence data to parameterize a reaction-diffusion model of the kinases PleC and DivJ and their cognate response regulator DivK. We then present a systematic computational analysis of the effects of changes in protein localization and abundance to determine whether PleC localization is required for correct developmental timing in Caulobacter. Our model predicts the developmental phenotypes of several localization mutants, and suggests that a novel strain with co-localization of PleC and DivJ could provide quantitative insight into the signaling threshold required for flagellar pole development. Our analysis indicates that normal development can be maintained through a wide range of localization phenotypes, and that developmental defects due to changes in PleC localization can be rescued by increased PleC expression. We also show that the system is remarkably robust to perturbation of the kinetic parameters, and while the localization of either PleC or DivJ is required for asymmetric development, the delocalization of one of these two components does not prevent

  9. Two outer membrane proteins contribute to cellular fitness in Caulobacter crescentus by preventing intracellular S-layer protein accumulation.

    Science.gov (United States)

    Overton, K Wesley; Park, Dan M; Yung, Mimi C; Dohnalkova, Alice C; Smit, John; Jiao, Yongqin

    2016-09-23

    Surface layers, or S-layers, are two-dimensional protein arrays that form the outermost layer of many bacteria and archaea. They serve several functions including physical protection of the cell from environmental threats. The high abundance of S-layer proteins necessitates a highly efficient export mechanism to transport S-layer protein from the cytoplasm to the cell exterior. Caulobacter crescentus is unique in that it has two homologous, seemingly redundant outer membrane proteins, RsaFa and RsaFb, that, together with other components, form a type I protein translocation pathway for S-layer export. These proteins have homology to E. coli TolC, the outer membrane channel of multidrug efflux pumps. Here we provide evidence that, unlike TolC, RsaFa and RsaFb are not involved in either the maintenance of membrane stability or the active export of antimicrobial compounds. Rather, RsaFa and RsaFb are required to prevent intracellular accumulation and aggregation of the S-layer protein RsaA; deletion of RsaFa and RsaFb led to a general growth defect and lowered cellular fitness. Using Western blotting, transmission electron microscopy, and RNA-seq, we show that loss of both RsaFa and RsaFb led to accumulation of insoluble RsaA in the cytoplasm, which in turn caused upregulation of a number of genes involved in protein mis-folding and degradation pathways. These findings provide new insight into the requirement for RsaFa and RsaFb in cellular fitness and tolerance to antimicrobial agents and further our understanding of the S-layer export mechanism on both the transcriptional and translational levels in C. crescentus IMPORTANCE: Decreased growth rate and reduced cell fitness are common side effects of protein production in overexpression systems. Inclusion bodies typically form inside the cell largely due to lack of sufficient export machinery to transport the overexpressed proteins to the extracellular environment. This phenomenon can conceivably also occur in natural

  10. A novel membrane anchor for FtsZ is linked to cell wall hydrolysis in Caulobacter crescentus.

    Science.gov (United States)

    Meier, Elizabeth L; Razavi, Shiva; Inoue, Takanari; Goley, Erin D

    2016-07-01

    In most bacteria, the tubulin-like GTPase FtsZ forms an annulus at midcell (the Z-ring) which recruits the division machinery and regulates cell wall remodeling. Although both activities require membrane attachment of FtsZ, few membrane anchors have been characterized. FtsA is considered to be the primary membrane tether for FtsZ in bacteria, however in Caulobacter crescentus, FtsA arrives at midcell after stable Z-ring assembly and early FtsZ-directed cell wall synthesis. We hypothesized that additional proteins tether FtsZ to the membrane and demonstrate that in C. crescentus, FzlC is one such membrane anchor. FzlC associates with membranes directly in vivo and in vitro and recruits FtsZ to membranes in vitro. As for most known membrane anchors, the C-terminal peptide of FtsZ is required for its recruitment to membranes by FzlC in vitro and midcell recruitment of FzlC in cells. In vivo, overproduction of FzlC causes cytokinesis defects whereas deletion of fzlC causes synthetic defects with dipM, ftsE and amiC mutants, implicating FzlC in cell wall hydrolysis. Our characterization of FzlC as a novel membrane anchor for FtsZ expands our understanding of FtsZ regulators and establishes a role for membrane-anchored FtsZ in the regulation of cell wall hydrolysis.

  11. The small protein MbiA interacts with MreB and modulates cell shape in Caulobacter crescentus.

    Science.gov (United States)

    Yakhnina, Anastasiya A; Gitai, Zemer

    2012-09-01

    In Caulobacter crescentus, the actin homologue MreB is critical for cell shape maintenance. Despite the central importance of MreB for cell morphology and viability, very little is known about MreB-interacting factors. Here, we use an overexpression approach to identify a novel MreB interactor, MbiA. MbiA interacts with MreB in both biochemical and genetic assays, colocalizes with MreB throughout the cell cycle, and relies on MreB for its localization. MbiA overexpression mimics the loss of MreB function, severely perturbing cell morphology, inhibiting growth and inducing cell lysis. Additionally, mbiA deletion shows a synthetic growth phenotype with a hypomorphic allele of the MreB interactor RodZ, suggesting that these two MreB-interacting proteins either have partially redundant functions or participate in the same functional complex. Our work thus establishes MbiA as a novel cell shape regulator that appears to function through regulating MreB, and opens avenues for discovery of more MreB-regulating factors by showing that overexpression screens are a valuable tool for uncovering potentially redundant cell shape effectors.

  12. Phosphotransferase-dependent accumulation of (p)ppGpp in response to glutamine deprivation in Caulobacter crescentus

    Science.gov (United States)

    Ronneau, Séverin; Petit, Kenny; De Bolle, Xavier; Hallez, Régis

    2016-01-01

    The alarmone (p)ppGpp is commonly used by bacteria to quickly respond to nutrient starvation. Although (p)ppGpp synthetases such as SpoT have been extensively studied, little is known about the molecular mechanisms stimulating alarmone synthesis upon starvation. Here, we describe an essential role of the nitrogen-related phosphotransferase system (PTSNtr) in controlling (p)ppGpp accumulation in Caulobacter crescentus. We show that cells sense nitrogen starvation by way of detecting glutamine deprivation using the first enzyme (EINtr) of PTSNtr. Decreasing intracellular glutamine concentration triggers phosphorylation of EINtr and its downstream components HPr and EIIANtr. Once phosphorylated, both HPr∼P and EIIANtr∼P stimulate (p)ppGpp accumulation by modulating SpoT activities. This burst of second messenger primarily impacts the non-replicative phase of the cell cycle by extending the G1 phase. This work highlights a new role for bacterial PTS systems in stimulating (p)ppGpp accumulation in response to metabolic cues and in controlling cell cycle progression and cell growth. PMID:27109061

  13. A dynamic complex of signaling proteins uses polar localization to regulate cell-fate asymmetry in Caulobacter crescentus.

    Science.gov (United States)

    Tsokos, Christos G; Perchuk, Barrett S; Laub, Michael T

    2011-03-15

    Cellular asymmetry is critical to metazoan development and the life cycle of many microbes. In Caulobacter, cell cycle progression and the formation of asymmetric daughter cells depend on the polarly-localized histidine kinase CckA. How CckA is regulated and why activity depends on localization are unknown. Here, we demonstrate that the unorthodox kinase DivL promotes CckA activity and that the phosphorylated regulator DivK inhibits CckA by binding to DivL. Early in the cell cycle, CckA is activated by the dephosphorylation of DivK throughout the cell. However, in later stages, when phosphorylated DivK levels are high, CckA activation relies on polar localization with a DivK phosphatase. Localization thus creates a protected zone for CckA within the cell, without the use of membrane-enclosed compartments. Our results reveal the mechanisms by which CckA is regulated in a cell-type-dependent manner. More generally, our findings reveal how cells exploit subcellular localization to orchestrate sophisticated regulatory processes.

  14. A DNA damage checkpoint in Caulobacter crescentus inhibits cell division through a direct interaction with FtsW.

    Science.gov (United States)

    Modell, Joshua W; Hopkins, Alexander C; Laub, Michael T

    2011-06-15

    Following DNA damage, cells typically delay cell cycle progression and inhibit cell division until their chromosomes have been repaired. The bacterial checkpoint systems responsible for these DNA damage responses are incompletely understood. Here, we show that Caulobacter crescentus responds to DNA damage by coordinately inducing an SOS regulon and inhibiting the master regulator CtrA. Included in the SOS regulon is sidA (SOS-induced inhibitor of cell division A), a membrane protein of only 29 amino acids that helps to delay cell division following DNA damage, but is dispensable in undamaged cells. SidA is sufficient, when overproduced, to block cell division. However, unlike many other regulators of bacterial cell division, SidA does not directly disrupt the assembly or stability of the cytokinetic ring protein FtsZ, nor does it affect the recruitment of other components of the cell division machinery. Instead, we provide evidence that SidA inhibits division by binding directly to FtsW to prevent the final constriction of the cytokinetic ring.

  15. DNA methylation by CcrM activates the transcription of two genes required for the division of Caulobacter crescentus.

    Science.gov (United States)

    Gonzalez, Diego; Collier, Justine

    2013-04-01

    DNA methylation regulates many processes, including gene expression, by superimposing secondary information on DNA sequences. The conserved CcrM enzyme, which methylates adenines in GANTC sequences, is essential to the viability of several Alphaproteobacteria. In this study, we find that Caulobacter crescentus cells lacking the CcrM enzyme accumulate low levels of the two conserved FtsZ and MipZ proteins, leading to a severe defect in cell division. This defect can be compensated by the expression of the ftsZ gene from an inducible promoter or by spontaneous suppressor mutations that promote FtsZ accumulation. We show that CcrM promotes the transcription of the ftsZ and mipZ genes and that the ftsZ and mipZ promoter regions contain a conserved CGACTC motif that is critical to their activities and to their regulation by CcrM. In addition, our results suggest that the ftsZ promoter has the lowest activity when the CGACTC motif is non-methylated, an intermediate activity when it is hemi-methylated and the highest activity when it is fully methylated. The regulation of ftsZ expression by DNA methylation may explain why CcrM is essential in a subset of Alphaproteobacteria.

  16. Crystal structure of Caulobacter crescentus polynucleotide phosphorylase reveals a mechanism of RNA substrate channelling and RNA degradosome assembly.

    Science.gov (United States)

    Hardwick, Steven W; Gubbey, Tobias; Hug, Isabelle; Jenal, Urs; Luisi, Ben F

    2012-04-01

    Polynucleotide phosphorylase (PNPase) is an exoribonuclease that cleaves single-stranded RNA substrates with 3'-5' directionality and processive behaviour. Its ring-like, trimeric architecture creates a central channel where phosphorolytic active sites reside. One face of the ring is decorated with RNA-binding K-homology (KH) and S1 domains, but exactly how these domains help to direct the 3' end of single-stranded RNA substrates towards the active sites is an unsolved puzzle. Insight into this process is provided by our crystal structures of RNA-bound and apo Caulobacter crescentus PNPase. In the RNA-free form, the S1 domains adopt a 'splayed' conformation that may facilitate capture of RNA substrates. In the RNA-bound structure, the three KH domains collectively close upon the RNA and direct the 3' end towards a constricted aperture at the entrance of the central channel. The KH domains make non-equivalent interactions with the RNA, and there is a marked asymmetry within the catalytic core of the enzyme. On the basis of these data, we propose that structural non-equivalence, induced upon RNA binding, helps to channel substrate to the active sites through mechanical ratcheting. Structural and biochemical analyses also reveal the basis for PNPase association with RNase E in the multi-enzyme RNA degradosome assembly of the α-proteobacteria.

  17. Cloning and characterization of the gene encoding the PepF endopeptidase from the aquatic bacterium Caulobacter crescentus

    Directory of Open Access Journals (Sweden)

    Braz Vânia S.

    2002-01-01

    Full Text Available The metallopeptidases have a very important role in bacteria, being involved in several processes that rely on protein turnover, such as nutrition, degradation of signal peptides, protein localization and virulence. We have cloned and characterized the gene of the metalloendopeptidase PepF from the aquatic bacterium Caulobacter crescentus. The gene upstream of pepF (orf1 encodes a conserved hypothetical protein found in Mycobacterium and Streptomyces. pepF is co-transcribed with the gene downstream (orf3, which encodes a protein that belongs to the ABC1 protein kinase family, suggesting that these two proteins may share a common function in the cell. The C. crescentus PepF protein possesses the conserved HEXGH motif present in zinc binding domains of PepF homologs. Disruption of the pepF gene by insertion of a vector sequence did not produced any growth defect, but the mutant strain possesses only 30% of the specific activity of endopeptidases present in the wild type strain. Deletions and point mutations in the regulatory region showed that there are two putative promoter regions, and the operon expression is independent of the transcription regulator CtrA. The results indicate that PepF is not essential for either growth or development of this bacterium using peptides as the sole carbon source, suggesting that other peptidases can be sharing this function.

  18. Osmolality-dependent relocation of penicillin-binding protein PBP2 to the division site in Caulobacter crescentus.

    Science.gov (United States)

    Hocking, Jason; Priyadarshini, Richa; Takacs, Constantin N; Costa, Teresa; Dye, Natalie A; Shapiro, Lucy; Vollmer, Waldemar; Jacobs-Wagner, Christine

    2012-06-01

    The synthesis of the peptidoglycan cell wall is carefully regulated in time and space. In nature, this essential process occurs in cells that live in fluctuating environments. Here we show that the spatial distributions of specific cell wall proteins in Caulobacter crescentus are sensitive to small external osmotic upshifts. The penicillin-binding protein PBP2, which is commonly branded as an essential cell elongation-specific transpeptidase, switches its localization from a dispersed, patchy pattern to an accumulation at the FtsZ ring location in response to osmotic upshifts as low as 40 mosmol/kg. This osmolality-dependent relocation to the division apparatus is initiated within less than a minute, while restoration to the patchy localization pattern is dependent on cell growth and takes 1 to 2 generations. Cell wall morphogenetic protein RodA and penicillin-binding protein PBP1a also change their spatial distribution by accumulating at the division site in response to external osmotic upshifts. Consistent with its ecological distribution, C. crescentus displays a narrow range of osmotolerance, with an upper limit of 225 mosmol/kg in minimal medium. Collectively, our findings reveal an unsuspected level of environmental regulation of cell wall protein behavior that is likely linked to an ecological adaptation.

  19. Shotgun proteomic analysis unveils survival and detoxification strategies by Caulobacter crescentus during exposure to uranium, chromium, and cadmium.

    Science.gov (United States)

    Yung, Mimi C; Ma, Jincai; Salemi, Michelle R; Phinney, Brett S; Bowman, Grant R; Jiao, Yongqin

    2014-04-01

    The ubiquitous bacterium Caulobacter crescentus holds promise to be used in bioremediation applications due to its ability to mineralize U(VI) under aerobic conditions. Here, cell free extracts of C. crescentus grown in the presence of uranyl nitrate [U(VI)], potassium chromate [Cr(VI)], or cadmium sulfate [Cd(II)] were used for label-free proteomic analysis. Proteins involved in two-component signaling and amino acid metabolism were up-regulated in response to all three metals, and proteins involved in aerobic oxidative phosphorylation and chemotaxis were down-regulated under these conditions. Clustering analysis of proteomic enrichment revealed that the three metals also induce distinct patterns of up- or down-regulated expression among different functional classes of proteins. Under U(VI) exposure, a phytase enzyme and an ABC transporter were up-regulated. Heat shock and outer membrane responses were found associated with Cr(VI), while efflux pumps and oxidative stress proteins were up-regulated with Cd(II). Experimental validations were performed on select proteins. We found that a phytase plays a role in U(VI) and Cr(VI) resistance and detoxification and that a Cd(II)-specific transporter confers Cd(II) resistance. Interestingly, analysis of promoter regions in genes associated with differentially expressed proteins suggests that U(VI) exposure affects cell cycle progression.

  20. Evaluating secretion and surface attachment of SapA, an S-layer-associated metalloprotease of Caulobacter crescentus.

    Science.gov (United States)

    Gandham, Lyngrace; Nomellini, John F; Smit, John

    2012-10-01

    Caulobacter crescentus is used to display foreign peptides at high density as insertions into the surface (S)-layer protein (RsaA). Many recombinant RsaA proteins, however, are cleaved by SapA, a 71-kDa metalloprotease, suggesting a role in maintaining S-layer integrity. When overexpressed on a multicopy plasmid SapA was detected on the surface by fluorescent antibody only if RsaA and the O-side chain of LPS that mediates S-layer attachment were removed by mutation, indicating an outer membrane location beneath the S-layer. Secretion was mediated by the RsaA type 1 transporter since secretion was eliminated in transporter deficient strains or by C-terminal deletions in SapA (the presumed location of type 1 secretion signals). Secretion was required to become an active protease; mass spectrometry suggested this might be due to N-terminal processing during secretion, a feature shared with other type 1-secreted proteases. Overexpression leads to additional processing C-terminal to the protease domain, producing a 45-kDa protein. This was demonstrated to be self-processing. Deletion analysis revealed the C-terminal 100 amino acids were sufficient for anchoring and secretion. When protein G was fused to the last 238 amino acids of SapA it was secreted, surface attached and bound immunoglobulin, indicating potential for foreign protein display.

  1. Identification of a U/Zn/Cu responsive global regulatory two-component system in Caulobacter crescentus.

    Science.gov (United States)

    Park, Dan M; Overton, K Wesley; Liou, Megan J; Jiao, Yongqin

    2016-12-30

    Despite the well-known toxicity of uranium (U) to bacteria, little is known about how cells sense and respond to U. The recent finding of a U-specific stress response in Caulobacter crescentus has provided a foundation for studying the mechanisms of U- perception in bacteria. To gain insight into this process, we used a forward genetic screen to identify the regulatory components governing expression of the urcA promoter (PurcA ) that is strongly induced by U. This approach unearthed a previously uncharacterized two-component system, named UzcRS, which is responsible for U-dependent activation of PurcA . UzcRS is also highly responsive to zinc and copper, revealing a broader specificity than previously thought. Using ChIP-seq, we found that UzcR binds extensively throughout the genome in a metal-dependent manner and recognizes a noncanonical DNA-binding site. Coupling the genome-wide occupancy data with RNA-seq analysis revealed that UzcR is a global regulator of transcription, predominately activating genes encoding proteins that are localized to the cell envelope; these include metallopeptidases, multidrug-resistant efflux (MDR) pumps, TonB-dependent receptors and many proteins of unknown function. Collectively, our data suggest that UzcRS couples the perception of U, Zn and Cu with a novel extracytoplasmic stress response.

  2. Bactofilins, a ubiquitous class of cytoskeletal proteins mediating polar localization of a cell wall synthase in Caulobacter crescentus.

    Science.gov (United States)

    Kühn, Juliane; Briegel, Ariane; Mörschel, Erhard; Kahnt, Jörg; Leser, Katja; Wick, Stephanie; Jensen, Grant J; Thanbichler, Martin

    2010-01-20

    The cytoskeleton has a key function in the temporal and spatial organization of both prokaryotic and eukaryotic cells. Here, we report the identification of a new class of polymer-forming proteins, termed bactofilins, that are widely conserved among bacteria. In Caulobacter crescentus, two bactofilin paralogues cooperate to form a sheet-like structure lining the cytoplasmic membrane in proximity of the stalked cell pole. These assemblies mediate polar localization of a peptidoglycan synthase involved in stalk morphogenesis, thus complementing the function of the actin-like cytoskeleton and the cell division machinery in the regulation of cell wall biogenesis. In other bacteria, bactofilins can establish rod-shaped filaments or associate with the cell division apparatus, indicating considerable structural and functional flexibility. Bactofilins polymerize spontaneously in the absence of additional cofactors in vitro, forming stable ribbon- or rod-like filament bundles. Our results suggest that these structures have evolved as an alternative to intermediate filaments, serving as versatile molecular scaffolds in a variety of cellular pathways.

  3. Phosphotransferase-dependent accumulation of (p)ppGpp in response to glutamine deprivation in Caulobacter crescentus.

    Science.gov (United States)

    Ronneau, Séverin; Petit, Kenny; De Bolle, Xavier; Hallez, Régis

    2016-04-25

    The alarmone (p)ppGpp is commonly used by bacteria to quickly respond to nutrient starvation. Although (p)ppGpp synthetases such as SpoT have been extensively studied, little is known about the molecular mechanisms stimulating alarmone synthesis upon starvation. Here, we describe an essential role of the nitrogen-related phosphotransferase system (PTS(Ntr)) in controlling (p)ppGpp accumulation in Caulobacter crescentus. We show that cells sense nitrogen starvation by way of detecting glutamine deprivation using the first enzyme (EI(Ntr)) of PTS(Ntr). Decreasing intracellular glutamine concentration triggers phosphorylation of EI(Ntr) and its downstream components HPr and EIIA(Ntr). Once phosphorylated, both HPr∼P and EIIA(Ntr)∼P stimulate (p)ppGpp accumulation by modulating SpoT activities. This burst of second messenger primarily impacts the non-replicative phase of the cell cycle by extending the G1 phase. This work highlights a new role for bacterial PTS systems in stimulating (p)ppGpp accumulation in response to metabolic cues and in controlling cell cycle progression and cell growth.

  4. Predicting aquatic macrophyte occurrence in soft-water oligotrophic lakes (Pyrenees mountain range

    Directory of Open Access Journals (Sweden)

    Cristina Pulido

    2014-08-01

    Full Text Available Distribution of aquatic macrophytes in lakes is related to geographical, morphological, catchment and water chemistry variables as well as human impacts, which modify the original environment. Here, we aim at building statistical models to establish the ecological niches of 11 aquatic macrophytes (10 different phanerogams and the genus Nitella from oligotrophic soft-water lakes and infer their ecological requirements and environmental constraints at the southernmost limit of their distribution. Macrophyte occurrence and environmental variables were obtained from 86 non-exploited oligotrophic soft-water lakes from the Pyrenees (Southern Europe; 42º50´N, 1º00´E; macrophytes inhabited 55 of these lakes. Optimum ranges and macrophyte occurrence were predicted in relation to 18 geographical, morphological, catchment and water chemistry variables using univariate and multivariate logistic models. Lakes at low altitude, in vegetated catchments and with low water concentration of NO3- and SO4-2, were the most suitable to host macrophytes. In general, individual species of aquatic macrophytes showed clear patterns of segregation along conductivity and pH gradients, although the specific combination of variables selected in the best models explaining their occurrence differed among species.  Based on the species response to pH and conductivity, we found Isoetes lacustris have its optimum in waters with low conductivity and pH (i.e. negative monotonic response. In contrast, Callitriche palustris, Ranunculus aquatilis, Subularia aquatica, Nitella spp., and Myriophyllum alterniflorum showed an optimum at intermediate values (i.e. unimodal response, whereas Potamogeton berchtoldii, Potamogeton alpinus, and Ranunculus trichophyllus as species had their optimum at relatively high water pH and conductivity (i.e. positive monotonic response. This pattern has been observed in other regions for the same species, although with different optima and tolerance

  5. Development of an HIV-1 specific microbicide using Caulobacter crescentus S-layer mediated display of CD4 and MIP1alpha.

    Directory of Open Access Journals (Sweden)

    John F Nomellini

    Full Text Available The development of alternative strategies to prevent HIV infection is a global public health priority. Initial efforts in anti-HIV microbicide development have met with poor success as the strategies have relied on a non-specific mechanism of action. Here, we report the development of a microbicide aimed at specifically blocking HIV entry by displaying molecular components of the HIV/host cell attachment complex on the surface of Caulobacter crescentus, a harmless aquatic bacterium. This bacterium can be readily manipulated to present heterologous proteins at high density on its surface by genetic insertion into its crystalline surface layer protein. In separate constructions, we generated bacteria displaying domain 1 of CD4 and MIP1alpha. Each moiety reacted with specific antibodies by Western immunoblot and immuno-fluorescence microscopy. Microbicide functionality was assessed using an HIV pseudotype virus assay system representing Clade B subtypes. Bacteria displaying MIP1alpha reduced infectivity by 35-78% depending on the specific subtype while CD4 display reduced infection by as much as 56%. Combinations of both constructs reduced infectivity by nearly 98%. We demonstrated that HIV infection could be inhibited using a strategy aimed at HIV-specific molecular interactions with Caulobacter surface protein display, and that sufficient protein folding and conformation could be mimicked to bind and block entry. Further, this is the first demonstration that Caulobacter surface protein display may be a useful approach to preventing HIV infection or other viruses as a microbicide. We propose that this harmless bacterium, which is inexpensive to produce and formulate, might be suitable for topical applications as a viable alternative in the search for effective microbicides to counteract the world wide incidence of HIV infection.

  6. Isolation and characterization of Caulobacter mutants impaired in adaptation to stationary phase Isolamento e caracterização de mutantes de Caulobacter deficientes na adaptação à fase estacionária

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    Valéria C. S. Italiani

    2003-04-01

    Full Text Available The entry into stationary phase causes a change in the pattern of gene expression of bacteria, when the cells must express a whole set of genes involved mainly with resistance to starvation and to environmental stresses. As an attempt to identify genes important for the survival of Caulobacter crescentus in stationary phase, we have screened a library of 5,000 clones generated by random transposon mutagenesis for mutants that showed reduced viability after prolonged growth. Four clones were selected, which displayed either lower viability or a longer time of recovery from stationary phase. The genes disrupted were identified, and the gene products were found to be mainly involved with amino acid metabolism (glutamate N-acetyltransferase, 4-hydroxyphenylpyruvate dioxygenase and L-aspartate oxidase or with recombination (exonuclease RecJ. Each mutant was tested for resistance to stresses, such as oxidative, saline, acidic, heat and UV exposure, showing different responses. Although the mutations obtained were not in genes involved specifically in stationary phase, our results suggest that amino acids metabolism may play an important role in keeping viability during this growth phase.A entrada em fase estacionária causa uma mudança no padrão de expressão gênica de bactérias, quando as células devem expressar um novo conjunto de genes envolvidos principalmente com resistência à carência alimentar e a estresses ambientais. Em uma tentativa de identificar genes importantes para a sobrevivência de Caulobacter crescentus em fase estacionária, nós varremos uma biblioteca de 5.000 clones gerados por transposição aleatória em busca de mutantes que mostrassem viabilidade reduzida após crescimento prolongado. Quatro clones foram selecionados, que mostraram menor viabilidade ou um maior tempo de recuperação da fase estacionária. Os genes interrompidos foram identificados, e os produtos gênicos mostraram-se estar envolvidos principalmente

  7. Turning off flagellum rotation requires the pleiotropic gene pleD: pleA, pleC, and pleD define two morphogenic pathways in Caulobacter crescentus.

    OpenAIRE

    Sommer, J M; Newton, A

    1989-01-01

    We have identified mutations in three pleiotropic genes, pleA, pleC, and pleD, that are required for differentiation in Caulobacter crescentus. pleA and pleC mutants were isolated in an extensive screen for strains defective in both motility and adsorption of polar bacteriophage phi CbK; using temperature-sensitive alleles, we determined the time at which the two genes act. pleA was required for a short period at 0.7 of the swarmer cell cycle for flagellum biosynthesis, whereas pleC was requi...

  8. Deposition, burial and sequestration of carbon in an oligotrophic, tropical lake

    Directory of Open Access Journals (Sweden)

    Javier Alcocer

    2014-02-01

    Full Text Available The amount of biogenic carbon that may be deposited, buried and eventually preserved (sequestered in the sediments of a tropical, oligotrophic lake, was evaluated based on i the temporal variation of the particulate organic carbon (POC concentration in the superficial sediments in the deep zone of lake Alchichica, Puebla, Mexico; and ii the POC accumulation and preservation in a 210Pb-dated sediment core from the lake. In lake Alchichica the POC concentration in the surficial sediments ranged between 12 and 60 mg POC g-1 (25 ± 12 mg POC g-1 dry weight. The magnitude of the sedimented POC in Alchichica was high and mostly of autochthonous origin. The POC concentrations recorded in the sediment core (16.6 to 31.6 mg g-1 dry weight were comparable to the concentration range observed in the surface sediment samples collected during the study period, which signaled a high POC preservation capacity in the sedimentary column of lake Alchichica. The POC fluxes, estimated from the 210Pb-dated sediment core, varied between 14.9 and 35.3 g m-2 year-1 within the past century; and the maximum POC losses through diagenesis during this period were estimated to be lower than 25%. This study concludes that deep tropical lakes, exemplified by lake Alchichica, accumulate and preserve most of the POC deposited, playing an important role in regional carbon balances.

  9. A study on an oligotrophic bacteria and its ecological characteristics in an arid desert area

    Institute of Scientific and Technical Information of China (English)

    PAN HuiXia; CHENG ZhengMing; ZHANG XueMei; MU ShuYong; QI XiaoLing; WANG Fang

    2007-01-01

    An oligotrophic bacterium was isolated from the biological soil crust underlayer in the Xinjiang Gurbantunggut Desert. It was numbered SGB-5, G+. Cell size is (0.328-0.746) μm×(0.171-0.240) μm.Raised colony is white, roundness and its diameter is 5 mm. The strain is a facultative aerobe. It was able to grow in conditions of 1 -15 mg·C·L-1 culture medium at 10-50℃. The strain's optimum growth temperature is 37℃. The range of its optimum growth pH is 8-9. A large amount of extracellular mucopolysaccharide was secreted during growth. The chemical composition of this mucopolysaccharide consists of arabinose, X sugar, glucose, galactose and mannitol. Mole ratio of these sugars is 1:14:19: 6:14. The viscosity of the mucopolysaccharide can reach 6300 mPa·s, when the strain is cultivated for 72 h. After the culture solution in which viscosity was 1500 mPa·s was sprinkled on the quicksand surface, 6 mm bacteria crust of conglutinating sand was formed. This crust could not only stabilize sand, but could also potentially slow the rate of the soil water evaporation.

  10. Bio-optical characterization in an ultra-oligotrophic region: the North central Red Sea

    KAUST Repository

    Kheireddine, Malika

    2015-04-01

    Until recently, satellite-derived ocean color observations have been the only means of evaluating optical variability of the Red Sea. During a cruise in autumn 2014, we investigated the variability of Inherent Optical Properties (IOPs) in the North Central Red Sea (NCRS) with a particular focus on the particulate backscattering coefficient, bbp, and colored dissolved organic matter, CDOM, absorption. To our knowledge, these are some of the measurements of these properties in the Red Sea. The IOPs are derived from the concentration and physical properties of suspended particles in the ocean. They provide a simple description of the influence of these particles on the light within the water column. Bio-optical relationships found for ultra-oligotrophic waters of the six stations sampled significantly depart from the mean standard relationships provided for the global ocean, showing the peculiar character of the Red Sea. These optical anomalies relate to the specific biological and environmental conditions occurring in the Red Sea ecosystem. Specifically, the surface specific phytoplankton absorption coefficients are lower than the values predicted from the global relationships due to a high proportion of relatively large sized phytoplankton. Conversely, bbp values are much higher than the mean standard values for a given chlorophyll-a concentration, Chl a. This presumably results from the influence of highly refractive submicrometer particles of Saharan origin in the surface layer of the water column.

  11. Barrenia, a new genus associated with roots of switchgrass and pine in the oligotrophic pine barrens.

    Science.gov (United States)

    Walsh, Emily; Luo, Jing; Naik, Abhishek; Preteroti, Thomas; Zhang, Ning

    2015-12-01

    A new genus Barrenia is described based on multi-gene phylogenetic analyses and phenotypic and ecological characters. Isolated from roots of switchgrass and pitch pine in the acidic and oligotrophic New Jersey Pine Barrens in this study, Barrenia likely has a wide distribution because its internal transcribed spacer (ITS) sequence has high similarity with a number of GenBank sequences from various ecological studies. The majority of these matching samples were from roots of plants in acidic, nutrient-poor environments, as well as from managed sugarcane plantations. Phylogenetic analyses based on ITS, LSU, and RPB1 sequence data strongly support that Barrenia is a monophyletic clade in Helotiales, distinct from any known taxa. Barrenia is phylogenetically close to Acidomelania, Loramyces, Mollisia, and Phialocephala fortinii - Acephala applanata species complex (PAC), the dark septate endophytes. Barrenia can be distinguished from Loramyces and Mollisia by its association with living plant roots. Taxa in PAC also are root endophytes but they have complex phialid arrangements that appear to be lacking in Barrenia. Plant-fungal interaction experiments showed that Barrenia panicia and Acidomelania panicicola significantly promoted root hair growth in switchgrass. Results from this work will facilitate ecological and evolutionary studies on root-associated fungi.

  12. The Biogeochemistry from the Oligotrophic to the Ultraoligotrophic Mediterranean (BOUM experiment

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    T. Moutin

    2011-08-01

    Full Text Available The overall goal of the BOUM (Biogeochemistry from the Oligotrophic to the Ultraoligotrophic Mediterranean experiment was to obtain a better representation of the interactions between planktonic organisms and the cycle of biogenic elements in the Mediterranean Sea (MS, in the context of global climate change and, more particularly, on the role of the ocean in carbon sequestration through biological processes. The BOUM experiment was organized around three main objectives which are: (1 to give a longitudinal description of the biogeochemistry and the biological diversity of the MS during the strongest stratified period, (2 to study processes at the centre of three anticyclonic eddies, and (3 to obtain a representation of the main biogeochemical fluxes and the dynamics of the planktonic trophic network. The international BOUM cruise took place between 16 June and 20 July 2008, involved 32 scientists on board, and covered around 3000 km in the MS from the South of Cyprus to Marseilles (France. This paper describes in detail the objectives of the BOUM experiment, the implementation plan of the cruise, the water masses and general biogeochemical trends encountered, and lays particular emphasis on description of the sections and the main physical characteristics of the three anticyclonic eddies studied, before concluding with first order biogeochemical budgets and a general overview of the 24 other papers published in this special issue.

  13. Atmospheric aerosol deposition influences marine microbial communities in oligotrophic surface waters of the western Pacific Ocean

    Science.gov (United States)

    Maki, Teruya; Ishikawa, Akira; Mastunaga, Tomoki; Pointing, Stephen B.; Saito, Yuuki; Kasai, Tomoaki; Watanabe, Koichi; Aoki, Kazuma; Horiuchi, Amane; Lee, Kevin C.; Hasegawa, Hiroshi; Iwasaka, Yasunobu

    2016-12-01

    Atmospheric aerosols contain particulates that are deposited to oceanic surface waters. These can represent a major source of nutrients, trace metals, and organic compounds for the marine environment. The Japan Sea and the western Pacific Ocean are particularly affected by aerosols due to the transport of desert dust and industrially derived particulate matter with aerodynamic diameter less than 2.5 μm (PM2.5) from continental Asia. We hypothesized that supplementing seawater with aerosol particulates would lead to measurable changes in surface water nutrient composition as well as shifts in the marine microbial community. Shipboard experiments in the Pacific Ocean involved the recovery of oligotrophic oceanic surface water and subsequent supplementation with aerosol particulates obtained from the nearby coastal mountains, to simulate marine particulate input in this region. Initial increases in nitrates due to the addition of aerosol particulates were followed by a decrease correlated with the increase in phytoplankton biomass, which was composed largely of Bacillariophyta (diatoms), including Pseudo-nitzschia and Chaetoceros species. This shift was accompanied by changes in the bacterial community, with apparent increases in the relative abundance of heterotrophic Rhodobacteraceae and Colwelliaceae in aerosol particulate treated seawater. Our findings provide empirical evidence revealing the impact of aerosol particulates on oceanic surface water microbiology by alleviating nitrogen limitation in the organisms.

  14. Polyextremotolerant black fungi: oligotrophism, adaptive potential and a link to lichen symbioses

    Directory of Open Access Journals (Sweden)

    Cene eGostinčar

    2012-11-01

    Full Text Available Black meristematic fungi can survive high doses of radiation and are resistant to desiccation. These adaptations help them to colonize harsh oligotrophic habitats, e.g. on the surface and subsurface of rocks. One of their most characteristic stress-resistance mechanisms is the accumulation of melanin in the cell walls. This, production of other protective molecules and a plastic morphology further contribute to ecological flexibility of black fungi. Increased growth rates of some species after exposure to ionizing radiation even suggest yet unknown mechanisms of energy production. Other unusual metabolic strategies may include harvesting UV or visible light or gaining energy by forming facultative lichen-like associations with algae or cyanobacteria. The latter is not entirely surprising, since certain black fungal lineages are phylogenetically related to clades of lichen-forming fungi. Similar to black fungi, lichen-forming fungi are adapted to growth on exposed surfaces with low availability of nutrients. They also efficiently use protective molecules to tolerate frequent periods of extreme stress. Traits shared by both groups of fungi may have been important in facilitating the evolution and radiation of lichen-symbioses.

  15. Seasonal variation of the protozooplanktonic community in a tropical oligotrophic environment (Ilha Solteira reservoir, Brazil

    Directory of Open Access Journals (Sweden)

    AS Mansano

    Full Text Available The seasonal variation of the protozooplanktonic community (ciliates and testate amoebae was studied in a tropical oligotrophic reservoir in Brazil, which was under the influence of two contrasting climatic seasons (rainy/warm and dry/cold. The aim of this study was to evaluate the effect of these climatic changes on physical, chemical and biological variables in the dynamic of this community. The highest mean density of total protozoans occurred in the rainy/warm season (5683.2 ind L−1, while the lowest was in the dry/cold (2016.0 ind L−1. Considering the seasonal variations, the protozoan groups that are truly planktonic, such as the oligotrichs (Spirotrichea, predominated in the dry season, whereas during the rainy season, due to the material input and resuspension of sediment, sessile protozoans of the Peritrichia group were the most important ones. The dominant protozoans were Urotricha globosa, Cothurnia annulata, Pseudodifflugia sp. and Halteria grandinella. The highest densities of H. grandinella were associated with more oxygenated and transparent water conditions, while the highest densities of C. annulata occurred in sites with high turbidity, pH and trophic state index (TSI. The study demonstrated that density and composition of protozooplanktonic species and groups of the reservoir suffered seasonal variation due to the environmental variables (mainly temperature, turbidity, water transparency, dissolved oxygen and TSI and the biological variables (e.g. morphological characteristics, eating habits and escape strategies from predation of the species.

  16. Cold-Active, Heterotrophic Bacteria from the Highly Oligotrophic Waters of Lake Vanda, Antarctica

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    Nicole A. Vander Schaaf

    2015-07-01

    Full Text Available The permanently ice-covered lakes of the McMurdo Dry Valleys, Antarctica are distinctive ecosystems that consist strictly of microbial communities. In this study, water samples were collected from Lake Vanda, a stratified Dry Valley lake whose upper waters (from just below the ice cover to nearly 60 m are highly oligotrophic, and used to establish enrichment cultures. Six strains of psychrotolerant, heterotrophic bacteria were isolated from lake water samples from a depth of 50 or 55 m. Phylogenetic analyses showed the Lake Vanda strains to be species of Nocardiaceae, Caulobacteraceae, Sphingomonadaceae, and Bradyrhizobiaceae. All Lake Vanda strains grew at temperatures near or below 0 °C, but optimal growth occurred from 18 to 24 °C. Some strains showed significant halotolerance, but no strains required NaCl for growth. The isolates described herein include cold-active species not previously reported from Dry Valley lakes, and their physiological and phylogenetic characterization broadens our understanding of these limnologically unique lakes.

  17. Ammonia-Oligotrophic and Diazotrophic Heavy Metal-Resistant Serratia liquefaciens Strains from Pioneer Plants and Mine Tailings.

    Science.gov (United States)

    Zelaya-Molina, Lily X; Hernández-Soto, Luis M; Guerra-Camacho, Jairo E; Monterrubio-López, Ricardo; Patiño-Siciliano, Alfredo; Villa-Tanaca, Lourdes; Hernández-Rodríguez, César

    2016-08-01

    Mine tailings are man-made environments characterized by low levels of organic carbon and assimilable nitrogen, as well as moderate concentrations of heavy metals. For the introduction of nitrogen into these environments, a key role is played by ammonia-oligotrophic/diazotrophic heavy metal-resistant guilds. In mine tailings from Zacatecas, Mexico, Serratia liquefaciens was the dominant heterotrophic culturable species isolated in N-free media from bulk mine tailings as well as the rhizosphere, roots, and aerial parts of pioneer plants. S. liquefaciens strains proved to be a meta-population with high intraspecific genetic diversity and a potential to respond to these extreme conditions. The phenotypic and genotypic features of these strains reveal the potential adaptation of S. liquefaciens to oligotrophic and nitrogen-limited mine tailings with high concentrations of heavy metals. These features include ammonia-oligotrophic growth, nitrogen fixation, siderophore and indoleacetic acid production, phosphate solubilization, biofilm formation, moderate tolerance to heavy metals under conditions of diverse nitrogen availability, and the presence of zntA, amtB, and nifH genes. The acetylene reduction assay suggests low nitrogen-fixing activity. The nifH gene was harbored in a plasmid of ∼60 kb and probably was acquired by a horizontal gene transfer event from Klebsiella variicola.

  18. Two Outer Membrane Proteins Contribute to Caulobacter crescentus Cellular Fitness by Preventing Intracellular S-Layer Protein Accumulation

    Energy Technology Data Exchange (ETDEWEB)

    Overton, K. Wesley; Park, Dan M.; Yung, Mimi C.; Dohnalkova, Alice C.; Smit, John; Jiao, Yongqin; Parales, R. E.

    2016-09-23

    ABSTRACT

    Surface layers, or S-layers, are two-dimensional protein arrays that form the outermost layer of many bacteria and archaea. They serve several functions, including physical protection of the cell from environmental threats. The high abundance of S-layer proteins necessitates a highly efficient export mechanism to transport the S-layer protein from the cytoplasm to the cell exterior.Caulobacter crescentusis unique in that it has two homologous, seemingly redundant outer membrane proteins, RsaFaand RsaFb, which together with other components form a type I protein translocation pathway for S-layer export. These proteins have homology toEscherichia coliTolC, the outer membrane channel of multidrug efflux pumps. Here we provide evidence that, unlike TolC, RsaFaand RsaFbare not involved in either the maintenance of membrane stability or the active export of antimicrobial compounds. Rather, RsaFaand RsaFbare required to prevent intracellular accumulation and aggregation of the S-layer protein RsaA; deletion of RsaFaand RsaFbled to a general growth defect and lowered cellular fitness. Using Western blotting, transmission electron microscopy, and transcriptome sequencing (RNA-seq), we show that loss of both RsaFaand RsaFbled to accumulation of insoluble RsaA in the cytoplasm, which in turn caused upregulation of a number of genes involved in protein misfolding and degradation pathways. These findings provide new insight into the requirement for RsaFaand RsaFbin cellular fitness and tolerance to antimicrobial agents and further our understanding of the S-layer export mechanism on both the transcriptional and translational levels in

  19. The β-sliding clamp directs the localization of HdaA to the replisome in Caulobacter crescentus.

    Science.gov (United States)

    Fernandez-Fernandez, Carmen; Grosse, Karin; Sourjik, Victor; Collier, Justine

    2013-11-01

    The initiation of chromosome replication is tightly regulated in bacteria to ensure that it takes place only once per cell cycle. In many proteobacteria, this process requires the ATP-bound form of the DnaA protein. The regulatory inactivation of DnaA (RIDA) facilitates the conversion of DnaA-ATP into replication-inactive DnaA-ADP, thereby preventing overinitiation. Homologues of the HdaA protein, together with the β-clamp of the DNA polymerase (DnaN), are required for this process. Here, we used fluorescence resonance energy transfer experiments to demonstrate that HdaA interacts with DnaN in live Caulobacter crescentus cells. We show that a QFKLPL motif in the N-terminal region of HdaA is required for this interaction and that this motif is also needed to recruit HdaA to the subcellular location occupied by the replisome during DNA replication. An HdaA mutant protein that cannot colocalize or interact with DnaN can also not support the essential function of HdaA. These results suggest that the recruitment of HdaA to the replisome is needed during RIDA in C. crescentus, probably as a means to sense whether chromosome replication has initiated before DnaA becomes inactivated. In addition, we show that a conserved R145 residue located in the AAA+ domain of HdaA is also needed for the function of HdaA, although it does not affect the interaction of HdaA with DnaN in vivo. The AAA+ domain of HdaA may therefore be required during RIDA after the initial recruitment of HdaA to the replisome by DnaN.

  20. Functional characterization of UDP-glucose:undecaprenyl-phosphate glucose-1-phosphate transferases of Escherichia coli and Caulobacter crescentus.

    Science.gov (United States)

    Patel, Kinnari B; Toh, Evelyn; Fernandez, Ximena B; Hanuszkiewicz, Anna; Hardy, Gail G; Brun, Yves V; Bernards, Mark A; Valvano, Miguel A

    2012-05-01

    Escherichia coli K-12 WcaJ and the Caulobacter crescentus HfsE, PssY, and PssZ enzymes are predicted to initiate the synthesis of colanic acid (CA) capsule and holdfast polysaccharide, respectively. These proteins belong to a prokaryotic family of membrane enzymes that catalyze the formation of a phosphoanhydride bond joining a hexose-1-phosphate with undecaprenyl phosphate (Und-P). In this study, in vivo complementation assays of an E. coli K-12 wcaJ mutant demonstrated that WcaJ and PssY can complement CA synthesis. Furthermore, WcaJ can restore holdfast production in C. crescentus. In vitro transferase assays demonstrated that both WcaJ and PssY utilize UDP-glucose but not UDP-galactose. However, in a strain of Salmonella enterica serovar Typhimurium deficient in the WbaP O antigen initiating galactosyltransferase, complementation with WcaJ or PssY resulted in O-antigen production. Gas chromatography-mass spectrometry (GC-MS) analysis of the lipopolysaccharide (LPS) revealed the attachment of both CA and O-antigen molecules to lipid A-core oligosaccharide (OS). Therefore, while UDP-glucose is the preferred substrate of WcaJ and PssY, these enzymes can also utilize UDP-galactose. This unexpected feature of WcaJ and PssY may help to map specific residues responsible for the nucleotide diphosphate specificity of these or similar enzymes. Also, the reconstitution of O-antigen synthesis in Salmonella, CA capsule synthesis in E. coli, and holdfast synthesis provide biological assays of high sensitivity to examine the sugar-1-phosphate transferase specificity of heterologous proteins.

  1. Extracytoplasmic function (ECF sigma factor σF is involved in Caulobacter crescentus response to heavy metal stress

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    Kohler Christian

    2012-09-01

    Full Text Available Abstract Background The α-proteobacterium Caulobacter crescentus inhabits low-nutrient environments and can tolerate certain levels of heavy metals in these sites. It has been reported that C. crescentus responds to exposure to various heavy metals by altering the expression of a large number of genes. Results In this work, we show that the ECF sigma factor σF is one of the regulatory proteins involved in the control of the transcriptional response to chromium and cadmium. Microarray experiments indicate that σF controls eight genes during chromium stress, most of which were previously described as induced by heavy metals. Surprisingly, σF itself is not strongly auto-regulated under metal stress conditions. Interestingly, σF-dependent genes are not induced in the presence of agents that generate reactive oxygen species. Promoter analyses revealed that a conserved σF-dependent sequence is located upstream of all genes of the σF regulon. In addition, we show that the second gene in the sigF operon acts as a negative regulator of σF function, and the encoded protein has been named NrsF (Negative regulator of sigma F. Substitution of two conserved cysteine residues (C131 and C181 in NrsF affects its ability to maintain the expression of σF-dependent genes at basal levels. Furthermore, we show that σF is released into the cytoplasm during chromium stress and in cells carrying point mutations in both conserved cysteines of the protein NrsF. Conclusion A possible mechanism for induction of the σF-dependent genes by chromium and cadmium is the inactivation of the putative anti-sigma factor NrsF, leading to the release of σF to bind RNA polymerase core and drive transcription of its regulon.

  2. Nitrogen cycling in the subsurface biosphere: nitrate isotopes in porewaters underlying the oligotrophic North Atlantic

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    S. D. Wankel

    2015-08-01

    Full Text Available Nitrogen (N is a key component of fundamental biomolecules. Hence, the cycling and availability of N is a central factor governing the extent of ecosystems across the Earth. In the organic-lean sediment porewaters underlying the oligotrophic ocean, where low levels of microbial activity persist despite limited organic matter delivery from overlying water, the extent and modes of nitrogen transformations have not been widely investigated. Here we use the N and oxygen (O isotopic composition of porewater nitrate (NO3− from a site in the oligotrophic North Atlantic (IODP to determine the extent and magnitude of microbial nitrate production (via nitrification and consumption (via denitrification. We find that NO3− accumulates far above bottom seawater concentrations (∼ 21 μM throughout the sediment column (up to ∼ 50 μM down to the oceanic basement as deep as 90 mbsf, reflecting the predominance of aerobic nitrification/remineralization within the deep marine sediments. Large changes in the δ15N and δ18O of nitrate, however, reveal variable influence of nitrate respiration across the three sites. We use an inverse porewater diffusion–reaction model, constrained by the N and O isotope systematics of nitrification and denitrification and the porewater NO3− isotopic composition, to estimate rates of nitrification and denitrification throughout the sediment column. Results indicate variability of reaction rates across and within the three boreholes that are generally consistent with the differential distribution of dissolved oxygen at this site, though not necessarily with the canonical view of how redox thresholds separate nitrate regeneration from dissimilative consumption spatially. That is, we provide isotope evidence for expanded zones of co-ocurring nitrification and denitrification. The isotope biogeochemical modeling also yielded estimates for the δ15N and δ18O of newly produced nitrate (δ15NNTR and δ18ONTR, as well as the

  3. Nitrogen cycling in the subsurface biosphere: nitrate isotopes in porewaters underlying the oligotrophic North Atlantic

    Science.gov (United States)

    Wankel, S. D.; Buchwald, C.; Ziebis, W.; Wenk, C. B.; Lehmann, M. F.

    2015-08-01

    Nitrogen (N) is a key component of fundamental biomolecules. Hence, the cycling and availability of N is a central factor governing the extent of ecosystems across the Earth. In the organic-lean sediment porewaters underlying the oligotrophic ocean, where low levels of microbial activity persist despite limited organic matter delivery from overlying water, the extent and modes of nitrogen transformations have not been widely investigated. Here we use the N and oxygen (O) isotopic composition of porewater nitrate (NO3-) from a site in the oligotrophic North Atlantic (IODP) to determine the extent and magnitude of microbial nitrate production (via nitrification) and consumption (via denitrification). We find that NO3- accumulates far above bottom seawater concentrations (∼ 21 μM) throughout the sediment column (up to ∼ 50 μM) down to the oceanic basement as deep as 90 mbsf, reflecting the predominance of aerobic nitrification/remineralization within the deep marine sediments. Large changes in the δ15N and δ18O of nitrate, however, reveal variable influence of nitrate respiration across the three sites. We use an inverse porewater diffusion-reaction model, constrained by the N and O isotope systematics of nitrification and denitrification and the porewater NO3- isotopic composition, to estimate rates of nitrification and denitrification throughout the sediment column. Results indicate variability of reaction rates across and within the three boreholes that are generally consistent with the differential distribution of dissolved oxygen at this site, though not necessarily with the canonical view of how redox thresholds separate nitrate regeneration from dissimilative consumption spatially. That is, we provide isotope evidence for expanded zones of co-ocurring nitrification and denitrification. The isotope biogeochemical modeling also yielded estimates for the δ15N and δ18O of newly produced nitrate (δ15NNTR and δ18ONTR), as well as the isotope effect for

  4. Nitrogen cycling in the deep sedimentary biosphere: nitrate isotopes in porewaters underlying the oligotrophic North Atlantic

    Science.gov (United States)

    Wankel, S. D.; Buchwald, C.; Ziebis, W.; Wenk, C. B.; Lehmann, M. F.

    2015-12-01

    Nitrogen (N) is a key component of fundamental biomolecules. Hence, its cycling and availability are central factors governing the extent of ecosystems across the Earth. In the organic-lean sediment porewaters underlying the oligotrophic ocean, where low levels of microbial activity persist despite limited organic matter delivery from overlying water, the extent and modes of nitrogen transformations have not been widely investigated. Here we use the N and oxygen (O) isotopic composition of porewater nitrate (NO3-) from a site in the oligotrophic North Atlantic (Integrated Ocean Drilling Program - IODP) to determine the extent and magnitude of microbial nitrate production (via nitrification) and consumption (via denitrification). We find that NO3- accumulates far above bottom seawater concentrations (~ 21 μM) throughout the sediment column (up to ~ 50 μM) down to the oceanic basement as deep as 90 m b.s.f. (below sea floor), reflecting the predominance of aerobic nitrification/remineralization within the deep marine sediments. Large changes in the δ15N and δ18O of nitrate, however, reveal variable influence of nitrate respiration across the three sites. We use an inverse porewater diffusion-reaction model, constrained by the N and O isotope systematics of nitrification and denitrification and the porewater NO3- isotopic composition, to estimate rates of nitrification and denitrification throughout the sediment column. Results indicate variability of reaction rates across and within the three boreholes that are generally consistent with the differential distribution of dissolved oxygen at this site, though not necessarily with the canonical view of how redox thresholds separate nitrate regeneration from dissimilative consumption spatially. That is, we provide stable isotopic evidence for expanded zones of co-occurring nitrification and denitrification. The isotope biogeochemical modeling also yielded estimates for the δ15N and δ18O of newly produced nitrate (

  5. Diversity of Picoeukaryotes at an Oligotrophic Site off the Northern Red Sea Coast

    KAUST Repository

    Espinosa, Francisco Jose Acosta

    2012-05-01

    Picoeukaryotes are protist 3 µm belonging to a wide diversity of taxonomic groups, and they are an important constituent of the ocean microbiota, performing essential ecological roles in marine trophic chains and in nutrient and carbon budgets. Despite this, the true extent of their diversity is currently unknown, and in the last decade molecular surveys have uncovered a substantial number of previously unknown groups from all taxonomic levels. No studies on this group have been done so far on the Red Sea, a unique marine environment characterized by oligotrophic conditions and high irradiance, salinity and water temperature. We sampled the surface waters of a site near the northern Red Sea coast, and analyzed the picoeukaryotic diversity through the construction of PCR clone libraries using the 18S ribosomal gene. The community captured by our library is dominated by three main groups, the alveolates (32%), chlorophytes (32%) and Stramenopiles (20.55%). Members of Radiolaria, Cercozoans and Haptophyta were also found, although in low abundances. Photosynthetic organisms are especially diverse and abundant in the sample, with heterotrophic organism mostly composed by the mostly parasitic novel alveolates and bacterivorous stramenopiles. Novel clades were detected among the Novel Alveolates- II and the photosynthetic stramenopiles taxa, which suggests that they may be part of a number of groups unique to the basin and adapted to the high salinity and temperature conditions. This is the first study done on the Red Sea focusing on the diversity of the complete picoeukaryotic fraction, and provides a stepping stone in the characterization of the picoeukaryotic component of the microbial diversity of the basin.

  6. Groups without cultured representatives dominate eukaryotic picophytoplankton in the oligotrophic South East Pacific Ocean.

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    Xiao Li Shi

    Full Text Available BACKGROUND: Photosynthetic picoeukaryotes (PPE with a cell size less than 3 microm play a critical role in oceanic primary production. In recent years, the composition of marine picoeukaryote communities has been intensively investigated by molecular approaches, but their photosynthetic fraction remains poorly characterized. This is largely because the classical approach that relies on constructing 18S rRNA gene clone libraries from filtered seawater samples using universal eukaryotic primers is heavily biased toward heterotrophs, especially alveolates and stramenopiles, despite the fact that autotrophic cells in general outnumber heterotrophic ones in the euphotic zone. METHODOLOGY/PRINCIPAL FINDINGS: In order to better assess the composition of the eukaryotic picophytoplankton in the South East Pacific Ocean, encompassing the most oligotrophic oceanic regions on earth, we used a novel approach based on flow cytometry sorting followed by construction of 18S rRNA gene clone libraries. This strategy dramatically increased the recovery of sequences from putative autotrophic groups. The composition of the PPE community appeared highly variable both vertically down the water column and horizontally across the South East Pacific Ocean. In the central gyre, uncultivated lineages dominated: a recently discovered clade of Prasinophyceae (IX, clades of marine Chrysophyceae and Haptophyta, the latter division containing a potentially new class besides Prymnesiophyceae and Pavlophyceae. In contrast, on the edge of the gyre and in the coastal Chilean upwelling, groups with cultivated representatives (Prasinophyceae clade VII and Mamiellales dominated. CONCLUSIONS/SIGNIFICANCE: Our data demonstrate that a very large fraction of the eukaryotic picophytoplankton still escapes cultivation. The use of flow cytometry sorting should prove very useful to better characterize specific plankton populations by molecular approaches such as gene cloning or metagenomics

  7. Light dependence of [3H]leucine incorporation in the oligotrophic North Pacific ocean.

    Science.gov (United States)

    Church, Matthew J; Ducklow, Hugh W; Karl, David M

    2004-07-01

    The influence of irradiance on bacterial incorporation of [(3)H]leucine was evaluated at Station ALOHA in the oligotrophic North Pacific subtropical gyre. Six experiments were conducted on three cruises to Station ALOHA to examine how [(3)H]leucine incorporation varied as a function of irradiance. Two experiments were also conducted to assess the photoautotrophic response to irradiance (based on photosynthetic uptake of [(14)C]bicarbonate) in both the upper and lower photic zones. Rates of [(3)H]leucine incorporation responded to irradiance in a photosynthesis-like manner, increasing sharply at low light and then saturating and sometimes declining with increasing light intensity. The influence of irradiance on bacterial growth was evaluated in both the well-lit (5 to 25 m) and dimly lit regions of the upper ocean (75 to 100 m) to determine whether the bacterial response to irradiance differed along the depth-dependent light gradient of the photic zone. [(3)H]leucine incorporation rates were analyzed with a photosynthesis-irradiance model for a quantitative description of the relationships between [(3)H]leucine incorporation and irradiance. Maximum rates of [(3)H]leucine incorporation in the upper photic zone increased 48 to 92% relative to those of dark-incubated samples, with [(3)H]leucine incorporation saturating at light intensities between 58 and 363 micromol of quanta m(-2) s(-1). Rates of [(3)H]leucine incorporation in the deep photic zone were photostimulated 53 to 114% and were susceptible to photoinhibition, with rates declining at light intensities of >100 micromol of quanta m(-2) s(-1). The results of these experiments revealed that sunlight directly influences bacterial growth in this open-ocean ecosystem.

  8. Trichodesmium genome maintains abundant, widespread noncoding DNA in situ, despite oligotrophic lifestyle

    Energy Technology Data Exchange (ETDEWEB)

    Walworth, Nathan G.; Pfreundt, Ulrike; Nelson, William C.; Mincer, Tracy; Heidelberg, John F.; Fu, Feixue; Waterbury, John B.; Glavina del Rio, T.; Goodwin, Lynne A.; Kyrpides, Nikos; Land, Miriam L.; Woyke, Tanja; Hutchins, David A.; Hess, Wolfgang R.; Webb, Eric A.

    2015-04-07

    Understanding the evolution of the free-living, cyanobacterial, diazotroph Trichodesmium is of great importance due to its critical role in oceanic biogeochemistry and primary production. Unlike the other >150 available genomes of free-living cyanobacteria, only 63.8% of the Trichodesmium erythraeum (strain IMS101) genome is predicted to encode protein, which is 20-25% less than the average for other cyanobacteria and non-pathogenic, free-living bacteria. We use distinctive isolates and metagenomic data to show that low coding density observed in IMS101 is a common feature of the Trichodesmium genus both in culture and in situ. Transcriptome analysis indicates that 86% of the non-coding space is expressed, although the function of these transcripts is unclear. The density of noncoding, possible regulatory elements predicted in Trichodesmium, when normalized per intergenic kilobase, was comparable and two fold higher than that found in the gene dense genomes of the sympatric cyanobacterial genera Synechococcus and Prochlorococcus, respectively. Conserved Trichodesmium ncRNA secondary structures were predicted between most culture and metagenomic sequences lending support to the structural conservation. Conservation of these intergenic regions in spatiotemporally separated Trichodesmium populations suggests possible genus-wide selection for their maintenance. These large intergenic spacers may have developed during intervals of strong genetic drift caused by periodic blooms of a subset of genotypes, which may have reduced effective population size. Our data suggest that transposition of selfish DNA, low effective population size, and high fidelity replication allowed the unusual ‘inflation’ of noncoding sequence observed in Trichodesmium despite its oligotrophic lifestyle.

  9. Enrichment experiment changes microbial interactions in an ultra-oligotrophic environment.

    Science.gov (United States)

    Ponce-Soto, Gabriel Y; Aguirre-von-Wobeser, Eneas; Eguiarte, Luis E; Elser, James J; Lee, Zarraz M-P; Souza, Valeria

    2015-01-01

    The increase of nutrients in water bodies, in particular nitrogen (N) and phosphorus (P) due to the recent expansion of agricultural and other human activities is accelerating environmental degradation of these water bodies, elevating the risk of eutrophication and reducing biodiversity. To evaluate the ecological effects of the influx of nutrients in an oligotrophic and stoichiometrically imbalanced environment, we performed a replicated in situ mesocosm experiment. We analyzed the effects of a N- and P-enrichment on the bacterial interspecific interactions in an experiment conducted in the Cuatro Cienegas Basin (CCB) in Mexico. This is a desert ecosystem comprised of several aquatic systems with a large number of microbial endemic species. The abundance of key nutrients in this basin exhibits strong stoichiometric imbalance (high N:P ratios), suggesting that species diversity is maintained mostly by competition for resources. We focused on the biofilm formation and antibiotic resistance of 960 strains of cultivated bacteria in two habitats, water and sediment, before and after 3 weeks of fertilization. The water habitat was dominated by Pseudomonas, while Halomonas dominated the sediment. Strong antibiotic resistance was found among the isolates at time zero in the nutrient-poor bacterial communities, but resistance declined in the bacteria isolated in the nutrient-rich environments, suggesting that in the nutrient-poor original environment, negative inter-specific interactions were important, while in the nutrient-rich environments, competitive interactions are not so important. In water, a significant increase in the percentage of biofilm-forming strains was observed for all treatments involving nutrient addition.

  10. Microbial methane production in oxygenated water column of an oligotrophic lake.

    Science.gov (United States)

    Grossart, Hans-Peter; Frindte, Katharina; Dziallas, Claudia; Eckert, Werner; Tang, Kam W

    2011-12-01

    The prevailing paradigm in aquatic science is that microbial methanogenesis happens primarily in anoxic environments. Here, we used multiple complementary approaches to show that microbial methane production could and did occur in the well-oxygenated water column of an oligotrophic lake (Lake Stechlin, Germany). Oversaturation of methane was repeatedly recorded in the well-oxygenated upper 10 m of the water column, and the methane maxima coincided with oxygen oversaturation at 6 m. Laboratory incubations of unamended epilimnetic lake water and inoculations of photoautotrophs with a lake-enrichment culture both led to methane production even in the presence of oxygen, and the production was not affected by the addition of inorganic phosphate or methylated compounds. Methane production was also detected by in-lake incubations of lake water, and the highest production rate was 1.8-2.4 nM⋅h(-1) at 6 m, which could explain 33-44% of the observed ambient methane accumulation in the same month. Temporal and spatial uncoupling between methanogenesis and methanotrophy was supported by field and laboratory measurements, which also helped explain the oversaturation of methane in the upper water column. Potentially methanogenic Archaea were detected in situ in the oxygenated, methane-rich epilimnion, and their attachment to photoautotrophs might allow for anaerobic growth and direct transfer of substrates for methane production. Specific PCR on mRNA of the methyl coenzyme M reductase A gene revealed active methanogenesis. Microbial methane production in oxygenated water represents a hitherto overlooked source of methane and can be important for carbon cycling in the aquatic environments and water to air methane flux.

  11. Trichodesmium genome maintains abundant, widespread noncoding DNA in situ, despite oligotrophic lifestyle.

    Science.gov (United States)

    Walworth, Nathan; Pfreundt, Ulrike; Nelson, William C; Mincer, Tracy; Heidelberg, John F; Fu, Feixue; Waterbury, John B; Glavina del Rio, Tijana; Goodwin, Lynne; Kyrpides, Nikos C; Land, Miriam L; Woyke, Tanja; Hutchins, David A; Hess, Wolfgang R; Webb, Eric A

    2015-04-07

    Understanding the evolution of the free-living, cyanobacterial, diazotroph Trichodesmium is of great importance because of its critical role in oceanic biogeochemistry and primary production. Unlike the other >150 available genomes of free-living cyanobacteria, only 63.8% of the Trichodesmium erythraeum (strain IMS101) genome is predicted to encode protein, which is 20-25% less than the average for other cyanobacteria and nonpathogenic, free-living bacteria. We use distinctive isolates and metagenomic data to show that low coding density observed in IMS101 is a common feature of the Trichodesmium genus, both in culture and in situ. Transcriptome analysis indicates that 86% of the noncoding space is expressed, although the function of these transcripts is unclear. The density of noncoding, possible regulatory elements predicted in Trichodesmium, when normalized per intergenic kilobase, was comparable and twofold higher than that found in the gene-dense genomes of the sympatric cyanobacterial genera Synechococcus and Prochlorococcus, respectively. Conserved Trichodesmium noncoding RNA secondary structures were predicted between most culture and metagenomic sequences, lending support to the structural conservation. Conservation of these intergenic regions in spatiotemporally separated Trichodesmium populations suggests possible genus-wide selection for their maintenance. These large intergenic spacers may have developed during intervals of strong genetic drift caused by periodic blooms of a subset of genotypes, which may have reduced effective population size. Our data suggest that transposition of selfish DNA, low effective population size, and high-fidelity replication allowed the unusual "inflation" of noncoding sequence observed in Trichodesmium despite its oligotrophic lifestyle.

  12. Photochemical production of ammonium in the oligotrophic Cyprus Gyre (Eastern Mediterranean

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    V. Kitidis

    2006-06-01

    Full Text Available We investigated the photoproduction of ammonium (NH4+ in surface waters of the Cyprus gyre in the central Eastern Mediterranean in May 2002, in 8 on deck irradiations with freshly collected, filtered samples. NH4+ photoproduction (photoammonification increased with time-integrated irradiance during the course of irradiations. Photoammonification rates around local noon were 0.4–2.9 nmol L−1 h−1. Normalised to time integrated irradiance, these rates were 0.9–3.8 pmol L−1 h−1/(W m−2 and were significantly correlated with Chromophoric Dissolved Organic Matter (CDOM absorbance at 300 nm normalised to Dissolved Organic Carbon (DOC. These results are consistent with the notion that successive CDOM photobleaching in the surface mixed layer results in decreased DOC-normalised light absorbance concurrent with decreased dissolved organic matter reactivity with regard to photochemical NH4+ release. Combining our experimental data with estimates of annual solar irradiance and water column light attenuation yields an annual photoammonification rate for the Cyprus Gyre of 40±17 mmol m−2 a−1, equivalent to ~12±5% of the previously estimated annual nitrogen requirement of new production in this region. Based on this analysis, NH4+ photoproduction makes a small, but significant contribution to the nitrogen budget of the euphotic zone in the oligotrophic Cyprus Gyre.

  13. Enrichment experiment changes microbial interactions in an ultra-oligotrophic environment

    Directory of Open Access Journals (Sweden)

    Gabriel Yaxal Ponce-Soto

    2015-04-01

    Full Text Available The increase of nutrients in water bodies, in particular nitrogen (N and phosphorus (P due to the recent expansion of agricultural and other human activities is accelerating environmental degradation of these water bodies, elevating the risk of eutrophication and reducing biodiversity. To evaluate the ecological effects of the influx of nutrients in an oligotrophic and stoichiometrically imbalanced environment, we performed a replicated in situ mesocosm experiment. We analyzed the effects of a N- and P-enrichment on the bacterial interspecific interactions in an experiment conducted in the Cuatro Cienegas Basin (CCB in Mexico. This is a desert ecosystem comprised of several aquatic systems with a large number of microbial endemic species. The abundance of key nutrients in this basin exhibits strong stoichiometric imbalance (high N:P ratios, suggesting that species diversity is maintained mostly by competition for resources. We focused on the biofilm formation and antibiotic resistance of 960 strains of cultivated bacteria in two habitats, water and sediment, before and after three weeks of fertilization. The water habitat was dominated by Pseudomonas, while Halomonas dominated the sediment. Strong antibiotic resistance was found among the isolates at time zero in the nutrient-poor bacterial communities, but resistance declined in the bacteria isolated in the nutrient-rich environments, suggesting that in the nutrient-poor original environment, negative inter-specific interactions were important, while in the nutrient-rich environments, competitive interactions are not so important. In water, a significant increase in the percentage of biofilm-forming strains was observed for all treatments involving nutrient addition.

  14. Tracking climate change in oligotrophic mountain lakes: Recent hydrology and productivity synergies in Lago de Sanabria (NW Iberian Peninsula).

    Science.gov (United States)

    Jambrina-Enríquez, Margarita; Recio, Clemente; Vega, José Carlos; Valero-Garcés, Blas

    2017-07-15

    Mountain lakes are particularly sensitive to global change as their oligotrophic conditions may be rapidly altered after reaching an ecological threshold, due to increasing human impact and climate change. Sanabria Lake, the largest mountain lake in the Iberian Peninsula and with a recent history of increased human impact in its watershed, provides an opportunity to investigate recent trends in an oligotrophic, hydrologically-open mountain lake, and their relationship with climate, hydrological variability and human pressure. We conducted the first systematic and detailed survey of stable isotope compositions of Sanabria Lake and Tera River together with limnological analyses during 2009-2011. δ(18)Olakewater and δDlakewater seasonal fluctuations are strongly linked to river discharges, and follow the monthly mean isotopic composition of precipitation, which is controlled by NAO dynamics. δ(13)CPOM and δ(13)CDIC revealed higher contribution of allochthonous organic matter in winter and spring due to higher river inflow and lower primary productivity. Increased phytoplankton biomass in late summer correlated significantly with higher pH and Chl-a, and higher nutrient input and lower river inflow. However, the small δ(13)CPOM seasonal amplitude underlines the stability of the oligotrophic conditions and the isotopic variation in POM and DIC reflect small seasonal fluctuations mostly as a consequence of strong throughflow. The stability of hydrology and productivity patterns is consistent with Holocene and last millennium reconstructions of past limnological changes in Sanabria Lake. The results of this study indicate that trophic state in this hydrologically-open mountain lake is strongly controlled by climate variability, but recent changes in human-land uses have increased sediment delivery and nutrients supply to the lake and have to be considered for management policies. Monitoring surveys including isotope techniques provide snapshots of modern isotope

  15. Diversity of cultivated and metabolically active aerobic anoxygenic phototrophic bacteria along an oligotrophic gradient in the Mediterranean Sea

    Science.gov (United States)

    Jeanthon, C.; Boeuf, D.; Dahan, O.; Le Gall, F.; Garczarek, L.; Bendif, E. M.; Lehours, A.-C.

    2011-07-01

    Aerobic anoxygenic phototrophic (AAP) bacteria play significant roles in the bacterioplankton productivity and biogeochemical cycles of the surface ocean. In this study, we applied both cultivation and mRNA-based molecular methods to explore the diversity of AAP bacteria along an oligotrophic gradient in the Mediterranean Sea in early summer 2008. Colony-forming units obtained on three different agar media were screened for the production of bacteriochlorophyll-a (BChl-a), the light-harvesting pigment of AAP bacteria. BChl-a-containing colonies represented a low part of the cultivable fraction. In total, 54 AAP strains were isolated and the phylogenetic analyses based on their 16S rRNA and pufM genes showed that they were all affiliated to the Alphaproteobacteria. The most frequently isolated strains belonged to Citromicrobium bathyomarinum, and Erythrobacter and Roseovarius species. Most other isolates were related to species not reported to produce BChl-a and/or may represent novel taxa. Direct extraction of RNA from seawater samples enabled the analysis of the expression of pufM, the gene coding for the M subunit of the reaction centre complex of aerobic anoxygenic photosynthesis. Clone libraries of pufM gene transcripts revealed that most phylotypes were highly similar to sequences previously recovered from the Mediterranean Sea and a large majority (~94 %) was affiliated to the Gammaproteobacteria. The most abundantly detected phylotypes occurred in the western and eastern Mediterranean basins. However, some were exclusively detected in the eastern basin, reflecting the highest diversity of pufM transcripts observed in this ultra-oligotrophic region. To our knowledge, this is the first study to document extensively the diversity of AAP isolates and to unveil the active AAP community in an oligotrophic marine environment. By pointing out the discrepancies between culture-based and molecular methods, this study highlights the existing gaps in the understanding

  16. Absorption and translocation of copper and arsenic in an aquatic macrophyte Myriophyllum alterniflorum DC. in oligotrophic and eutrophic conditions.

    Science.gov (United States)

    Krayem, Maha; Baydoun, Mohamad; Deluchat, Véronique; Lenain, Jean-Francois; Kazpard, Véronique; Labrousse, Pascal

    2016-06-01

    The aim of this study is to evaluate copper and arsenic accumulation and translocation at a concentration of 100 μg/L of a submersed macrophyte Myriophyllum alterniflorum. The trophic level (eutrophic and oligotrophic conditions) of the medium was also considered. To achieve this goal, plants were incubated for 21 days in the presence of 100 μg/L of Cu or AsV. The heavy metal transfers from the contaminated medium to plants and into plant tissues was discussed in terms of the bioconcentration factor (BCF) and the translocation factor (TF). Malondialdehyde (MDA) content in tissues was analyzed in order to study the toxicity of these two contaminants. Our results show that copper was more accumulated in shoots, than roots, whereas the opposite trend was observed for arsenic. In addition, the two contaminants were more accumulated in oligotrophic than eutrophic medium. The BCF of copper in shoots was 1356 in oligotrophic condition, while that of arsenic was higher in roots about 620 in the same condition. The TF was less than 1 for arsenic, and higher than 1 for copper, indicating that watermilfoil restrains the translocation of arsenic to shoots, while it has a low capacity to control the translocation of an essential micronutrient like copper. An increase in MDA content was observed under Cu and As stress. On the basis of this experiment, M. alterniflorum has a higher accumulation potential of copper and arsenic, and therefore, it can be a good candidate for the phytofiltration of these two contaminants from water.

  17. The most oligotrophic subtropical zones of the global ocean: similarities and differences in terms of chlorophyll and yellow substance

    Science.gov (United States)

    Morel, A.; Claustre, H.; Gentili, B.

    2010-10-01

    The cores of the subtropical anticyclonic gyres are characterized by their oligotrophic status and minimal chlorophyll concentration, compared to that of the whole ocean. These zones are unambiguously detected by space borne ocean color sensors thanks to their typical spectral reflectance, which is that of extremely clear and deep blue waters. Not only the low chlorophyll (denoted [Chl]) level, but also a reduced amount of colored dissolved organic matter (CDOM or "yellow substance") account for this clarity. The oligotrophic waters of the North and South Pacific gyres, the North and South Atlantic gyres, and the South Indian gyre have been comparatively studied with respect to both [Chl] and CDOM contents, by using 10-year data (1998-2007) of the Sea-viewing Wide field-of-view Sensor (SeaWiFS, NASA). Albeit similar these oligotrophic zones are not identical regarding their [Chl] and CDOM contents, as well as their seasonal cycles. According to the zone, the averaged [Chl] value varies from 0.026 to 0.059 mg m-3, whereas the ay(443) average (the absorption coefficient due to CDOM at 443 nm) is between 0.0033 and 0.0072 m-1. The CDOM-to-[Chl] relative proportions also differ between the zones. The clearest waters, corresponding to the lowest [Chl] and CDOM concentrations, are found near Easter Island and near Mariana Islands in the western part of the North Pacific Ocean. In spite of its low [Chl], the Sargasso Sea presents the highest CDOM content amongst the six zones studied. Except in the North Pacific gyre (near Mariana and south of Hawaii islands), a conspicuous seasonality appears to be the rule in the other 4 gyres and affects both [Chl] and CDOM; both quantities vary in a ratio of about 2 (maximum-to-minimum). Coinciding [Chl] and CDOM peaks occur just after the local winter solstice, which is also the period of the maximal mixed layer depth in these latitudes. It is hypothesized that the vertical transport of unbleached CDOM from the subthermocline layers

  18. Quantifying nitrate dynamics in an oligotrophic lake using Δ17O

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    A. Tanaka

    2011-03-01

    Full Text Available The stable isotopic compositions of nitrate, including the 17O anomalies (Δ17O, were determined twice in 1 yr (June and August 2007 in the oligotrophic water column of Lake Mashu, Japan. These data were then used to quantify the geochemical dynamics of nitrate in the lake, by using the deposition rate of the atmospheric nitrate onto the entire catchment area of the lake. The total amount of nitrate in the lake water decreased from 4.2 to 2.1 Mmol during the period between the observations, while the average Δ17O values remained uniform at +2.5‰. The Δ17O values corresponded to an small and uniform mixing ratio of atmospheric nitrate to total nitrate of 9.7 ± 0.8%. These results indicate that 0.52 ± 0.34 Mmol of the remineralized nitrate was fed into the water column through nitrification, while 2.6 ± 0.4 Mmol of nitrate was simultaneously removed from the water column by assimilation, during the period between the observations. The lake water dissolved nitrate was characterized by rapid removal through assimilation during summer until it was almost completely removed from the euphotic layer, as well as continuous feeding into the lake through nitrification (3.2 ± 0.3 Mmol a−1 and deposition (0.35 ± 0.2 Mmol a−1, regardless of the seasons. The 15N-depleted nitrogen isotopic compositions of nitrate were as low as −6.5‰ in June, which also indicates that in-lake nitrification is the major source of nitrate in the lake and suggests that there is low potential for denitrification in and around the lake. Atmospheric nitrate deposited into the lake will be assimilated quickly, having a mean residence time of 1.2 ± 0.1 yr. In addition, more than 90% of the assimilated nitrate will be remineralized to nitrate and re-assimilated via active nitrogen cycling in the lake.

  19. Biodiversity patterns of crustacean suprabenthic assemblages along an oligotrophic gradient in the bathyal Mediterranean Sea

    Science.gov (United States)

    Almeida, Mariana; Frutos, Inmaculada; Tecchio, Samuele; Lampadariou, Nikolaos; Company, Joan B.; Ramirez-Llodra, Eva; Cunha, Marina R.

    2017-03-01

    Crustacean suprabenthic abundance, community structure, α-diversity (both taxonomic and trophic) and β-diversity were studied along a West-East gradient of oligotrophy in the deep Mediterranean Sea. The assemblages were sampled with a suprabenthic sledge in three regions (western, central and eastern basins) at three water depths (1200, 2000 and 3000 m) in May-June 2009. Environmental data were obtained at each sampling location including sediment properties, oceanographic variables near the seafloor and in the water column, and proxies of epipelagic productivity at the surface. Our results, concerning the crustacean component of the suprabenthos, showed complex trends in community structure and biodiversity across different spatial scales (longitudinal, bathymetric, and near-bottom distribution). A decrease in the number of species and abundance, accompanied by changes in the trophic structure of the assemblages were observed from West to East. In the eastern region the assemblages were impoverished in number of trophic guilds and trophic diversity. The West-East oligotrophic gradient was identified as the main driver in community structure as shown by the significant correlation with trophic environmental variables. Differences in community structure across regions were more marked at greater depths, while at the shallower sites assemblages were more similar. Within each basin, abundance, number of species and number of trophic groups decreased with depth, showing high turnover rates between 1200 and 2000 m depths. The small-scale (0-150 cm) vertical distribution of the suprabenthos was interpreted in relation to the species' functional traits (e.g. swimming activity, migratory behaviour, bottom dependence, feeding habits). Bottom-dependent and more mobile components of the suprabenthos were apparently responding differently to the various environmental challenges imposed by the large-scale longitudinal and bathymetric gradients. We propose that the bathyal

  20. Bio-active trace elements (cd, cu, fe, ni) in the oligotrophic south china sea

    Science.gov (United States)

    Wen, L.-S.; Jiann, K.-T.; Liu, K.-K.

    2003-04-01

    Bio-active trace elements (Cu, Ni, Cd, Fe) in seawater play a critical role in regulating oceanic phytoplankton growth and, hence, may influence global carbon cycle. However, their in-situ speciation and bio-reactivity are poorly understood. Dissolved copper and nickel are believed to be present in seawater predominantly as low molecular weight soluble organic complexes which are readily available to marine organism and immune from particle scavenging. Dissolved iron is believed to exist predominantly as high molecular weight colloidal species. Using ultraclean ultrafiltration and ion exchange/affinity chelating chemistry, we demonstrate that in the oligotrophic ocean waters, these four bio-active elements have distinctive characteristics of speciation and reactivity, even though they display similar nutrient-type distributions. For dissolved Cu, the concentration increased from 0.9 nM in the surface water to 3 nM at depths below 500 m; for dissolved Ni, 2˜9 nM; for dissolved Cd, 0.01˜0.9 nM; for dissolved Fe, 0.1˜0.6 nM. All four elements showed a subsurface minimum around 60 m deep, which corresponded to the subsurface Chl a maximum, indicating strong biological interactions with these elements. Detailed analysis revealed distinct size distribution and chemical reactivity for each element. For Cu, more than 50% in surface water was in smaller than 1kDa labile forms; the strongly complexed inert form increased from 28% at surface to 50% below 500 meter; the colloidal form Cu decreased from 12% at surface to a minimum of 6% at 60 meter, and then gradually increased to 16% in deeper water. For Ni, more than 80% was in smaller than 1kDa labile form, and very small fraction (˜5%) in colloidal from. For Cd, almost all dissolved fraction was in smaller than 1kDa labile form. As for Fe, its dynamic nature in water column caused by complicated bio-interactions was evident. This study indicated that, with preferential uptake of trace elements by different phytoplankton

  1. Differential Assimilation of Inorganic Carbon and Leucine by Prochlorococcus in the Oligotrophic North Pacific Subtropical Gyre.

    Science.gov (United States)

    Björkman, Karin M; Church, Matthew J; Doggett, Joseph K; Karl, David M

    2015-01-01

    The light effect on photoheterotrophic processes in Prochlorococcus, and primary and bacterial productivity in the oligotrophic North Pacific Subtropical Gyre was investigated using (14)C-bicarbonate and (3)H-leucine. Light and dark incubation experiments were conducted in situ throughout the euphotic zone (0-175 m) on nine expeditions to Station ALOHA over a 3-year period. Photosynthetrons were also used to elucidate rate responses in leucine and inorganic carbon assimilation as a function of light intensity. Taxonomic group and cell-specific rates were assessed using flow cytometric sorting. The light:dark assimilation rate ratios of leucine in the top 150 m were ∼7:1 for Prochlorococcus, whereas the light:dark ratios for the non-pigmented bacteria (NPB) were not significant different from 1:1. Prochlorococcus assimilated leucine in the dark at per cell rates similar to the NPB, with a contribution to the total community bacterial production, integrated over the euphotic zone, of approximately 20% in the dark and 60% in the light. Depth-resolved primary productivity and leucine incorporation showed that the ratio of Prochlorococcus leucine:primary production peaked at 100 m then declined steeply below the deep chlorophyll maximum (DCM). The photosynthetron experiments revealed that, for Prochlorococcus at the DCM, the saturating irradiance (E k) for leucine incorporation was reached at approximately half the light intensity required for light saturation of (14)C-bicarbonate assimilation. Additionally, high and low red fluorescing Prochlorococcus populations (HRF and LRF), co-occurring at the DCM, had similar E k values for their respective substrates, however, maximum assimilation rates, for both leucine and inorganic carbon, were two times greater for HRF cells. Our results show that Prochlorococcus contributes significantly to bacterial production estimates using (3)H-leucine, whether or not the incubations are conducted in the dark or light, and this should

  2. Profundal benthic invertebrates in an oligotrophic tropical lake: different strategies for coping with anoxia

    Directory of Open Access Journals (Sweden)

    María del Carmen Hernández

    2014-05-01

    Full Text Available The deep benthic communities of tropical lakes are poorly understood with respect to their composition, abundance, biomass and regulatory factors. Whereas the hypolimnia of temperate oligotrophic lakes remain oxygenated, the higher temperatures in tropical lakes frequently lead to the rapid development of hypolimnetic anoxia independent of trophic status. The deep benthic communities of tropical lakes must therefore develop strategies to respond to anoxic conditions. The dynamics of the deep benthic community of Lake Alchichica were studied over 15 months. We hypothesized that the sedimentation of the winter diatom bloom constitutes an input of high-quality food that contributes to the establishment and development of the deep benthic community. However, the remineralization of this organic matter leads to the prompt development of hypolimnetic anoxia, thus limiting the establishment and/or persistence of the deep benthic community. In contrast with the diverse littoral benthic community (50 taxa in Lake Alchichica, only two species constitute its deep benthic community, the ostracod Candona cf. patzcuaro and the chironomid Chironomus cf. austini, which combined exhibit a low density (1197±1976 ind m-2 and biomass (16.13±30.81 mg C m-2. C. patzcuaro is dominant and is present throughout the year, whereas Ch. austini is recorded only when the bottom water of the lake is oxygenated. A comparison with the analogous but temperate Lake Mergozzo in Italy illustrates the role that anoxia plays in tropical lakes by diminishing not only taxonomic richness (13 versus 2 spp. in temperate versus tropical lakes, respectively but also abundance (1145 versus 287 ind m-2, respectively. C. patzcuaro is found throughout the annual cycle of the lake’s profundal zone, entering into diapause during the anoxic period and recovering as soon as the profundal zone reoxygenates. Ch. austini has adjusted its life cycle to use the habitat and available resources while

  3. Spatial contrast in phytoplankton, bacteria and microzooplankton grazing between the eutrophic Yellow Sea and the oligotrophic South China Sea

    Science.gov (United States)

    Zhang, Yafeng; Wang, Xutao; Yin, Kedong

    2017-06-01

    Three cruises were conducted to investigate the distributions of nutrients, chlorophyll a (Chl-a), new and regenerated primary production, bacterial abundance and production, and microzooplankton grazing rates in the Yellow Sea (YS) and the South China Sea (SCS) during March and May. As the water column moved from low to high temperature, weak to strong stratification and high to low nutrients from the YS to the SCS, Chl-a, primary production and bacterial biomass decreased. In contrast, bacterial production, microzooplankton grazing and size preference increased from the YS to the SCS. The increasing grazing activity and decreasing f-ratio from the YS to the SCS suggest roles of regenerated nutrients in the supporting the community increased and more bacteria played important roles in the carbon flow in the oligotrophic SCS than in the eutrophic YS. These variabilities force the classical food chain dominated community in the eutrophic waters into the microbial loop, which is dominant in oligotrophic waters. As nutrients decrease, temperature and grazing activity increase from the YS to the SCS. The increasing ratio of integrated bacterial production to integrated primary production indicates that communities change from autotrophy to heterotrophy and waters change from a carbon sink to a carbon source.

  4. Synergistic effects of UVR and simulated stratification on commensalistic algal-bacterial relationship in two optically contrasting oligotrophic Mediterranean lakes

    Science.gov (United States)

    Carrillo, P.; Medina-Sánchez, J. M.; Durán, C.; Herrera, G.; Villafañe, V. E.; Helbling, E. W.

    2014-08-01

    An indirect effect of global warming is the shallowing epilimnion, causing organisms to be exposed to higher levels of ultraviolet (UVR, 280-400 nm) and photosynthetically active radiation (PAR, 400-700 nm), which could affect primary and bacterial production as well as the commensalistic algal-bacterial relationship. The combined effects of UVR and reduction in the depth of the upper mixed layer (UML) were assessed on variables related to the metabolism of algae and bacteria, during in situ experiments performed with natural microplanktonic communities from two oligotrophic lakes with contrasting UVR-transparency (clear vs. opaque) of southern Spain. The negative UVR effects on epilimnetic primary production (PP) and on heterotrophic bacterial production (HBP), intensified by high mean irradiances, were higher in the UVR-opaque than in the UVR-clear lake, and stronger on the algae than on the heterotrophic bacterial communities. Under UVR and high mean irradiance, the algal-bacterial relationship was strengthened in the UVR-clear lake, where excreted organic carbon (EOC) rates exceeded the bacterial carbon demand (BCD). This did not occur in the UVR-opaque lake. The greater UVR damage to algae and bacteria and the weakening of their commensalistic interaction found in the UVR-opaque lake indicates that these ecosystems would be especially vulnerable to stressors related to global change. Thus, our findings may have important implications for the carbon cycle in oligotrophic lakes of the Mediterranean region.

  5. Synergistic effects of UVR and simulated stratification on commensalistic phytoplankton-bacteria relationship in two optically contrasting oligotrophic Mediterranean lakes

    Science.gov (United States)

    Carrillo, P.; Medina-Sánchez, J. M.; Durán, C.; Herrera, G.; Villafañe, V. E.; Helbling, E. W.

    2015-02-01

    An indirect effect of global warming is a reduction in the depth of the upper mixed layer (UML) causing organisms to be exposed to higher levels of ultraviolet (UVR, 280-400 nm) and photosynthetically active radiation (PAR, 400-700 nm). This can affect primary and bacterial production as well as the commensalistic phytoplankton-bacteria relationship. The combined effects of UVR and reduction in the depth of the UML were assessed on variables related to the metabolism of phytoplankton and bacteria, during in situ experiments performed with natural pico- and nanoplankton communities from two oligotrophic lakes with contrasting UVR transparency (high-UVR versus low-UVR waters) of southern Spain. The negative UVR effects on epilimnetic primary production (PP) and on heterotrophic bacterial production (HBP), intensified under increased stratification, were higher in the low-UVR than in the high-UVR lake, and stronger on the phytoplanktonic than on the heterotrophic bacterial communities. Under UVR and increased stratification, the commensalistic phytoplankton-bacteria relationship was strengthened in the high-UVR lake where excretion of organic carbon (EOC) rates exceeded the bacterial carbon demand (BCD; i.e., BCD : EOC(%) ratio 100). The greater UVR damage to phytoplankton and bacteria and the weakening of their commensalistic interaction found in the low-UVR lake indicates that these ecosystems would be especially vulnerable to UVR and increased stratification as stressors related to global climate change. Thus, our findings may have important implications for the carbon cycle in oligotrophic lakes of the Mediterranean region.

  6. Differential responses of gut microbiota to the same prebiotic formula in oligotrophic and eutrophic batch fermentation systems.

    Science.gov (United States)

    Long, Wenmin; Xue, Zhengsheng; Zhang, Qianpeng; Feng, Zhou; Bridgewater, Laura; Wang, Linghua; Zhao, Liping; Pang, Xiaoyan

    2015-08-25

    The same prebiotics have produced inconsistent effects on microbiota when evaluated in different batch fermentation studies. To understand the reasons behind these discrepancies, we compared impact of one prebiotic formula on the same inoculated fecal microbiota in two frequently used batch systems: phosphate-buffered saline (PBS, oligotrophic) and basal culture medium (BCM, eutrophic). The microbiota was monitored using 454 pyrosequencing. Negative controls (no prebiotic) of both systems showed significant shifts in the microbiota during fermentation, although their pH remained relatively stable, especially in BCM, with increases in Bilophila and Escherichia/Shigella but a decrease in Faecalibacterium. We identified prebiotic responders via redundancy analysis by including both baseline and negative controls. The key positive and negative responders in the two systems were very different, with only 8 consistently modulated OTUs (7 of the 28 positive responders and 1 of the 35 negative responders). Moreover, some OTUs within the same genus responded to the prebiotic in opposite ways. Therefore, to obtain a complete in vitro evaluation of the modulatory effects of a prebiotic on microbiota, it is necessary to use both oligotrophic and eutrophic systems, compare treatment groups with both baseline and negative controls, and analyze the microbiota changes down to the OTU level.

  7. Palaeolimnological assessment of environmental change over the last two centuries in oligotrophic Lake Nohipalu Valgjärv, southern Estonia

    Directory of Open Access Journals (Sweden)

    Heinsalu, Atko

    2009-06-01

    Full Text Available The main objectives of this study were to reconstruct the environmental conditions for a small oligotrophic lake during the last two centuries, to determine if the environment of the lake was anthropogenically mediated, and to assess the pre-impact reference conditions with palaeolimnological techniques. A short sediment core from Lake Nohipalu Valgjärv was analysed in detail for diatom assemblages as well as for loss-on-ignition measurements. Accurate chronology of the sediment core was established and evaluated by different independent approaches – 210Pb, 137Cs, and 241Am dating, and the distribution of spheroidal fly-ash particles in sediments. Quantitative inference models based on sedimentary diatoms were applied to reconstruct changes in past lake water pH. Before the mid-19th century, Nohipalu Valgjärv was an oligotrophic lake with clear water continuously transparent down to the bottom and with rich benthic diatom flora. Since the early second half of the 19th century, presumably as a result of forest logging around the lake, water transparency decreased and benthic diatom productivity diminished, and the lake did not recover any more to natural baseline conditions. Due to peat mining activities in the Meenikunno bog, the quality of lake water has changed during the last two decades. The lowered lake level, deteriorated light climate, and decreased pH are the most important environmental variables that have influenced the lake ecosystem.

  8. Identification of the in vivo function of the high-efficiency D-mannonate dehydratase in Caulobacter crescentus NA1000 from the enolase superfamily.

    Science.gov (United States)

    Wichelecki, Daniel J; Graff, Dylan C; Al-Obaidi, Nawar; Almo, Steven C; Gerlt, John A

    2014-07-01

    The d-mannonate dehydratase (ManD) subgroup of the enolase superfamily contains members with varying catalytic activities (high-efficiency, low-efficiency, or no activity) that dehydrate d-mannonate and/or d-gluconate to 2-keto-3-deoxy-d-gluconate [Wichelecki, D. J., et al. (2014) Biochemistry 53, 2722-2731]. Despite extensive in vitro characterization, the in vivo physiological role of a ManD has yet to be established. In this study, we report the in vivo functional characterization of a high-efficiency ManD from Caulobacter crescentus NA1000 (UniProt entry B8GZZ7) by in vivo discovery of its essential role in d-glucuronate metabolism. This in vivo functional annotation may be extended to ~50 additional proteins.

  9. Immobilization of bacterial S-layer proteins from Caulobacter crescentus on iron oxide-based nanocomposite: synthesis and spectroscopic characterization of zincite-coated Fe₂O₃ nanoparticles.

    Science.gov (United States)

    Habibi, Neda

    2014-05-05

    Zinc oxide was coated on Fe2O3 nanoparticles using sol-gel spin-coating. Caulobacter crescentus have a crystalline surface layer (S-layer), which consist of one protein or glycoprotein species. The immobilization of bacterial S-layers obtained from C. crescentus on zincite-coated nanoparticles of iron oxide was investigated. The SDS PAGE results of S-layers isolated from C. crescentus showed the weight of 50 KDa. Nanoparticles of the Fe2O3 and zinc oxide were synthesized by a sol-gel technique. Fe2O3 nanoparticles with an average size of 50 nm were successfully prepared by the proper deposition of zinc oxide onto iron oxide nanoparticles surface annealed at 450 °C. The samples were characterized by field-emission scanning electron microscope (FESEM), atomic force microscopy (AFM), powder X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FT-IR).

  10. Three-dimensional super-resolution imaging of the midplane protein FtsZ in live Caulobacter crescentus cells using astigmatism.

    Science.gov (United States)

    Biteen, Julie S; Goley, Erin D; Shapiro, Lucy; Moerner, W E

    2012-03-01

    Single-molecule super-resolution imaging provides a non-invasive method for nanometer-scale imaging and is ideally suited to investigations of quasi-static structures within live cells. Here, we extend the ability to image subcellular features within bacteria cells to three dimensions based on the introduction of a cylindrical lens in the imaging pathway. We investigate the midplane protein FtsZ in Caulobacter crescentus with super-resolution imaging based on fluorescent-protein photoswitching and the natural polymerization/depolymerization dynamics of FtsZ associated with the Z-ring. We quantify these dynamics and determine the FtsZ depolymerization time to be divisional stage.

  11. The Caulobacter crescentus ctrA P1 promoter is essential for the coordination of cell cycle events that prevent the overinitiation of DNA replication.

    Science.gov (United States)

    Schredl, Alexander T; Perez Mora, Yannet G; Herrera, Anabel; Cuajungco, Math P; Murray, Sean R

    2012-10-01

    The master regulator CtrA oscillates during the Caulobacter cell cycle due to temporally regulated proteolysis and transcription. It is proteolysed during the G1-S transition and reaccumulates in predivisional cells as a result of transcription from two sequentially activated promoters, P1 and P2. CtrA reinforces its own synthesis by directly mediating the activation of P2 concurrently with repression of P1. To explore the role of P1 in cell cycle control, we engineered a mutation into the native ctrA locus that prevents transcription from P1 but not P2. As expected, the ctrA P1 mutant exhibits striking growth, morphological and DNA replication defects. Unexpectedly, we found CtrA and its antagonist SciP, but not DnaA, GcrA or CcrM accumulation to be dramatically reduced in the ctrA P1 mutant. SciP levels closely paralleled CtrA accumulation, suggesting that CtrA acts as a rheostat to modulate SciP abundance. Furthermore, the reappearance of CtrA and CcrM in predivisional cells was delayed in the P1 mutant by 0.125 cell cycle unit in synchronized cultures. High levels of ccrM transcription despite low levels of CtrA and increased transcription of ctrA P2 in the ctrA P1 mutant are two examples of robustness in the cell cycle. Thus, Caulobacter can adjust regulatory pathways to partially compensate for reduced and delayed CtrA accumulation in the ctrA P1 mutant.

  12. Organic geochemical evidence for nitrogen-limited oligotrophic seas in the Late Ordovician

    Science.gov (United States)

    Rohrssen, M.; Love, G. D.

    2011-12-01

    .5-2.0. H/St ratios reach a maximum below the main pulse of the HICE, and are at the lowest values during the maximum carbon isotope excursion. We propose that the biomarker distribution prior to the HICE reflects a nitrogen-limited oligotrophic ecosystem in which warm sea temperatures and nutrient limitation allowed marine bacterial communities to thrive relative to eukaryotes. Lower H/St ratios and declining 3β-methylhopane abundances during the main pulse of the HICE interval likely reflect microbial community response to cooler water temperature, contraction of OMZs, and increased ocean ventilation. Integration of the Anticosti record with data from the Hirnantian Vinini formation of Nevada, USA and Katyan formations from the Midcontinent region, USA, extends the temporal, spatial, and facies range of stratigraphic biomarker records of the response of marine microbial communities during the Late Ordovician transition from hothouse with extensive OMZs to ice-house with reduced OMZs.

  13. Diversity of cultivated and metabolically active aerobic anoxygenic phototrophic bacteria along an oligotrophic gradient in the Mediterranean Sea

    Directory of Open Access Journals (Sweden)

    C. Jeanthon

    2011-07-01

    Full Text Available Aerobic anoxygenic phototrophic (AAP bacteria play significant roles in the bacterioplankton productivity and biogeochemical cycles of the surface ocean. In this study, we applied both cultivation and mRNA-based molecular methods to explore the diversity of AAP bacteria along an oligotrophic gradient in the Mediterranean Sea in early summer 2008. Colony-forming units obtained on three different agar media were screened for the production of bacteriochlorophyll-a (BChl-a, the light-harvesting pigment of AAP bacteria. BChl-a-containing colonies represented a low part of the cultivable fraction. In total, 54 AAP strains were isolated and the phylogenetic analyses based on their 16S rRNA and pufM genes showed that they were all affiliated to the Alphaproteobacteria. The most frequently isolated strains belonged to Citromicrobium bathyomarinum, and Erythrobacter and Roseovarius species. Most other isolates were related to species not reported to produce BChl-a and/or may represent novel taxa. Direct extraction of RNA from seawater samples enabled the analysis of the expression of pufM, the gene coding for the M subunit of the reaction centre complex of aerobic anoxygenic photosynthesis. Clone libraries of pufM gene transcripts revealed that most phylotypes were highly similar to sequences previously recovered from the Mediterranean Sea and a large majority (~94 % was affiliated to the Gammaproteobacteria. The most abundantly detected phylotypes occurred in the western and eastern Mediterranean basins. However, some were exclusively detected in the eastern basin, reflecting the highest diversity of pufM transcripts observed in this ultra-oligotrophic region. To our knowledge, this is the first study to document extensively the diversity of AAP isolates and to unveil the active AAP community in an oligotrophic marine environment. By pointing out the discrepancies

  14. Diverse gene cassettes in class 1 integrons of facultative oligotrophic bacteria of River Mahananda,West Bengal, India.

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    Ranadhir Chakraborty

    Full Text Available BACKGROUND: In this study a large random collection (n=2188 of facultative oligotrophic bacteria, from 90 water samples gathered in three consecutive years (2007-2009 from three different sampling sites of River Mahananda in Siliguri, West Bengal, India, were investigated for the presence of class 1 integrons and sequences of the amplification products. METHODOLOGY/PRINCIPAL FINDINGS: Replica plating method was employed for determining the antibiotic resistance profile of the randomly assorted facultative oligotrophic isolates. Genomic DNA from each isolate was analyzed by PCR for the presence of class 1 integron. Amplicons were cloned and sequenced. Numerical taxonomy and 16S rRNA gene sequence analyses were done to ascertain putative genera of the class 1 integron bearing isolates. Out of 2188 isolates, 1667 (76.19% were antibiotic-resistant comprising of both single-antibiotic resistance (SAR and multiple-antibiotic resistant (MAR, and 521 (23.81% were sensitive to all twelve different antibiotics used in this study. Ninety out of 2188 isolates produced amplicon(s of varying sizes from 0.15 to 3.45 KB. Chi-square (χ(2 test revealed that the possession of class 1 integron in sensitive, SAR and MAR is not equally probable at the 1% level of significance. Diverse antibiotic-resistance gene cassettes, aadA1, aadA2, aadA4, aadA5, dfrA1, dfrA5, dfrA7, dfrA12, dfrA16, dfrA17, dfrA28, dfrA30, dfr-IIe, blaIMP-9, aacA4, Ac-6'-Ib, oxa1, oxa10 and arr2 were detected in 64 isolates. The novel cassettes encoding proteins unrelated to any known antibiotic resistance gene function were identified in 26 isolates. Antibiotic-sensitive isolates have a greater propensity to carry gene cassettes unrelated to known antibiotic-resistance genes. The integron-positive isolates under the class Betaproteobacteria comprised of only two genera, Comamonas and Acidovorax of family Comamonadaceae, while isolates under class Gammaproteobacteria fell under the families

  15. Modeling wave-like dynamics of oligotrophic and copiotrophic bacteria along wheat roots in response to nutrient input from a growing root tip

    NARCIS (Netherlands)

    Zelenev, V.V.; Bruggen, van A.H.C.; Semenov, A.M.

    2005-01-01

    Dynamics of oligotrophic bacteria (OB) have not been modeled in soil nor along roots. We extended a spatial¿temporal model ¿BACWAVE¿, describing wave-like dynamics of copiotrophic bacteria (CB) isolated on C-rich media to include dynamics of OB isolated on C-poor media and broad-range bacteria (BRB)

  16. Specific growth rate plays a critical role in hydrogen peroxide resistance of the marine oligotrophic ultramicrobacterium Sphingomonas alaskensis strain RB2256

    NARCIS (Netherlands)

    Ostrowski, M; Cavicchioli, R; Gottschal, JC; Blaauw, Maarten

    2001-01-01

    The marine oligotrophic ultramicrobacterium Sphingomonas alaskensis RB2256 has a physiology that is distinctly different from that of typical copiotrophic marine bacteria, such as Vibrio angustum S14. This includes a high level of inherent stress resistance and the absence of starvation-induced stre

  17. The most oligotrophic subtropical zones of the global ocean: similarities and differences in terms of chlorophyll and yellow substance

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    A. Morel

    2010-07-01

    Full Text Available The cores of the subtropical anticyclonic gyres are characterized by their oligotrophic status and minimal chlorophyll concentration, compared to that of the whole ocean. These zones are unambiguously detected by space borne ocean color sensors thanks to their typical spectral reflectance, which is that of extremely clear and deep blue waters. Not only the low chlorophyll (denoted [Chl] level, but also a reduced amount of colored dissolved organic matter (CDOM or "yellow substance" account for this clarity. The oligotrophic waters of the North and South Pacific gyres, the North and South Atlantic gyres, and the South Indian gyre have been comparatively studied with respect to both [Chl] and CDOM contents, by using 10-year data (1998–2007 of the Sea-viewing Wide field-of-view Sensor (SeaWiFS, NASA. Albeit similar these oligotrophic zones are not identical regarding their [Chl] and CDOM contents, as well as their seasonal cycles. According to the zone, the averaged [Chl] value varies from 0.026 to 0.059 mg m−3, whereas the ay(443 average (the absorption coefficient due to CDOM at 443 nm is comprised between 0.0033 and 0.0072 m−1. The CDOM-to-[Chl] relative proportions also differ between the zones. The clearest waters, corresponding to the lowest [Chl] and CDOM concentrations, are found near Easter Island and near Mariana Islands in the western part of the North Pacific Ocean. In spite of its low [Chl], the Sargasso Sea presents the highest CDOM content amongst the six zones studied. Except in the North Pacific gyre (near Mariana and south of Hawaii islands, a conspicuous seasonality appears to be the rule in the other 4 gyres and affects both [Chl] and CDOM; both quantities vary in a ratio of about 2 (maximum-to-minimum. Coinciding [Chl] and CDOM peaks occur just after the local winter solstice, which is also the period of the maximal mixed layer depth in these latitudes. It is hypothesized

  18. Large clean mesocosms and simulated dust deposition: a new methodology to investigate responses of marine oligotrophic ecosystems to atmospheric inputs

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    C. Guieu

    2010-04-01

    Full Text Available Intense Saharan dust deposition occurs over large oligotrophic areas in the Mediterranean Sea and the Tropical Atlantic and its impact on the biogeochemical functioning of such oligotrophic ecosystems needs to be understood. However, due to the logistical difficulties to investigate in-situ natural dust events and due to the inherent limitations of microcosm laboratory experiments, new experimental approaches need to be developed. In this paper, we present a new experimental set up based on large clean mesocoms deployed in the frame of the DUNE (a DUst experiment in a low-Nutrient, low-chlorophyll Ecosystem project. We demonstrate that these tools are highly relevant and provide a powerful new strategy to in situ study the response of an oligotrophic ecosystem to chemical forcing by atmospheric deposition of African dust. First, we describe how to cope with the large amount of dust aerosol needed to conduct the seeding experiments, by producing an analogue from soil collection in a source area and performing subsequent appropriate physico-chemical treatment in the laboratory including an eventual processing by simulated cloud water. The comparison of physico-chemical characteristics of produced dust analogues with the literature confirms that our experimental simulations are representative of dust, ageing during atmospheric transport, and subsequent deposition to the Mediterranean. Second, we demonstrate the feasibility in coastal area to installing in situ, a series of large (6×52 m3 mesocosms without perturbing the local ecosystem. All the setup, containing no metallic part and with as less as possible induced perturbation during the sampling sequence, allows working with the required conditions for biogeochemical studies in oligotrophic environments where nutrient and micronutrients are at nano- or subnano-molar levels. Two distinct "seeding experiments" were conducted by deploying mesocosms in triplicates: three mesocosms serving

  19. Large clean mesocosms and simulated dust deposition: a new methodology to investigate responses of marine oligotrophic ecosystems to atmospheric inputs

    Directory of Open Access Journals (Sweden)

    C. Guieu

    2010-09-01

    Full Text Available Intense Saharan dust deposition occurs over large oligotrophic areas in the Mediterranean Sea and in the Tropical Atlantic, and its impact on the biogeochemical functioning of such oligotrophic ecosystems needs to be understood. However, due to the logistical difficulties of investigating in situ natural dust events, and due to the inherent limitations of microcosm laboratory experiments, new experimental approaches need to be developed. In this paper, we present a new experimental setup based on large, clean mesocoms deployed in the frame of the DUNE (a DUst experiment in a low-Nutrient, low-chlorophyll Ecosystem project. We demonstrate that these tools are highly relevant and provide a powerful new strategy to in situ studies of the response of an oligotrophic ecosystem to chemical forcing by atmospheric deposition of African dust. First, we describe how to cope with the large amount of dust aerosol needed to conduct the seeding experiments by producing an analogue from soil collected in a source area and by performing subsequent appropriate physico-chemical treatments in the laboratory, including an eventual processing by simulated cloud water. The comparison of the physico-chemical characteristics of produced dust analogues with the literature confirms that our experimental simulations are representative of dust, aging during atmospheric transport, and subsequent deposition to the Mediterranean. Second, we demonstrate the feasibility in coastal areas to installing, in situ, a series of large (6 × 52 m3 mesocosms without perturbing the local ecosystem. The setup, containing no metallic parts and with the least possible induced perturbation during the sampling sequence, provides an approach for working with the required conditions for biogeochemical studies in oligotrophic environments, where nutrient and micronutrients are at nano- or subnano-molar levels. Two, distinct "seeding experiments" were conducted by deploying three

  20. The most oligotrophic subtropical zones of the global ocean: similarities and differences in terms of chlorophyll and yellow substance

    Directory of Open Access Journals (Sweden)

    A. Morel

    2010-10-01

    Full Text Available The cores of the subtropical anticyclonic gyres are characterized by their oligotrophic status and minimal chlorophyll concentration, compared to that of the whole ocean. These zones are unambiguously detected by space borne ocean color sensors thanks to their typical spectral reflectance, which is that of extremely clear and deep blue waters. Not only the low chlorophyll (denoted [Chl] level, but also a reduced amount of colored dissolved organic matter (CDOM or "yellow substance" account for this clarity. The oligotrophic waters of the North and South Pacific gyres, the North and South Atlantic gyres, and the South Indian gyre have been comparatively studied with respect to both [Chl] and CDOM contents, by using 10-year data (1998–2007 of the Sea-viewing Wide field-of-view Sensor (SeaWiFS, NASA. Albeit similar these oligotrophic zones are not identical regarding their [Chl] and CDOM contents, as well as their seasonal cycles. According to the zone, the averaged [Chl] value varies from 0.026 to 0.059 mg m−3, whereas the ay(443 average (the absorption coefficient due to CDOM at 443 nm is between 0.0033 and 0.0072 m−1. The CDOM-to-[Chl] relative proportions also differ between the zones. The clearest waters, corresponding to the lowest [Chl] and CDOM concentrations, are found near Easter Island and near Mariana Islands in the western part of the North Pacific Ocean. In spite of its low [Chl], the Sargasso Sea presents the highest CDOM content amongst the six zones studied. Except in the North Pacific gyre (near Mariana and south of Hawaii islands, a conspicuous seasonality appears to be the rule in the other 4 gyres and affects both [Chl] and CDOM; both quantities vary in a ratio of about 2 (maximum-to-minimum. Coinciding [Chl] and CDOM peaks occur just after the local winter solstice, which is also the period of the maximal mixed layer depth in these latitudes. It is hypothesized that the vertical

  1. Dense microbial community on a ferromanganese nodule from the ultra-oligotrophic South Pacific Gyre: Implications for biogeochemical cycles

    Science.gov (United States)

    Shiraishi, Fumito; Mitsunobu, Satoshi; Suzuki, Katsuhiko; Hoshino, Tatsuhiko; Morono, Yuki; Inagaki, Fumio

    2016-08-01

    During Integrated Ocean Drilling Program (IODP) Expedition 329, a deep-sea ferromanganese nodule and surrounding sediment were collected from the South Pacific Gyre, the most oligotrophic oceanic environment on earth. Using a combination of cryo-sectioning and fluorescence-based cell counting techniques, we determined that the microbial cell density at the very surface of the nodule was ∼108 cells cm-3, three orders of magnitude higher than that in the surrounding sediment. Analysis of bacterial and archaeal 16S rRNA gene sequences (∼1400 bp) indicated that the taxonomic composition of the nodule-associated community differed markedly from that of the sediment-associated community. Members of Marine Group I (MGI) Thaumarchaeota are potentially crucial for sustaining the high cell density because both ammonia and Cu were available on the nodule surface, making it suitable for ammonia-oxidizing chemolithoautotrophy mediated by copper enzymes. Combined cryo-sectioning and synchrotron analysis of the nodule surface revealed both hexagonal birnessite resembling δ-MnO2 and triclinic birnessite, minerals characteristic of biogenic oxide and its secondary product, respectively. Regardless of these possible biogenic features, only one gene sequence exhibited some similarity to previously identified manganese-oxidizing bacteria. On the other hand, MGI Thaumarchaeota were assumed as potential candidate of manganese oxidizers because they have multi-copper oxidase that is utilized by most known manganese oxidizers. Therefore, this archaeal group is considered to play a significant ecological role as a primary producer in biogeochemical elemental cycles in the ultra-oligotrophic abyssal plain.

  2. Variable nutrient stoichiometry (carbon:nitrogen:phosphorus) across trophic levels determines community and ecosystem properties in an oligotrophic mangrove system

    Science.gov (United States)

    Scharler, U.M.; Ulanowicz, Robert E.; Fogel, M.L.; Wooller, M.J.; Jacobson-Meyers, M.E.; Lovelock, C.E.; Feller, I.C.; Frischer, M.; Lee, R.; Mckee, Karen L.; Romero, I.C.; Schmit, J.P.; Shearer, C.

    2015-01-01

    Our study investigated the carbon:nitrogen:phosphorus (C:N:P) stoichiometry of mangrove island of the Mesoamerican Barrier Reef (Twin Cays, Belize). The C:N:P of abiotic and biotic components of this oligotrophic ecosystem was measured and served to build networks of nutrient flows for three distinct mangrove forest zones (tall seaward fringing forest, inland dwarf forests and a transitional zone). Between forest zones, the stoichiometry of primary producers, heterotrophs and abiotic components did not change significantly, but there was a significant difference in C:N:P, and C, N, and P biomass, between the functional groups mangrove trees, other primary producers, heterotrophs, and abiotic components. C:N:P decreased with increasing trophic level. Nutrient recycling in the food webs was highest for P, and high transfer efficiencies between trophic levels of P and N also indicated an overall shortage of these nutrients when compared to C. Heterotrophs were sometimes, but not always, limited by the same nutrient as the primary producers. Mangrove trees and the primary tree consumers were P limited, whereas the invertebrates consuming leaf litter and detritus were N limited. Most compartments were limited by P or N (not by C), and the relative depletion rate of food sources was fastest for P. P transfers thus constituted a bottleneck of nutrient transfer on Twin Cays. This is the first comprehensive ecosystem study of nutrient transfers in a mangrove ecosystem, illustrating some mechanisms (e.g. recycling rates, transfer efficiencies) which oligotrophic systems use in order to build up biomass and food webs spanning various trophic levels.

  3. Nitrogen Removal Characteristics of a Newly Isolated Indigenous Aerobic Denitrifier from Oligotrophic Drinking Water Reservoir, Zoogloea sp. N299

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    Ting-Lin Huang

    2015-05-01

    Full Text Available Nitrogen is considered to be one of the most widespread pollutants leading to eutrophication of freshwater ecosystems, especially in drinking water reservoirs. In this study, an oligotrophic aerobic denitrifier was isolated from drinking water reservoir sediment. Nitrogen removal performance was explored. The strain was identified by 16S rRNA gene sequence analysis as Zoogloea sp. N299. This species exhibits a periplasmic nitrate reductase gene (napA. Its specific growth rate was 0.22 h−1. Obvious denitrification and perfect nitrogen removal performances occurred when cultured in nitrate and nitrite mediums, at rates of 75.53% ± 1.69% and 58.65% ± 0.61%, respectively. The ammonia removal rate reached 44.12% ± 1.61% in ammonia medium. Zoogloea sp. N299 was inoculated into sterilized and unsterilized reservoir source waters with a dissolved oxygen level of 5–9 mg/L, pH 8–9, and C/N 1.14:1. The total nitrogen removal rate reached 46.41% ± 3.17% (sterilized and 44.88% ± 4.31% (unsterilized. The cell optical density suggested the strain could survive in oligotrophic drinking water reservoir water conditions and perform nitrogen removal. Sodium acetate was the most favorable carbon source for nitrogen removal by strain N299 (p < 0.05. High C/N was beneficial for nitrate reduction (p < 0.05. The nitrate removal efficiencies showed no significant differences among the tested inoculums dosage (p > 0.05. Furthermore, strain N299 could efficiently remove nitrate at neutral and slightly alkaline and low temperature conditions. These results, therefore, demonstrate that Zoogloea sp. N299 has high removal characteristics, and can be used as a nitrogen removal microbial inoculum with simultaneous aerobic nitrification and denitrification in a micro-polluted reservoir water ecosystem.

  4. Variable nutrient stoichiometry (carbon:nitrogen:phosphorus) across trophic levels determines community and ecosystem properties in an oligotrophic mangrove system.

    Science.gov (United States)

    Scharler, U M; Ulanowicz, R E; Fogel, M L; Wooller, M J; Jacobson-Meyers, M E; Lovelock, C E; Feller, I C; Frischer, M; Lee, R; McKee, K; Romero, I C; Schmit, J P; Shearer, C

    2015-11-01

    Our study investigated the carbon:nitrogen:phosphorus (C:N:P) stoichiometry of mangrove island of the Mesoamerican Barrier Reef (Twin Cays, Belize). The C:N:P of abiotic and biotic components of this oligotrophic ecosystem was measured and served to build networks of nutrient flows for three distinct mangrove forest zones (tall seaward fringing forest, inland dwarf forests and a transitional zone). Between forest zones, the stoichiometry of primary producers, heterotrophs and abiotic components did not change significantly, but there was a significant difference in C:N:P, and C, N, and P biomass, between the functional groups mangrove trees, other primary producers, heterotrophs, and abiotic components. C:N:P decreased with increasing trophic level. Nutrient recycling in the food webs was highest for P, and high transfer efficiencies between trophic levels of P and N also indicated an overall shortage of these nutrients when compared to C. Heterotrophs were sometimes, but not always, limited by the same nutrient as the primary producers. Mangrove trees and the primary tree consumers were P limited, whereas the invertebrates consuming leaf litter and detritus were N limited. Most compartments were limited by P or N (not by C), and the relative depletion rate of food sources was fastest for P. P transfers thus constituted a bottleneck of nutrient transfer on Twin Cays. This is the first comprehensive ecosystem study of nutrient transfers in a mangrove ecosystem, illustrating some mechanisms (e.g. recycling rates, transfer efficiencies) which oligotrophic systems use in order to build up biomass and food webs spanning various trophic levels.

  5. Effects of olive oil wastes on river basins and an oligotrophic coastal marine ecosystem: a case study in Greece.

    Science.gov (United States)

    Pavlidou, A; Anastasopoulou, E; Dassenakis, M; Hatzianestis, I; Paraskevopoulou, V; Simboura, N; Rousselaki, E; Drakopoulou, P

    2014-11-01

    This work aims to contribute to the knowledge of the impacts of olive oil waste discharge to freshwater and oligotrophic marine environments, since the ecological impact of olive oil wastes in riverine and coastal marine ecosystems, which are the final repositories of the pollutants, is a great environmental problem on a global scale, mostly concerning all the Mediterranean countries with olive oil production. Messinia, in southwestern Greece, is one of the greatest olive oil production areas in Europe. During the last decade around 1.4×10(6)tons of olive oil mill wastewater has been disposed in the rivers of Messinia and finally entered the marine ecosystem of Messiniakos gulf. The pollution from olive oil mill wastewater in the main rivers of Messinia and the oligotrophic coastal zone of Messiniakos gulf and its effects on marine organisms were evaluated, before, during and after the olive oil production period. Elevated amounts of phenols (36.2-178 mg L(-1)) and high concentrations of ammonium (7.29-18.9 mmol L(-1)) and inorganic phosphorus (0.5-7.48 mmol L(-1)) were measured in small streams where the liquid disposals from several olive oil industries were gathered before their discharge in the major rivers of Messinia. The large number of olive oil units has downgraded the riverine and marine ecosystems during the productive period and a period more than five months is needed for the recovery of the ecosystem. Statistical analysis showed that the enrichment of freshwater and the coastal zone of Messiniakos gulf in ammonia, nitrite, phenols, total organic carbon, copper, manganese and nickel was directly correlated with the wastes from olive oil. Toxicity tests using 24h LC50 Palaemonidae shrimp confirm that olive mill wastewater possesses very high toxicity in the aquatic environment.

  6. Dynamical Localization of DivL and PleC in the Asymmetric Division Cycle of Caulobacter crescentus: A Theoretical Investigation of Alternative Models.

    Science.gov (United States)

    Subramanian, Kartik; Paul, Mark R; Tyson, John J

    2015-07-01

    Cell-fate asymmetry in the predivisional cell of Caulobacter crescentus requires that the regulatory protein DivL localizes to the new pole of the cell where it up-regulates CckA kinase, resulting in a gradient of CtrA~P across the cell. In the preceding stage of the cell cycle (the "stalked" cell), DivL is localized uniformly along the cell membrane and maintained in an inactive form by DivK~P. It is unclear how DivL overcomes inhibition by DivK~P in the predivisional cell simply by changing its location to the new pole. It has been suggested that co-localization of DivL with PleC phosphatase at the new pole is essential to DivL's activity there. However, there are contrasting views on whether the bifunctional enzyme, PleC, acts as a kinase or phosphatase at the new pole. To explore these ambiguities, we formulated a mathematical model of the spatiotemporal distributions of DivL, PleC and associated proteins (DivJ, DivK, CckA, and CtrA) during the asymmetric division cycle of a Caulobacter cell. By varying localization profiles of DivL and PleC in our model, we show how the physiologically observed spatial distributions of these proteins are essential for the transition from a stalked cell to a predivisional cell. Our simulations suggest that PleC is a kinase in predivisional cells, and that, by sequestering DivK~P, the kinase form of PleC enables DivL to be reactivated at the new pole. Hence, co-localization of PleC kinase and DivL is essential to establishing cellular asymmetry. Our simulations reproduce the experimentally observed spatial distribution and phosphorylation status of CtrA in wild-type and mutant cells. Based on the model, we explore novel combinations of mutant alleles, making predictions that can be tested experimentally.

  7. Dynamical Localization of DivL and PleC in the Asymmetric Division Cycle of Caulobacter crescentus: A Theoretical Investigation of Alternative Models.

    Directory of Open Access Journals (Sweden)

    Kartik Subramanian

    2015-07-01

    Full Text Available Cell-fate asymmetry in the predivisional cell of Caulobacter crescentus requires that the regulatory protein DivL localizes to the new pole of the cell where it up-regulates CckA kinase, resulting in a gradient of CtrA~P across the cell. In the preceding stage of the cell cycle (the "stalked" cell, DivL is localized uniformly along the cell membrane and maintained in an inactive form by DivK~P. It is unclear how DivL overcomes inhibition by DivK~P in the predivisional cell simply by changing its location to the new pole. It has been suggested that co-localization of DivL with PleC phosphatase at the new pole is essential to DivL's activity there. However, there are contrasting views on whether the bifunctional enzyme, PleC, acts as a kinase or phosphatase at the new pole. To explore these ambiguities, we formulated a mathematical model of the spatiotemporal distributions of DivL, PleC and associated proteins (DivJ, DivK, CckA, and CtrA during the asymmetric division cycle of a Caulobacter cell. By varying localization profiles of DivL and PleC in our model, we show how the physiologically observed spatial distributions of these proteins are essential for the transition from a stalked cell to a predivisional cell. Our simulations suggest that PleC is a kinase in predivisional cells, and that, by sequestering DivK~P, the kinase form of PleC enables DivL to be reactivated at the new pole. Hence, co-localization of PleC kinase and DivL is essential to establishing cellular asymmetry. Our simulations reproduce the experimentally observed spatial distribution and phosphorylation status of CtrA in wild-type and mutant cells. Based on the model, we explore novel combinations of mutant alleles, making predictions that can be tested experimentally.

  8. Role of core promoter sequences in the mechanism of swarmer cell-specific silencing of gyrB transcription in Caulobacter crescentus

    Directory of Open Access Journals (Sweden)

    Gober James W

    2005-05-01

    Full Text Available Abstract Background Each Caulobacter crescentus cell division yields two distinct cell types: a flagellated swarmer cell and a non-motile stalked cell. The swarmer cell is further distinguished from the stalked cell by an inability to reinitiate DNA replication, by the physical properties of its nucleoid, and its discrete program of gene expression. Specifically, with regard to the latter feature, many of the genes involved in DNA replication are not transcribed in swarmer cells. Results We show that for one of these genes involved in DNA replication, gyrB, its pattern of temporal expression depends upon an 80 base pair promoter region with strong resemblance to the Caulobacter crescentus σ73 consensus promoter sequence; regulation does not appear to be affected by the general strength of the promoter activity, as mutations that increased its conformity with the consensus did not affect its cell-cycle expression pattern. Transcription from the gyrB promoter in vitro required only the presence of the σ73 RNA polymerase (from E. coli and the requisite nucleoside triphosphates, although a distinct binding activity, present in crude whole-cell extracts, formed a complex gyrB promoter DNA. We also assayed the effect on gyrB expression in strains containing mutations in either smc or dps, two genes encoding proteins that condense DNA. However we found there was no change in the temporal pattern of gyrB transcription in strains containing deletions in either of these genes. Conclusion These experiments demonstrate that gyrB transcription does not require any auxiliary factors, suggesting that temporal regulation is not dependent upon an activator protein. Swarmer-specific silencing may not be attributable to the observed physical difference in the swarmer cell nucleoid, since mutations in either smc or dps, two genes encoding proteins that condense DNA, did not alter the temporal pattern of gyrB transcription in strains containing deletions in either

  9. CtrA response regulator binding to the Caulobacter chromosome replication origin is required during nutrient and antibiotic stress as well as during cell cycle progression.

    Science.gov (United States)

    Bastedo, D Patrick; Marczynski, Gregory T

    2009-04-01

    The Caulobacter crescentus chromosome replication origin (Cori) has five binding sites for CtrA, an OmpR/PhoB family 'response regulator'. CtrA is degraded in replicating 'stalked' cells but is abundant in the non-replicating 'swarmer' cells, where it was proposed to repress replication by binding to Cori. We systematically mutated all Cori CtrA binding sites, and examined their consequences in the contexts of autonomous Cori-plasmid replication and in the natural chromosome locus. Remarkably, the C. crescentus chromosome tolerates severe mutations in all five CtrA binding sites, demonstrating that CtrA is not essential for replication. Further physiological and cell cycle experiments more rigorously supported the original hypothesis that CtrA represses replication. However, our experiments argued against another hypothesis that residual and/or replenished CtrA protein in stalked cells might prevent extra or unscheduled chromosome replication before cell division. Surprisingly, we also demonstrated that Cori CtrA binding sites are very advantageous and can become essential when cells encounter nutrients and antibiotics. Therefore, the CtrA cell cycle regulator co-ordinates replication with viable cell growth in stressful and rapidly changing environments. We argue that this new role for CtrA provided the primary selective pressure for evolving control by CtrA.

  10. A dual binding site for integration host factor and the response regulator CtrA inside the Caulobacter crescentus replication origin.

    Science.gov (United States)

    Siam, Rania; Brassinga, Ann Karen C; Marczynski, Gregory T

    2003-09-01

    The response regulator CtrA controls chromosome replication by binding to five sites, a, b, c, d, and e, inside the Caulobacter crescentus replication origin (Cori). In this study, we demonstrate that integration host factor (IHF) binds Cori over the central CtrA binding site c. Surprisingly, IHF and CtrA share DNA recognition sequences. Rather than promoting cooperative binding, IHF binding hinders CtrA binding to site c and nearby site d. Unlike other CtrA binding sites, DNA mutations in the CtrA c/IHF site uniquely impair autonomous Cori plasmid replication. These mutations also alter transcription from distant promoters more than 100 bp away. When the CtrA c/IHF site was deleted from the chromosome, these cells grew slowly and became selectively intolerant to a CtrA phosphor-mimic allele (D51E). Since CtrA protein concentration decreases during the cell cycle as IHF protein concentration increases, we propose a model in which IHF displaces CtrA in order to bend Cori and promote efficient chromosome replication.

  11. A protein critical for cell constriction in the Gram-negative bacterium Caulobacter crescentus localizes at the division site through its peptidoglycan-binding LysM domains.

    Science.gov (United States)

    Poggio, Sebastian; Takacs, Constantin N; Vollmer, Waldemar; Jacobs-Wagner, Christine

    2010-07-01

    During division of Gram-negative bacteria, invagination of the cytoplasmic membrane and inward growth of the peptidoglycan (PG) are followed by the cleavage of connective septal PG to allow cell separation. This PG splitting process requires temporal and spatial regulation of cell wall hydrolases. In Escherichia coli, LytM factors play an important role in PG splitting. Here we identify and characterize a member of this family (DipM) in Caulobacter crescentus. Unlike its E. coli counterparts, DipM is essential for viability under fast-growth conditions. Under slow-growth conditions, the DeltadipM mutant displays severe defects in cell division and FtsZ constriction. Consistent with its function in division, DipM colocalizes with the FtsZ ring during the cell cycle. Mutagenesis suggests that the LytM domain of DipM is essential for protein function, despite being non-canonical. DipM also carries two tandems of the PG-binding LysM domain that are sufficient for FtsZ ring localization. Localization and fluorescence recovery after photobleaching microscopy experiments suggest that DipM localization is mediated, at least in part, by the ability of the LysM tandems to distinguish septal, multilayered PG from non-septal, monolayered PG.

  12. Activation and polar sequestration of PopA, a c-di-GMP effector protein involved in Caulobacter crescentus cell cycle control.

    Science.gov (United States)

    Ozaki, Shogo; Schalch-Moser, Annina; Zumthor, Ludwig; Manfredi, Pablo; Ebbensgaard, Anna; Schirmer, Tilman; Jenal, Urs

    2014-11-01

    When Caulobacter crescentus enters S-phase the replication initiation inhibitor CtrA dynamically positions to the old cell pole to be degraded by the polar ClpXP protease. Polar delivery of CtrA requires PopA and the diguanylate cyclase PleD that positions to the same pole. Here we present evidence that PopA originated through gene duplication from its paralogue response regulator PleD and subsequent co-option as c-di-GMP effector protein. While the C-terminal catalytic domain (GGDEF) of PleD is activated by phosphorylation of the N-terminal receiver domain, functional adaptation has reversed signal transduction in PopA with the GGDEF domain adopting input function and the receiver domain serving as regulatory output. We show that the N-terminal receiver domain of PopA specifically interacts with RcdA, a component required for CtrA degradation. In contrast, the GGDEF domain serves to target PopA to the cell pole in response to c-di-GMP binding. In agreement with the divergent activation and targeting mechanisms, distinct markers sequester PleD and PopA to the old cell pole upon S-phase entry. Together these data indicate that PopA adopted a novel role as topology specificity factor to help recruit components of the CtrA degradation pathway to the protease specific old cell pole of C. crescentus.

  13. A two-component system, an anti-sigma factor and two paralogous ECF sigma factors are involved in the control of general stress response in Caulobacter crescentus.

    Science.gov (United States)

    Lourenço, Rogério F; Kohler, Christian; Gomes, Suely L

    2011-06-01

    The extracytoplasmic function sigma factor σ(T) is the master regulator of general stress response in Caulobacter crescentus and controls the expression of its paralogue σ(U). In this work we showed that PhyR and NepR act, respectively, as positive and negative regulators of σ(T) expression and function. Biochemical data also demonstrated that NepR directly binds σ(T) and the phosphorylated form of PhyR. We also described the essential role of the histidine kinase gene CC3474, here denominated phyK, for expression of σ(T)-dependent genes and for resistance to stress conditions. Additionally, in vivo evidence of PhyK-dependent phosphorylation of PhyR is presented. This study also identified a conserved cysteine residue (C95) located in the periplasmic portion of PhyK that is crucial for the function of the protein. Furthermore, we showed that PhyK, PhyR and σ(T) regulate the same set of genes and that σ(T) apparently directly controls most of its regulon. In contrast, σ(U) seems to have a very modest contribution to the expression of a subset of σ(T)-dependent genes. In conclusion, this report describes the molecular mechanism involved in the control of general stress response in C. crescentus.

  14. Suppression of amber codons in Caulobacter crescentus by the orthogonal Escherichia coli histidyl-tRNA synthetase/tRNAHis pair.

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    Ko, Jae-hyeong; Llopis, Paula Montero; Heinritz, Jennifer; Jacobs-Wagner, Christine; Söll, Dieter

    2013-01-01

    While translational read-through of stop codons by suppressor tRNAs is common in many bacteria, archaea and eukaryotes, this phenomenon has not yet been observed in the α-proteobacterium Caulobacter crescentus. Based on a previous report that C. crescentus and Escherichia coli tRNA(His) have distinctive identity elements, we constructed E. coli tRNA(His) CUA, a UAG suppressor tRNA for C. crescentus. By examining the expression of three UAG codon- containing reporter genes (encoding a β-lactamase, the fluorescent mCherry protein, or the C. crescentus xylonate dehydratase), we demonstrated that the E. coli histidyl-tRNA synthetase/tRNA(His) CUA pair enables in vivo UAG suppression in C. crescentus. E. coli histidyl-tRNA synthetase (HisRS) or tRNA(His) CUA alone did not achieve suppression; this indicates that the E. coli HisRS/tRNA(His) CUA pair is orthogonal in C. crescentus. These results illustrate that UAG suppression can be achieved in C. crescentus with an orthogonal aminoacyl-tRNA synthetase/suppressor tRNA pair.

  15. Copper-zinc superoxide dismutase of Caulobacter crescentus: Cloning, sequencing, and mapping of the gene and periplasmic location of the enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Steinman, H.M. (Albert Einstein College of Medicine, Bronx, NY (USA)); Ely, B. (Univ. of South Carolina, Columbia (USA))

    1990-06-01

    To investigate the function of the copper-zinc form of superoxide dismutase (CuZnSOD) (and its structural relationship to the eucaryotic CuZnSoDs) in the freshwater bacterium Caulobacter crecentus, the gene encoding CuZnSOD (sodC) of C. crescentus CB15 was cloned and sequenced. By hybridization to pulsed-field electrophoresis gels, sodC was mapped near cysE in the C. crescentus chromosome. Through analysis of spheroplasts, the two SODs of C. crescentus were shown to be differently localized, CuZnSOD in the periplasm and FeSOD in the cytoplasm. In its natural habitat, C. crescentus is frequently associated with blue-green algae (cyanobacteria). The oxygen evolved by these photosynthetic algae may create an extracellular oxidative stress against which the periplasmic CuZnSOD may defend more effectively than the cytoplasmic FeSOD. Amino acid sequence alignments of C. crescentus CuZnSOD with eucaryotic CuZnSODs and with CuZnSOD of Photobacterium leiognathi (the only other bacterium from which CuZnSOD has been isolated and sequenced) suggest similar supersecondary structures for bacterial and eucaryotic CuZnSODs but reveal four novel substitutions in C. crescentus CuZnSOD: a phenylalanine critical to intrasubunit hydrophobic bonding replaced by alanine, a histidine ligand of zinc replaced by aspartate, and substitutions of two other previously invariant residues that stabilize zinc or both copper and zinc.

  16. Suppression of amber codons in Caulobacter crescentus by the orthogonal Escherichia coli histidyl-tRNA synthetase/tRNAHis pair.

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    Jae-hyeong Ko

    Full Text Available While translational read-through of stop codons by suppressor tRNAs is common in many bacteria, archaea and eukaryotes, this phenomenon has not yet been observed in the α-proteobacterium Caulobacter crescentus. Based on a previous report that C. crescentus and Escherichia coli tRNA(His have distinctive identity elements, we constructed E. coli tRNA(His CUA, a UAG suppressor tRNA for C. crescentus. By examining the expression of three UAG codon- containing reporter genes (encoding a β-lactamase, the fluorescent mCherry protein, or the C. crescentus xylonate dehydratase, we demonstrated that the E. coli histidyl-tRNA synthetase/tRNA(His CUA pair enables in vivo UAG suppression in C. crescentus. E. coli histidyl-tRNA synthetase (HisRS or tRNA(His CUA alone did not achieve suppression; this indicates that the E. coli HisRS/tRNA(His CUA pair is orthogonal in C. crescentus. These results illustrate that UAG suppression can be achieved in C. crescentus with an orthogonal aminoacyl-tRNA synthetase/suppressor tRNA pair.

  17. Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2.

    Science.gov (United States)

    Milam, Sara L; Erickson, Harold P

    2013-08-16

    FtsZ from most bacteria assembles rapidly in vitro, reaching a steady-state plateau in 5-10 s after addition of GTP. A recent study used a novel dynamic light-scattering technique to assay the assembly of FtsZ from Caulobacter crescentus (CcFtsZ) and reported that assembly required 10 min, ∼100 times slower than for related bacteria. Previous studies had indicated normal, rapid assembly of CcFtsZ. We have reinvestigated the assembly kinetics using a mutant L72W, where assembly of subunits into protofilaments results in a significant increase in tryptophan fluorescence. We found that assembly reached a plateau in 5-10 s and showed no change in the following 10 min. This was confirmed by 90° light scattering and negative-stain electron microscopy. The very slow kinetics in the dynamic light-scattering study may be related to a refractory state induced when the FtsZ protein is stored without nucleotide, a phenomenon that we had observed in a previous study of EcFtsZ. We conclude that CcFtsZ is not an outlier, but shows rapid assembly kinetics similar to FtsZ from related bacteria.

  18. A multiple antibiotic and serum resistant oligotrophic strain, Klebsiella pneumoniae MB45 having novel dfrA30, is sensitive to ZnO QDs

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    Chakrabarti Pinak

    2011-05-01

    Full Text Available Abstract Background The aim of this study was to describe a novel trimethoprim resistance gene cassette, designated dfrA30, within a class 1 integron in a facultatively oligotrophic, multiple antibiotic and human serum resistant test strain, MB45, in a population of oligotrophic bacteria isolated from the river Mahananda; and to test the efficiency of surface bound acetate on zinc oxide quantum dots (ZnO QDs as bactericidal agent on MB45. Methods Diluted Luria broth/Agar (10-3 media was used to cultivate the oligotrophic bacteria from water sample. Multiple antibiotic resistant bacteria were selected by employing replica plate method. A rapid assay was performed to determine the sensitivity/resistance of the test strain to human serum. Variable region of class 1 integron was cloned, sequenced and the expression of gene coding for antibiotic resistance was done in Escherichia coli JM 109. Identity of culture was determined by biochemical phenotyping and 16S rRNA gene sequence analyses. A phylogenetic tree was constructed based on representative trimethoprim resistance-mediating DfrA proteins retrieved from GenBank. Growth kinetic studies for the strain MB45 were performed in presence of varied concentrations of ZnO QDs. Results and conclusions The facultatively oligotrophic strain, MB45, resistant to human serum and ten antibiotics trimethoprim, cotrimoxazole, ampicillin, gentamycin, netilmicin, tobramycin, chloramphenicol, cefotaxime, kanamycin and streptomycin, has been identified as a new strain of Klebsiella pneumoniae. A novel dfr gene, designated as dfrA30, found integrated in class 1 integron was responsible for resistance to trimethoprim in Klebsiella pneumoniae strain MB45. The growth of wild strain MB45 was 100% arrested at 500 mg/L concentration of ZnO QDs. To our knowledge this is the first report on application of ZnO quantum dots to kill multiple antibiotics and serum resistant K. pneumoniae strain.

  19. Species- and habitat-specific bioaccumulation of total mercury and methylmercury in the food web of a deep oligotrophic lake.

    Science.gov (United States)

    Arcagni, Marina; Juncos, Romina; Rizzo, Andrea; Pavlin, Majda; Fajon, Vesna; Arribére, María A; Horvat, Milena; Ribeiro Guevara, Sergio

    2017-09-08

    Niche segregation between introduced and native fish in Lake Nahuel Huapi, a deep oligotrophic lake in Northwest Patagonia (Argentina), occurs through the consumption of different prey. Therefore, in this work we analyzed total mercury [THg] and methylmercury [MeHg] concentrations in top predator fish and in their main prey to test whether their feeding habits influence [Hg]. Results indicate that [THg] and [MeHg] varied by foraging habitat and they increased with greater percentage of benthic diet and decreased with pelagic diet in Lake Nahuel Huapi. This is consistent with the fact that the native creole perch, a mostly benthivorous feeder, which shares the highest trophic level of the food web with introduced salmonids, had higher [THg] and [MeHg] than the more pelagic feeder rainbow trout and bentho-pelagic feeder brown trout. This differential THg and MeHg bioaccumulation observed in native and introduced fish provides evidence to the hypothesis that there are two main Hg transfer pathways from the base of the food web to top predators: a pelagic pathway where Hg is transferred from water, through plankton (with Hg in inorganic species mostly), forage fish to salmonids, and a benthic pathway, as Hg is transferred from the sediments (where Hg methylation occurs mostly), through crayfish (with higher [MeHg] than plankton), to native fish, leading to one fold higher [Hg]. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Diurnal variation in bacterioplankton composition and DNA damage in the microbial community from an Andean oligotrophic lake.

    Science.gov (United States)

    Fernández-Zenoff, María V; Estévez, María C; Farías, María E

    2014-01-01

    Laguna Azul is an oligotrophic lake situated at 4,560 m above sea level and subject to a high level of solar radiation. Bacterioplankton community composition (BCC) was analysed by denaturing gradient gel electrophoresis and the impact of solar ultraviolet radiation was assessed by measuring cyclobutane pyrimidine dimers (CPD). Furthermore, pure cultures of Acinetobacter johnsonii A2 and Rhodococcus sp. A5 were exposed simultaneously and CPD accumulation was studied. Gel analyses generated a total of 7 sequences belonging to Alpha-proteobacteria (1 band), Beta-proteobacteria (1 band), Bacteroidetes (2 bands), Actinobacteria (1 band), and Firmicutes (1 band). DGGE profiles showed minimal changes in BCC and no CPD was detected even though a high level of damage was found in biodosimeters. A. johnsonii A2 showed low level of DNA damage while Rhodococcus sp. A5 exhibited high resistance since no CPD were detected under natural UV-B exposure, suggesting that the bacterial community is well adapted to this highly solar irradiated environment. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España. All rights reserved.

  1. Re-oligotrophication as a challenge for tropical reservoir management with reference to Itaparica Reservoir, São Francisco, Brazil.

    Science.gov (United States)

    Gunkel, G; Sobral, M

    2013-01-01

    The process of reservoir eutrophication has been recognised as a central problem in tropical reservoir environmental quality. Effects of eutrophication are complex interactions involving a decrease in water quality, especially loss of aquatic biodiversity, occurrence of undesired species such as cyanobacteria with its cyanotoxins, mass development of macrophytes such as Egeria densa with its mechanical impact on turbines, and an increase in greenhouse gas emissions, mainly of methane. The eutrophication process can be described by the OECD critical load concept or related models. The phosphorus use efficiency is given by the Chl a-P - relationship, indicating eutrophic conditions by only 10 μg L(-1) P in Itaparica Reservoir, Brazil. Eutrophication of the reservoir is quantified for internal phosphorus sources (inflow, mineralisation of inundated soils and vegetation, net cage aquaculture) and external ones (agriculture, emissions of natural caatinga vegetation and rural communities) The actual internal P load is calculated to be 0.40 g m(-2) a(-1), and the critical P load is given with 1.20 g m(-2) a(-1). The external P load amounts about 1.16 g m(-2) a(-1) and thus exceeds the critical export rate of 7.1 kg km(-2) a(-1) by 50%, thus a bundling of measurements has to be considered when attempting to promote re-oligotrophication.

  2. Impacts of light shading and nutrient enrichment geo-engineering approaches on the productivity of a stratified, oligotrophic ocean ecosystem.

    Science.gov (United States)

    Hardman-Mountford, Nick J; Polimene, Luca; Hirata, Takafumi; Brewin, Robert J W; Aiken, Jim

    2013-12-06

    Geo-engineering proposals to mitigate global warming have focused either on methods of carbon dioxide removal, particularly nutrient fertilization of plant growth, or on cooling the Earth's surface by reducing incoming solar radiation (shading). Marine phytoplankton contribute half the Earth's biological carbon fixation and carbon export in the ocean is modulated by the actions of microbes and grazing communities in recycling nutrients. Both nutrients and light are essential for photosynthesis, so understanding the relative influence of both these geo-engineering approaches on ocean ecosystem production and processes is critical to the evaluation of their effectiveness. In this paper, we investigate the relationship between light and nutrient availability on productivity in a stratified, oligotrophic subtropical ocean ecosystem using a one-dimensional water column model coupled to a multi-plankton ecosystem model, with the goal of elucidating potential impacts of these geo-engineering approaches on ecosystem production. We find that solar shading approaches can redistribute productivity in the water column but do not change total production. Macronutrient enrichment is able to enhance the export of carbon, although heterotrophic recycling reduces the efficiency of carbon export substantially over time. Our results highlight the requirement for a fuller consideration of marine ecosystem interactions and feedbacks, beyond simply the stimulation of surface blooms, in the evaluation of putative geo-engineering approaches.

  3. No detectable effect of ocean acidification on plankton metabolism in the NW oligotrophic Mediterranean Sea: Results from two mesocosm studies

    Science.gov (United States)

    Maugendre, L.; Gattuso, J.-P.; Poulton, A. J.; Dellisanti, W.; Gaubert, M.; Guieu, C.; Gazeau, F.

    2017-02-01

    Oligotrophic areas account for about 30% of oceanic primary production and are projected to expand in a warm, high-CO2 world. Changes in primary production in these areas could have important impacts on future global carbon cycling. To assess the response of primary production and respiration of plankton communities to increasing partial pressure of CO2 (pCO2) levels in Low Nutrient Low Chorophyll areas, two mesocosm experiments were conducted in the Bay of Calvi (Corsica, France) and in the Bay of Villefranche (France) in June-July 2012 and February-March 2013 under different trophic state, temperature and irradiance conditions. Nine mesocosms of 50 m3 were deployed for 20 and 12 days, respectively, and were subjected to seven pCO2 levels (3 control and 6 elevated levels). The metabolism of the community was studied using several methods based on in situ incubations (oxygen light-dark, 18O and 14C uptake). Increasing pCO2 had no significant effect on gross primary production, net community production, particulate and dissolved carbon production, as well as on community respiration. These two mesocosm experiments, the first performed under maintained low nutrient and low chlorophyll, suggest that in large areas of the ocean, increasing pCO2 levels may not lead to a significant change in plankton metabolic rates and sea surface biological carbon fixation.

  4. High abundances of potentially active ammonia-oxidizing bacteria and archaea in oligotrophic, high-altitude lakes of the Sierra Nevada, California, USA.

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    Curtis J Hayden

    Full Text Available Nitrification plays a central role in the nitrogen cycle by determining the oxidation state of nitrogen and its subsequent bioavailability and cycling. However, relatively little is known about the underlying ecology of the microbial communities that carry out nitrification in freshwater ecosystems--and particularly within high-altitude oligotrophic lakes, where nitrogen is frequently a limiting nutrient. We quantified ammonia-oxidizing archaea (AOA and bacteria (AOB in 9 high-altitude lakes (2289-3160 m in the Sierra Nevada, California, USA, in relation to spatial and biogeochemical data. Based on their ammonia monooxygenase (amoA genes, AOB and AOA were frequently detected. AOB were present in 88% of samples and were more abundant than AOA in all samples. Both groups showed >100 fold variation in abundance between different lakes, and were also variable through time within individual lakes. Nutrient concentrations (ammonium, nitrite, nitrate, and phosphate were generally low but also varied across and within lakes, suggestive of active internal nutrient cycling; AOB abundance was significantly correlated with phosphate (r(2 = 0.32, p<0.1, whereas AOA abundance was inversely correlated with lake elevation (r(2 = 0.43, p<0.05. We also measured low rates of ammonia oxidation--indicating that AOB, AOA, or both, may be biogeochemically active in these oligotrophic ecosystems. Our data indicate that dynamic populations of AOB and AOA are found in oligotrophic, high-altitude, freshwater lakes.

  5. Microbial food web dynamics in response to a Saharan dust event: results from a mesocosm study in the oligotrophic Mediterranean Sea

    Science.gov (United States)

    Pulido-Villena, E.; Baudoux, A.-C.; Obernosterer, I.; Landa, M.; Caparros, J.; Catala, P.; Georges, C.; Harmand, J.; Guieu, C.

    2014-10-01

    The significant impact of dust deposition on heterotrophic bacterial dynamics in the surface oligotrophic ocean has recently been evidenced. Considering the central role of bacteria in the microbial loop, it is likely that dust deposition also affects the structure and the functioning of the whole microbial food web. In the frame of the DUNE project, aiming to estimate the impact of dust deposition on the oligotrophic Mediterranean Sea through mesocosm experiments, the main goal of the present paper was to assess how two successive dust deposition events affect the dynamics of the microbial food web. The first dust seeding delivered new P and N to the amended mesocosms and resulted in a pronounced stimulation of bacterial respiration. It also induced pronounced, but transient, changes in the bacterial community composition. No significant effects were observed on the abundances of viruses and heterotrophic nanoflagellates. The second dust seeding also delivered new P and N to the amended mesocosms, but the effect on the microbial food web was very different. Bacterial respiration remained constant and bacterial abundance decreased. Compositional changes following the second seeding were minor compared to the first one. The decrease in bacterial abundance coincided with an increase in virus abundance, resulting in higher virus:bacteria ratios throughout the second seeding period. Our study shows that dust deposition to the surface oligotrophic ocean may involve important modifications of the trophic links among the components of the microbial food web with presumed consequences on C and nutrient cycling.

  6. Primary marine aerosol emissions from the Mediterranean Sea during pre-bloom and oligotrophic conditions: correlations to seawater chlorophyll a from a mesocosm study

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    A. N. Schwier

    2014-10-01

    Full Text Available The effect of ocean acidification and changing water conditions on primary marine aerosol emissions is not well understood on a regional or a global scale. To investigate this effect as well as the indirect effect on aerosol that changing biogeochemical parameters can have, ~52 m3 pelagic mesocosms were deployed for several weeks in the Mediterranean Sea during both winter pre-bloom and summer oligotrophic conditions and were subjected to various levels of CO2 to simulate the conditions foreseen in this region for the coming decades. After seawater sampling, primary bubble-bursting aerosol experiments were performed using a plunging water jet system to test both chemical and physical aerosol parameters. Comparing results obtained during pre-bloom and oligotrophic conditions, we find the same four log-normal modal diameters (18.5, 37.5, 91.5, 260 nm describing the aerosol size distribution during both campaigns, yet pre-bloom conditions significantly increased the number fraction of the second (Aitken mode, with an amplitude correlated to virus-like particles, heterotrophic prokaryotes, TEPs, chlorophyll a and other pigments. Organic fractions determined from κ closure calculations for Dp ~50 nm were much larger during the pre-bloom period (64% than during the oligotrophic period (38%, and the organic fraction increased as the particle size decreased. Combining data from both campaigns together, strong positive correlations were found between the organic fraction of the aerosol and chlorophyll a concentrations, heterotrophic and autotrophic bacteria abundance, and dissolved organic carbon (DOC concentrations. As a consequence of the changes in the organic fraction and the size distributions between pre-bloom and oligotrophic periods, we find that the ratio of cloud condensation nuclei (CCN to condensation nuclei (CN slightly decreased during the pre-bloom period. The enrichment of the seawater samples with microlayer samples did not have any

  7. Analysis of the Caulobacter crescentus Zur regulon reveals novel insights in zinc acquisition by TonB-dependent outer membrane proteins.

    Science.gov (United States)

    Mazzon, Ricardo Ruiz; Braz, Vânia Santos; da Silva Neto, José Freire; do Valle Marques, Marilis

    2014-08-28

    Intracellular zinc concentration needs to be maintained within strict limits due to its toxicity at high levels, and this is achieved by a finely regulated balance between uptake and efflux. Many bacteria use the Zinc Uptake Regulator Zur to orchestrate zinc homeostasis, but little is known regarding the transport of this metal across the bacterial outer membrane. In this work we determined the Caulobacter crescentus Zur regulon by global transcriptional and in silico analyses. Among the genes directly repressed by Zur in response to zinc availability are those encoding a putative high affinity ABC uptake system (znuGHI), three TonB-dependent receptors (znuK, znuL and znuM) and one new putative transporter of a family not yet characterized (zrpW). Zur is also directly involved in the activation of a RND and a P-type ATPase efflux systems, as revealed by β-galactosidase and site-directed mutagenesis assays. Several genes belonging to the Fur regulon were also downregulated in the zur mutant, suggesting a putative cross-talk between Zur and Fur regulatory networks. Interestingly, a phenotypic analysis of the znuK and znuL mutants has shown that these genes are essential for growth under zinc starvation, suggesting that C. crescentus uses these TonB-dependent outer membrane transporters as key zinc scavenging systems. The characterization of the C. crescentus Zur regulon showed that this regulator coordinates not only uptake, but also the extrusion of zinc. The uptake of zinc by C. crescentus in conditions of scarcity of this metal is highly dependent on TonB-dependent receptors, and the extrusion is mediated by an RND and P-type ATPase transport systems. The absence of Zur causes a disturbance in the dynamic equilibrium of zinc intracellular concentration, which in turn can interfere with other regulatory networks as seen for the Fur regulon.

  8. Cold shock genes cspA and cspB from Caulobacter crescentus are posttranscriptionally regulated and important for cold adaptation.

    Science.gov (United States)

    Mazzon, Ricardo R; Lang, Elza A S; Silva, Carolina A P T; Marques, Marilis V

    2012-12-01

    Cold shock proteins (CSPs) are nucleic acid binding chaperones, first described as being induced to solve the problem of mRNA stabilization after temperature downshift. Caulobacter crescentus has four CSPs: CspA and CspB, which are cold induced, and CspC and CspD, which are induced only in stationary phase. In this work we have determined that the synthesis of both CspA and CspB reaches the maximum levels early in the acclimation phase. The deletion of cspA causes a decrease in growth at low temperature, whereas the strain with a deletion of cspB has a very subtle and transient cold-related growth phenotype. The cspA cspB double mutant has a slightly more severe phenotype than that of the cspA mutant, suggesting that although CspA may be more important to cold adaptation than CspB, both proteins have a role in this process. Gene expression analyses were carried out using cspA and cspB regulatory fusions to the lacZ reporter gene and showed that both genes are regulated at the transcriptional and posttranscriptional levels. Deletion mapping of the long 5'-untranslated region (5'-UTR) of each gene identified a common region important for cold induction, probably via translation enhancement. In contrast to what was reported for other bacteria, these cold shock genes have no regulatory regions downstream from ATG that are important for cold induction. This work shows that the importance of CspA and CspB to C. crescentus cold adaptation, mechanisms of regulation, and pattern of expression during the acclimation phase apparently differs in many aspects from what has been described so far for other bacteria.

  9. Development of an HIV-1 Microbicide Based on Caulobacter crescentus: Blocking Infection by High-Density Display of Virus Entry Inhibitors.

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    Christina Farr

    Full Text Available The HIV/AIDS pandemic remains an enormous global health concern. Despite effective prevention options, 2.6 million new infections occur annually, with women in developing countries accounting for more than half of these infections. New prevention strategies that can be used by women are urgently needed. Topical microbicides specific for HIV-1 represent a promising prevention strategy. Conceptually, using harmless bacteria to display peptides or proteins capable of blocking entry provides an inexpensive approach to microbicide development. To avoid the potential pitfalls of engineering commensal bacteria, our strategy is to genetically display infection inhibitors on a non-native bacterium and rely on topical application of stabilized bacteria before potential virus exposure. Due to the high density cell-surface display capabilities and the inherent low toxicity of the bacterium, the S-layer mediated protein display capabilities of the non-pathogenic bacterium Caulobacter crescentus has been exploited for this approach. We have demonstrated that C. crescentus displaying MIP1α or CD4 interfered with the virus entry pathway and provided significant protection from HIV-1 pseudovirus representing clade B in a standard single cycle infection assay. Here we have expanded our C. crescentus based microbicide approach with additional and diverse classes of natural and synthetic inhibitors of the HIV-1 entry pathway. All display constructs provided variable but significant protection from HIV-1 infection; some with protection as high as 70%. Further, we describe protection from infection with additional viral clades. These findings indicate the significant potential for engineering C. crescentus to be an effective and readily adaptable HIV-1 microbicide platform.

  10. The LovK-LovR two-component system is a regulator of the general stress pathway in Caulobacter crescentus.

    Science.gov (United States)

    Foreman, Robert; Fiebig, Aretha; Crosson, Sean

    2012-06-01

    A conserved set of regulators control the general stress response in Caulobacter crescentus, including σ(T), its anti-σ factor NepR, the anti-anti-σ factor PhyR, and the transmembrane sensor kinase PhyK. We report that the soluble histidine kinase LovK and the single-domain response regulator LovR also function within the C. crescentus general stress pathway. Our genetic data support a model in which LovK-LovR functions upstream of σ(T) by controlling the phosphorylation state and thus anti-anti-σ activity of PhyR. Transcription of lovK and lovR is independently activated by stress through a mechanism that requires sigT and phyR. Conversely, lovK and lovR function together to repress transcription of the general stress regulon. Concordant with a functional role of the LovK-LovR two-component system as a negative regulator of the general stress pathway, lovK-lovR-null mutants exhibit increased cell survival after osmotic stress, while coordinate overexpression of lovK and lovR attenuates cell survival relative to that of the wild type. Notably, lovK can complement the transcriptional and cell survival defects of a phyK-null mutant when lovR is deleted. Moreover, in this same genetic background, σ(T)-dependent transcription is activated in response to osmotic stress. This result suggests that flavin-binding LOV (light, oxygen, or voltage) histidine kinases are competent to perceive cytoplasmic signals in addition to the environmental signal blue light. We conclude that the PhyK-PhyR and LovK-LovR two-component signaling systems coordinately regulate stress physiology in C. crescentus.

  11. (p)ppGpp modulates cell size and the initiation of DNA replication in Caulobacter crescentus in response to a block in lipid biosynthesis.

    Science.gov (United States)

    Stott, Kristina V; Wood, Shannon M; Blair, Jimmy A; Nguyen, Bao T; Herrera, Anabel; Mora, Yannet G Perez; Cuajungco, Math P; Murray, Sean R

    2015-03-01

    Stress conditions, such as a block in fatty acid synthesis, signal bacterial cells to exit the cell cycle. Caulobacter crescentus FabH is a cell-cycle-regulated β-ketoacyl-acyl carrier protein synthase that initiates lipid biosynthesis and is essential for growth in rich media. To explore how C. crescentus responds to a block in lipid biosynthesis, we created a FabH-depletion strain. We found that FabH depletion blocks lipid biosynthesis in rich media and causes a cell cycle arrest that requires the alarmone (p)ppGpp for adaptation. Notably, basal levels of (p)ppGpp coordinate both a reduction in cell volume and a block in the over-initiation of DNA replication in response to FabH depletion. The gene ctrA encodes a master transcription factor that directly regulates 95 cell-cycle-controlled genes while also functioning to inhibit the initiation of DNA replication. Here, we demonstrate that ctrA transcription is (p)ppGpp-dependent during fatty acid starvation. CtrA fails to accumulate when FabH is depleted in the absence of (p)ppGpp due to a substantial reduction in ctrA transcription. The (p)ppGpp-dependent maintenance of ctrA transcription during fatty acid starvation initiated from only one of the two ctrA promoters. In the absence of (p)ppGpp, the majority of FabH-depleted cells enter a viable but non-culturable state, with multiple chromosomes, and are unable to recover from the miscoordination of cell cycle events. Thus, basal levels of (p)ppGpp facilitate C. crescentus' re-entry into the cell cycle after termination of fatty acid starvation.

  12. DNA binding of the cell cycle transcriptional regulator GcrA depends on N6-adenosine methylation in Caulobacter crescentus and other Alphaproteobacteria.

    Science.gov (United States)

    Fioravanti, Antonella; Fumeaux, Coralie; Mohapatra, Saswat S; Bompard, Coralie; Brilli, Matteo; Frandi, Antonio; Castric, Vincent; Villeret, Vincent; Viollier, Patrick H; Biondi, Emanuele G

    2013-05-01

    Several regulators are involved in the control of cell cycle progression in the bacterial model system Caulobacter crescentus, which divides asymmetrically into a vegetative G1-phase (swarmer) cell and a replicative S-phase (stalked) cell. Here we report a novel functional interaction between the enigmatic cell cycle regulator GcrA and the N6-adenosine methyltransferase CcrM, both highly conserved proteins among Alphaproteobacteria, that are activated early and at the end of S-phase, respectively. As no direct biochemical and regulatory relationship between GcrA and CcrM were known, we used a combination of ChIP (chromatin-immunoprecipitation), biochemical and biophysical experimentation, and genetics to show that GcrA is a dimeric DNA-binding protein that preferentially targets promoters harbouring CcrM methylation sites. After tracing CcrM-dependent N6-methyl-adenosine promoter marks at a genome-wide scale, we show that these marks recruit GcrA in vitro and in vivo. Moreover, we found that, in the presence of a methylated target, GcrA recruits the RNA polymerase to the promoter, consistent with its role in transcriptional activation. Since methylation-dependent DNA binding is also observed with GcrA orthologs from other Alphaproteobacteria, we conclude that GcrA is the founding member of a new and conserved class of transcriptional regulators that function as molecular effectors of a methylation-dependent (non-heritable) epigenetic switch that regulates gene expression during the cell cycle.

  13. DNA binding of the cell cycle transcriptional regulator GcrA depends on N6-adenosine methylation in Caulobacter crescentus and other Alphaproteobacteria.

    Directory of Open Access Journals (Sweden)

    Antonella Fioravanti

    2013-05-01

    Full Text Available Several regulators are involved in the control of cell cycle progression in the bacterial model system Caulobacter crescentus, which divides asymmetrically into a vegetative G1-phase (swarmer cell and a replicative S-phase (stalked cell. Here we report a novel functional interaction between the enigmatic cell cycle regulator GcrA and the N6-adenosine methyltransferase CcrM, both highly conserved proteins among Alphaproteobacteria, that are activated early and at the end of S-phase, respectively. As no direct biochemical and regulatory relationship between GcrA and CcrM were known, we used a combination of ChIP (chromatin-immunoprecipitation, biochemical and biophysical experimentation, and genetics to show that GcrA is a dimeric DNA-binding protein that preferentially targets promoters harbouring CcrM methylation sites. After tracing CcrM-dependent N6-methyl-adenosine promoter marks at a genome-wide scale, we show that these marks recruit GcrA in vitro and in vivo. Moreover, we found that, in the presence of a methylated target, GcrA recruits the RNA polymerase to the promoter, consistent with its role in transcriptional activation. Since methylation-dependent DNA binding is also observed with GcrA orthologs from other Alphaproteobacteria, we conclude that GcrA is the founding member of a new and conserved class of transcriptional regulators that function as molecular effectors of a methylation-dependent (non-heritable epigenetic switch that regulates gene expression during the cell cycle.

  14. The CcrM DNA methyltransferase is widespread in the alpha subdivision of proteobacteria, and its essential functions are conserved in Rhizobium meliloti and Caulobacter crescentus.

    Science.gov (United States)

    Wright, R; Stephens, C; Shapiro, L

    1997-09-01

    The Caulobacter crescentus DNA methyltransferase CcrM (M.CcrMI) methylates the adenine residue in the sequence GANTC. The CcrM DNA methyltransferase is essential for viability, but it does not appear to be part of a DNA restriction-modification system. CcrM homologs are widespread in the alpha subdivision of gram-negative bacteria. We have amplified and sequenced a 258-bp region of the cerM gene from several of these bacteria, including Rhizobium meliloti, Brucella abortus, Agrobacterium tumefaciens, and Rhodobacter capsulatus. Alignment of the deduced amino acid sequences revealed that these proteins constitute a highly conserved DNA methyltransferase family. Isolation of the full-length ccrM genes from the aquatic bacterium C. crescentus, the soil bacterium R. meliloti, and the intracellular pathogen B. abortus showed that this sequence conservation extends over the entire protein. In at least two alpha subdivision bacteria, R. meliloti and C. crescentus, CcrM-mediated methylation has important cellular functions. In both organisms, CcrM is essential for viability. Overexpression of CcrM in either bacterium results in defects in cell division and cell morphology and in the initiation of DNA replication. Finally, the C. crescentus and R. meliloti ccrM genes are functionally interchangeable, as the complemented strains are viable and the chromosomes are methylated. Thus, in both R. meliloti and C. crescentus, CcrM methylation is an integral component of the cell cycle. We speculate that CcrM-mediated DNA methylation is likely to have similar roles among alpha subdivision bacteria.

  15. Localization of the outer membrane protein OmpA2 in Caulobacter crescentus depends on the position of the gene in the chromosome.

    Science.gov (United States)

    Ginez, Luis David; Osorio, Aurora; Poggio, Sebastian

    2014-08-01

    The outer membrane of Gram-negative bacteria is an essential structure involved in nutrient uptake, protection against harmful substances, and cell growth. Different proteins keep the outer membrane from blebbing out by simultaneously interacting with it and with the cell wall. These proteins have been mainly studied in enterobacteria, where OmpA and the Braun and Pal lipoproteins stabilize the outer membrane. Some degree of functional redundancy exists between these proteins, since none of them is essential but the absence of two of them results in a severe phenotype. Caulobacter crescentus has a different strategy to maintain its outer membrane, since it lacks the Braun lipoprotein and Pal is essential. In this work, we characterized OmpA2, an OmpA-like protein, in this bacterium. Our results showed that this protein is required for normal stalk growth and that it plays a minor role in the stability of the outer membrane. An OmpA2 fluorescent fusion protein showed that the concentration of this protein decreases from the stalk to the new pole. This localization pattern is important for its function, and it depends on the position of the gene locus in the chromosome and, as a consequence, in the cell. This result suggests that little diffusion occurs from the moment that the gene is transcribed until the mature protein attaches to the cell wall in the periplasm. This mechanism reveals the integration of different levels of information from protein function down to genome arrangement that allows the cell to self-organize.

  16. Sources and sinks of nitrogen and phosphorus to a deep, oligotrophic lake, Lake Crescent, Olympic National Park, Washington

    Science.gov (United States)

    Moran, P.W.; Cox, S.E.; Embrey, S.S.; Huffman, R.L.; Olsen, T.D.; Fradkin, S.C.

    2012-01-01

    Lake Crescent, in Olympic National Park in the northwest corner of Washington State is a deep-water lake renowned for its pristine water quality and oligotrophic nature. To examine the major sources and sinks of nutrients (as total nitrogen, total phosphorus, and dissolved nitrate), a study was conducted in the Lake Crescent watershed. The study involved measuring five major inflow streams, the Lyre River as the major outflow, recording weather and climatic data, coring lake bed sediment, and analyzing nutrient chemistry in several relevant media over 14 months. Water samples for total nitrogen, total phosphorous, and dissolved nitrate from the five inflow streams, the outlet Lyre River, and two stations in the lake were collected monthly from May 2006 through May 2007. Periodic samples of shallow water from temporary sampling wells were collected at numerous locations around the lake. Concentrations of nutrients detected in Lake Crescent and tributaries were then applied to the water budget estimates to arrive at monthly and annual loads from various environmental components within the watershed. Other sources, such as leaf litter, pollen, or automobile exhaust were estimated from annual values obtained from various literature sources. This information then was used to construct a nutrient budget for total nitrogen and total phosphorus. The nitrogen budget generally highlights vehicle traffic-diesel trucks in particular-along U.S. Highway 101 as a potential major anthropogenic source of nitrogen compounds in the lake. In contrast, contribution of nitrogen compounds from onsite septic systems appears to be relatively minor related to the other sources identified.

  17. An experimental study on the effects of nutrient enrichment on organic carbon storage in western Pacific oligotrophic gyre

    Directory of Open Access Journals (Sweden)

    J. Liu

    2014-02-01

    Full Text Available Carbon sequestration in the ocean is of great concern with respect to the mitigation of global warming. How to hold the fixed organic carbon in the presence of tremendous heterotrophic microorganisms in marine environments is the central issue. We have previously hypothesized that excessive nutrients would ultimately decrease the storage of organic carbon in marine environments. To test it out, a series of in situ nutrient enrichment incubation experiments were conducted at a site (17.59° N, 127.00° E within the Western Pacific oligotrophic gyre. Five treatments were employed: glucose or algal exudation organic material (EOM and nitrate and phosphate were added alone or in combination to approximate final concentrations of 10 μmol C kg−1, 1 μmol N kg−1 and 0.11 μmol P kg−1 respectively. The results showed that the dissolved organic carbon (DOC consumption rates and bacterial community specific growth rates were enhanced by inorganic nutrients enrichment treatments during the initial 48 h incubation. At the end of 14 days incubation, about 1/3 (average 3.29 μmol C kg−1 more organic carbon was respired from the glucose enriched incubation with addition of inorganic nutrients compared to that without addition of inorganic nutrients. In the case no essential nutrients were available, even glucose could not be efficiently used by bacteria and thus remained in the environment. These results suggest that repletion of inorganic nutrients has negative impacts on carbon preservation, presumably due to elevated nutrient-stimulated bacterial metabolism and respiration, which is meaningful for potential coastal water management and worth for further studies.

  18. Effects of increasing temperatures on methane concentrations and methanogenesis during experimental incubation of sediments from oligotrophic and mesotrophic lakes

    Science.gov (United States)

    Fuchs, Andrea; Lyautey, Emilie; Montuelle, Bernard; Casper, Peter

    2016-05-01

    Global warming is expected to raise temperatures in freshwater lakes, which have been acknowledged to contribute up to 10% of the atmospheric methane concentrations. Increasing temperature enhances methane production and oxidation rates, but few studies have considered the balance between both processes at experimentally higher temperatures within lake sediments. The temperature dependence of methane concentrations, methane production rates, and methanogenic (mcrA) and methanotrophic (pmoA) community size was investigated in intact sediment cores incubated with aerobic hypolimnion water at 4, 8, and 12°C over 3 weeks. Sediment cores of 25 cm length were collected at two temperate lakes—Lake Stechlin (Germany; mesotrophic-oligotrophic, maximum depth 69.5 m) and Lake Geneva (France/Switzerland; mesotrophic, maximum depth 310 m). While methane production rates in Lake Stechlin sediments did not change with increasing temperatures, methane concentrations decreased significantly. In contrast, methane production rates increased in 20-25 cm in Lake Geneva sediments with increasing temperatures, but methane concentrations did not differ. Real-time PCR demonstrated the methanogenic and methanotrophic community size remained stable independently of the incubation temperature. Methane concentrations as well as community sizes were 1-2 orders of magnitude higher in Lake Stechlin than in Lake Geneva, while potential methane production rates after 24 h were similar in both lakes, with on average 2.5 and 1.9 nmol g-1 DW h-1, respectively. Our results suggest that at higher temperatures methane oxidation could balance, and even exceed, methane production. This suggests that anaerobic methane oxidation could be involved in the methane balance at a more important rate than previously anticipated.

  19. Plankton ecosystem functioning and nitrogen fluxes in the most oligotrophic waters of the Beaufort Sea, Arctic Ocean: a modeling study

    Directory of Open Access Journals (Sweden)

    V. Le Fouest

    2012-10-01

    Full Text Available The Arctic Ocean (AO undergoes profound changes of its physical and biotic environments due to climate change. The greater light exposure and stratification alter its plankton ecosystem structure, functioning and productivity promoting oligotrophy in some areas as the Beaufort Sea. A one-dimension (1-D physical-biological coupled model based on the large multiparametric database of the Malina project in the Beaufort Sea was used (i to infer the functioning and nitrogen fluxes within the summer plankton ecosystem and (ii to assess the model sensitivity to key light-associated processes involved in nutrient recycling and phytoplankton growth. The coupled model suggested that ammonium photochemically produced from photosensitive dissolved organic nitrogen (i.e. photoammonification process was a necessary nitrogen source to achieve the observed levels of microbial biomass and production. It contributed to ca. two-thirds and one-third of the simulated surface (0–10 m and depth-integrated primary and bacterial production, respectively. The model also suggested that carbon to chlorophyll ratios for small (< 5 μm phytoplankton (ca. 15–45 g g−1 lower than those commonly used in biogeochemical models applied to the AO were required to simulate the observed herbivorous versus microbial food web competition and realistic nitrogen fluxes in the Beaufort Sea oligotrophic waters. In face of accelerating Arctic warming, more attention should be paid in the future to the mechanistic processes involved in food webs and functional groups competition, nutrient recycling and primary production in poorly productive waters of the AO as they are expected to expand rapidly.

  20. Constraining the sources and cycling of dissolved organic carbon in a large oligotrophic lake using radiocarbon analyses

    Science.gov (United States)

    Zigah, Prosper K.; Minor, Elizabeth C.; McNichol, Ann P.; Xu, Li; Werne, Josef P.

    2017-07-01

    We measured the concentrations and isotopic compositions of solid phase extracted (SPE) dissolved organic carbon (DOC) and high molecular weight (HMW) DOC and their constituent organic components in order to better constrain the sources and cycling of DOC in a large oligotrophic lacustrine system (Lake Superior, North America). SPE DOC constituted a significant proportion (41-71%) of the lake DOC relative to HMW DOC (10-13%). Substantial contribution of 14C-depleted components to both SPE DOC (Δ14C = 25-43‰) and HMW DOC (Δ14C = 22-32‰) was evident during spring mixing, and depressed their radiocarbon values relative to the lake dissolved inorganic carbon (DIC; Δ14C ∼ 59‰). There was preferential removal of 14C-depleted (older) and thermally recalcitrant components from HMW DOC and SPE DOC in the summer. Contemporary photoautotrophic addition to HMW DOC was observed during summer stratification in contrast to SPE DOC, which decreased in concentration during stratification. Serial thermal oxidation radiocarbon analysis revealed a diversity of sources (both contemporary and older) within the SPE DOC, and also showed distinct components within the HMW DOC. The thermally labile components of HMW DOC were 14C-enriched and are attributed to heteropolysaccharides (HPS), peptides/amide and amino sugars (AMS) relative to the thermally recalcitrant components reflecting the presence of older material, perhaps carboxylic-rich alicyclic molecules (CRAM). The solvent extractable lipid-like fraction of HMW DOC was very 14C-depleted (as old as 1270-2320 14C years) relative to the carbohydrate-like and protein-like substances isolated by acid hydrolysis of HMW DOC. Our data constrain relative influences of contemporary DOC and old DOC, and DOC cycling in a modern freshwater ecosystem.

  1. Sources and sinks of nitrogen and phosphorus to a deep, oligotrophic lake, Lake Crescent, Olympic National Park, Washington

    Science.gov (United States)

    Moran, P.W.; Cox, S.E.; Embrey, S.S.; Huffman, R.L.; Olsen, T.D.; Fradkin, S.C.

    2012-01-01

    Lake Crescent, in Olympic National Park in the northwest corner of Washington State is a deep-water lake renowned for its pristine water quality and oligotrophic nature. To examine the major sources and sinks of nutrients (as total nitrogen, total phosphorus, and dissolved nitrate), a study was conducted in the Lake Crescent watershed. The study involved measuring five major inflow streams, the Lyre River as the major outflow, recording weather and climatic data, coring lake bed sediment, and analyzing nutrient chemistry in several relevant media over 14 months. Water samples for total nitrogen, total phosphorous, and dissolved nitrate from the five inflow streams, the outlet Lyre River, and two stations in the lake were collected monthly from May 2006 through May 2007. Periodic samples of shallow water from temporary sampling wells were collected at numerous locations around the lake. Concentrations of nutrients detected in Lake Crescent and tributaries were then applied to the water budget estimates to arrive at monthly and annual loads from various environmental components within the watershed. Other sources, such as leaf litter, pollen, or automobile exhaust were estimated from annual values obtained from various literature sources. This information then was used to construct a nutrient budget for total nitrogen and total phosphorus. The nitrogen budget generally highlights vehicle traffic-diesel trucks in particular-along U.S. Highway 101 as a potential major anthropogenic source of nitrogen compounds in the lake. In contrast, contribution of nitrogen compounds from onsite septic systems appears to be relatively minor related to the other sources identified.

  2. Relative importance of P and N in macrophyte and epilithic algae biomass in a wastewater-impacted oligotrophic river.

    Science.gov (United States)

    Taube, Nadine; He, Jianxun; Ryan, M Cathryn; Valeo, Caterina

    2016-08-01

    The role of nutrient loading on biomass growth in wastewater-impacted rivers is important in order to effectively optimize wastewater treatment to avoid excessive biomass growth in the receiving water body. This paper directly relates wastewater treatment plant (WWTP) effluent nutrients (including ammonia (NH3-N), nitrate (NO3-N) and total phosphorus (TP)) to the temporal and spatial distribution of epilithic algae and macrophyte biomass in an oligotrophic river. Annual macrophyte biomass, epilithic algae data and WWTP effluent nutrient data from 1980 to 2012 were statistically analysed. Because discharge can affect aquatic biomass growth, locally weighted scatterplot smoothing (LOWESS) was used to remove the influence of river discharge from the aquatic biomass (macrophytes and algae) data before further analysis was conducted. The results from LOWESS indicated that aquatic biomass did not increase beyond site-specific threshold discharge values in the river. The LOWESS-estimated biomass residuals showed a variable response to different nutrients. Macrophyte biomass residuals showed a decreasing trend concurrent with enhanced nutrient removal at the WWTP and decreased effluent P loading, whereas epilithic algae biomass residuals showed greater response to enhanced N removal. Correlation analysis between effluent nutrient concentrations and the biomass residuals (both epilithic algae and macrophytes) suggested that aquatic biomass is nitrogen limited, especially by NH3-N, at most sampling sites. The response of aquatic biomass residuals to effluent nutrient concentrations did not change with increasing distance to the WWTP but was different for P and N, allowing for additional conclusions about nutrient limitation in specific river reaches. The data further showed that the mixing process between the effluent and the river has an influence on the spatial distribution of biomass growth.

  3. Zooplanktivory and nutrient regeneration by invertebrate (Mysis relicta) and vertebrate (Oncorhynchus nerka) planktivores: Implications for trophic interactions in oligotrophic lakes

    Science.gov (United States)

    Chipps, S.R.; Bennett, D.H.

    2000-01-01

    We investigated zooplanktivory and nutrient regeneration by the opossum shrimp Mysis relicta and kokanee Oncorhynchus nerka to assess the relative roles of these planktivores in oligotrophic food webs. Using bioenergetic models and clearance rate estimates, we quantified phosphorus (P) excretion rates and consumption of cladoceran prey by Mysis and kokanees in Lake Pend Oreille, Idaho, from 1995 to 1996. Consumption of cladoceran prey by Mysis was 186 kg ?? ha-1 ?? year-1, whereas consumption by kokanees was less than one quarter as much, at 45 kg ?? ha-1 ?? year-1. Similarly, Mysis excreted approximately 0.250 kg P ?? ha-1 ?? year-1 during nighttime migrations into the upper water column, whereas P excretion by kokanees was less than one third as much, at approximately 0.070 kg P ?? ha-1 ?? year-1. On a volumetric basis, nocturnal excretion by Mysis ranged from 0.002 to 0.007 ??g P ?? L-1 ?? d-1 and accounted for less than 1% of the soluble reactive P typically measured in the upper water column of the lake. Hence, nutrient recycling by Mysis may be limited in the upper water column because of the nocturnal feeding habitats that constrain Mysis to deeper strata for much of the day. In spring and autumn months, low abundance of cladoceran prey coincided with high seasonal energy requirements of the Mysis population that were linked to timing of annual Mysis brood release and abundance of age-0 Mysis. Predation by Mysis accounted for 5-70% of daily cladoceran standing stock, supporting the notion that seasonal availability of cladocerans may be regulated by Mysis predation. In lakes where Mysis experience little predation mortality, they likely play a dominant role in food web interactions (e.g., trophic cascades) relative to planktivorous fishes. Biotic mechanisms, such as successful predator-avoidance behavior, omnivorous feeding habits, and seasonal variation in Mysisbiomass, enhance the ability of Mysis to influence food web interactions from an intermediate

  4. Abiotic variables affect STX concentration in a meso-oligotrophic subtropical coastal lake dominated by Cylindrospermopsis raciborskii (Cyanophyceae).

    Science.gov (United States)

    Brentano, Débora Monteiro; Giehl, Eduardo L Hettwer; Petrucio, Maurício Mello

    2016-06-01

    The cyanobacterium Cylindrospermopsis raciborskii is capable of producing toxins including saxitoxin (STX). Few studies have verified the influence of environmental variables on the production of STX and most have only been studied in the laboratory. The goal of this work was to identify the abiotic variables related to STX concentration in situ. The relationship among STX concentration and the physical variables, nutrients and chlorophyll-a (chl-a) concentration was examined in a meso-oligotrophic subtropical coastal lake dominated by C. raciborskii. A generalized linear model was developed, incorporating all variables measured monthly over a 45-month monitoring period. Conductivity and dissolved inorganic nitrogen (DIN) concentration provided the greatest explanatory power for STX concentration in situ. Previous studies suggested that C. raciborskii cells exposed to stress associated with higher ionic concentrations appear to activate the biosynthesis of STX suggesting that STX can elicit changes cell permeability and may contribute to the homeostasis of this organism. An increase of DIN concentration results in a higher concentration of STX which may be related to a reduced metabolic demand, since the uptake of inorganic nitrogen requires less energy than N2-fixation. Thus, increased DIN can favor the growth of C. raciborskii population or improve cellular homeostasis, both potentially increasing STX concentration in the aquatic system, which was observed through a delayed response pattern. The developed model, while providing only a moderate predictive power, can assist in the understanding of the environmental variables associated with increases in STX concentration, and in monitoring and minimizing the risks of toxic blooms.

  5. İzmir İlinde Çeşitli Su Kaynaklarındaki Caulobacter'lerin Karakterizasyonu ve Antibiyotik Dirençlerinin İncelenmesi.

    Directory of Open Access Journals (Sweden)

    Aslı Kaçar

    2015-12-01

    Full Text Available Caulobacter, uzantılı bakteriler grubunun farklı bir genusudur. Bu bakteriler, hayat döngüleri boyunca tek flajelli yüzücü hücre ve bir sapa sahip hareketsiz hücre formlarını sırayla oluşturmaktadır. Denizden ve kıyı bölgelerinden olduğu gibi topraktan ve tatlısulardan izole edilebilirler. Bu çalışmada, İzmir ilindeki 10 farklı su kaynağından alınan örneklerden 29 Caulobacter genusu üyesi izole edilmiştir. Yapılan mikroskobik incelemeler ve biyokimyasal testler sonucunda, 9 izolatın C. leidyi, 14’ünün C. variabilis, 6’sının C. intermedius olduğu belirlenmiş ve 7 farklı antibiyotiğin bu bakterilerin büyümesi üzerine etkisi tespit edilmiştir

  6. Nitrogen- and irradiance-dependent variations of the maximum quantum yield of carbon fixation in eutrophic, mesotrophic and oligotrophic marine systems

    Science.gov (United States)

    Babin, Marcel; Morel, André; Claustre, Hervé; Bricaud, Annick; Kolber, Zbigniew; Falkowski, Paul G.

    1996-08-01

    Natural variability of the maximum quantum yield of carbon fixation ( φC max), as determined from the initial slope of the photosynthesis-irradiance curve and from light absorption measurements, was studied at three sites in the northeast tropical Atlantic representing typical eutrophic, mesotrophic and oligotrophic regimes. At the eutrophic and mesotrophic sites, where the mixed layer extended deeper than the euphotic layer, all photosynthetic parameters were nearly constant with depth, and φC max averaged between 0.05 and 0.03 molC (mol quanta absorbed) -1, respectively. At the oligotrophic site, a deep chlorophyll maximum (DCM) existed and φC max varied from ca 0.005 in the upper nutrient-depleted mixed layer to 0.063 below the DCM in stratified waters. firstly, φC max was found roughly to covary with nitrate concentration between sites and with depth at the oligotrophic site, and secondly, it was found to decrease with increasing relative concentrations of non-photosynthetic pigments. The extent of φC max variations directly related to nitrate concentration was inferred from variations in the fraction of functional PS2 reaction centers ( f), measured using fast repetition rate fluorometry. Covariations between f and nitrate concentration indicate that the latter factor may be responsible for a 2-fold variation in φC max. Moreover, partitioning light absorption between photosynthetic and non-photosynthetic pigments suggests that the variable contribution of the non-photosynthetic absorption may explain a 3-fold variation in φC max, as indicated by variations in the effective absorption cross-section of photosystem 2 ( σPS2). Results confirm the role of nitrate in φC max variation, and emphasize those of light and vertical mixing.

  7. Decade-long time delays in nutrient and plant species dynamics during eutrophication and re-oligotrophication of Lake Fure 1900–2015

    DEFF Research Database (Denmark)

    Sand-Jensen, Kaj; Bruun, Hans Henrik; Båstrup-Spohr, Lars

    2017-01-01

    sediment release. Fifty years of eutrophication led to a reduction in aquatic macrophyte richness from 36 species to 12. Species’ responses were closely related to their growth strategy and depth distribution. Deep-growing mosses, charophytes and short angiosperms disappeared, while tall angiosperms...... survived and pollution-tolerant macroalgae colonized and spread. Subsequently, 45 years of oligotrophication led to clearer waters, macrophyte richness recovering to 28 species and some charophytes and short angiosperms reappearing. Dominance of pollution-tolerant macroalgae persisted, however. Change...

  8. Carbon and nitrogen stable isotope ratios of pelagic zooplankton elucidate ecohydrographic features in the oligotrophic Red Sea

    Science.gov (United States)

    Kürten, Benjamin; Al-Aidaroos, Ali M.; Kürten, Saskia; El-Sherbiny, Mohsen M.; Devassy, Reny P.; Struck, Ulrich; Zarokanellos, Nikolaos; Jones, Burton H.; Hansen, Thomas; Bruss, Gerd; Sommer, Ulrich

    2016-01-01

    Although zooplankton occupy key roles in aquatic biogeochemical cycles, little is known about the pelagic food web and trophodynamics of zooplankton in the Red Sea. Natural abundance stable isotope analysis (SIA) of carbon (δ13C) and N (δ15N) is one approach to elucidating pelagic food web structures and diet assimilation. Integrating the combined effects of ecological processes and hydrography, ecohydrographic features often translate into geographic patterns in δ13C and δ15N values at the base of food webs. This is due, for example, to divergent 15N abundances in source end-members (deep water sources: high δ15N, diazotrophs: low δ15N). Such patterns in the spatial distributions of stable isotope values were coined isoscapes. Empirical data of atmospheric, oceanographic, and biological processes, which drive the ecohydrographic gradients of the oligotrophic Red Sea, are under-explored and some rather anticipated than proven. Specifically, five processes underpin Red Sea gradients: (a) monsoon-related intrusions of nutrient-rich Indian Ocean water; (b) basin scale thermohaline circulation; (c) mesoscale eddy activity that causes up-welling of deep water nutrients into the upper layer; (d) the biological fixation of atmospheric nitrogen (N2) by diazotrophs; and (e) the deposition of dust and aerosol-derived N. This study assessed relationships between environmental samples (nutrients, chlorophyll a), oceanographic data (temperature, salinity, current velocity [ADCP]), particulate organic matter (POM), and net-phytoplankton, with the δ13C and δ15N values of zooplankton collected in spring 2012 from 16°28‧ to 26°57‧N along the central axis of the Red Sea. The δ15N of bulk POM and most zooplankton taxa increased from North (Duba) to South (Farasan). The potential contribution of deep water nutrient-fueled phytoplankton, POM, and diazotrophs varied among sites. Estimates suggested higher diazotroph contributions in the North, a greater contribution of

  9. Landslide-induced iron mobilisation shapes benthic accumulation of nutrients, trace metals and REE fractionation in an oligotrophic alpine stream

    Science.gov (United States)

    Johnston, Scott G.; Rose, Andrew L.; Burton, Edward D.; Webster-Brown, Jenny

    2015-01-01

    Large alpine landslides that entrain substantial organic material below the water table and create suspended floodplains may have long-term consequences for the mobilisation of redox sensitive elements, such as Fe, into streamwaters. In turn, the cycling of iron in aquatic systems can influence the fate of nutrients, alter primary productivity, enhance accumulation of trace metals and induce fractionation of rare earth elements (REE). In this study we examine a reach of a pristine oligotrophic alpine stream bracketing a 30 year-old landslide and explore the consequences of landslide-induced Fe mobilisation for aqueous geochemistry and the composition of benthic stream cobble biofilm. Elevated Fe2+ and Mn in landslide zone stream waters reflect inputs of circumneutral groundwater from the landslide debris-zone floodplain. Geochemical characteristics are consistent with reductive dissolution being a primary mechanism of Fe2+ and Mn mobilisation. Stream cobble biofilm in the landslide zone is significantly (P extractable; Fe(III)Ab). While the landslide zone accounts for less than ∼9% of the total stream length, we estimate it is responsible for approximately 60-80% of the stream's benthic biofilm load of poorly crystalline Fe(III) and Mn. Biofilm Fe(III) precipitates are comprised mainly of ferrihydrite, lepidocrocite and an organic-Fe species, while precipitate samples collected proximal to hyporheic seeps contain abundant sheath structures characteristic of the neutrophilic Fe(II)-oxidising bacteria Leptothrix spp. Stream-cobble Fe(III)-rich biofilm is accumulating PO43- (∼3-30 times background) and behaving as a preferential substrate for photosynthetic periphyton, with benthic PO43-, chlorophyll a, organic carbonHCl and total N all significantly positively correlated with Fe(III)Ab and significantly elevated within the landslide zone (P transformed basis. Stream cobble biofilm also exhibits distinct REE fractionation along the flow path, with light REE (La

  10. Preliminary investigations on picoplankton-related precipitation of alkaline-earth metal carbonates in meso-oligotrophic lake Geneva (Switzerland

    Directory of Open Access Journals (Sweden)

    Jean-Michel Jaquet

    2013-10-01

    Full Text Available In the course of a routine water-quality survey in meso-oligotrophic lake Geneva (Switzerland, suspended matter was collected by filtration on 0.2 μm membranes in July and August 2012 at the depth of maximal chlorophyll a (Chl a concentration (2 mg m–3. Examination by scanning electron microscopy revealed the presence of numerous dark and gelatinous patches occluding the pores of the membranes, containing high numbers of picoplanktonic cells and, in places, clusters of high-reflectance smooth microspheres (1-2 μm in diameter. Their chemical composition, determined by semi-quantitative, energy-dispersive X ray spectroscopy (EDS showed magnesium (Mg, calcium (Ca, strontium (Sr and barium (Ba (alkaline earth metals to be the dominant cations. Among the anions, phosphorus (P and carbon (C were present, but only the latter is considered here (as carbonate. The microspheres were subdivided into four types represented in a Ca-Sr-Ba ternary space. All types are confined within a domain bound by Ca>45, Sr<10 and Ba<50 (in mole %. Type I, the most frequent, displays a broad variability in Ba/Ca, even within a given cluster. Types II and III are devoid of Ba, but may incorporate P. Type IV contains only Ca. The Type I composition resembles that of benstonite, a Group IIA carbonate that was recently found as intracellular granules in a cyanobacterium from alkaline lake Alchichica (Mexico.Lake Geneva microspheres are solid, featureless and embedded in a mucilage-looking substance in the vicinity of, but seemingly not inside, picoplanktonic cells morphologically similar to Chlorella and Synechococcus. In summer 2012, the macroscopic physico-chemical conditions in lake Geneva epilimnion were such as to allow precipitation of Ca but not of Sr and Ba carbonates. Favourable conditions did exist, though, in the micro-environment provided by the combination of active picoplankton and a mucilaginous envelope. Further studies are ongoing to investigate the

  11. Digestive tract morphology of the Neotropical piscivorous fish Cichla kelberi(Perciformes: Cichlidae introduced into an oligotrophic Brazilian reservoir

    Directory of Open Access Journals (Sweden)

    Alejandra Filippo Gonzalez Neves dos Santos

    2011-09-01

    Full Text Available Despite being one of the most well-known cichlid fish of importance to artisanal and sport fishing, and among the largest fishes in the Neotropics, data on digestive tract anatomy of peacock basses (Cichla spp. are largely lacking, especially for non-native populations. in this paper, we describe for the first time the digestive tract morphology of Cichla kelberi, a voracious piscivore that was introduced in the 1950s into an oligotrophic and physically low-complex impoundment in Brazil. Peacock basses were collected between 1994 and 2002 in Lajes Reservoir, through gillnets (25 to 55mm mesh; 20-50x2m, seines (10x2.5m; 8.0mm mesh, cast nets and angling. All the fishes were kept on ice in the field and then transferred to the laboratory, where they were identified, measured, weighed and dissected for digestive tract analyses. The index of Relative importance-IRI was calculated for diet characterization while linear and non-linear regressions were performed to assess growth patterns of four morphological characters related to feeding (e.g. mouth width, mouth height, stomach length and intestine length and the number of gill rakers during the C. kelberi ontogeny. Most digestive tract structures were directly related to the piscivorous diet of C. kelberi, indicating that peacock bass is a diurnal, bathypelagic and gape-size limited predator that feeds largely on shallow-water prey species within the littoral zone. Mouth width and height grew allometrically (b>1 with the size of peacock bass, broadening the size range in which prey can be eaten, but especially for predators smaller than ~400mm of total length. Differently, stomach and intestine lengths increased isometrically (b=1, which could constrain prey consumption for adult C. kelberi, especially those at advanced stages of gonadal maturation. The presence of longer-drawn, sharp and furcated gill rakers in C. kelberi may be related to increased prey retention in the resource-limited Lajes

  12. Bacterial diversity in shallow oligotrophic marine benthos and overlying waters: effects of virus infection, containment, and nutrient enrichment.

    Science.gov (United States)

    Hewson, I; Vargo, G A; Fuhrman, J A

    2003-10-01

    Little is known of the factors shaping sediment bacterial communities, despite their high abundance and reports of high diversity. Two factors hypothesized to shape bacterial communities in the water column are nutrient (resource) availability and virus infection. The role these factors play in benthic bacterial diversity was assessed in oligotrophic carbonate-based sediments of Florida Bay (USA). Sediment-water mesocosm enclosures were made from 1-m diameter clear polycarbonate cylinders which were pushed into sediments to approximately 201 cm sediment depth enclosing approximately 80 L of water. Mesocosms were amended each day for 14 d with 10 microM NH4+ and 1 microM PO4(3-). In a second experiment, viruses from a benthic flocculent layer were concentrated and added back to flocculent layer samples which were collected near the mesocosm enclosures. Photosynthesis by microalgae in virus-amended incubations was monitored by pulse-amplitude modulated (PAM) fluorescence. In both experiments, bacterial diversity was estimated using automated rRNA intergenic spacer analysis (ARISA), a high-resolution fingerprinting approach. Initial sediment bacterial operational taxonomic unit (OTU) richness (236 +/- 3) was higher than in the water column (148 +/- 9), where an OTU was detectable when its amplified DNA represented >0.09% of the total amplified DNA. Effects on bacterial diversity and operational taxonomic unit (OTU) richness in nutrient-amended mesocosms may have been masked by the effects of containment, which stimulated OTU richness in the water column, but depressed OTU richness and diversity in sediments. Nutrient addition significantly elevated virus abundance and the ratio of viruses to bacteria (p < 0.05 for both) in the sediments, concomitant with elevated bacterial diversity. However, water column bacterial diversity (in unamended controls) was not affected by nutrient amendments, which may be due to rapid nutrient uptake by sediment organisms or adsorption of

  13. Carbon and nitrogen stable isotope ratios of pelagic zooplankton elucidate ecohydrographic features in the oligotrophic Red Sea

    KAUST Repository

    Kürten, Benjamin

    2015-11-10

    Although zooplankton occupy key roles in aquatic biogeochemical cycles, little is known about the pelagic food web and trophodynamics of zooplankton in the Red Sea. Natural abundance stable isotope analysis (SIA) of carbon (δ13C) and N (δ15N) is one approach to elucidating pelagic food web structures and diet assimilation Integrating the combined effects of ecological processes and hydrography, ecohydrographic features often translate into geographic patterns in δ13C and δ15N values at the base of food webs. This is due, for example, to divergent 15N abundances in source end-members (deep water sources: high δ15N, diazotrophs: low δ15N). Such patterns in the spatial distributions of stable isotope values were coined isoscapes. Empirical data of atmospheric, oceanographic, and biological processes, which drive the ecohydrographic gradients of the oligotrophic Red Sea, are under-explored and some rather anticipated than proven. Specifically, five processes underpin Red Sea gradients: a) monsoon-related intrusions of nutrient-rich Indian Ocean water; b) basin scale thermohaline circulation; c) mesoscale eddy activity that causes up-welling of deep water nutrients into the upper layer; d) the biological fixation of atmospheric nitrogen (N2) by diazotrophs; and e) the deposition of aerosol-derived N. This study assessed relationships between environmental samples (nutrients, chlorophyll a), oceanographic data (temperature, salinity, current velocity [ADCP]), particulate organic matter (POM), and net-phytoplankton, with the δ13C and δ15N values of zooplankton collected in spring 2012 from 16°28’ to 26°57’N along the central axis of the Red Sea. The δ15N of bulk POM and most zooplankton taxa increased from North (Duba) to South (Farasan). The potential contribution of deep water nutrient-fueled phytoplankton, POM, and diazotrophs varied among sites. Estimates suggested higher diazotroph contributions in the North, a greater contribution of POM in the South

  14. Crystallization and X-ray diffraction analysis of an l-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii and a d-xylonate dehydratase from Caulobacter crescentus

    Science.gov (United States)

    Rahman, Mohammad Mubinur; Andberg, Martina; Koivula, Anu; Rouvinen, Juha; Hakulinen, Nina

    2016-01-01

    l-Arabinonate dehydratase (EC 4.2.1.25) and d-xylonate dehydratase (EC 4.2.1.82) are two enzymes that are involved in a nonphosphorylative oxidation pathway of pentose sugars. l-Arabinonate dehydratase converts l-arabinonate into 2-dehydro-3-deoxy-l-arabinonate, and d-xylonate dehydratase catalyzes the dehydration of d-xylonate to 2-dehydro-3-deoxy-d-xylonate. l-Arabinonate and d-xylonate dehydratases belong to the IlvD/EDD family, together with 6-phosphogluconate dehydratases and dihydroxyacid dehydratases. No crystal structure of any l-arabinonate or d-xylonate dehydratase is available in the PDB. In this study, recombinant l-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii (RlArDHT) and d-xylonate dehydratase from Caulobacter crescentus (CcXyDHT) were heterologously expressed in Escherichia coli and purified by the use of affinity chromatography followed by gel-filtration chromatography. The purified proteins were crystallized using the hanging-drop vapour-diffusion method at 293 K. Crystals of RlArDHT that diffracted to 2.40 Å resolution were obtained using sodium formate as a precipitating agent. They belonged to space group P21, with unit-cell parameters a = 106.07, b = 208.61, c = 147.09 Å, β = 90.43°. Eight RlArDHT molecules (two tetramers) in the asymmetric unit give a V M value of 3.2 Å3 Da−1 and a solvent content of 62%. Crystals of CcXyDHT that diffracted to 2.66 Å resolution were obtained using sodium formate and polyethylene glycol 3350. They belonged to space group C2, with unit-cell parameters a = 270.42, b = 236.13, c = 65.17 Å, β = 97.38°. Four CcXyDHT molecules (a tetramer) in the asymmetric unit give a V M value of 4.0 Å3 Da−1 and a solvent content of 69%. PMID:27487924

  15. Bi-modal distribution of the second messenger c-di-GMP controls cell fate and asymmetry during the caulobacter cell cycle.

    Directory of Open Access Journals (Sweden)

    Sören Abel

    Full Text Available Many bacteria mediate important life-style decisions by varying levels of the second messenger c-di-GMP. Behavioral transitions result from the coordination of complex cellular processes such as motility, surface adherence or the production of virulence factors and toxins. While the regulatory mechanisms responsible for these processes have been elucidated in some cases, the global pleiotropic effects of c-di-GMP are poorly understood, primarily because c-di-GMP networks are inherently complex in most bacteria. Moreover, the quantitative relationships between cellular c-di-GMP levels and c-di-GMP dependent phenotypes are largely unknown. Here, we dissect the c-di-GMP network of Caulobacter crescentus to establish a global and quantitative view of c-di-GMP dependent processes in this organism. A genetic approach that gradually reduced the number of diguanylate cyclases identified novel c-di-GMP dependent cellular processes and unraveled c-di-GMP as an essential component of C. crescentus cell polarity and its bimodal life cycle. By varying cellular c-di-GMP concentrations, we determined dose response curves for individual c-di-GMP-dependent processes. Relating these values to c-di-GMP levels modeled for single cells progressing through the cell cycle sets a quantitative frame for the successive activation of c-di-GMP dependent processes during the C. crescentus life cycle. By reconstructing a simplified c-di-GMP network in a strain devoid of c-di-GMP we defined the minimal requirements for the oscillation of c-di-GMP levels during the C. crescentus cell cycle. Finally, we show that although all c-di-GMP dependent cellular processes were qualitatively restored by artificially adjusting c-di-GMP levels with a heterologous diguanylate cyclase, much higher levels of the second messenger are required under these conditions as compared to the contribution of homologous c-di-GMP metabolizing enzymes. These experiments suggest that a common c-di-GMP pool

  16. Crystallization and X-ray diffraction analysis of an l-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii and a d-xylonate dehydratase from Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Rahman, Mohammad Mubinur [University of Eastern Finland, Joensuu Campus, PO Box 111, FIN-80101 Joensuu (Finland); Andberg, Martina; Koivula, Anu [VTT Technical Research Centre of Finland Ltd, PO Box 1000, FIN-02044 VTT Espoo (Finland); Rouvinen, Juha; Hakulinen, Nina, E-mail: nina.hakulinen@uef.fi [University of Eastern Finland, Joensuu Campus, PO Box 111, FIN-80101 Joensuu (Finland)

    2016-07-13

    l-Arabinonate dehydratase and d-xylonate dehydratase from the IlvD/EDD family were crystallized by the vapour-diffusion method. Diffraction data sets were collected to resolutions of 2.40 and 2.66 Å from crystals of l-arabinonate dehydratase and d-xylonate dehydratase, respectively. l-Arabinonate dehydratase (EC 4.2.1.25) and d-xylonate dehydratase (EC 4.2.1.82) are two enzymes that are involved in a nonphosphorylative oxidation pathway of pentose sugars. l-Arabinonate dehydratase converts l-arabinonate into 2-dehydro-3-deoxy-l-arabinonate, and d-xylonate dehydratase catalyzes the dehydration of d-xylonate to 2-dehydro-3-deoxy-d-xylonate. l-Arabinonate and d-xylonate dehydratases belong to the IlvD/EDD family, together with 6-phosphogluconate dehydratases and dihydroxyacid dehydratases. No crystal structure of any l-arabinonate or d-xylonate dehydratase is available in the PDB. In this study, recombinant l-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii (RlArDHT) and d-xylonate dehydratase from Caulobacter crescentus (CcXyDHT) were heterologously expressed in Escherichia coli and purified by the use of affinity chromatography followed by gel-filtration chromatography. The purified proteins were crystallized using the hanging-drop vapour-diffusion method at 293 K. Crystals of RlArDHT that diffracted to 2.40 Å resolution were obtained using sodium formate as a precipitating agent. They belonged to space group P2{sub 1}, with unit-cell parameters a = 106.07, b = 208.61, c = 147.09 Å, β = 90.43°. Eight RlArDHT molecules (two tetramers) in the asymmetric unit give a V{sub M} value of 3.2 Å{sup 3} Da{sup −1} and a solvent content of 62%. Crystals of CcXyDHT that diffracted to 2.66 Å resolution were obtained using sodium formate and polyethylene glycol 3350. They belonged to space group C2, with unit-cell parameters a = 270.42, b = 236.13, c = 65.17 Å, β = 97.38°. Four CcXyDHT molecules (a tetramer) in the asymmetric unit give a V{sub M

  17. Crystallization and X-ray diffraction analysis of an L-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii and a D-xylonate dehydratase from Caulobacter crescentus.

    Science.gov (United States)

    Rahman, Mohammad Mubinur; Andberg, Martina; Koivula, Anu; Rouvinen, Juha; Hakulinen, Nina

    2016-08-01

    L-Arabinonate dehydratase (EC 4.2.1.25) and D-xylonate dehydratase (EC 4.2.1.82) are two enzymes that are involved in a nonphosphorylative oxidation pathway of pentose sugars. L-Arabinonate dehydratase converts L-arabinonate into 2-dehydro-3-deoxy-L-arabinonate, and D-xylonate dehydratase catalyzes the dehydration of D-xylonate to 2-dehydro-3-deoxy-D-xylonate. L-Arabinonate and D-xylonate dehydratases belong to the IlvD/EDD family, together with 6-phosphogluconate dehydratases and dihydroxyacid dehydratases. No crystal structure of any L-arabinonate or D-xylonate dehydratase is available in the PDB. In this study, recombinant L-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii (RlArDHT) and D-xylonate dehydratase from Caulobacter crescentus (CcXyDHT) were heterologously expressed in Escherichia coli and purified by the use of affinity chromatography followed by gel-filtration chromatography. The purified proteins were crystallized using the hanging-drop vapour-diffusion method at 293 K. Crystals of RlArDHT that diffracted to 2.40 Å resolution were obtained using sodium formate as a precipitating agent. They belonged to space group P21, with unit-cell parameters a = 106.07, b = 208.61, c = 147.09 Å, β = 90.43°. Eight RlArDHT molecules (two tetramers) in the asymmetric unit give a VM value of 3.2 Å(3) Da(-1) and a solvent content of 62%. Crystals of CcXyDHT that diffracted to 2.66 Å resolution were obtained using sodium formate and polyethylene glycol 3350. They belonged to space group C2, with unit-cell parameters a = 270.42, b = 236.13, c = 65.17 Å, β = 97.38°. Four CcXyDHT molecules (a tetramer) in the asymmetric unit give a VM value of 4.0 Å(3) Da(-1) and a solvent content of 69%.

  18. Analysis of the xynB5 gene encoding a multifunctional GH3-BglX β-glucosidase-β-xylosidase-α-arabinosidase member in Caulobacter crescentus.

    Science.gov (United States)

    Justo, Priscila Innocenti; Corrêa, Juliana Moço; Maller, Alexandre; Kadowaki, Marina Kimiko; da Conceição-Silva, José Luis; Gandra, Rinaldo Ferreira; Simão, Rita de Cássia Garcia

    2015-10-01

    The Caulobacter crescentus (NA1000) xynB5 gene (CCNA_03149) encodes a predicted β-glucosidase-β-xylosidase enzyme that was amplified by polymerase chain reaction; the product was cloned into the blunt ends of the pJet1.2 plasmid. Analysis of the protein sequence indicated the presence of conserved glycosyl hydrolase 3 (GH3), β-glucosidase-related glycosidase (BglX) and fibronectin type III-like domains. After verifying its identity by DNA sequencing, the xynB5 gene was linked to an amino-terminal His-tag using the pTrcHisA vector. A recombinant protein (95 kDa) was successfully overexpressed from the xynB5 gene in E. coli Top 10 and purified using pre-packed nickel-Sepharose columns. The purified protein (BglX-V-Ara) demonstrated multifunctional activities in the presence of different substrates for β-glucosidase (pNPG: p-nitrophenyl-β-D-glucoside) β-xylosidase (pNPX: p-nitrophenyl-β-D-xyloside) and α-arabinosidase (pNPA: p-nitrophenyl-α-L-arabinosidase). BglX-V-Ara presented an optimal pH of 6 for all substrates and optimal temperature of 50 °C for β-glucosidase and α-L-arabinosidase and 60 °C for β-xylosidase. BglX-V-Ara predominantly presented β-glucosidase activity, with the highest affinity for its substrate and catalytic efficiency (Km 0.24 ± 0.0005 mM, Vmax 0.041 ± 0.002 µmol min(-1) mg(-1) and Kcat/Km 0.27 mM(-1) s(-1)), followed by β-xylosidase (Km 0.64 ± 0.032 mM, Vmax 0.055 ± 0.002 µmol min(-1) mg(-1) and Kcat/Km 0.14 mM(-1)s(-1)) and finally α-L-arabinosidase (Km 1.45 ± 0.05 mM, Vmax 0.091 ± 0.0004 µmol min(-1) mg(-1) and Kcat/Km 0.1 mM(-1) s(-1)). To date, this is the first report to demonstrate the characterization of a GH3-BglX family member in C. crescentus that may have applications in biotechnological processes (i.e., the simultaneous saccharification process) because the multifunctional enzyme could play an important role in bacterial hemicellulose degradation.

  19. Cloning and expression of the xynA1 gene encoding a xylanase of the GH10 group in Caulobacter crescentus.

    Science.gov (United States)

    Graciano, Luciana; Corrêa, Juliana Moço; Vieira, Fabíola Giovanna Nesello; Bosetto, Adilson; Loth, Eduardo Alexandre; Kadowaki, Marina Kimiko; Gandra, Rinaldo Ferreira; Simão, Rita de Cássia Garcia

    2015-04-01

    Caulobacter crescentus (NA1000 strain) are aquatic bacteria that can live in environments of low nutritional quality and present numerous genes that encode enzymes involved in plant cell wall deconstruction, including five genes for β-xylosidases (xynB1-xynB5) and three genes for xylanases (xynA1-xynA3). The overall activity of xylanases in the presence of different agro-industrial residues was evaluated, and it was found that the residues from the processing of corn were the most efficient in inducing bacterial xylanases. The xynA1 gene (CCNA_02894) encoding a predicted xylanase of group 10 of glyco-hydrolases (GH10) that was efficiently overexpressed in Escherichia coli LMG194 using 0.02 % arabinose, after cloning into the vector pJet1.2blunt and subcloning into the expression vector pBAD/gIII, provided a fusion protein that contained carboxy-terminal His-tags, named XynA1. The characterization of pure XynA1 showed an enzymatic activity of 18.26 U mL(-1) and a specific activity of 2.22 U mg-(1) in the presence of xylan from beechwood as a substrate. XynA1 activity was inhibited by EDTA and metal ions such as Cu(2+) and Mg(2+). By contrast, β-mercaptoethanol, dithiothreitol (DTT), and Ca(2+) induced recombinant enzyme activity. Kinetic data for XynA1 revealed K M and V max values of 3.77 mg mL-(1) and 10.20 μM min-(1), respectively. Finally, the enzyme presented an optimum pH of 6 and an optimum temperature of 50 °C. In addition, 80 % of the activity of XynA1 was maintained at 50 °C for 4 h of incubation, suggesting a thermal stability for the biotechnological processes. This work is the first study concerning the cloning, overexpression, and enzymatic characterization of C. crescentus xylanase.

  20. Bathypelagic particle flux signatures from a suboxic eddy in the oligotrophic tropical North Atlantic: production, sedimentation and preservation

    Science.gov (United States)

    Fischer, Gerhard; Karstensen, Johannes; Romero, Oscar; Baumann, Karl-Heinz; Donner, Barbara; Hefter, Jens; Mollenhauer, Gesine; Iversen, Morten; Fiedler, Björn; Monteiro, Ivanice; Körtzinger, Arne

    2016-06-01

    Particle fluxes at the Cape Verde Ocean Observatory (CVOO) in the eastern tropical North Atlantic for the period December 2009 until May 2011 are discussed based on bathypelagic sediment trap time-series data collected at 1290 and 3439 m water depth. The typically oligotrophic particle flux pattern with weak seasonality is modified by the appearance of a highly productive and low oxygen (minimum concentration below 2 µmol kg-1 at 40 m depth) anticyclonic modewater eddy (ACME) in winter 2010. The eddy passage was accompanied by unusually high mass fluxes of up to 151 mg m-2 d-1, lasting from December 2009 to May 2010. Distinct biogenic silica (BSi) and organic carbon flux peaks of ˜ 15 and 13.3 mg m-2 d-1, respectively, were observed in February-March 2010 when the eddy approached the CVOO. The flux of the lithogenic component, mostly mineral dust, was well correlated with that of organic carbon, in particular in the deep trap samples, suggesting a tight coupling. The lithogenic ballasting obviously resulted in high particle settling rates and, thus, a fast transfer of epi-/meso-pelagic signatures to the bathypelagic traps. We suspect that the two- to three-fold increase in particle fluxes with depth as well as the tight coupling of mineral dust and organic carbon in the deep trap samples might be explained by particle focusing processes within the deeper part of the eddy. Molar C : N ratios of organic matter during the ACME passage were around 18 and 25 for the upper and lower trap samples, respectively. This suggests that some productivity under nutrient (nitrate) limitation occurred in the euphotic zone of the eddy in the beginning of 2010 or that a local nitrogen recycling took place. The δ15N record showed a decrease from 5.21 to 3.11 ‰ from January to March 2010, while the organic carbon and nitrogen fluxes increased. The causes of enhanced sedimentation from the eddy in February/March 2010 remain elusive, but nutrient depletion and/or an increased

  1. 寡营养细菌对古尔班通古特沙漠土壤环境的影响%Effect of Oligotrophic Bacteria on Soil Environment in the Gurbantunggut Desert,Xinjiang,China

    Institute of Scientific and Technical Information of China (English)

    邱东; 程争鸣; 张元明; 吴楠; 牟书勇; 齐晓玲; 潘惠霞

    2012-01-01

    利用从古尔班通古特沙漠生物结皮中分离的1株寡营养细菌制成的菌剂,喷洒在流沙表面,分析寡营养细菌对沙漠土壤微生物、酶活性、土壤养分及生物结皮形成的影响。结果表明:①喷洒寡营养细菌菌剂,提高各层土壤主要微生物数量,其中显著(P〈0.05)提高了0~2 cm土层的细菌和放线菌数量及2~5 cm层的寡营养细菌数量;②对不同土壤养分的影响有所差异,提高了土壤中全磷、全钾、有效氮和有效磷含量,但降低了有机质和全氮含量;③对不同土壤酶活性的影响也表现出一定的差异,提高了过氧化氢酶和多酚氧化酶活性,但降低了蔗糖酶和碱性磷酸酶活性。相关性分析表明,寡营养细菌与放线菌为极显著正相关,与其他因子相关性不显著。总体来讲,寡营养细菌的施加改变了土壤原有微生物和养分格局,促进了微生物和藻结皮的存在与发展。研究将为有效恢复生物结皮、修复受损沙地提供新的方法。%The oligotrophic bacteria isolated from the biotic crust in the Gurbantunggut Desert were made into solvent,and two spraying levels were set to compare the effects of oligotrophic bacteria on soil microorganisms,enzyme activities,soil nutrients and formation and regeneration of biotic soil crust.The results are as follows:(1) It could improve the numbers of bacteria,actinomycetes and oligotrophic bacteria at two soil layers(0-2 cm and 2-5 cm in depth) to spray the oligotrophic bacteria,and the numbers of bacteria and actinomycetes at 0-2 cm layer and of oligotrophic bacteria at 2-5 cm layer were more than that without spraying oligotrophic bacteria;(2) The effects of spraying the oligotrophic bacteria on soil nutrients were different,the contents of total P,total K,available N and available P were effectively improved,but the contents of soil organic matter and total N were decreased;(3) There were also the difference

  2. Immobilization of bacterial S-layer proteins from Caulobacter crescentus on iron oxide-based nanocomposite: Synthesis and spectroscopic characterization of zincite-coated Fe2O3 nanoparticles

    Science.gov (United States)

    Habibi, Neda

    Zinc oxide was coated on Fe2O3 nanoparticles using sol-gel spin-coating. Caulobacter crescentus have a crystalline surface layer (S-layer), which consist of one protein or glycoprotein species. The immobilization of bacterial S-layers obtained from C. crescentus on zincite-coated nanoparticles of iron oxide was investigated. The SDS PAGE results of S-layers isolated from C. crescentus showed the weight of 50 KDa. Nanoparticles of the Fe2O3 and zinc oxide were synthesized by a sol-gel technique. Fe2O3 nanoparticles with an average size of 50 nm were successfully prepared by the proper deposition of zinc oxide onto iron oxide nanoparticles surface annealed at 450 °C. The samples were characterized by field-emission scanning electron microscope (FESEM), atomic force microscopy (AFM), powder X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FT-IR).

  3. AnÃlise da expressÃo da β-Xilosidade II da bactÃria aquÃtica Caulobacter crescentus e seu papel no aproveitamento de resÃduos agroindustriais.

    OpenAIRE

    Juliana MoÃo CorrÃa

    2011-01-01

    Materiais lignocelulÃsicos sÃo abundantes em resÃduos agroindustriais e subprodutos da agroindÃstria e podem ser usados para produÃÃo de combustÃveis e outros quÃmicos de interesse comercial. Uma alternativa aos mÃtodos fÃsicos e quÃmicos para bioconversÃo de material lignocelulÃsico à o uso de enzimas produzidas por micro-organismos. A bactÃria aquÃtica Gram negativa Caulobacter crescentus apresenta potencial biotecnolÃgico para o uso destes resÃduos por conter em seu genoma v...

  4. Variability of rRNA Operon Copy Number and Growth Rate Dynamics of Bacillus Isolated from an Extremely Oligotrophic Aquatic Ecosystem.

    Science.gov (United States)

    Valdivia-Anistro, Jorge A; Eguiarte-Fruns, Luis E; Delgado-Sapién, Gabriela; Márquez-Zacarías, Pedro; Gasca-Pineda, Jaime; Learned, Jennifer; Elser, James J; Olmedo-Alvarez, Gabriela; Souza, Valeria

    2015-01-01

    The ribosomal RNA (rrn) operon is a key suite of genes related to the production of protein synthesis machinery and thus to bacterial growth physiology. Experimental evidence has suggested an intrinsic relationship between the number of copies of this operon and environmental resource availability, especially the availability of phosphorus (P), because bacteria that live in oligotrophic ecosystems usually have few rrn operons and a slow growth rate. The Cuatro Ciénegas Basin (CCB) is a complex aquatic ecosystem that contains an unusually high microbial diversity that is able to persist under highly oligotrophic conditions. These environmental conditions impose a variety of strong selective pressures that shape the genome dynamics of their inhabitants. The genus Bacillus is one of the most abundant cultivable bacterial groups in the CCB and usually possesses a relatively large number of rrn operon copies (6-15 copies). The main goal of this study was to analyze the variation in the number of rrn operon copies of Bacillus in the CCB and to assess their growth-related properties as well as their stoichiometric balance (N and P content). We defined 18 phylogenetic groups within the Bacilli clade and documented a range of from six to 14 copies of the rrn operon. The growth dynamic of these Bacilli was heterogeneous and did not show a direct relation to the number of operon copies. Physiologically, our results were not consistent with the Growth Rate Hypothesis, since the copies of the rrn operon were decoupled from growth rate. However, we speculate that the diversity of the growth properties of these Bacilli as well as the low P content of their cells in an ample range of rrn copy number is an adaptive response to oligotrophy of the CCB and could represent an ecological mechanism that allows these taxa to coexist. These findings increase the knowledge of the variability in the number of copies of the rrn operon in the genus Bacillus and give insights about the

  5. Present status and changes of the phytoplankton community after invasion of Neosalanx taihuensis since 1982 in a deep oligotrophic plateau lake, Lake Fuxian in the subtropical China

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xia; XIE Ping; CHEN Fei-zhou; LI Yan-ling; LI Si-xin; GUO Ni-chun; QIN Jian-hui

    2005-01-01

    Phytoplankton assemblages in the subtrophical oligotrophic Lake Fuxian, the second deepest lake in China, were investigated monthly from September 2002 to August 2003. A total of 113 species belonging to seven phyla were identified, among them, a filamentous green alga, Mougeotia sp., dominated almost throughout the study period and comprised most of the total phytoplankton biomass.Mougeotia sp. has made a substantial development during the past decades: it was absent in 1957, only occasionally present in 1983,increased substantially in 1993, and became predominant in 2002-2003. It is likely that natural invasion of the Taihu Lake noodlefish (Neosalanx taihuensis) has led to a change of dominant herbivorous zooplankton from small to large calanoid, which has increased grazing pressure on small edible algae, and thus has indirectly favored the development of the inedible filamentous Mougeotia sp.

  6. The use of the stable isotope, oxygen-18, as a tracer to measure gross primary production in coastal and oligotrophic waters and in monoclonal cultures of marine phytoplankton

    Energy Technology Data Exchange (ETDEWEB)

    Grande, K.D.

    1988-01-01

    A new technique was developed to measure the in vitro rates of gross oxygen production in planktonic communities, in which water is enriched with the stable isotope {sup 18}O, and photosynthetic evolution of {sup 18}O{sup 16}O is measured. In order to calculate gross oxygen production, they correct for the {sup 18}O fractionation due to respiration. The standard technique for measuring production in the oceans, {sup 14}C-bicarbonate uptake, has recently been questioned due to discrepancies with other estimates of water column production and suspected intrinsic problems with the {sup 14}C technique. They compared rates of {sup 18}O gross production and {sup 14}C production in oligotrophic and coastal sites. Samples were generally incubated under natural lighting with neutral density screening. Some oligotrophic bottles were incubated in situ at the depth of collection. Rates of {sup 14}C production were 60-100% of {sup 18}O gross production in both coastal and oceanic communities. Assuming a PQ of 1.0 to 1.5, these comparisons suggest that {sup 14}C production rates are not seriously underestimating actual in vitro rates of production. They also measured rates of light respiration in cultures and natural populations by the {sup 18}O technique. The rates of light respiration in algal cultures were generally greater than dark rates by a factor of two to ten. The increased respiration in the light could be due to increased rates of mitochondrial respiration, photorespiration or Mehler respiration. They measured the enhanced {sup 14}C production under reduced (O{sub 2}) conditions (the Warburg effect), as an estimate of the rate of photorespiration. In three clones, rates of photorespiration were a significant fraction of light respiration. In five other clones examined, photorespiration was not implicated as a source of light respiration.

  7. Shifts in microbial community structure and activity in the ultra-oligotrophic Eastern Mediterranean Sea driven by the deposition of Saharan dust and European aerosols

    Directory of Open Access Journals (Sweden)

    Cui Guo

    2016-09-01

    Full Text Available The atmospheric deposition of gases and particulates from the Sahara Desert and European landmass is an important source of nutrients for the Mediterranean Sea. In this study, we investigated how such atmospheric input might affect bacterial metabolic activities and community dynamics in the ultra-oligotrophic Eastern Mediterranean Sea. Thus a mesocosm simulation experiment was conducted using pure Saharan dust (SD and mixed aerosols (A, polluted and desert origin. The cell specific bacterial production (BP was stimulated soon after the addition of SD and A, with a higher degree of stimulation being observed in the activity of Alphaproteobacteria than in Gammaproteobacteria, and this lead to significant increases in community BP. Subsequently, a shift between these two dominating classes was observed (such that the proportion of Gammaproteobacteria increased while that of Alphaproteobacteria decreased, along with significant increases in bacterial abundance and chlorophyll a concentration. After a few days, although the abundance of bacteria was still significantly higher in the SD- or A-treated groups, differences in the active community composition between the treatment and control groups were reduced. The altered activity of the two dominating Proteobacteria classes observed, might reflect their different strategies in responding to external nutrient input: with Alphaproteobacteria being more responsive to the direct dust input, whereas Gammaproteobacteria seemed to benefit more from the increase in phytoplankton biomass. In addition, the input of A had a stronger immediate effect and longer lasting influence on changing the active bacterial community composition than did that of SD. Our findings show that episodic atmospheric deposition events might affect the microbial community with regards to their abundance, activity and composition over a short period of time, and thus regulate the function of the microbial community and carbon cycling

  8. Limited impact of ocean acidification on phytoplankton community structure and carbon export in an oligotrophic environment: Results from two short-term mesocosm studies in the Mediterranean Sea

    Science.gov (United States)

    Gazeau, F.; Sallon, A.; Pitta, P.; Tsiola, A.; Maugendre, L.; Giani, M.; Celussi, M.; Pedrotti, M. L.; Marro, S.; Guieu, C.

    2017-02-01

    Modifications in the strength of the biological pump as a consequence of ocean acidification, whether positive or negative, have the potential to impact atmospheric CO2 and therefore climate. So far, most plankton community perturbation studies have been performed in nutrient-rich areas although there are some indications that CO2-dependent growth could differ in nutrient-replete vs. -limited regions and with different community compositions. Two in situ mesocosm experiments were performed in the NW Mediterranean Sea during two seasons with contrasted environmental conditions: summer oligotrophic stratified waters in the Bay of Calvi vs. winter mesotrophic well-mixed waters in the Bay of Villefranche. Nine mesocosms were deployed for 20 and 12 d, respectively, and subjected to seven CO2 levels (3 controls, 6 elevated levels). Both phytoplankton assemblages were dominated by pico- and nano-phytoplankton cells. Although haptophyceae and dinoflagellates benefited from short-term CO2 enrichment in summer, their response remained small with no consequences on organic matter export due to strong environmental constraints (nutrient availability). In winter, most of the plankton growth and associated nutrient consumption occurred during the 4-day acidification period (before the experimental phase). During the remaining experimental period, characterized by low nutrient availability, plankton growth was minimal and no clear CO2-dependency was found for any of the tested parameters. While there is a strong confidence on the absence of significant effect of short-term CO2 addition under oligotrophic conditions, more investigations are needed to assess the response of plankton communities in winter when vertical mixing and weather conditions are major factors controlling plankton dynamics.

  9. Digestive tract morphology of the Neotropical piscivorous fish Cichla kelberi(Perciformes: Cichlidae introduced into an oligotrophic Brazilian reservoir

    Directory of Open Access Journals (Sweden)

    Alejandra Filippo Gonzalez Neves dos Santos

    2011-09-01

    Full Text Available Despite being one of the most well-known cichlid fish of importance to artisanal and sport fishing, and among the largest fishes in the Neotropics, data on digestive tract anatomy of peacock basses (Cichla spp. are largely lacking, especially for non-native populations. in this paper, we describe for the first time the digestive tract morphology of Cichla kelberi, a voracious piscivore that was introduced in the 1950s into an oligotrophic and physically low-complex impoundment in Brazil. Peacock basses were collected between 1994 and 2002 in Lajes Reservoir, through gillnets (25 to 55mm mesh; 20-50x2m, seines (10x2.5m; 8.0mm mesh, cast nets and angling. All the fishes were kept on ice in the field and then transferred to the laboratory, where they were identified, measured, weighed and dissected for digestive tract analyses. The index of Relative importance-IRI was calculated for diet characterization while linear and non-linear regressions were performed to assess growth patterns of four morphological characters related to feeding (e.g. mouth width, mouth height, stomach length and intestine length and the number of gill rakers during the C. kelberi ontogeny. Most digestive tract structures were directly related to the piscivorous diet of C. kelberi, indicating that peacock bass is a diurnal, bathypelagic and gape-size limited predator that feeds largely on shallow-water prey species within the littoral zone. Mouth width and height grew allometrically (b>1 with the size of peacock bass, broadening the size range in which prey can be eaten, but especially for predators smaller than ~400mm of total length. Differently, stomach and intestine lengths increased isometrically (b=1, which could constrain prey consumption for adult C. kelberi, especially those at advanced stages of gonadal maturation. The presence of longer-drawn, sharp and furcated gill rakers in C. kelberi may be related to increased prey retention in the resource-limited Lajes

  10. How do Bacteria Adapt to the Red Sea? Cultivation and Genomic and Physiological Characterization of Oligotrophic Bacteria of the PS1, OM43, and SAR11 Clades

    KAUST Repository

    Jimenez Infante, Francy M.

    2015-05-01

    Given the high salinity, prevailing annual high temperatures, and ultra-oligotrophic conditions in the Red Sea isolation and characterization of important microbial groups thriving in this environment is important in understanding the ecological significance and metabolic capabilities of these communities. By using a high-­throughput cultivation technique in natural seawater amended with minute amounts of nutrients, members of the rare biosphere (PS1), methylotrophic Betaproteobacteria (OM43), and the ubiquitous and abundant SAR11 group (Pelagibacterales), were isolated in pure culture. Phylogenetic analyses of Red Sea isolates along with comparative genomics with close representatives from disparate provinces revealed ecotypes and genomic differentiation among the groups. Firstly, the PS1 alphaproteobacterial clade was found to be present in very low abundance in several metagenomic datasets form divergent environments. While strain RS24 (Red Sea) harbored genomic islands involved in polymer degradation, IMCC14465 (East (Japan) Sea) contained unique genes for degradation of aromatic compounds. Secondly, methylotrophic OM43 bacteria from the Red Sea (F5, G12 and H7) showed higher similarities with KB13 isolate from Hawaii, forming a ‘H-­RS’ (Hawaii-­Red Sea) cluster separate from HTCC2181 (Oregon isolate). HTCC2181 members were shown to prevail in cold, productive coastal environments and had an nqrA-­F system for energy generation by sodium motive force. On the contrary, H-­RS cluster members may be better adapted to warm and oligotrophic environments, and seem to generate energy by using a proton-­translocating NADH:Quinone oxidoreductase (complex I; nuoA-­N subunits). Moreover, F5, G12, and H7 had unique proteins related to resistance to UV, temperature and salinity, in addition to a heavy metal ‘resistance island’ as adaptive traits to cope with the environmental conditions in the Red Sea. Finally, description of the Red Sea Pelagibacterales

  11. Spatial variations in microbial community composition in surface seawater from the ultra-oligotrophic center to rim of the South Pacific Gyre.

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    Qi Yin

    Full Text Available Surface seawater in the South Pacific Gyre (SPG is one of the cleanest oceanic environments on earth, and the photosynthetic primary production is extremely low. Despite the ecological significance of the largest aquatic desert on our planet, microbial community composition in the ultra-oligotrophic seawater remain largely unknown. In this study, we collected surface seawater along a southern transect of the SPG during the Integrated Ocean Drilling Program (IODP Expedition 329. Samples from four distinct sites (Sites U1368, U1369, U1370 and U1371 were examined, representing ~5400 kilometers of transect line from the gyre heart to the edge area. Real-time PCR analysis showed 16S rRNA gene abundance in the gyre seawater, ranging from 5.96×10(5 to 2.55×10(6 copies ml(-1 for Bacteria and 1.17×10(3 to 1.90×10(4 copies ml(-1 for Archaea. The results obtained by statistic analyses of 16S rRNA gene clone libraries revealed the community composition in the southern SPG area: diversity richness estimators in the gyre center (Sites U1368 & U1369 are generally lower than those at sites in the gyre edge (Sites U1370 & U1371 and their community structures are clearly distinguishable. Phylogenetic analysis showed the predominance of Proteobacteria (especially Alphaproteobacteria and Cyanobacteria in bacterial 16S rRNA gene clone libraries, whereas phylotypes of Betaproteobacteria were only detected in the central gyre. Archaeal 16S rRNA genes in the clone libraries were predominated by the sequences of Marine Group II within the Euryarchaeota, and the Crenarchaeota sequences were rarely detected, which is consistent with the real-time PCR data (only 9.9 to 22.1 copies ml(-1. We also performed cultivation of heterotrophic microbes onboard, resulting in 18.9% of phylogenetically distinct bacterial isolates at least at the species level. Our results suggest that the distribution and diversity of microbial communities in the SPG surface seawater are closely

  12. Microfungal oasis in an oligotrophic desert: diversity patterns and community structure in three freshwater systems of Cuatro Ciénegas, Mexico

    Directory of Open Access Journals (Sweden)

    Patricia Velez

    2016-06-01

    Full Text Available The Cuatro Ciénegas Basin (CCB comprises several oligotrophic aquatic ecosystems limited by phosphorus. These aquatic systems are dominated by a high prokaryotic diversity, shaped by the stress of low nutrient supplies and interspecific competition. Although fungi constitute a diverse and important component of microbial diversity, the microfungal diversity in the CCB remains to be unveiled. With the aim to explore microfungal diversity and ecological patterns in this area, we present the first investigation analyzing cultivable taxa from sediment and water, as well as lignocellulolytic taxa obtained from incubated submerged plant debris, and wood panels in three contrasting freshwater systems in the CCB: Churince, Becerra and Pozas Rojas. We chose a culture-based approach to analyze sediment and water samples in order to obtain fungal cultures, providing opportunities for a posteriori studies, and the possibility of ex situ preservation of the diversity. We evaluated sequence data from the nuclear ribosomal internal transcribed spacer including the 5.8 rDNA region for 126 isolates, revealing 37 OTUs. These OTUs were phylogenetically affiliated to several genera in the fungal phyla: Zygomycota, Basidiomycota, and Ascomycota. We recorded two OTUs with saline affinity, agreeing with previous findings on the prokaryotic communities with ancestral marine resemblances. All the studied systems showed moderate diversity levels, however discrepancies among the diversity indexes were observed, due to the occurrence of abundant taxa in the samples. Our results indicated that lignocellulolytic microfungal communities are dominated by transient fungal taxa, as resident species were not recorded perhaps as a result of the long-term strong competition with the highly adapted prokaryotic community. Moreover, the obtained microfungal taxa occurred mostly on the resident plant debris, rather than submerged wood panels, perhaps as a result of the high adaptation

  13. Microfungal oasis in an oligotrophic desert: diversity patterns and community structure in three freshwater systems of Cuatro Ciénegas, Mexico.

    Science.gov (United States)

    Velez, Patricia; Gasca-Pineda, Jaime; Rosique-Gil, Edmundo; Eguiarte, Luis E; Espinosa-Asuar, Laura; Souza, Valeria

    2016-01-01

    The Cuatro Ciénegas Basin (CCB) comprises several oligotrophic aquatic ecosystems limited by phosphorus. These aquatic systems are dominated by a high prokaryotic diversity, shaped by the stress of low nutrient supplies and interspecific competition. Although fungi constitute a diverse and important component of microbial diversity, the microfungal diversity in the CCB remains to be unveiled. With the aim to explore microfungal diversity and ecological patterns in this area, we present the first investigation analyzing cultivable taxa from sediment and water, as well as lignocellulolytic taxa obtained from incubated submerged plant debris, and wood panels in three contrasting freshwater systems in the CCB: Churince, Becerra and Pozas Rojas. We chose a culture-based approach to analyze sediment and water samples in order to obtain fungal cultures, providing opportunities for a posteriori studies, and the possibility of ex situ preservation of the diversity. We evaluated sequence data from the nuclear ribosomal internal transcribed spacer including the 5.8 rDNA region for 126 isolates, revealing 37 OTUs. These OTUs were phylogenetically affiliated to several genera in the fungal phyla: Zygomycota, Basidiomycota, and Ascomycota. We recorded two OTUs with saline affinity, agreeing with previous findings on the prokaryotic communities with ancestral marine resemblances. All the studied systems showed moderate diversity levels, however discrepancies among the diversity indexes were observed, due to the occurrence of abundant taxa in the samples. Our results indicated that lignocellulolytic microfungal communities are dominated by transient fungal taxa, as resident species were not recorded perhaps as a result of the long-term strong competition with the highly adapted prokaryotic community. Moreover, the obtained microfungal taxa occurred mostly on the resident plant debris, rather than submerged wood panels, perhaps as a result of the high adaptation to specific

  14. Estimates of vertical turbulent mixing used to determine a vertical gradient in net and gross oxygen production in the oligotrophic South Pacific Gyre

    Science.gov (United States)

    Haskell, W. Z.; Prokopenko, M. G.; Stanley, R. H. R.; Knapp, A. N.

    2016-07-01

    Mixed layer (ML) gross (GOP) and net (NOP) oxygen production rates based on in situ mass balances of triple oxygen isotopes (TOI) and O2/Ar are influenced by vertical transport from below, a term traditionally difficult to constrain. Here we present a new approach to estimate vertical eddy diffusivity (Kz) based on density gradients in the upper thermocline and wind speed-based rates of turbulent shear at the ML depth. As an example, we use this Kz, verified by an independent 7Be-based estimate, in an O2/TOI budget at a site in the oligotrophic South Pacific Gyre. NOP equaled 0.31 ± 0.16 mmol m-2 d-1 in the ML (~55-65 m depth) and 1.2 ± 0.4 mmol m-2 d-1 (80%) beneath the ML, while GOP equaled 74 ± 27 mmol m-2 d-1 (86%) in the ML and 12 ± 4 mmol m-2 d-1 (14%) below, revealing a vertical gradient in production rates unquantifiable without the Kz estimate.

  15. Modeling plankton ecosystem functioning and nitrogen fluxes in the oligotrophic waters of the Beaufort Sea, Arctic Ocean: a focus on light-driven processes

    Directory of Open Access Journals (Sweden)

    V. Le Fouest

    2013-07-01

    Full Text Available The Arctic Ocean (AO undergoes profound changes of its physical and biotic environments due to climate change. In some areas of the Beaufort Sea, the stronger haline stratification observed in summer alters the plankton ecosystem structure, functioning and productivity, promoting oligotrophy. A one-dimension (1-D physical–biological coupled model based on the large multiparametric database of the Malina project in the Beaufort Sea was used (i to infer the plankton ecosystem functioning and related nitrogen fluxes and (ii to assess the model sensitivity to key light-driven processes involved in nutrient recycling and phytoplankton growth. The coupled model suggested that ammonium photochemically produced from photosensitive dissolved organic nitrogen (i.e., photoammonification process was a necessary nitrogen source to achieve the observed levels of microbial biomass and production. Photoammonification directly and indirectly (by stimulating the microbial food web activity contributed to 70% and 18.5% of the 0–10 m and whole water column, respectively, simulated primary production (respectively 66% and 16% for the bacterial production. The model also suggested that variable carbon to chlorophyll ratios were required to simulate the observed herbivorous versus microbial food web competition and realistic nitrogen fluxes in the Beaufort Sea oligotrophic waters. In face of accelerating Arctic warming, more attention should be paid in the future to the mechanistic processes involved in food webs and functional group competition, nutrient recycling and primary production in poorly productive waters of the AO, as they are expected to expand rapidly.

  16. Identification of temporal and small-scale spatial variations of phosphate concentration in the near-shore groundwater of an oligotrophic lake

    Science.gov (United States)

    Pöschke, Franziska; Schlichting, Hendrik; Lewandowski, Jörg

    2016-04-01

    Lake Stechlin is one of the last oligotrophic lakes in the German North-Eastern Lake District. In recent years there was some worry over a small but continuous increase of phosphate concentrations in the open water body. The reasons remain unclear. Since the lake obtains its water only from groundwater and precipitation there is the assumption that the former can be a significant source of phosphate inputs into the lake. In the present study, three different groundwater sampling settings on different scales in time and space were used to investigate the phosphate concentration in the near-shore groundwater. A multi-level sampling grid of twelve samplers and 60 sampling ports was installed to study the temporal small-scale fluctuations of P concentration in the groundwater and the interstitial water. Furthermore, a one-time sampling campaign of shallow near-shore groundwater was conducted every 500 m along the lake shore. Additionally, nests of permanent groundwater wells were sampled monthly for one year to identify concentration patterns in the deeper aquifer. The results indicate a large spatial and small temporal heterogeneity of P concentrations. The range of P concentration is shore village Neuglobsow. Since the groundwater catchment belongs since 1938 to a natural protected area other anthropogenic impacts are quite unlikely. Hence, the main source for phosphate is probably the decomposition of naturally present organic material under anaerobic and warm conditions.

  17. Coupled dynamics of iron and phosphorus in sediments of an oligotrophic coastal basin and the impact of anaerobic oxidation of methane.

    Directory of Open Access Journals (Sweden)

    Caroline P Slomp

    Full Text Available Studies of phosphorus (P dynamics in surface sediments of lakes and coastal seas typically emphasize the role of coupled iron (Fe, sulfur (S and P cycling for sediment P burial and release. Here, we show that anaerobic oxidation of methane (AOM also may impact sediment P cycling in such systems. Using porewater and sediment profiles for sites in an oligotrophic coastal basin (Bothnian Sea, we provide evidence for the formation of Fe-bound P (possibly vivianite; Fe3(PO42(·8H2O below the zone of AOM with sulfate. Here, dissolved Fe(2+ released from oxides is no longer scavenged by sulfide and high concentrations of both dissolved Fe(2+ (>1 mM and PO4 in the porewater allow supersaturation with respect to vivianite to be reached. Besides formation of Fe(II-P, preservation of Fe-oxide bound P likely also contributes to permanent burial of P in Bothnian Sea sediments. Preliminary budget calculations suggest that the burial of Fe-bound P allows these sediments to act as a major sink for P from the adjacent eutrophic Baltic Proper.

  18. 嗜麦芽寡养单胞菌临床感染诱因的调查与分析%Stenotrophomonas oligotrophic incentives Aeromonas Clinical Investigation and Analysis of infection

    Institute of Scientific and Technical Information of China (English)

    王景德

    2012-01-01

      Objective Stenotrophomonas oligotrophic incentive Aeromonas infection.Method 23 patients in our hospital unit cel oligotrophic maltophilia respiratory infection cases were retrospectively analyzed and as a group Ⅰ.Replacement of the 23 cases were compared to group Ⅱ.Results In group Ⅰ higher than the rate of use of third generation cephalosporin group Ⅱ, there are significant differences.Conclusion The excessive use of third generation cephalosporin is Stenotrophomonas oligotrophic positive rate of Aeromonas isolated one reason for the increase.%  目的探讨嗜麦芽寡养单胞菌的感染诱因。方法对我院23例嗜麦芽寡养单胞菌感染的呼吸系统的病例进行回顾性分析并以此作为Ⅰ组。另选23例为Ⅱ组进行对照分析。结果Ⅰ组使用三代头孢的比率高于Ⅱ组,有显著性差异。结论过度使用三代头孢是嗜麦芽寡养单胞菌分离阳性率增加的原因之一。

  19. Multivariate and phylogenetic analyses assessing the response of bacterial mat communities from an ancient oligotrophic aquatic ecosystem to different scenarios of long-term environmental disturbance.

    Science.gov (United States)

    Pajares, Silvia; Souza, Valeria; Eguiarte, Luis E

    2015-01-01

    Understanding the response of bacterial communities to environmental change is extremely important in predicting the effect of biogeochemical modifications in ecosystem functioning. The Cuatro Cienegas Basin is an ancient oasis in the Mexican Chihuahuan desert that hosts a wide diversity of microbial mats and stromatolites that have survived in extremely oligotrophic pools with nearly constant conditions. However, thus far, the response of these unique microbial communities to long-term environmental disturbances remains unexplored. We therefore studied the compositional stability of these bacterial mat communities by using a replicated (3x) mesocosm experiment: a) Control; b) Fluct: fluctuating temperature; c) 40C: increase to 40 ºC; d) UVplus: artificial increase in UV radiation; and f) UVmin: UV radiation protection. In order to observe the changes in biodiversity, we obtained 16S rRNA gene clone libraries from microbial mats at the end of the experiment (eight months) and analyzed them using multivariate and phylogenetic tools. Sequences were assigned to 13 major lineages, among which Cyanobacteria (38.8%) and Alphaproteobacteria (25.5%) were the most abundant. The less extreme treatments (Control and UVmin) had a more similar composition and distribution of the phylogenetic groups with the natural pools than the most extreme treatments (Fluct, 40C, and UVplus), which showed drastic changes in the community composition and structure, indicating a different community response to each environmental disturbance. An increase in bacterial diversity was found in the UVmin treatment, suggesting that protected environments promote the establishment of complex bacterial communities, while stressful environments reduce diversity and increase the dominance of a few Cyanobacterial OTUs (mainly Leptolyngbya sp) through environmental filtering. Mesocosm experiments using complex bacterial communities, along with multivariate and phylogenetic analyses of molecular data, can

  20. Multivariate and phylogenetic analyses assessing the response of bacterial mat communities from an ancient oligotrophic aquatic ecosystem to different scenarios of long-term environmental disturbance.

    Directory of Open Access Journals (Sweden)

    Silvia Pajares

    Full Text Available Understanding the response of bacterial communities to environmental change is extremely important in predicting the effect of biogeochemical modifications in ecosystem functioning. The Cuatro Cienegas Basin is an ancient oasis in the Mexican Chihuahuan desert that hosts a wide diversity of microbial mats and stromatolites that have survived in extremely oligotrophic pools with nearly constant conditions. However, thus far, the response of these unique microbial communities to long-term environmental disturbances remains unexplored. We therefore studied the compositional stability of these bacterial mat communities by using a replicated (3x mesocosm experiment: a Control; b Fluct: fluctuating temperature; c 40C: increase to 40 ºC; d UVplus: artificial increase in UV radiation; and f UVmin: UV radiation protection. In order to observe the changes in biodiversity, we obtained 16S rRNA gene clone libraries from microbial mats at the end of the experiment (eight months and analyzed them using multivariate and phylogenetic tools. Sequences were assigned to 13 major lineages, among which Cyanobacteria (38.8% and Alphaproteobacteria (25.5% were the most abundant. The less extreme treatments (Control and UVmin had a more similar composition and distribution of the phylogenetic groups with the natural pools than the most extreme treatments (Fluct, 40C, and UVplus, which showed drastic changes in the community composition and structure, indicating a different community response to each environmental disturbance. An increase in bacterial diversity was found in the UVmin treatment, suggesting that protected environments promote the establishment of complex bacterial communities, while stressful environments reduce diversity and increase the dominance of a few Cyanobacterial OTUs (mainly Leptolyngbya sp through environmental filtering. Mesocosm experiments using complex bacterial communities, along with multivariate and phylogenetic analyses of molecular

  1. The combined impact of land use change and aquaculture on sediment and water quality in oligotrophic Lake Rupanco (North Patagonia, Chile, 40.8°S).

    Science.gov (United States)

    León-Muñoz, Jorge; Echeverría, Cristian; Marcé, Rafael; Riss, Wolfgang; Sherman, Bradford; Iriarte, Jose Luis

    2013-10-15

    Water and sediment quality in North Patagonia's large, oligotrophic lakes are expected to suffer as native forest continues to be fragmented and degraded by its conversion to cropping and pasture land uses. These changes in land use are expected to increase diffuse nutrient loads to the region's lakes. In addition, these lakes are home to the world's second largest salmon aquaculture industry which provides additional point sources of nutrients within the lakes. We studied the combined influences of land use change and salmon farming on the nutrient concentrations in a North Patagonian lake (Lake Rupanco, 233 km(2) water surface, 163 m average depth) in four sub-watersheds ranging in disturbance from near-pristine forest to 53% converted to cropping and pasture. Nitrogen exports from the tributary sub-watersheds increased from 33 kg TN/km(2)/y to 621 kg TN/km(2)/y as the proportion of crop and pasture land increased. The combined nutrient load from land use change and salmon farming has led to significant differences in the nitrogen concentrations of the lake's water column and sediments in the near-shore zones across the lake. Total nitrogen concentrations in the sediments varied from 37 ± 18 mg/kg in near-pristine sub-watersheds without salmon farming to 6400 ± 698 mg/kg where the sub-watershed was dominated by crop and pasture lands combined with the presence of salmon farming. These results demonstrate the importance of considering the impacts of both salmon farming and land use on water and sediment quality for future environmental planning, management and decision making.

  2. Influence of dissolved organic matter character on mercury incorporation by planktonic organisms: an experimental study using oligotrophic water from Patagonian lakes

    Science.gov (United States)

    Diéguez, Maria C.; Queimaliños, Claudia P.; Guevara, Sergio Ribeiro; Marvin-DiPasquale, Mark C.; Cárdenas, Carolina Soto; Arribére, María A.

    2013-01-01

    Ligands present in dissolved organic matter (DOM) form complexes with inorganic divalent mercury (Hg2+) affecting its bioavailability in pelagic food webs. This investigation addresses the influence of a natural gradient of DOM present in Patagonian lakes on the bioaccumulation of Hg2+ (the prevailing mercury species in the water column of these lakes) by the algae Cryptomonas erosa and the zooplankters Brachionus calyciflorus and Boeckella antiqua. Hg2+ accumulation was studied through laboratory experiments using natural water of four oligotrophic Patagonian lakes amended with197Hg2+. The bioavailability of Hg2+ was affected by the concentration and character of DOM. The entrance of Hg2+ into pelagic food webs occurs mostly through passive and active accumulation. The incorporation of Hg2+ by Cryptomonas, up to 27% of the Hg2+ amended, was found to be rapid and dominated by passive adsorption, and was greatest when low molecular weight compounds with protein-like or small phenolic signatures prevailed in the DOM. Conversely, high molecular weight compounds with a humic or fulvic signature kept Hg2+ in the dissolved phase, resulting in the lowest Hg2+ accumulation in this algae. In Brachionus and Boeckella the direct incorporation of Hg from the aqueous phase was up to 3% of the Hg2+ amended. The dietary incorporation of Hg2+ by Boeckella exceeded the direct absorption of this metal in natural water, and was remarkably similar to the Hg2+ adsorbed in their prey. Overall, DOM concentration and character affected the adsorption of Hg2+ by algae through competitive binding, while the incorporation of Hg2+ into the zooplankton was dominated by trophic or dietary transfer.

  3. Effects of low pH and raised temperature on egg production, hatching and metabolic rates of a Mediterranean copepod species (Acartia clausi under oligotrophic conditions

    Directory of Open Access Journals (Sweden)

    S. ZERVOUDAKI

    2013-09-01

    Full Text Available This study includes the first information on the combined effect of low pH and raised temperature on egg production rate (EP, hatching success (HS, excretion and respiration of the Mediterranean copepod Acartia clausi. Adult individuals of A. clausi and fresh surface seawater were collected at a coastal station in Saronikos Gulf during April 2012. Four different conditions were applied: two different pH levels (present: 8.09 and future: 7.83 at two temperature values (present: 16°C and present+4 °C= 20°C. EP and HS success decreased significantly over the duration of exposure at future pH at both temperature conditions. However, the analysis of the combined effect of pH, T, chlorophyll α and the duration of the experiments on EP and HS revealed that ocean acidification had no discernible effect, whereas warming; food and the duration of exposure were more significant for the reproductive output of A. clausi. Temperature appeared to have a positive effect on respiration and excretion. Acidification had no clear effect on respiration, but a negative effect on the A. clausi excretion was observed. Acidification and warming resulted in the increase of the excretion rate and the increase was higher than that observed by warming only. Our findings showed that a direct effect of ocean acidification on copepod’s vital rates was not obvious, except maybe in the case of excretion. Therefore, the combination of acidification with the ambient oligotrophic conditions and the warming could result in species being less able to allocate resources for coping with multiple stressors.

  4. Seasonal and vertical changes of molecular size distribution of lead-210 and polonium-210 in an oligotrophic lake, lake Towada in Japan

    Energy Technology Data Exchange (ETDEWEB)

    Ohtsuka, Y.; Takaku, Y.; Kakiuchi, H.; Hisamatsu, S.; Inaba, J.; Yamamoto, M. [Institute for Environmental Sciences (IES), Dept. of radioecology, okkasho-mura, Aomori Prefecture (Japan)

    2004-07-01

    Lead-210 and {sup 210}Po are naturally occurring long-lived radionuclides in the {sup 238}U decay series. These nuclides are easily measurable, and have been used as tracers of non-conservative material cycling in aquatic systems because of reactivity with particles in water. To deepen our understandings of the behaviors and cycling of these nuclides in lake systems, it is important to know the existing forms (speciation) of these nuclides in the water and seasonal variations of the speciation, including their material balances. We report here a detailed study of molecular size distributions and their seasonal variations for {sup 210}Pb and {sup 210}Po in the water column of an oligo-trophic lake, Lake Towada in the northern area of Japan, Aomori Prefecture. The samples were separated to four fractions; particulate phase (>0.45 {mu}m), 0.45 {mu}m - 100 kDa, 100 10 kDa and <10 kDa. Dissolved phases (<0.45 {mu}m) were separated to the three fractions by using a tangential flow ultra-filtering system. The concentrations of particulate {sup 210}Pb and {sup 210}Po in the epi-limnion over thermocline (20 - 25 m) were higher than those in the hypo-limnion. Fluctuations of total concentrations of both nuclides in water could attribute to concentration variation of them in particulate phase. The pattern of seasonal variation of particulate {sup 210}Pb were reverse to that of the <10 kDa fraction in both epi-limnion and hypo-limnion. However, the variation pattern of <10 kDa fraction of the {sup 210}Po was similar to that of the particulate phase. (author)

  5. Complexity of diatom response to Lateglacial and Holocene climate and environmental change in ancient, deep, and oligotrophic Lake Ohrid (Macedonia/Albania

    Directory of Open Access Journals (Sweden)

    X. S. Zhang

    2015-09-01

    Full Text Available Lake Ohrid (Macedonia/Albania is a rare example of a deep, ancient Mediterranean lake and is a key site for palaeoclimate research in the northeastern Mediterranean region. This study conducts the first high-resolution diatom analysis during the Lateglacial and Holocene in Lake Ohrid. It demonstrates a complex diatom response to temperature change, with a direct response to temperature-induced productivity and an indirect response to temperature-related stratification/mixing regime and epilimnetic nutrient availability. During the Lateglacial (ca. 12 300–11 800 cal yr BP, the low-diversity dominance of hypolimnetic Cyclotella fottii indicates low temperature-dependent lake productivity. During the earliest Holocene (ca. 11 800–10 600 cal yr BP, although the slight increase in small, epilimnetic C. minuscula suggests climate warming and enhanced thermal stratification, diatom concentration remains very low as during the Lateglacial, indicating that temperature increase was muted. The early Holocene (ca. 10 600–8200 cal yr BP marked a sustained increase in epilimnetic taxa, with mesotrophic C. ocellata indicating high temperature-induced lake productivity between ca. 10 600–10 200 cal yr BP and between ca. 9500–8200 cal yr BP, and with C. minuscula in response to low nutrient availability in the epilimnion between ca. 10 200–9500 cal yr BP. During the mid Holocene (ca. 8200–2600 cal yr BP, when sedimentological and geochemical proxies provide evidence for high temperature, anomalously low C. ocellata abundance is probably a response to epilimnetic nutrient limitation, almost mimicking the Lateglacial flora apart from mesotrophic Stephanodiscus transylvanicus indicative of high temperature-induced productivity in the hypolimnion. During the late Holocene (ca. 2600–0 cal yr BP, high abundance and fluctuating composition of epilimnetic taxa is largely a response to enhanced anthropogenic nutrient input. In this deep, oligotrophic lake

  6. Short-term dynamics of nutrients influenced by upwelling in a small oligotrophic coastal ecosystem,Gan Bay,in the northwest Philippines

    Institute of Scientific and Technical Information of China (English)

    Yanping Xu; Minhan Dai; Weidong Zhai; Dongxing Yuan; Jinwen Liu; Zhenyu Sun; Hua Lin; Jingyu Wi

    2009-01-01

    We present a time-series analysis of nutrient and pCO2 (partial pressure of CO2) levels in an oligotrophic coastal ecosystem (Gan Bay),which was likely to be influenced by upwelled subsurface water.Gan Bay is off Currimao Harbor,in the northwest Philippines and is located at the boundary of the South China Sea (SCS).This 42-h time-series observation was conducted in December 2006.In addition to continuous observations of dissolved oxygen (DO) and pCO2,discrete samples were collected at a depth of 5 m every 3 h for measurements of nutrients,including soluble reactive phosphorus (SRP) and inorganic nitrogen (NO3- + NO2-) in order to examine their dynamics and possible physical and biological controls.We observed remarkably large short-term variations in the surface water,spanning a 10-fold change for SRP (32-330 nM) and from <0.3 μM to 4.3 μM for NO3- + NO2-.DO also varied substantially from a lower end of 171 to 205 μM O2.Surface water pCO2 changed from an equilibrium stage with the atmosphere (~386 μatm) to a stage where it was a significant source for the atmospheric CO2 (seawater pCO2~469 μatm).We found that the variation of nutrients was driven neither by tidal mixing nor by biological activities,as was suggested by the variations in the total bacterial abundance and chlorophyll a.Instead,our inverse T-S relationship suggested a two end-member mixing process during the observation period.The N:P ratio throughout the observation period was ~13.2,which is characteristic of SCS subsurface and deep waters.Moreover,pCO2 was correlated inversely with the sea surface temperature.It is likely,therefore,that an upwelled subsurface cold water with high nutrients,low-temperature and high-pCO2 existed.It should be noted that this upwelled cold water did not appear to impact the entire observation period (approximately 35 h of 42 h),which might suggest an extremely dynamic nature for this upwelled cold water mass.(C) 2009 National Natural Science Foundation of

  7. A Lab-On-Chip Phosphate Analyzer for Long-term In Situ Monitoring at Fixed Observatories: Optimization and Performance Evaluation in Estuarine and Oligotrophic Coastal Waters

    Directory of Open Access Journals (Sweden)

    Maxime M. Grand

    2017-08-01

    Full Text Available The development of phosphate sensors suitable for long-term in situ deployments in natural waters, is essential to improve our understanding of the distribution, fluxes, and biogeochemical role of this key nutrient in a changing ocean. Here, we describe the optimization of the molybdenum blue method for in situ work using a lab-on-chip (LOC analyzer and evaluate its performance in the laboratory and at two contrasting field sites. The in situ performance of the LOC sensor is evaluated using hourly time-series data from a 56-day trial in Southampton Water (UK, as well as a month-long deployment in the subtropical oligotrophic waters of Kaneohe Bay (Hawaii, USA. In Kaneohe Bay, where phosphate concentrations were characteristic of the dry season (0.13 ± 0.03 μM, n = 704, the in situ sensor accuracy was 16 ± 12% and a potential diurnal cycle in phosphate concentrations was observed. In Southampton Water, the sensor data (1.02 ± 0.40 μM, n = 1,267 were accurate to ±0.10 μM relative to discrete reference samples. Hourly in situ monitoring revealed striking tidal and storm derived fluctuations in phosphate concentrations in Southampton Water that would not have been captured via discrete sampling. We show the impact of storms on phosphate concentrations in Southampton Water is modulated by the spring-neap tidal cycle and that the 10-fold decline in phosphate concentrations observed during the later stages of the deployment was consistent with the timing of a spring phytoplankton bloom in the English Channel. Under controlled laboratory conditions in a 250 L tank, the sensor demonstrated an accuracy and precision better than 10% irrespective of the salinity (0–30, turbidity (0–100 NTU, colored dissolved organic matter (CDOM concentration (0–10 mg/L, and temperature (5–20°C of the water (0.3–13 μM phosphate being analyzed. This work demonstrates that the LOC technology is mature enough to quantify the influence of stochastic events on

  8. Contribution and pathways of diazotroph-derived nitrogen to zooplankton during the VAHINE mesocosm experiment in the oligotrophic New Caledonia lagoon

    Science.gov (United States)

    Hunt, Brian P. V.; Bonnet, Sophie; Berthelot, Hugo; Conroy, Brandon J.; Foster, Rachel A.; Pagano, Marc

    2016-05-01

    In oligotrophic tropical and subtropical oceans, where strong stratification can limit the replenishment of surface nitrate, dinitrogen (N2) fixation by diazotrophs can represent a significant source of nitrogen (N) for primary production. The VAHINE (VAriability of vertical and tropHIc transfer of fixed N2 in the south-wEst Pacific) experiment was designed to examine the fate of diazotroph-derived nitrogen (DDN) in such ecosystems. In austral summer 2013, three large ( ˜ 50 m3) in situ mesocosms were deployed for 23 days in the New Caledonia lagoon, an ecosystem that typifies the low-nutrient, low-chlorophyll environment, to stimulate diazotroph production. The zooplankton component of the study aimed to measure the incorporation of DDN into zooplankton biomass, and assess the role of direct diazotroph grazing by zooplankton as a DDN uptake pathway. Inside the mesocosms, the diatom-diazotroph association (DDA) het-1 predominated during days 5-15 while the unicellular diazotrophic cyanobacteria UCYN-C predominated during days 15-23. A Trichodesmium bloom was observed in the lagoon (outside the mesocosms) towards the end of the experiment. The zooplankton community was dominated by copepods (63 % of total abundance) for the duration of the experiment. Using two-source N isotope mixing models we estimated a mean ˜ 28 % contribution of DDN to zooplankton nitrogen biomass at the start of the experiment, indicating that the natural summer peak of N2 fixation in the lagoon was already contributing significantly to the zooplankton. Stimulation of N2 fixation in the mesocosms corresponded with a generally low-level enhancement of DDN contribution to zooplankton nitrogen biomass, but with a peak of ˜ 73 % in mesocosm 1 following the UCYN-C bloom. qPCR analysis targeting four of the common diazotroph groups present in the mesocosms (Trichodesmium, het-1, het-2, UCYN-C) demonstrated that all four were ingested by copepod grazers, and that their abundance in copepod

  9. Stability of toxin gene proportion in red-pigmented populations of the cyanobacterium Planktothrix during 29 years of re-oligotrophication of Lake Zürich

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    Ostermaier Veronika

    2012-12-01

    Full Text Available Abstract Background Harmful algal blooms deteriorate the services of aquatic ecosystems. They are often formed by cyanobacteria composed of genotypes able to produce a certain toxin, for example, the hepatotoxin microcystin (MC, but also of nontoxic genotypes that either carry mutations in the genes encoding toxin synthesis or that lost those genes during evolution. In general, cyanobacterial blooms are favored by eutrophication. Very little is known about the stability of the toxic/nontoxic genotype composition during trophic change. Results Archived samples of preserved phytoplankton on filters from aquatic ecosystems that underwent changes in the trophic state provide a so far unrealized possibility to analyze the response of toxic/nontoxic genotype composition to the environment. During a period of 29 years of re-oligotrophication of the deep, physically stratified Lake Zürich (1980 to 2008, the population of the stratifying cyanobacterium Planktothrix was at a minimum during the most eutrophic years (1980 to 1984, but increased and dominated the phytoplankton during the past two decades. Quantitative polymerase chain reaction revealed that during the whole observation period the proportion of the toxic genotype was strikingly stable, that is, close to 100%. Inactive MC genotypes carrying mutations within the MC synthesis genes never became abundant. Unexpectedly, a nontoxic genotype, which lost its MC genes during evolution, and which could be shown to be dominant under eutrophic conditions in shallow polymictic lakes, also co-occurred in Lake Zürich but was never abundant. As it is most likely that this nontoxic genotype contains relatively weak gas vesicles unable to withstand the high water pressure in deep lakes, it is concluded that regular deep mixing selectively reduced its abundance through the destruction of gas vesicles. Conclusions The stability in toxic genotype dominance gives evidence for the adaptation to deep mixing of a

  10. Characterization of the bio-optical anomaly and diurnal variability of the particulate matter, as seen from the scattering and backscattering coefficients, in ultra-oligotrophic eddies of the Mediterranean Sea

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    H. Loisel

    2011-08-01

    Full Text Available The variability of the inherent optical properties is investigated in the ultra-oligotrophic waters of the Mediterranean Sea sampled during the BOUM experiment performed during the early summer 2008. Bio-optical relationships found for the ultra-oligotrophic waters of the three anticyclonic gyres sampled significantly depart from the mean standard relationships provided for the global ocean, confirming the particular character of these Mediterranean waters. These optical anomalies are diversely related to the specific biological and environmental conditions occurring in the studied ecosystem. Specifically, the surface specific phytoplankton absorption coefficient exhibits values lower than those expected from the general relationships mainly in relation with a high contribution of relatively large sized phytoplankton. Conversely, the particulate backscattering coefficient, bbp, values are much higher than the mean standard values for a given chlorophyll-a concentration, TChl-a. This feature can presumably be related to the relevant influence of highly refractive submicrometer particles of Saharan origin in the surface layer of the water column. The present measurements also show that the Mediterranean Sea is greener than TChl-a alone indicates, as already stressed in previous studies. This color anomaly is partly explained by the estimated colored dissolved organic matter and submicrometer particles absorption coefficients, and to a greater extent by the high bbp/TChl-a values assuming that these particles backscatter light similarly in the green and blue parts of the visible spectrum. The diel variation of both the particulate matter attenuation and backscattering coefficients were also investigated specifically. Despite some differences in the timing and the magnitude of the daily oscillations found for these optical parameters, potential for the backscattering coefficient daily

  11. Water Quality and Hydrology of Whitefish (Bardon) Lake, Douglas County, Wisconsin, With Special Emphasis on Responses of an Oligotrophic Seepage Lake to Changes in Phosphorus Loading and Water Level

    Science.gov (United States)

    Robertson, Dale M.; Rose, William J.; Juckem, Paul F.

    2009-01-01

    Whitefish Lake, which is officially named Bardon Lake, is an oligotrophic, soft-water seepage lake in northwestern Wisconsin, and classified by the Wisconsin Department of Natural Resources as an Outstanding Resource Water. Ongoing monitoring of the lake demonstrated that its water quality began to degrade (increased phosphorus and chlorophyll a concentrations) around 2002 following a period of high water level. To provide a better understanding of what caused the degradation in water quality, and provide information to better understand the lake and protect it from future degradation, the U.S. Geological Survey did a detailed study from 2004 to 2008. The goals of the study were to describe the past and present water quality of the lake, quantify water and phosphorus budgets for the lake, simulate the potential effects of changes in phosphorus inputs on the lake's water quality, analyze changes in the water level in the lake since 1900, and relate the importance of changes in climate and changes in anthropogenic (human-induced) factors in the watershed to the water quality of the lake. Since 1998, total phosphorus concentrations increased from near the 0.005-milligrams per liter (mg/L) detection limit to about 0.010 mg/L in 2006, and then decreased slightly in 2007-08. During this time, chlorophyll a concentrations and Secchi depths remained relatively stable at about 1.5 micrograms per liter (ug/L) and 26 feet, respectively. Whitefish Lake is typically classified as oligotrophic. Because the productivity in Whitefish Lake is limited by phosphorus, phosphorus budgets were constructed for the lake. Because it was believed that much of its phosphorus comes from the atmosphere, phosphorus deposition was measured in this study. Phosphorus input from the atmosphere was greater than computed based on previously reported wetfall phosphorus concentrations. The concentrations and deposition rates can be used to estimate atmospheric loading in future lake studies. The

  12. Application of a Three-Dimensional Water Quality Model as a Decision Support Tool for the Management of Land-Use Changes in the Catchment of an Oligotrophic Lake

    Science.gov (United States)

    Trolle, Dennis; Spigel, Bob; Hamilton, David P.; Norton, Ned; Sutherland, Donna; Plew, David; Allan, Mathew G.

    2014-09-01

    While expansion of agricultural land area and intensification of agricultural practices through irrigation and fertilizer use can bring many benefits to communities, intensifying land use also causes more contaminants, such as nutrients and pesticides, to enter rivers, lakes, and groundwater. For lakes such as Benmore in the Waitaki catchment, South Island, New Zealand, an area which is currently undergoing agricultural intensification, this could potentially lead to marked degradation of water clarity as well as effects on ecological, recreational, commercial, and tourism values. We undertook a modeling study to demonstrate science-based options for consideration of agricultural intensification in the catchment of Lake Benmore. Based on model simulations of a range of potential future nutrient loadings, it is clear that different areas within Lake Benmore may respond differently to increased nutrient loadings. A western arm (Ahuriri) could be most severely affected by land-use changes and associated increases in nutrient loadings. Lake-wide annual averages of an eutrophication indicator, the trophic level index (TLI) were derived from simulated chlorophyll a, total nitrogen, and total phosphorus concentrations. Results suggest that the lake will shift from oligotrophic (TLI = 2-3) to eutrophic (TLI = 4-5) as external loadings are increased eightfold over current baseline loads, corresponding to the potential land-use intensification in the catchment. This study provides a basis for use of model results in a decision-making process by outlining the environmental consequences of a series of land-use management options, and quantifying nutrient load limits needed to achieve defined trophic state objectives.

  13. Ecomicrobiology and microbial assimilative capacity of the oligotrophic Andean Lake Laja, Chile Ecomicrobiología y capacidad asimilativa microbiana del lago oligotrófico andino Laja, Chile

    Directory of Open Access Journals (Sweden)

    BERNHARD KARRASCH

    2011-09-01

    Full Text Available A strong socio-economic development pressure in South Chile will more and more cause an impact to the present lakes and rivers. Nevertheless, our knowledge concerning the ecological structure and the microbial self-purification capabilities of these lacustrine water bodies is scant but essential for a future sustainable development of land and water use. We studied Lake Laja, a lake already heavily impacted by water diversions for hydropower generation and irrigation. Typical for the Andean region Lake Laja is an oligotrophic water body, limited by nitrogen nutrients. Only very low chlorophyll a and particulate organic carbon (POC concentrations and a small abundance and biomass of bacteria (mainly ultramicrobacteria and heterotrophic flagellates were encountered. Weak trophic interrelations were derived from a high bacteria-to-heterotrophic flagellate ratio. For the ten investigated extracellular enzymes (alkaline phosphatase, leucine, arginine-, glycine and tyrosine-aminopeptidase, α-, β-D-glucosidase, α-, β-D-galactosidase, N-acetyl-β-D-glucosaminidase, with the exception of α-D-glucosidase, a broad range of organic matter degradation activities was proven. Probably, due to the N-limitation, organic nitrogen hydrolysing extracellular enzymes reached activities on average of 45 % compared to other studies in oligotrophic waters. The possible effect of N-limitation on extracellular enzyme activities was more pronounced by cell specific extracellular enzymatic activity rates, which exceeded those of other oligotrophic water bodies on average by factor 2. The overall activities of all microbial extracellular enzymes studied proved to be dominated by the dissolved free external enzymes (up to 98 % over the ectoenzymes which are associated with particulate organic matter (bacterial cell walls, particles, and aggregates. It is concluded that future socio-economic changes, dealing with watershed human intervention (accelerating inorganic and

  14. Diatomáceas epilíticas de águas oligotróficas e ácidas do Sudeste do Brasil Epilithic diatoms from acidic and oligotrophic waters in Southeast Brazil

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    Luís Gustavo de Castro Canani

    2011-03-01

    Full Text Available Poucos estudos tem abordado a flora de diatomáceas de sistemas oligotróficos do Brasil. São apresentados os resultados da composição taxonômica e distribuição espacial e temporal de diatomáceas no rio do Salto, situado no Parque Estadual do Ibitipoca, Minas Gerais (21º40'-21º44'S e 43º52'-43º55'W. As águas desse rio possuem pH entre 4,2 e 5,5 e concentração de ortofosfato entre 1,37 e 10,98 µg.L-1. O estudo baseia-se na análise de 80 amostras coletadas em oito estações no leito rochoso do rio, em janeiro e julho de 2004, períodos de chuva e estiagem, respectivamente. Um total de 15 espécies e duas variedades taxonômicas, distribuídas em oito gêneros e oito famílias foram descritas e ilustradas, com destaque para o gênero Eunotia Ehrenberg. O estudo sobre diatomáceas bentônicas em ambientes lóticos é pioneiro no estado de Minas Gerais, sendo registradas 14 novas citações, das quais Fragilariforma stevensonii Metzeltin & Lange-Bertalot, Eunotia pirarucu Metzeltin & Lange Betalot, Nupela giluwensis Vyverman & Compère e Sellaphora blackfordensis Mann & Droop são primeiros registros para o Brasil.Few studies have been done on the diatom flora of acidic and oligotrophic environments from Brazil. We present the taxonomic composition and distribution of diatoms of Salto River, situated at Ibitipoca State Park, Minas Gerais (21º40'-21º44'S; 43º52'-43º55'W. The pH values ranged from 4.2 to 5.5 and orthophosphate was between 1.37 and 10.98 µg.L-1. The study is based on the analysis of 80 samples collected at eight stations in the rocky stream bed, in January and July 2004, rainy and dry seasons, respectively. A total of 15 species and two taxonomic varieties distributed in eight genera and eight families were described and illustrated, notably the Eunotia Ehrenberg genus. The study of benthic diatoms in lotic systems is pioneer in Minas Gerais state; 14 new records were recorded of which Fragilariforma stevensonii

  15. MBR和CAS工艺污泥在贫营养培养条件下的微生物群落结构研究%Microbial community structure of sludge in MBR and CAS under oligotrophic environment

    Institute of Scientific and Technical Information of China (English)

    胡怡杉; 孙宝盛; 王盛勇

    2011-01-01

    采用聚合酶链式-变性梯度凝胶电泳(PCR-DGGE)技术,研究了膜生物反应器(MBR)和传统活性污泥工艺(CAS)反应器中微生物在贫营养条件下的总细菌群落结构.结果表明,在培养过程中,污泥的微生物种群经历了一个比较明显的变化过程,且以CAS污泥微生物种群的变化更为明显,演替过程中既有原始优势种群的消亡,又有新的优势种群的增强,故优势种群的功能地位处于动态变化中.两种污泥的来源、工艺进水水质和运行条件的不同造成了活性污泥中菌群结构的差异.两种污泥中主要存在的优势菌种大部分为未经培养菌种(uncultured%Total microbial community structure in membrane bio-reactor(MBR) and conventional activated sludge(CAS) processes operated under oligotrophic environment was investigated using PCR-denaturing gradient gel electrophoresis(PCR-DGGE).Investigation results show that microbial population of the two types of sludge had significant changes during cultivation process,especially for the sludge in CAS process.In the evolvement of microbial population,the original dominant species was replaced by new species gradually and the original dominant species was even died out from the system.The state of the dominant populations was dynamically changing.Different sludge sources,influent water quality and operating conditions of the two systems lead to the differences of microbial community structure in activated sludge.The main dominant bacteria in the two types of sludge are uncultured bacterium,e.g.Arcobacter,Pseudomonas,Bacillus and gamma proteobacterium bacteria.Moreover,there also exist some pathogenic microorganisms,such as Arcobacter,Serratia and Klebsiella species.

  16. Ocean fertilization and its potential for fishery development in the oligotrophic South China Sea%海洋施肥及其在南海外海渔业开发方面的应用前景

    Institute of Scientific and Technical Information of China (English)

    邱永松

    2013-01-01

    well as disputes over its application were also reviewed. Examination of the debate over a low fraction of new primary production sinking as particulate organic carbon leads to the suggestion that the most flows through the food web and ocean fertilization may have a greater poten-tial for increasing sustainable fish catch. The potential of macronutrient nourishment for fishery development in the oligotrophic South China Sea was further explored. In the central South China Sea basin, there is abundant heat and sunlight in the euphotic zone, but the phytoplankton primary productivity is very low. The biological productivity is strongly limited by available macronutrients because of permanent stratification. It was suggested that intentional introduction of macronutrient elements could help to increase new primary production and hence sustainable fish catch. The enriched patch with a bloom of dietary plankton may serve to attract and concentrate fish schools. This would increase fishing efficiency for the dispersing oceanic species. Fish stocks in the coastal and shelf waters of the northern South China Sea are in a status of depletion and overfishing, respectively. Ocean fertilization in the oligotrophic South China Sea represents a potential opportunity to relieve heavy fishing pres-sure on the coastal and shelf stocks. Assuming continuation of the current policy of strict limits on the number of fishing licenses and total fishing horsepower, developing a fishery in the open sea with ocean fertilization would lead to partial relocation of fishing effort away from the heavily fished waters. In addition to the enhanced fish production in the open sea, this would correspond to a reduction in fishing pressure and a recovery growth in fish harvest in the coastal and shelf waters.

  17. Oligotrophic patterns in southern Chilean lakes: the relevance of nutrients and mixing depth Patrones oligotróficos en lagos del sur de Chile: relevancia de los nutrientes y de la profundidad de mezcla

    Directory of Open Access Journals (Sweden)

    DORIS SOTO

    2002-06-01

    Full Text Available Southern Chilean monomictic, temperate lakes are generally oligotrophic with high water transparency, low productivity and some apparent resistance to disturbances such as phosphorus additions. This paper attempts to explain low chlorophyll-a values using descriptive and experimental approaches. Three different scales are used, (a a micro scale both in space and time, with experimental manipulations of N, P and total light in 24 enclosures in lake Llanquihue, (b a longer time scale analysing 18 months of data in several sites within lake Llanquihue grouped as salmon farming sites, town bays and control sites, and (c a broader time scale (9 years by monitoring lakes Puyehue, Rupanco and Llanquihue, all in the Araucanian lake region and Yelcho lake in the north Patagonian region. In the Llanquihue in-lake sampling, total phosphorus values varied between 1 and 12 mug L-1 between sites, showing marginal site effects, P = 0.09 (salmon sites had greater values, however, chlorophyll a (Chl-a concentrations were generally low (Los lagos monomicticos templados del sur de Chile son generalmente oligotróficos de gran transparencia, baja productividad y aparente resistencia a perturbaciones tales como adiciones de fósforo. Este trabajo intenta explicar los bajos valores de clorofila de acuerdo a aproximaciones descriptivas y experimentales. Se utilizan aquí tres escalas de aproximación; (a una microescala en espacio y tiempo con manipulaciones experimentales de nitrógeno (N, fósforo (P y luz en 24 limnocorrales ubicados en una bahía del lago Llanquihue, (b una escala mayor analizando 18 meses de muestreo en 10 sitios del lago Llanquihue agrupados como sitios de cultivo de salmones, bahías de ciudades y sitios de control, (c una escala aún mayor en tiempo y espacio monitoreando semestralmente por 9 años los lagos Puyehue, Rupanco, Llanquihue en la región de los Lagos Araucanos y el lago Yelcho en la región nor- patagónica. En el muestreo intra

  18. Analyses of mitogenome sequences revealed that Asian citrus psyllid (Diaphorina citri) from California was related to those from Florida but different from those in Southern China

    Science.gov (United States)

    Asian citrus psyllid (ACP, Diaphorina citri Kuwayama; Hemiptera: Liviidae) transmits “Candidatus Liberibacter asiaticus” (CLas), an unculturable alpha-proteobacterium associated with citrus Huanglongbing (HLB, yellow shoot disease, also called citrus greening disease). HLB is threatening citrus prod...

  19. New criteria for selecting the origin of DNA replication in Wolbachia and closely related bacteria

    Directory of Open Access Journals (Sweden)

    Baldo Laura

    2007-06-01

    Full Text Available Abstract Background The annotated genomes of two closely related strains of the intracellular bacterium Wolbachia pipientis have been reported without the identifications of the putative origin of replication (ori. Identifying the ori of these bacteria and related alpha-Proteobacteria as well as their patterns of sequence evolution will aid studies of cell replication and cell density, as well as the potential genetic manipulation of these widespread intracellular bacteria. Results Using features that have been previously experimentally verified in the alpha-Proteobacterium Caulobacter crescentus, the origin of DNA replication (ori regions were identified in silico for Wolbachia strains and eleven other related bacteria belonging to Ehrlichia, Anaplasma, and Rickettsia genera. These features include DnaA-, CtrA- and IHF-binding sites as well as the flanking genes in C. crescentus. The Wolbachia ori boundary genes were found to be hemE and COG1253 protein (CBS domain protein. Comparisons of the putative ori region among related Wolbachia strains showed higher conservation of bases within binding sites. Conclusion The sequences of the ori regions described here are only similar among closely related bacteria while fundamental characteristics like presence of DnaA and IHF binding sites as well as the boundary genes are more widely conserved. The relative paucity of CtrA binding sites in the ori regions, as well as the absence of key enzymes associated with DNA replication in the respective genomes, suggest that several of these obligate intracellular bacteria may have altered replication mechanisms. Based on these analyses, criteria are set forth for identifying the ori region in genome sequencing projects.

  20. Quantification of ploidy in proteobacteria revealed the existence of monoploid, (mero-oligoploid and polyploid species.

    Directory of Open Access Journals (Sweden)

    Vito Pecoraro

    Full Text Available Bacteria are generally assumed to be monoploid (haploid. This assumption is mainly based on generalization of the results obtained with the most intensely studied model bacterium, Escherichia coli (a gamma-proteobacterium, which is monoploid during very slow growth. However, several species of proteobacteria are oligo- or polyploid, respectively. To get a better overview of the distribution of ploidy levels, genome copy numbers were quantified in four species of three different groups of proteobacteria. A recently developed Real Time PCR approach, which had been used to determine the ploidy levels of halophilic archaea, was optimized for the quantification of genome copy numbers of bacteria. Slow-growing (doubling time 103 minutes and fast-growing (doubling time 25 minutes E. coli cultures were used as a positive control. The copy numbers of the origin and terminus region of the chromosome were determined and the results were in excellent agreement with published data. The approach was also used to determine the ploidy levels of Caulobacter crescentus (an alpha-proteobacterium and Wolinella succinogenes (an epsilon-proteobacterium, both of which are monoploid. In contrast, Pseudomonas putida (a gamma-proteobacterium contains 20 genome copies and is thus polyploid. A survey of the proteobacteria with experimentally-determined genome copy numbers revealed that only three to four of 11 species are monoploid and thus monoploidy is not typical for proteobacteria. The ploidy level is not conserved within the groups of proteobacteria, and there are no obvious correlations between the ploidy levels with other parameters like genome size, optimal growth temperature or mode of life.

  1. Genetic and computational identification of a conserved bacterial metabolic module.

    Directory of Open Access Journals (Sweden)

    Cara C Boutte

    2008-12-01

    Full Text Available We have experimentally and computationally defined a set of genes that form a conserved metabolic module in the alpha-proteobacterium Caulobacter crescentus and used this module to illustrate a schema for the propagation of pathway-level annotation across bacterial genera. Applying comprehensive forward and reverse genetic methods and genome-wide transcriptional analysis, we (1 confirmed the presence of genes involved in catabolism of the abundant environmental sugar myo-inositol, (2 defined an operon encoding an ABC-family myo-inositol transmembrane transporter, and (3 identified a novel myo-inositol regulator protein and cis-acting regulatory motif that control expression of genes in this metabolic module. Despite being encoded from non-contiguous loci on the C. crescentus chromosome, these myo-inositol catabolic enzymes and transporter proteins form a tightly linked functional group in a computationally inferred network of protein associations. Primary sequence comparison was not sufficient to confidently extend annotation of all components of this novel metabolic module to related bacterial genera. Consequently, we implemented the Graemlin multiple-network alignment algorithm to generate cross-species predictions of genes involved in myo-inositol transport and catabolism in other alpha-proteobacteria. Although the chromosomal organization of genes in this functional module varied between species, the upstream regions of genes in this aligned network were enriched for the same palindromic cis-regulatory motif identified experimentally in C. crescentus. Transposon disruption of the operon encoding the computationally predicted ABC myo-inositol transporter of Sinorhizobium meliloti abolished growth on myo-inositol as the sole carbon source, confirming our cross-genera functional prediction. Thus, we have defined regulatory, transport, and catabolic genes and a cis-acting regulatory sequence that form a conserved module required for myo

  2. Regulatory Response to Carbon Starvation in Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Britos, Leticia C.; Abeliuk, Eduardo; Taverner, Thomas; Lipton, Mary S.; McAdams, Harley; Shapiro, Lucy

    2011-04-11

    Bacteria adapt to shifts from rapid to slow growth, and have developed strategies for long-term survival during prolonged starvation and stress conditions. We report the regulatory response of C. crescentus to carbon starvation, based on combined high-throughput proteome and transcriptome analyses. Our results identify cell cycle changes in gene expression in response to carbon starvation that involve the prominent role of the FixK FNR/CAP family transcription factor and the CtrA cell cycle regulator. Notably, the SigT ECF sigma factor mediates the carbon starvation-induced degradation of CtrA, while activating a core set of general starvation-stress genes that respond to carbon starvation, osmotic stress, and exposure to heavy metals. Comparison of the response of swarmer cells and stalked cells to carbon starvation revealed four groups of genes that exhibit different expression profiles. Also, cell pole morphogenesis and initiation of chromosome replication normally occurring at the swarmer-to-stalked cell transition are uncoupled in carbon-starved cells.

  3. Regulatory response to carbon starvation in Caulobacter crescentus.

    Directory of Open Access Journals (Sweden)

    Leticia Britos

    Full Text Available Bacteria adapt to shifts from rapid to slow growth, and have developed strategies for long-term survival during prolonged starvation and stress conditions. We report the regulatory response of C. crescentus to carbon starvation, based on combined high-throughput proteome and transcriptome analyses. Our results identify cell cycle changes in gene expression in response to carbon starvation that involve the prominent role of the FixK FNR/CAP family transcription factor and the CtrA cell cycle regulator. Notably, the SigT ECF sigma factor mediates the carbon starvation-induced degradation of CtrA, while activating a core set of general starvation-stress genes that respond to carbon starvation, osmotic stress, and exposure to heavy metals. Comparison of the response of swarmer cells and stalked cells to carbon starvation revealed four groups of genes that exhibit different expression profiles. Also, cell pole morphogenesis and initiation of chromosome replication normally occurring at the swarmer-to-stalked cell transition are uncoupled in carbon-starved cells.

  4. Acoplamiento pelágico-bentónico: respuesta de la zona bentónica profunda a la sedimentación del florecimiento invernal de diatomeas en el lago oligotrófico Alchichica, Puebla, México Pelagic-benthic coupling: deep benthic zone response to winter diatom bloom sinking in oligotrophic Lake Alchichica, Puebla, Mexico

    Directory of Open Access Journals (Sweden)

    Javier Alcocer

    2008-08-01

    Full Text Available El objetivo del presente estudio es reconocer la existencia de un acoplamiento pelágico-bentónico en el lago oligotrófico tropical Alchichica evaluando la respuesta de la zona bentónica profunda a la sedimentación del florecimiento invernal de diatomeas. Se midió la biomasa fitoplanctónica en la columna de agua a lo largo de un ciclo anual, al igual que la concentración de clorofila a sedimentaria. Alchichica es un lago monomíctico cálido con un periodo de circulación invernal y estratificación el resto del año. La presencia de turbulencia y nutrimentos durante el periodo de circulación favorecen el desarrollo de un florecimiento de diatomeas compuesto por especies de talla grande (p. e., Cyclotella alchichicana, las cuales se sedimentan al no ser consumidas en su totalidad. La zona bentónica profunda del lago responde a la sedimentación del florecimiento invernal de diatomeas con el desarrollo y permanencia por un periodo prolongado de anoxia hipolimnética, lo que a su vez, impide el establecimiento y desarrollo de fauna bentónica en el Lago Alchichica, favorece la pérdida interna de nitrógeno por desnitrificación y consecuentemente, conlleva a que el nitrógeno sea el elemento que más frecuentemente límite el crecimiento fitoplanctónico.The aim of this study is to recognize the existence of a pelagic-benthic coupling in the oligotrophic, tropical Lake Alchichica through analysis of the response of the deep benthic zone to the winter diatom bloom deposition. The water column phytoplankton biomass and the sedimentary chlorophyll a were analyzed along an annual cycle. Alchichica is a warm monomictic lake circulating in winter and stratified over the rest of the year. The presence of turbulence and nutrient availability during the mixing period, favor the development of a diatom bloom composed by large species (e. g., Cyclotella alchichicana which are not totally consumed and settle down. The deep benthic zone responds to

  5. Algal response to nutrient enrichment in forested oligotrophic stream

    NARCIS (Netherlands)

    Veraart, A.J.; Romaní, A.M.; Tornés, E.; Sabater, S.

    2008-01-01

    Nutrient input in streams alters the density and species composition of attached algal communities in open systems. However, in forested streams, the light reaching the streambed (rather than the local nutrient levels) may limit the growth of these communities. A nutrient-enrichment experiment in a

  6. Algal response to nutrient enrichment in a forested oligotrophic stream

    NARCIS (Netherlands)

    Veraart, A.J.; Romani, A.M.; Tornes, E.; Sabater, S.

    2008-01-01

    Nutrient input in streams alters the density and species composition of attached algal communities in open systems. However, in forested streams, the light reaching the streambed (rather than the local nutrient levels) may limit the growth of these communities. A nutrient-enrichment experiment in a

  7. Algal response to nutrient enrichment in forested oligotrophic stream

    NARCIS (Netherlands)

    Veraart, A.J.; Romaní, A.M.; Tornés, E.; Sabater, S.

    2008-01-01

    Nutrient input in streams alters the density and species composition of attached algal communities in open systems. However, in forested streams, the light reaching the streambed (rather than the local nutrient levels) may limit the growth of these communities. A nutrient-enrichment experiment in a

  8. Carbon and phosphorus regulating bacterial metabolism in oligotrophic boreal lakes

    DEFF Research Database (Denmark)

    Vidal, L. O.; Graneli, W.; Daniel, C. B.;

    2011-01-01

    -P and glucose-C alone or in combination (0.01 and 0.3 mg L(-1), respectively) was added to 1.0 mu m filtered lake water and incubated in darkness at 20 degrees C. Additions of glucose (C) and phosphorus (P) alone did not lead to changes in the rates of bacterial metabolic processes, whereas bacterial...... respiration and bacterial production responded positively to C + P enrichment for most of the lakes sampled. Bacterial growth efficiency showed a wide range (2.5-28.7%) and low mean value (12%). These variations were not correlated with the DOC concentration. Our results show that heterotrophic bacterial...

  9. Silica cycling in the ultra-oligotrophic Eastern Mediterranean Sea

    Directory of Open Access Journals (Sweden)

    M. D. Krom

    2014-03-01

    Full Text Available Although silica is a key plant nutrient there have been few studies aimed at understanding the Si cycle in the Eastern Mediterranean Sea (EMS. Here we use a combination of new measurements and literature values to explain the silicic acid distribution across the basin and to calculate a silica budget to identify the key controlling processes. The surface water concentration of ~ 1 μM, which is unchanging seasonally across the basin was due to the inflow of Western Mediterranean Sea (WMS water at the Straits of Sicily. It does not change seasonally because there is only a sparse population of diatoms due to the low nutrient (N and P supply to the photic zone in the EMS. The concentration of silicic acid in the deep water of the western Ionian Sea (6.3 μM close to the S. Adriatic area of formation was due to the preformed silicic acid (3 μM plus biogenic silica (BSi from the dissolution of diatoms from the winter phytoplankton bloom (3.2 μM. The increase of 4.4 μM across the deep water of the EMS was due to silicic acid formed from in-situ diagenetic weathering of alumina-silicate minerals fluxing out of the sediment. The major inputs to the EMS are silicic acid and BSi inflowing from the western Mediterranean (121 × 109 mol Si year−1 silicic acid and 16 × 109 mol Si year−1 BSi, silicic acid fluxing from the sediment (54 × 109 mol Si year−1, riverine (27 × 109 mol Si year−1 and subterranean ground water (9.7 × 109 mol Si year−1 inputs, with only a minor direct input from dissolution of dust in the water column (1 × 109 mol Si year−1. This budget shows the importance of rapidly dissolving BSi and in-situ weathering of alumino-silicate minerals as sources of silica to balance the net export of silicic acid at the Straits of Sicily. Future measurements to improve the accuracy of this preliminary budget have been identified.

  10. Chlorophyll Variability in the Oligotrophic Gyres: Mechanisms, Seasonality and Trends

    Directory of Open Access Journals (Sweden)

    Sergio Romano Signorini

    2015-02-01

    Full Text Available A 16-year (1998-2013 analysis of trends and seasonal patterns was conducted for the five subtropical ocean gyres using satellite data: chlorophyll-a (Chl-a retrievals from ocean color, sea surface temperature (SST, and sea-level anomaly (SLA. Trend analysis was also performed on mixed-layer data derived from ocean model gridded temperature and salinity profiles (1998-2010. The Chl-a monthly composites were constructed from the Sea-viewing Wide Field-of-view Sensor (SeaWiFS and Moderate-resolution Imaging Spectroradiometer (MODIS on Aqua using two different algorithms: the standard algorithm (STD that has been in use since the start of the SeaWiFS mission in 1997, and a more recently developed Ocean Color Index (OCI algorithm with improved accuracy in low Chl-a waters. Trends were obtained for all gyres using both STD and OCI algorithms, which demonstrated generally consistent results. The North Pacific, Indian Ocean, North Atlantic and South Atlantic gyres showed significant downward trends in Chl-a, while the South Pacific gyre has a much weaker upward trend with no statistical significance. Time series of satellite-derived net primary production (NPP showed downward trends for all the gyres, while all five gyres exhibited positive trends in SST and SLA. The seasonal variability of Chl-a in each gyre is tightly coupled to the variability in mixed layer depth (MLD with peak values in winter in both hemispheres when vertical mixing is more vigorous, reaching depths approaching the nutricline. On a seasonal basis, Chl-a concentrations increase when the MLD approaches or is deeper than the nutricline depth, in agreement with the concept that vertical mixing is the major driving mechanism for phytoplankton photosynthesis in the interior of the gyres. The combination of surface warming trends and biomass reduction over the 16-year period has the potential to reduce atmospheric CO2 uptake by the gyres and therefore influence the global carbon cycle.

  11. Oligotrophic bacteria and root disease suppression in organically managed soils

    NARCIS (Netherlands)

    Senechkin, I.V.

    2013-01-01

      The objective of this thesis was to obtain a better understanding of soil health in terms of microbial and chemical characteristics as well as suppression of soil borne plant pathogens. Organic soils were chosen as an appropriate model for studying soil health. Four different organic amendme

  12. From Endosymbiont to Host-Controlled Organelle: The Hijacking of Mitochondrial Protein Synthesis and Metabolism

    NARCIS (Netherlands)

    Gabaldon, T.; Huynen, M.A.

    2007-01-01

    Mitochondria are eukaryotic organelles that originated from the endosymbiosis of an alpha-proteobacterium. To gain insight into the evolution of the mitochondrial proteome as it proceeded through the transition from a free-living cell to a specialized organelle, we compared a reconstructed ancestral

  13. From endosymbiont to host-controlled organelle: the hijacking of mitochondrial protein synthesis and metabolism.

    NARCIS (Netherlands)

    Gabaldon, T.; Huynen, M.A.

    2007-01-01

    Mitochondria are eukaryotic organelles that originated from the endosymbiosis of an alpha-proteobacterium. To gain insight into the evolution of the mitochondrial proteome as it proceeded through the transition from a free-living cell to a specialized organelle, we compared a reconstructed ancestral

  14. Scanning electron microscopy and in vitro cultivation of endophytic bacteria from potato tubers related to Zebra Chip disease

    Science.gov (United States)

    Zebra chip disease (ZCD) drastically reduces the quality and market value of potatoes in North America. The disease is associated with a phloem-limited alpha-proteobacterium, “Candidatus Liberibacter solanacearum”. No effective measure is currently available to control ZCD. It is known that endoph...

  15. Scanning electron microscopy of “Candidatus Liberibacter solanacearum” in infected tomato phloem tissue

    Science.gov (United States)

    “Candidatus Liberibacter solanacearum” is an alpha-proteobacterium associated with potato zebra-chip disease. The bacterium is currently non-cultureable. Very little is known about the bacterial morphology, an important characteristic of a complete bacterial description. In this study, “Ca. L. so...

  16. Unusual five copies and dual forms of nrdB in “Candidatus Liberibacter asiaticus”: biological implications and PCR detection

    Science.gov (United States)

    “Candidatus Liberibacter asiaticus” (CLas), an alpha-proteobacterium, is associated with citrus Huanglongbing (HLB, yellow shoot disease), which is currently threatening citrus production worldwide. Research in CLas biology is challenging because the bacterium cannot be cultivated in vitro. In this ...

  17. From Endosymbiont to Host-Controlled Organelle: The Hijacking of Mitochondrial Protein Synthesis and Metabolism

    NARCIS (Netherlands)

    Gabaldon, T.; Huynen, M.A.

    2007-01-01

    Mitochondria are eukaryotic organelles that originated from the endosymbiosis of an alpha-proteobacterium. To gain insight into the evolution of the mitochondrial proteome as it proceeded through the transition from a free-living cell to a specialized organelle, we compared a reconstructed ancestral

  18. From endosymbiont to host-controlled organelle: the hijacking of mitochondrial protein synthesis and metabolism.

    NARCIS (Netherlands)

    Gabaldon, T.; Huynen, M.A.

    2007-01-01

    Mitochondria are eukaryotic organelles that originated from the endosymbiosis of an alpha-proteobacterium. To gain insight into the evolution of the mitochondrial proteome as it proceeded through the transition from a free-living cell to a specialized organelle, we compared a reconstructed ancestral

  19. Replication initiator DnaA binds at the Caulobacter centromere and enables chromosome segregation.

    Science.gov (United States)

    Mera, Paola E; Kalogeraki, Virginia S; Shapiro, Lucy

    2014-11-11

    During cell division, multiple processes are highly coordinated to faithfully generate genetically equivalent daughter cells. In bacteria, the mechanisms that underlie the coordination of chromosome replication and segregation are poorly understood. Here, we report that the conserved replication initiator, DnaA, can mediate chromosome segregation independent of replication initiation. It does so by binding directly to the parS centromere region of the chromosome, and mutations that alter this interaction result in cells that display aberrant centromere translocation and cell division. We propose that DnaA serves to coordinate bacterial DNA replication with the onset of chromosome segregation.

  20. Localizing the Subunit Pool for the Temporally Regulated Polar Pili of Caulobacter crescentus.

    Science.gov (United States)

    1987-01-01

    Roosendaal, B., P.M.P. Van Bergen en Henegouwen and F.K. De Graaf (1986) Subcellular localization of K99 fimbrial subunits and effect of temperature on...ORGANIZAT A4A97 University of California Office of Naval Research 6c. ADDRE(t Silage , a tory 7b AJDlS jity, State, and ZIP Code) 2’r~aaulsciences Laborat...colloidal gold conjugate, diluted 1:8 in OLG, and incubated 1 1/2 hr at room temperature . A squirt wash with high salt buffer and then water followed. The

  1. Disease: H01148 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available H01148 Caulobacter infection The genus Caulobacter includes gram-negative bacteria characterized by asymmetr...ic cell division and stalk. Although infection with Caulobacter species is rare, it

  2. Microbial food web in an oligotrophic high mountain lake (Jöri Lake III, Switzerland

    Directory of Open Access Journals (Sweden)

    Ferdinand SCHANZ

    1999-08-01

    Full Text Available Jöri Lake III (2512 m a.s.l., zm = 22 m, A = 57.81 ´ 103 m2, V = 601.1 ´ 103 m3 is situated in the Vereina region in the eastern part of the Swiss Alps. We studied microbial grazing on bacteria and bacterial productivity during the ice-free period. The lake normally gets thermally stratified for two months between July and September. In 1996, chlorophyll-a concentrations varied from 0.5 to 2.0 μg l-1 with maximum values just below the thermocline (6 m depth, in 1997, they were between 0.6 and 5.0 μg l-1 with maximum values at 10 m depth – several meters below the thermocline. Bacterial densities varied between 0.7 and 1.7 ´ 106 ml-1 with maxima in the thermocline, one to two meters above the chlorophyll maximum. The areal bacterial biomass (volume beneath 1 m2 to a depth of 8 m was 10 μg C l-1 which remained more or less constant for the periods investigated. In 1997, bacterial growth rate and production rates were determined using [3H]-thymidine incorporation. The rates were as low as 0.002 to 0.006 h-1 and 0.01 to 0.03 μg C l-1 h-1, respectively. We found a carbon ratio of bacteria, phytoplankton, and autotrophic picoplancton (APP of 1.5:1.1:1 which shows a rather high abundance of bacteria and autotrophic picoplankton (APP compared to larger phytoplankton. Bacterial growth followed a temperature dependence similar to the one observed for bacteria from Lake Zürich, a prealpine and mesotrophic lake which was studied for comparison. Microbial food web in Jöri Lake III was not top down controlled during the periods of our study and mixotrophic algae like Dinobryon cylindricum var. alpinum and autotrophic nanoflagellates (ANF were the dominant bacterial grazers observed.

  3. Theoretical analysis of the cost of antagonistic activity for aquatic bacteria in oligotrophic environments

    Directory of Open Access Journals (Sweden)

    Eneas eAguirre-Von-Wobeser

    2015-05-01

    Full Text Available Many strains of bacteria produce antagonistic substances that restrain the growth of others, and potentially give them a competitive advantage. These substances are commonly released to the surrounding environment, involving metabolic costs in terms of energy and nutrients. The rate at which these molecules need to be produced to maintain a certain amount of them close to the producing cell before they are diluted into the environment has not been explored so far. To understand the potential cost of production of antagonistic substances in water environments, we used two different theoretical approaches. Using a probabilistic model, we determined the rate at which a cell needs to produce individual molecules in order to keep on average a single molecule in its vicinity at all times. For this minimum protection, a cell would need to invest 3.92X10-22 kg s-1 of organic matter, which is 9 orders of magnitude lower than the estimated expense for growth. Next, we used a continuous model, based on Fick’s laws, to explore the production rate needed to sustain minimum inhibitory concentrations around a cell, which would provide much more protection from competitors. In this scenario, cells would need to invest 1.20X10-11 kg s-1, which is 2 orders of magnitude higher than the estimated expense for growth, and thus not sustainable. We hypothesize that the production of antimicrobial compounds by bacteria in aquatic environments lies between these two extremes.

  4. Theoretical analysis of the cost of antagonistic activity for aquatic bacteria in oligotrophic environments.

    Science.gov (United States)

    Aguirre-von-Wobeser, Eneas; Eguiarte, Luis E; Souza, Valeria; Soberón-Chávez, Gloria

    2015-01-01

    Many strains of bacteria produce antagonistic substances that restrain the growth of others, and potentially give them a competitive advantage. These substances are commonly released to the surrounding environment, involving metabolic costs in terms of energy and nutrients. The rate at which these molecules need to be produced to maintain a certain amount of them close to the producing cell before they are diluted into the environment has not been explored so far. To understand the potential cost of production of antagonistic substances in water environments, we used two different theoretical approaches. Using a probabilistic model, we determined the rate at which a cell needs to produce individual molecules in order to keep on average a single molecule in its vicinity at all times. For this minimum protection, a cell would need to invest 3.92 × 10(-22) kg s(-1) of organic matter, which is 9 orders of magnitude lower than the estimated expense for growth. Next, we used a continuous model, based on Fick's laws, to explore the production rate needed to sustain minimum inhibitory concentrations around a cell, which would provide much more protection from competitors. In this scenario, cells would need to invest 1.20 × 10(-11) kg s(-1), which is 2 orders of magnitude higher than the estimated expense for growth, and thus not sustainable. We hypothesize that the production of antimicrobial compounds by bacteria in aquatic environments lies between these two extremes.

  5. Agricultural land-use change in a Mexican oligotrophic desert depletes ecosystem stability

    Directory of Open Access Journals (Sweden)

    Natali Hernández-Becerra

    2016-08-01

    Full Text Available Background Global demand for food has led to increased land-use change, particularly in dry land ecosystems, which has caused several environmental problems due to the soil degradation. In the Cuatro Cienegas Basin (CCB, alfalfa production irrigated by flooding impacts strongly on the soil. Methods In order to analyze the effect of such agricultural land-use change on soil nutrient dynamics and soil bacterial community composition, this work examined an agricultural gradient within the CCB which was comprised of a native desert grassland, a plot currently cultivated with alfalfa and a former agricultural field that had been abandoned for over 30 years. For each site, we analyzed C, N and P dynamic fractions, the activity of the enzyme phosphatase and the bacterial composition obtained using 16S rRNA clone libraries. Results The results showed that the cultivated site presented a greater availability of water and dissolved organic carbon, these conditions promoted mineralization processes mediated by heterotrophic microorganisms, while the abandoned land was limited by water and dissolved organic nitrogen. The low amount of dissolved organic matter promoted nitrification, which is mediated by autotrophic microorganisms. The microbial N immobilization process and specific phosphatase activity were both favored in the native grassland. As expected, differences in bacterial taxonomical composition were observed among sites. The abandoned site exhibited similar compositions than native grassland, while the cultivated site differed. Discussion The results suggest that the transformation of native grassland into agricultural land induces drastic changes in soil nutrient dynamics as well as in the bacterial community. However, with the absence of agricultural practices, some of the soil characteristics analyzed slowly recovers their natural state.

  6. Microbial food webs and metabolic state across oligotrophic waters of the Mediterranean Sea during summer

    Directory of Open Access Journals (Sweden)

    U. Christaki

    2011-07-01

    Full Text Available The abundance and activity of the major members of the heterotrophic microbial community – from viruses to ciliates – were studied along a longitudinal transect across the Mediterranean Sea in the summer of 2008. The Mediterranean Sea is characterized by a west to-east gradient of deepening of DCM (deep chlorophyll maximum and increasing oligotrophy reflected in gradients of biomass and production. However, within this well documented longitudinal trend, hydrological mesoscale features exist and likely influence microbial dynamics. Here we present data from a W-E transect of 17 stations during the period of summer stratification. Along the transect the production and fate of organic matter was investigated at three selected sites each one located in the centre of an anticyclonic eddy: in the Algero-Provencal Basin (St. A, the Ionian Basin (St. B, and the Levantine Basin (St. C. The 3 geographically distant eddies showed low values of the different heterotrophic compartments of the microbial food web, and except for viruses in site C, all integrated (0–150 m stocks were higher in reference stations located in the same basin outside the eddies. During our study the 3 eddies showed equilibrium between GPP (Gross Primary Production and DCR (Dark Community Respiration. Integrated PPp (Particulate Primary Production values at A, B and C varied from ~140 to ~190 mg C m−2.

  7. Long-term atmospheric deposition of nitrogen, phosphorus and sulfate in a large oligotrophic lake.

    Science.gov (United States)

    Ellis, Bonnie K; Craft, James A; Stanford, Jack A

    2015-01-01

    We documented significantly increasing trends in atmospheric loading of ammonium (NH4) and nitrate/nitrite (NO2/3) and decreasing trends in total phosphorus (P) and sulfate (SO4) to Flathead Lake, Montana, from 1985 to 2004. Atmospheric loading of NO2/3 and NH4 increased by 48 and 198% and total P and SO4 decreased by 135 and 39%. The molar ratio of TN:TP also increased significantly. Severe air inversions occurred periodically year-round and increased the potential for substantial nutrient loading from even small local sources. Correlations between our loading data and various measures of air quality in the basin (e.g., particulate matter dust and smoke are important sources. Ammonium was the primary form of N in atmospheric deposition, whereas NO3 was the primary N form in tributary inputs. Atmospheric loading of NH4 to Flathead Lake averaged 44% of the total load and on some years exceeded tributary loading. Primary productivity in the lake is colimited by both N and P most of the year; and in years of high atmospheric loading of inorganic N, deposition may account for up to 6.9% of carbon converted to biomass.

  8. Nutrient Limitation in Surface Waters of the Oligotrophic Eastern Mediterranean Sea: an Enrichment Microcosm Experiment

    KAUST Repository

    Tsiola, A.

    2015-12-01

    The growth rates of planktonic microbes in the pelagic zone of the Eastern Mediterranean Sea are nutrient limited, but the type of limitation is still uncertain. During this study, we investigated the occurrence of N and P limitation among different groups of the prokaryotic and eukaryotic (pico-, nano-, and micro-) plankton using a microcosm experiment during stratified water column conditions in the Cretan Sea (Eastern Mediterranean). Microcosms were enriched with N and P (either solely or simultaneously), and the PO4 turnover time, prokaryotic heterotrophic activity, primary production, and the abundance of the different microbial components were measured. Flow cytometric and molecular fingerprint analyses showed that different heterotrophic prokaryotic groups were limited by different nutrients; total heterotrophic prokaryotic growth was limited by P, but only when both N and P were added, changes in community structure and cell size were detected. Phytoplankton were N and P co-limited, with autotrophic pico-eukaryotes being the exception as they increased even when only P was added after a 2-day time lag. The populations of Synechococcus and Prochlorococcus were highly competitive with each other; Prochlorococcus abundance increased during the first 2 days of P addition but kept increasing only when both N and P were added, whereas Synechococcus exhibited higher pigment content and increased in abundance 3 days after simultaneous N and P additions. Dinoflagellates also showed opportunistic behavior at simultaneous N and P additions, in contrast to diatoms and coccolithophores, which diminished in all incubations. High DNA content viruses, selective grazing, and the exhaustion of N sources probably controlled the populations of diatoms and coccolithophores.

  9. Biology of the fire blight pathogen Erwinia amylovora in oligotrophic environments: survival responses and virulence

    OpenAIRE

    Delgado Santander, Ricardo

    2016-01-01

    Erwinia amylovora es una bacteria fitopatógena de la familia Enterobacteriaceae, responsable del fuego bacteriano de las rosáceas. Los efectos destructivos de este patógeno sobre frutos, flores y prácticamente todos los órganos de las plantas hospedadoras afectadas constituyen una amenaza importante para la producción de pera y manzana, y suponen graves pérdidas económicas anuales en todo el mundo. E. amylovora está clasificada como un organismo de cuarentena en la Unión Europea y en otros pa...

  10. Patterns of ecological specialization among microbial populations in the Red Sea and diverse oligotrophic marine environments

    KAUST Repository

    Thompson, Luke R

    2013-05-11

    Large swaths of the nutrient-poor surface ocean are dominated numerically by cyanobacteria (Prochlorococcus), cyanobacterial viruses (cyanophage), and alphaproteobacteria (SAR11). How these groups thrive in the diverse physicochemical environments of different oceanic regions remains poorly understood. Comparative metagenomics can reveal adaptive responses linked to ecosystem-specific selective pressures. The Red Sea is well-suited for studying adaptation of pelagic-microbes, with salinities, temperatures, and light levels at the extreme end for the surface ocean, and low nutrient concentrations, yet no metagenomic studies have been done there. The Red Sea (high salinity, high light, low N and P) compares favorably with the Mediterranean Sea (high salinity, low P), Sargasso Sea (low P), and North Pacific Subtropical Gyre (high light, low N). We quantified the relative abundance of genetic functions among Prochlorococcus, cyanophage, and SAR11 from these four regions. Gene frequencies indicate selection for phosphorus acquisition (Mediterranean/Sargasso), DNA repair and high-light responses (Red Sea/Pacific Prochlorococcus), and osmolyte C1 oxidation (Red Sea/Mediterranean SAR11). The unexpected connection between salinity-dependent osmolyte production and SAR11 C1 metabolism represents a potentially major coevolutionary adaptation and biogeochemical flux. Among Prochlorococcus and cyanophage, genes enriched in specific environments had ecotype distributions similar to nonenriched genes, suggesting that inter-ecotype gene transfer is not a major source of environment-specific adaptation. Clustering of metagenomes using gene frequencies shows similarities in populations (Red Sea with Pacific, Mediterranean with Sargasso) that belie their geographic distances. Taken together, the genetic functions enriched in specific environments indicate competitive strategies for maintaining carrying capacity in the face of physical stressors and low nutrient availability. 2013 The Authors. Ecology and Evolution.

  11. Single-cell genomics reveal metabolic strategies for microbial growth and survival in an oligotrophic aquifer

    Energy Technology Data Exchange (ETDEWEB)

    Wilkins, Michael J.; Kennedy, David W.; Castelle, Cindy; Field, Erin; Stepanauskas, Ramunas; Fredrickson, Jim K.; Konopka, Allan

    2014-02-09

    Bacteria from the genus Pedobacter are a major component of microbial assemblages at Hanford Site and have been shown to significantly change in abundance in response to the subsurface intrusion of Columbia River water. Here we employed single cell genomics techniques to shed light on the physiological niche of these microorganisms. Analysis of four Pedobacter single amplified genomes (SAGs) from Hanford Site sediments revealed a chemoheterotrophic lifestyle, with the potential to exist under both aerobic and microaerophilic conditions via expression of both aa3­-type and cbb3-type cytochrome c oxidases. These SAGs encoded a wide-range of both intra-and extra­-cellular carbohydrate-active enzymes, potentially enabling the degradation of recalcitrant substrates such as xylan and chitin, and the utilization of more labile sugars such as mannose and fucose. Coupled to these enzymes, a diversity of transporters and sugar-binding molecules were involved in the uptake of carbon from the extracellular local environment. The SAGs were enriched in TonB-dependent receptors (TBDRs), which play a key role in uptake of substrates resulting from degradation of recalcitrant carbon. CRISPR-Cas mechanisms for resisting viral infections were identified in all SAGs. These data demonstrate the potential mechanisms utilized for persistence by heterotrophic microorganisms in a carbon-limited aquifer, and hint at potential linkages between observed Pedobacter abundance shifts within the 300 Area subsurface and biogeochemical shifts associated with Columbia River water intrusion.

  12. Monitoring of biofilm formation and activity in drinking water distribution networks under oligotrophic conditions

    DEFF Research Database (Denmark)

    Boe-Hansen, Rasmus; Martiny, Adam Camillo; Arvin, Erik

    2003-01-01

    In this study, the construction a model distribution system suitable for studies of attached and suspended microbial activity in drinking water under controlled circumstances is outlined. The model system consisted of two loops connected in series with a total of 140 biofilm sampling points...

  13. Autonomous, high-resolution observations of particle flux in the oligotrophic ocean

    Directory of Open Access Journals (Sweden)

    M. L. Estapa

    2013-01-01

    Full Text Available Observational gaps limit our understanding of particle flux attenuation through the upper mesopelagic because available measurements (sediment traps and radiochemical tracers have limited temporal resolution, are labor-intensive, and require ship support. Here, we conceptually evaluate an autonomous, optical proxy-based method for high-resolution observations of particle flux. We present four continuous records of particle flux collected with autonomous, profiling floats in the western Sargasso Sea and the subtropical North Pacific, as well as one shorter record of depth-resolved particle flux near the Bermuda Atlantic Timeseries Study (BATS and Oceanic Flux Program (OFP sites. These observations illustrate strong variability in particle flux over very short (~1 day timescales, but at longer timescales they reflect patterns of variability previously recorded during sediment trap timeseries. While particle flux attenuation at BATS/OFP agreed with the canonical power-law model when observations were averaged over a month, flux attenuation was highly variable on timescales of 1–3 days. Particle fluxes at different depths were decoupled from one another and from particle concentrations and chlorophyll fluorescence in the immediately-overlying surface water, consistent with horizontal advection of settling particles. We finally present an approach for calibrating this optical proxy in units of carbon flux, discuss in detail the related, inherent physical and optical assumptions, and look forward toward the requirements for the quantitative application of this method in highly time-resolved studies of particle export and flux attenuation.

  14. The Impact of Host Rock Geochemistry on Bacterial Community Structure in Oligotrophic Cave Environments.

    Directory of Open Access Journals (Sweden)

    Hazel A. Barton

    2007-07-01

    Full Text Available Despite extremely starved conditions, caves contain surprisingly diverse microbial communities. Our research is geared toward understanding what ecosystems drivers are responsible for this high diversity. To asses the effect of rock fabric and mineralogy, we carried out a comparative geomicrobiology study within Carlsbad Cavern, New Mexico, USA. Samples were collected from two different geologic locations within the cave: WF1 in the Massive Member of the Capitan Formation and sF88 in the calcareous siltstones of the Yates Formation. We examined the organic content at each location using liquid chromatography mass spectroscopy and analyzed microbial community structure using molecular phylogenetic analyses. In order to assess whether microbial activity was leading to changes in the bedrock at each location, the samples were also examined by petrology, X-ray diffraction (XRD and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX. Our results suggest that on the chemically complex Yates Formation (sF88, the microbial community was significantly more diverse than on the limestone surfaces of the Capitan (WF1, despite a higher total number of cells on the latter. Further, the broader diversity of bacterial species at sF88 reflected a larger range of potential metabolic capabilities, presumably due to opportunities to use ions within the rock as nutrients and for chemolithotrophic energy production. The use of these ions at sF88 is supported by the formation of a corrosion residue, presumably through microbial scavenging activities. Our results suggest that rock fabric and mineralogy may be an important driver of ecosystem function and should be carefully reviewed when carrying out microbial community analysis in cave environments.

  15. Seasonal variability in nutrient regeneration by mussel Mytilus edulis rope culture in oligotrophic systems

    NARCIS (Netherlands)

    Jansen, H.M.; Strand, O.; Strohmeier, T.; Krogness, C.; Verdegem, M.C.J.; Smaal, A.C.

    2011-01-01

    Blue mussel Mytilus edulis cultures contribute to nutrient cycling in coastal ecosystems. Mussel populations filter particulate nutrients from the water column and inorganic nutrients are regenerated by excretion of metabolic wastes and decomposition of (pseudo-)faeces. The objective of this study w

  16. Trophic ecology and persistence of invasive silver carp Hypophthalmichthys molitrix in an oligotrophic South African impoundment

    CSIR Research Space (South Africa)

    Lübcker, N

    2016-12-01

    Full Text Available to assess the trophic ecology of H. molitrix, which was then compared with indigenous Mozambique tilapia Oreochromis mossambicus, on a seasonal basis during 2011. Hypophthalmichthys molitrix are generalist filter feeders, with a diet consisting primarily...

  17. Elucidating the biogeochemical roles of novel microorganisms from oligotrophic Lake Superior sediments

    Science.gov (United States)

    Sheik, C.

    2016-12-01

    Lake Superior (LS) is Earth's largest lake in surface area and fifth largest by volume, yet little is known of the microbial life that drives carbon, nitrogen and sulfur cycles, especially in sediments. LS sediments are typically low in organic matter due to low concentrations of phosphorus in overlying water that constrain primary productivity. Yet, LS sediments are biogeochemically active and have been shown to have oxygen consumption rates akin to the pelagic ocean. Furthermore, microbial activity, potentially from sediments has been suggested for increasing nitrate concentrations in overlying waters. Together, LS is an interesting contrast to highly productive freshwater and coastal marine and less productive pelagic ocean sediments. Here I will present a mixture of results, both spatially and with sediment depth, from cores taken from six stations in the western arm of LS in July 2015. Overall, station locations were quite different in total carbon, nitrogen and microbial community structure. The variability observed likely stems from land influence, as some stations are more coastal, thus have higher rates of sedimentation, allochthonous carbon inputs and productivity than pelagic superior sites. Pelagic stations show a steady decline in both carbon and nitrogen with depth. Microbial communities, from the pelagic sediments are quite different from coastal communities. However, more interesting is that microbial community structure changes abruptly with depth in the sediment, suggesting that microbial membership is changing with geochemistry. At these pelagic stations, geochemical evidence of denitrification via traditional and ANAMMOX pathways and a pronounced sulfate methane transition zone have been shown previously. We see evidence of both denitrification processes in metagenomes and 16S rRNA gene libraries. Additionally, methanogen abundance generally increases with depth, while sulfate reducing bacteria peak and decline mid-core. Similarly, recovered genome bin's coverage follow similar patterns. Together, LS sediments offer a unique glimpse into microbial metabolism in carbon limited environments, and show that microbial biogeochemical cycling is, in part, performed by novel microorganisms.

  18. Resource utilization and trophic niche width in sandy beach macrobenthos from an oligotrophic coast

    Science.gov (United States)

    Ortega-Cisneros, Kelly; de Lecea, Ander M.; Smit, Albertus J.; Schoeman, David S.

    2017-01-01

    One of the paradigms underlying sandy beach ecology is the overriding control by physical processes; thus, biological interactions (i.e. food availability, competition and predation) are believed to play a role structuring macrofaunal communities only in benign habitats such as dissipative beaches. Moreover, sandy beaches are characterized by low in-situ productivity, so their food webs rely heavily on marine inputs. Studies have shown that estuarine organic matter plays a key role in influencing the dynamics of marine ecosystems. However, very few studies have tested the role of estuarine input on sandy beaches. Here, we aim to determine the impact of estuarine input on the food web of a sandy beach macrobenthic community. To this end, particulate organic matter (POM) samples from the marine environment and the estuary, as well as macrobenthic samples from the beach, were analysed for their stable isotope (SI) signature. Our results indicated that the POM SI signatures were not different along the beach, but differences were recorded between marine and estuarine sources. Bayesian mixing models indicated that the organisms did not make use of the estuarine POM at the beginning of the wet season, but relied more heavily on this resource towards the end of the wet season. This leads to the conclusion that changes in estuarine flow throughout the wet season can impact the trophic structure of macrobenthos communities, confirming a link between lotic and marine communities. Moreover, SI signatures suggest that the species collected here exhibit overlapping trophic niches, indicating high level of inter-specific competition. This highlights that species in low-productivity areas, such as the one studied here, can experience high levels of competition even in physically controlled environments such as sandy beaches.

  19. Patterns of ecological specialization among microbial populations in the Red Sea and diverse oligotrophic marine environments.

    Science.gov (United States)

    Thompson, Luke R; Field, Chris; Romanuk, Tamara; Ngugi, David; Siam, Rania; El Dorry, Hamza; Stingl, Ulrich

    2013-06-01

    Large swaths of the nutrient-poor surface ocean are dominated numerically by cyanobacteria (Prochlorococcus), cyanobacterial viruses (cyanophage), and alphaproteobacteria (SAR11). How these groups thrive in the diverse physicochemical environments of different oceanic regions remains poorly understood. Comparative metagenomics can reveal adaptive responses linked to ecosystem-specific selective pressures. The Red Sea is well-suited for studying adaptation of pelagic-microbes, with salinities, temperatures, and light levels at the extreme end for the surface ocean, and low nutrient concentrations, yet no metagenomic studies have been done there. The Red Sea (high salinity, high light, low N and P) compares favorably with the Mediterranean Sea (high salinity, low P), Sargasso Sea (low P), and North Pacific Subtropical Gyre (high light, low N). We quantified the relative abundance of genetic functions among Prochlorococcus, cyanophage, and SAR11 from these four regions. Gene frequencies indicate selection for phosphorus acquisition (Mediterranean/Sargasso), DNA repair and high-light responses (Red Sea/Pacific Prochlorococcus), and osmolyte C1 oxidation (Red Sea/Mediterranean SAR11). The unexpected connection between salinity-dependent osmolyte production and SAR11 C1 metabolism represents a potentially major coevolutionary adaptation and biogeochemical flux. Among Prochlorococcus and cyanophage, genes enriched in specific environments had ecotype distributions similar to nonenriched genes, suggesting that inter-ecotype gene transfer is not a major source of environment-specific adaptation. Clustering of metagenomes using gene frequencies shows similarities in populations (Red Sea with Pacific, Mediterranean with Sargasso) that belie their geographic distances. Taken together, the genetic functions enriched in specific environments indicate competitive strategies for maintaining carrying capacity in the face of physical stressors and low nutrient availability.

  20. Monitoring of biofilm formation and activity in drinking water distribution networks under oligotrophic conditions

    DEFF Research Database (Denmark)

    Boe-Hansen, Rasmus; Martiny, Adam Camillo; Arvin, Erik

    2003-01-01

    In this study, the construction a model distribution system suitable for studies of attached and suspended microbial activity in drinking water under controlled circumstances is outlined. The model system consisted of two loops connected in series with a total of 140 biofilm sampling points....... The biofilm from the system was studied using 11 different microbial methods and the results were compared and discussed. The methods were used for biomass quantification (AODC, HPC and ATP determination), visualisation of structure (CLSM), activity measurement (leucine incorporation, AOC removal rate...

  1. Seasonal and vertical distribution of putative ammonia-oxidizing thaumarchaeotal communities in an oligotrophic lake

    NARCIS (Netherlands)

    Vissers, E.W.; Blaga, C.I.; Bodelier, P.L.E.; Muyzer, G.; Schleper, C.; Sinninghe Damsté, J.S.; Tourna, M.; Laanbroek, H.J.

    2013-01-01

    The discovery of Archaea carrying an amoA gene coding for the A-subunit of ammonia monooxygenase gave a boost to studies aimed at detecting this gene under diverse conditions. Despite numerous studies describing the archaeal amoA gene abundance and richness in different habitats, the understanding o

  2. Autonomous, high-resolution observations of particle flux in the oligotrophic ocean

    Directory of Open Access Journals (Sweden)

    M. L. Estapa

    2013-08-01

    Full Text Available Observational gaps limit our understanding of particle flux attenuation through the upper mesopelagic because available measurements (sediment traps and radiochemical tracers have limited temporal resolution, are labor-intensive, and require ship support. Here, we conceptually evaluate an autonomous, optical proxy-based method for high-resolution observations of particle flux. We present four continuous records of particle flux collected with autonomous profiling floats in the western Sargasso Sea and the subtropical North Pacific, as well as one shorter record of depth-resolved particle flux near the Bermuda Atlantic Time-series Study (BATS and Oceanic Flux Program (OFP sites. These observations illustrate strong variability in particle flux over very short (~1-day timescales, but at longer timescales they reflect patterns of variability previously recorded during sediment trap time series. While particle flux attenuation at BATS/OFP agreed with the canonical power-law model when observations were averaged over a month, flux attenuation was highly variable on timescales of 1–3 days. Particle fluxes at different depths were decoupled from one another and from particle concentrations and chlorophyll fluorescence in the immediately overlying surface water, consistent with horizontal advection of settling particles. We finally present an approach for calibrating this optical proxy in units of carbon flux, discuss in detail the related, inherent physical and optical assumptions, and look forward toward the requirements for the quantitative application of this method in highly time-resolved studies of particle export and flux attenuation.

  3. Evidence for significant photochemical production of carbon monoxide by particles in coastal and oligotrophic marine waters

    Science.gov (United States)

    Xie, Huixiang; Zafiriou, Oliver C.

    2009-12-01

    Carbon monoxide (CO) photoproduction from particulate and chromophoric dissolved organic matter (CDOM) was determined in seawater from open-ocean and coastal areas. In confirmatory tests, poisoned or non-poisoned filtered and unfiltered blue-water samples, were exposed to sunlight. CO photoproduction was 21-42% higher in the unfiltered than in the filtered samples. In a more thorough study utilizing concentrated particles prepared by 0.2-μm cross-flow filtration, samples containing varying levels of particles were irradiated under simulated solar radiation. Their CO photoproduction rates increased linearly with particle concentration factor. Particulate CO production was 11-35% of CDOM-based CO production. On an absorbed-photons basis, the former was 30-108% more efficient than the latter. This study suggests that in both coastal and blue waters these new-found particulate photoprocesses are of similar biogeochemical importance to the well-known CDOM photoproduction term.

  4. Microbial biogeochemistry of Boiling Springs Lake: a physically dynamic, oligotrophic, low-pH geothermal ecosystem.

    Science.gov (United States)

    Siering, P L; Wolfe, G V; Wilson, M S; Yip, A N; Carey, C M; Wardman, C D; Shapiro, R S; Stedman, K M; Kyle, J; Yuan, T; Van Nostrand, J D; He, Z; Zhou, J

    2013-07-01

    Boiling Springs Lake (BSL) in Lassen Volcanic National Park, California, is North America's largest hot spring, but little is known about the physical, chemical, and biological features of the system. Using a remotely operated vessel, we characterized the bathymetry and near-surface temperatures at sub-meter resolution. The majority of the 1.2 ha, pH 2.2 lake is 10 m deep and 50-52 °C, but temperatures reach 93 °C locally. We extracted DNA from water and sediments collected from warm (52 °C) and hot (73-83 °C) sites separated by 180 m. Gene clone libraries and functional gene microarray (GeoChip 3.0) were used to investigate the BSL community, and uptake of radiolabeled carbon sources was used to assess the relative importance of heterotrophic vs. autotrophic production. Microbial assemblages are similar in both sites despite the strong temperature differential, supporting observations of a dynamic, convectively mixed system. Bacteria in the Actinobacteria and Aquificales phyla are abundant in the water column, and Archaea distantly related to known taxa are abundant in sediments. The functional potential appears similar across a 5-year time span, indicating a stable community with little inter-annual variation, despite the documented seasonal temperature cycle. BSL water-derived DNA contains genes for complete C, N, and S cycles, and low hybridization to probes for N and S oxidation suggests that reductive processes dominate. Many of the detected genes for these processes were from uncultivated bacteria, suggesting novel organisms are responsible for key ecosystem services. Selection imposed by low nutrients, low pH, and high temperature appear to result in low diversity and evenness of genes for key functions involved in C, N, and S cycling. Conversely, organic degradation genes appear to be functionally redundant, and the rapid assimilation of radiolabeled organic carbon into BSL cells suggests the importance of allochthonous C fueling heterotrophic production in the BSL C cycle.

  5. Sphingomonas alaskensis sp nov., a dominant bacterium from a marine oligotrophic environment

    NARCIS (Netherlands)

    Vancanneyt, M; Schut, F; Snauwaert, C; Goris, J; Swings, J; Gottschal, JC

    2001-01-01

    Seven Gram-negative strains, isolated in 1990 from a 10(6)-fold dilution series of seawater from Resurrection Bay, a deep fjord of the Gulf of Alaska, were identified in a polyphasic taxonomic study. Analysis of 16S rDNA sequences and DNA-homology studies confirmed the phylogenetic position of all s

  6. Polycyclic aromatic hydrocarbons alter the structure of oceanic and oligotrophic microbial food webs

    KAUST Repository

    Cerezo, Maria Isabel

    2015-11-01

    One way organic pollutants reach remote oceanic regions is by atmospheric transport. During the Malaspina-2010 expedition, across the Atlantic, Indian, and Pacific Oceans, we analyzed the polycyclic aromatic hydrocarbon (PAH) effects on oceanic microbial food webs. We performed perturbation experiments adding PAHs to classic dilution experiments. The phytoplankton growth rates were reduced by more than 5 times, being Prochlorococcus spp. the most affected. 62% of the experiments showed a reduction in the grazing rates due to the presence of PAHs. For the remaining experiments, grazing usually increased likely due to cascading effects. We identified changes in the slope of the relation between the growth rate and the dilution fraction induced by the pollutants, moving from no grazing to V-shape, or to negative slope, indicative of grazing increase by cascade effects and alterations of the grazers\\' activity structure. Our perturbation experiments indicate that PAHs could influence the structure oceanic food-webs structure.

  7. Selective uptake of prokaryotic picoplankton by a marine sponge ( Callyspongia sp.) within an oligotrophic coastal system

    Science.gov (United States)

    Hanson, Christine E.; McLaughlin, M. James; Hyndes, Glenn A.; Strzelecki, Joanna

    2009-09-01

    Marine sponges are key players in the transfer of carbon from the pelagic microbial food web into the benthos. Selective uptake of prokaryotic picoplankton (bacteria and autotrophic Synechococcus cyanobacteria identified and enumerated by flow cytometry. Callyspongia sp. demonstrated high filtration efficiencies, particularly for high DNA (HDNA) bacteria (up to 85.3% in summer 2008) and Synechococcus (up to 91.1% in autumn 2007), however efficiency varied non-uniformly with time and food type ( p bacteria (40 ± 17.2%), except during winter 2007 ( p = 0.14) when ambient Synechococcus concentrations were lowest. When compared to ambient abundances of the different food types, Callyspongia sp. exhibited consistently negative selectivity for LDNA bacteria and positive selectivity for Synechococcus, while HDNA bacteria was generally a neutral or positive selection. The total carbon removal rate (sum of all prokaryotic picoplankton cells), calculated on a per unit area basis, varied significantly with time ( p food webs of southwestern Australia, and support the conclusion that sponges actively select food particles that optimise their nutritional intake.

  8. Experimental evidence of dust-induced shaping of surface dissolved organic matter in the oligotrophic ocean

    Science.gov (United States)

    Pulido-Villena, Elvira; Djaoudi, Kahina; Barani, Aude; Charrière, Bruno; Delmont, Anne; Hélias-Nunige, Sandra; Marc, Tedetti; Wambeke France, Van

    2016-04-01

    Recent research has shown that dust deposition may impact the functioning of the microbial loop. On one hand, it enhances bacterial mineralization of dissolved organic matter (DOM), and so may limit the carbon export. On the other hand, the interaction between heterotrophic bacteria and DOM in the surface ocean can increase the residence time of DOM, promoting its export and sequestration in the deep ocean. The main goal of this study was to experimentally assess whether the bacterial response to dust deposition is prone to have an effect on the residence time of the DOM pool by modifying its bioavailability. The bacterial degradation of DOM was followed on dust-amended and control treatments during long-term incubations. Dissolved organic carbon concentration decreased by 9 μmol L-1 over the course of the experiment in both control and dust-enriched conditions, with no significant differences between treatments. However, significant differences in DOM optical properties appeared at the latest stage of the incubations suggesting an accumulation of DOM of high molecular weight in the dust-amended treatment. At the end of the incubations, the remaining water was filtered and re-used as a new culture medium for a bacterial natural assemblage. Bacterial abundance and production was lower in the treatment previously submitted to dust enrichment, suggesting a decrease in DOM lability after a dust deposition event. These preliminary results point to a new link between dust and ocean carbon cycle through the modification of the residence time of the DOM pool.

  9. Biology of the fire blight pathogen Erwinia amylovora in oligotrophic environments: survival responses and virulence

    OpenAIRE

    Delgado Santander, Ricardo

    2016-01-01

    Erwinia amylovora es una bacteria fitopatógena de la familia Enterobacteriaceae, responsable del fuego bacteriano de las rosáceas. Los efectos destructivos de este patógeno sobre frutos, flores y prácticamente todos los órganos de las plantas hospedadoras afectadas constituyen una amenaza importante para la producción de pera y manzana, y suponen graves pérdidas económicas anuales en todo el mundo. E. amylovora está clasificada como un organismo de cuarentena en la Unión Europea y en otros pa...

  10. Infestation of zooplankton with Triaenophorus and Proteocephalus procercoids (Cestoda in a deep oligotrophic lake

    Directory of Open Access Journals (Sweden)

    Peter Anegg

    2014-07-01

    Full Text Available In spring 2004, a massive infestation of the whitefish population in the Austrian Lake Achensee with Triaenophorus crassus was observed. Procercoids, the larval stage of parasitic cestodes, infest copepods as their first intermediate host. Therefore, in spring 2011, zooplankton samples were taken weekly at two sampling sites and depth ranges to determine the abundances of crustaceans as well as percentages of infected copepods and temporal occurrence of parasites. In addition, whitefish (Coregonus lavaretus stomach contents were analysed for food spectrum and parasite infestation. From the end of June to mid-August, procercoids of Triaenophorus spp. were detected in Cyclops abyssorum, the only first intermediate host for this parasite in Lake Achensee. Highest percentages of infected copepods were reached in mid-July (prevalence: 0.38%. Furthermore, an infestation of Proteocephalus sp. was observed in this copepod species, which occurred earlier until the end of the sampling period (prevalence: 1.34%. Besides C. abyssorum, also Eudiaptomus gracilis was occasionally infected with Proteocephalus (prevalence: 0.05%. The procercoids were found in both depth ranges, with no clear vertical infestation preference. More female C. abyssorum were Triaenophorus-infected than males, while the opposite was observed for Proteocephalus infection. The whitefish stomachs contained large numbers of Proteocephalus and Triaenophorus procercoids, coinciding with the occurrence of these parasites in the copepods.

  11. Diversity and bioprospection of fungal community present in oligotrophic soil of continental Antarctica

    Science.gov (United States)

    The diversity of fungal communities from different substrates in Antarctica were studied and their capability to produce bioactive compounds. A one hundred and one fungal isolates were identified by molecular analysis in 35 different fungal taxa from 20 genera. Pseudogymnoascus sp. 3, Pseudogymnoasc...

  12. Uptake of inorganic phosphorus by the aquatic plant Isoetes australis inhabiting oligotrophic vernal rock pools

    DEFF Research Database (Denmark)

    Christiansen, Nina Høj; Pulido, Cristina; Pedersen, Ole

    2017-01-01

    experiment revealed high amounts of Pi translocation internally in the plant which seemed to go from roots and oldest leaves to younger leaves. As a result of the high root to shoot ratio, high surface area, root uptake kinetics, and sediment Pi availability, roots accounted for 87% of plant Pi uptake...

  13. Upward nitrate transport by phytoplankton in oceanic waters: balancing nutrient budgets in oligotrophic seas

    Directory of Open Access Journals (Sweden)

    Tracy A. Villareal

    2014-03-01

    Full Text Available In oceanic subtropical gyres, primary producers are numerically dominated by small (1–5 µm diameter pro- and eukaryotic cells that primarily utilize recycled nutrients produced by rapid grazing turnover in a highly efficient microbial loop. Continuous losses of nitrogen (N to depth by sinking, either as single cells, aggregates or fecal pellets, are balanced by both nitrate inputs at the base of the euphotic zone and N2-fixation. This input of new N to balance export losses (the biological pump is a fundamental aspect of N cycling and central to understanding carbon fluxes in the ocean. In the Pacific Ocean, detailed N budgets at the time-series station HOT require upward transport of nitrate from the nutricline (80–100 m into the surface layer (∼0–40 m to balance productivity and export needs. However, concentration gradients are negligible and cannot support the fluxes. Physical processes can inject nitrate into the base of the euphotic zone, but the mechanisms for transporting this nitrate into the surface layer across many 10s of m in highly stratified systems are unknown. In these seas, vertical migration by the very largest (102–103 µm diameter phytoplankton is common as a survival strategy to obtain N from sub-euphotic zone depths. This vertical migration is driven by buoyancy changes rather than by flagellated movement and can provide upward N transport as nitrate (mM concentrations in the cells. However, the contribution of vertical migration to nitrate transport has been difficult to quantify over the required basin scales. In this study, we use towed optical systems and isotopic tracers to show that migrating diatom (Rhizosolenia mats are widespread in the N. Pacific Ocean from 140°W to 175°E and together with other migrating phytoplankton (Ethmodiscus, Halosphaera, Pyrocystis, and solitary Rhizosolenia can mediate time-averaged transport of N (235 µmol N m-2 d-1 equivalent to eddy nitrate injections (242 µmol NO3− m-2 d-1. This upward biotic transport can close N budgets in the upper 250 m of the central Pacific Ocean and together with diazotrophy creates a surface zone where biological nutrient inputs rather than physical processes dominate the new N flux. In addition to these numerically rare large migrators, there is evidence in the literature of ascending behavior in small phytoplankton that could contribute to upward flux as well. Although passive downward movement has dominated models of phytoplankton flux, there is now sufficient evidence to require a rethinking of this paradigm. Quantifying these fluxes is a challenge for the future and requires a reexamination of individual phytoplankton sinking rates as well as methods for capturing and enumerating ascending phytoplankton in the sea.

  14. Carbon dynamics modelization and biological community sensitivity to temperature in an oligotrophic freshwater Antarctic lake

    DEFF Research Database (Denmark)

    Antonio Villaescusa, Juan; Jorgensen, Sven Erik; Rochera, Carlos

    2016-01-01

    Lake Limnopolar, located in one of the areas on Earth experiencing the strongest local warming, has been studied as a maritime Antarctic lake model by the Limnopolar Research Team during the last decade. Data collected during this period revealed the existence of an important meteorological inter...

  15. Relevance of bacterioplankton abundance and production in the oligotrophic equatorial Indian Ocean

    Digital Repository Service at National Institute of Oceanography (India)

    Fernandes, V.; Rodrigues, V.; Ramaiah, N.; Paul, J.T.

    degrees N to 5 degrees S along 83 degrees E. The average bacterial abundance was 0.52 plus or minus 0.29, 0.62 plus or minus 0.33 and 0.46 plus or minus 0.19 (x 10 sup(8) cells l sup(–1)), respectively during NEM, SWM and SpIM in the top 100 m. In the deep...

  16. Reconstruction of annual carbon dynamics and balance for an oligotrophic pine fen

    Energy Technology Data Exchange (ETDEWEB)

    Alm, J.; Silvola, J.; Aaltonen, H. [Joensuu Univ. (Finland). Dept. of Biology; Talanov, A.; Ikkonen, E. [Karelian Research Centre of Russian Academy of Sciences (Russian Federation). Inst. of Biology; Nykaenen, H.; Martikainen, P.J. [National Public Health Inst. Kuopio (Finland). Dept. of Environmental Microbiology

    1996-12-31

    Atmospheric carbon dioxide (CO{sub 2}) is bound by mire vegetation in photosynthesis during the growing season, and is re-released by respiration of plants, soil animals and microorganisms consuming dead organic matter. A small proportion of annual primary production may fall below the water table to anoxic conditions and thus escapes the oxidative decomposition. Also from anoxic peat, carbon is released with clear seasonal and spatial variation as methane (CH{sub 4}.). The rate of carbon accumulation in peat depends on the annual inbalance of plant production and litter decomposition. Exchange of CO{sub 2} and CH{sub 4} between peat, vegetation and the atmosphere thus reflects the dynamics of carbon flows in the ecosystem. Net ecosystem CO{sub 2} exchange (PN), total CO{sub 2} release (RTOT) and CH{sub 4} release (D) from different treeless surfaces of low-sedge Sphagnum papillosum pine fen was studied in eastern Finland. (8 refs.)

  17. CelR, an Ortholog of the Diguanylate Cyclase PleD of Caulobacter, Regulates Cellulose Synthesis in Agrobacterium tumefaciens

    OpenAIRE

    Barnhart, D. Michael; Su, Shengchang; Baccaro, Brenna E.; Banta, Lois M.; Farrand, Stephen K.

    2013-01-01

    Cellulose fibrils play a role in attachment of Agrobacterium tumefaciens to its plant host. While the genes for cellulose biosynthesis in the bacterium have been identified, little is known concerning the regulation of the process. The signal molecule cyclic di-GMP (c-di-GMP) has been linked to the regulation of exopolysaccharide biosynthesis in many bacterial species, including A. tumefaciens. In this study, we identified two putative diguanylate cyclase genes, celR (atu1297) and atu1060, th...

  18. CelR, an ortholog of the diguanylate cyclase PleD of Caulobacter, regulates cellulose synthesis in Agrobacterium tumefaciens.

    Science.gov (United States)

    Barnhart, D Michael; Su, Shengchang; Baccaro, Brenna E; Banta, Lois M; Farrand, Stephen K

    2013-12-01

    Cellulose fibrils play a role in attachment of Agrobacterium tumefaciens to its plant host. While the genes for cellulose biosynthesis in the bacterium have been identified, little is known concerning the regulation of the process. The signal molecule cyclic di-GMP (c-di-GMP) has been linked to the regulation of exopolysaccharide biosynthesis in many bacterial species, including A. tumefaciens. In this study, we identified two putative diguanylate cyclase genes, celR (atu1297) and atu1060, that influence production of cellulose in A. tumefaciens. Overexpression of either gene resulted in increased cellulose production, while deletion of celR, but not atu1060, resulted in decreased cellulose biosynthesis. celR overexpression also affected other phenotypes, including biofilm formation, formation of a polar adhesion structure, plant surface attachment, and virulence, suggesting that the gene plays a role in regulating these processes. Analysis of celR and Δcel mutants allowed differentiation between phenotypes associated with cellulose production, such as biofilm formation, and phenotypes probably resulting from c-di-GMP signaling, which include polar adhesion, attachment to plant tissue, and virulence. Phylogenetic comparisons suggest that species containing both celR and celA, which encodes the catalytic subunit of cellulose synthase, adapted the CelR protein to regulate cellulose production while those that lack celA use CelR, called PleD, to regulate specific processes associated with polar localization and cell division.

  19. Unusually negative nitrogen isotopic compositions (δ15N of mangroves and lichens in an oligotrophic, microbially-influenced ecosystem

    Directory of Open Access Journals (Sweden)

    I. Romero

    2008-12-01

    Full Text Available Extremes in δ15N values in mangrove tissues and lichens (range =+4 to −22‰ were measured from a mangrove forest ecosystem located on Twin Cays, offshore islands in Belize, Central America. The N isotopic compositions and concentrations of NH4+/NH3 in porewater, rainwater, and atmospheric ammonia, and the δ15N of lichens, mangrove leaves, roots, stems, and wood were examined to study the biogeochemical processes important for establishing these unusual N isotopic ratios. Dwarfed Rhizophora mangle trees had the most negative δ15N, whereas fringing Rhizophora trees, the most positive δ15N values. Porewater ammonium concentrations had little relationship to N isotopic fractionation in mangrove tissues. In dwarfed mangroves, the δ15N of fine and coarse roots were 6–9‰ more positive than leaf tissue from the same tree, indicating different sources of N for root and leaf tissues. When P was added to dwarfed mangrove trees without added N, δ15N increased within one year from −12‰ to −2‰, approaching the δ15N of porewater ammonium (δ15N=+4‰. Isotopically depleted ammonia in the atmosphere (δ15N=−19‰ and in rainwater (δ15N=−10‰ were found on Twin Cays. We propose that foliar uptake of these atmospheric sources by P-stressed, dwarfed mangrove trees and lichens can explain their very negative δ15N values. In environments where P is limiting for growth, uptake of atmospheric N by Rhizophora mangle may be an important adaptive strategy.

  20. Autotrophic and heterotrophic metabolism of microbial planktonic communities in an oligotrophic coastal marine ecosystem: seasonal dynamics and episodic events

    Directory of Open Access Journals (Sweden)

    O. Bonilla-Findji

    2010-11-01

    Full Text Available A 18 month study was performed in the Bay of Villefranche to assess the episodic and seasonal variation of autotrophic and heterotrophic ecosystem processes. A typical spring bloom was encountered, where maximum of gross primary production (GPP was followed by maxima of bacterial respiration (BR and production (BP. The trophic balance (heterotrophy vs. autotrophy of the system did not exhibit any seasonal trend although a strong intra-annual variability was observed. On average, the community tended to be net heterotrophic with a GPP threshold for a balanced metabolism of 1.1 μmol O2 l−1 d−1. Extended forest fires in summer 2003 and a local episodic upwelling in July 2003 likely supplied orthophosphate and nitrate into the system. These events were associated with an enhanced bacterioplankton production (up to 2.4-fold, respiration (up to 4.5-fold and growth efficiency (up to 2.9-fold but had no effect on GPP. A Sahara dust wet deposition event in February 2004 stimulated bacterial abundance, production and growth efficiency but not GPP. Our study suggests that short-term disturbances such as wind-driven upwelling, forest fires and Sahara dust depositions can have a significant but previously not sufficiently considered influence on phytoplankton- and bacterioplankton-mediated ecosystem functions and can modify or even mask the seasonal dynamics. The study also indicates that atmospheric deposition of nutrients and particles not only impacts phytoplankton but also bacterioplankton and could, at times, also shift systems stronger towards net heterotrophy.

  1. Surface distribution of dissolved trace metals in the oligotrophic ocean and their influence on phytoplankton biomass and productivity

    KAUST Repository

    Pinedo-González, Paulina

    2015-10-25

    The distribution of bioactive trace metals has the potential to enhance or limit primary productivity and carbon export in some regions of the world ocean. To study these connections, the concentrations of Cd, Co, Cu, Fe, Mo, Ni, and V were determined for 110 surface water samples collected during the Malaspina 2010 Circumnavigation Expedition (MCE). Total dissolved Cd, Co, Cu, Fe, Mo, Ni, and V concentrations averaged 19.0 ± 5.4 pM, 21.4 ± 12 pM, 0.91 ± 0.4 nM, 0.66 ± 0.3 nM, 88.8 ± 12 nM, 1.72 ± 0.4 nM, and 23.4 ± 4.4 nM, respectively, with the lowest values detected in the Central Pacific and increased values at the extremes of all transects near coastal zones. Trace metal concentrations measured in surface waters of the Atlantic Ocean during the MCE were compared to previously published data for the same region. The comparison revealed little temporal changes in the distribution of Cd, Co, Cu, Fe, and Ni over the last 30 years. We utilized a multivariable linear regression model to describe potential relationships between primary productivity and the hydrological, biological, trace nutrient and macronutrient data collected during the MCE. Our statistical analysis shows that primary productivity in the Indian Ocean is best described by chlorophyll a, NO3, Ni, temperature, SiO4, and Cd. In the Atlantic Ocean, primary productivity is correlated with chlorophyll a, NO3, PO4, mixed layer depth, Co, Fe, Cd, Cu, V, and Mo. The variables salinity, temperature, SiO4, NO3, PO4, Fe, Cd, and V were found to best predict primary productivity in the Pacific Ocean. These results suggest that some of the lesser studied trace elements (e.g., Ni, V, Mo, and Cd) may play a more important role in regulating oceanic primary productivity than previously thought and point to the need for future experiments to verify their potential biological functions.

  2. Natural origin arsenic in aquatic organisms from a deep oligotrophic lake under the influence of volcanic eruptions.

    Science.gov (United States)

    Juncos, Romina; Arcagni, Marina; Rizzo, Andrea; Campbell, Linda; Arribére, María; Guevara, Sergio Ribeiro

    2016-02-01

    Volcanic eruptions are recognized sources of toxic elements to freshwater, including arsenic (As). In order to study the short term changes in the bioaccumulation of naturally occurring As by aquatic organisms in Lake Nahuel Huapi (Argentina), located close to the Puyehue-Cordón Caulle volcanic complex (PCCVC), we described As concentrations at different trophic levels and food web transfer patterns in three sites of the lake prior to the last PCCVC eruption (June 2011), and compared As concentrations in biota before and after the eruption. The highest As concentrations and greater variations both between sites and position in the water column, were observed in phytoplankton (3.9-64.8 µg g(-1) dry weight, DW) and small zooplankton (4.3-22.3 µg g(-1) DW). The pattern of As accumulation in aquatic organisms (whole body or muscle) was: primary producers (phytoplankton) > scrapper mollusks (9.3-15.3 µg g(-1) DW) > filter feeding mollusks (5.4-15.6 µg g(-1) DW) > omnivorous invertebrates (0.4-9.2 µg g(-1) DW) > zooplankton (1.2-3.5 µg g(-1) DW) > fish (0.2-1.9 µg g(-1) DW). We observed As biodilution in the whole food web, and in salmonids food chains, feeding on fish prey; but biomagnification in the food chain of creole perch, feeding on benthic crayfish. The impact of the 2011 PCCVC eruption on the As levels of biota was more evident in pelagic-associated organisms (zooplankton and planktivorous fish), but only in the short term, suggesting a brief high bioavailability of As in water after ash deposition. In benthic organisms As variations likely responded to shift in diet due to coverage of the littoral zone with ashes.

  3. Developmental Strategy For Effective Sampling To Detect Possible Nutrient Fluxes In Oligotrophic Coastal Reef Waters In The Caribbean

    Science.gov (United States)

    Mendoza, W. G.; Corredor, J. E.; Ko, D.; Zika, R. G.; Mooers, C. N.

    2008-05-01

    The increasing effort to develop the coastal ocean observing system (COOS) in various institutions has gained momentum due to its high value to climate, environmental, economic, and health issues. The stress contributed by nutrients to the coral reef ecosystem is among many problems that are targeted to be resolved using this system. Traditional nutrient sampling has been inadequate to resolve issues on episodic nutrient fluxes in reef regions due to temporal and spatial variability. This paper illustrates sampling strategy using the COOS information to identify areas that need critical investigation. The area investigated is within the Puerto Rico subdomain (60-70oW, 15-20oN), and Caribbean Time Series (CaTS), World Ocean Circulation Experiment (WOCE), Intra-America Sea (IAS) ocean nowcast/forecast system (IASNFS), and other COOS-related online datasets are utilized. Nutrient profile results indicate nitrate is undetectable in the upper 50 m apparently due to high biological consumption. Nutrients are delivered in Puerto Rico particularly in the CaTS station either via a meridional jet formed from opposing cyclonic and anticyclonic eddies or wind-driven upwelling. The strong vertical fluctuation in the upper 50 m demonstrates a high anomaly in temperature and salinity and a strong cross correlation signal. High chlorophyll a concentration corresponding to seasonal high nutrient influx coincides with higher precipitation accumulation rates and apparent riverine input from the Amazon and Orinoco Rivers during summer (August) than during winter (February) seasons. Non-detectability of nutrients in the upper 50 m is a reflection of poor sampling frequency or the absence of a highly sensitive nutrient analysis method to capture episodic events. Thus, this paper was able to determine the range of depths and concentrations that need to be critically investigated to determine nutrient fluxes, nutrient sources, and climatological factors that can affect nutrient delivery. It also provides some insight into needed sampling rates and temporal and spatial domain choices. Finally, it demonstrates a scientific reconnaissance for a field study that is now possible with online in-situ and remote sensing observations and numerical simulations, as a consequence of IOOS in general and COOS in particular.

  4. Two-role model of an interaction network of free-living γ-proteobacteria from an oligotrophic environment.

    Science.gov (United States)

    Aguirre-von-Wobeser, Eneas; Soberón-Chávez, Gloria; Eguiarte, Luis E; Ponce-Soto, Gabriel Yaxal; Vázquez-Rosas-Landa, Mirna; Souza, Valeria

    2014-05-01

    Antagonistic interactions are frequently observed among bacteria in the environment and result in complex networks, which could promote co-existence, and therefore promote biodiversity. We analysed interactions of aquatic bacteria isolated by their ability to grow in Pseudomonas isolation agar from Churince, Cuatro Ciénegas, Mexico. In the resulting network, highly antagonistic and highly sensitive strains could be distinguished, forming a largely hierarchical structure. Most of the highly antagonistic strains belonged to the genus Pseudomonas. The network was sender-determined, which means that the antagonist strains had a larger influence on its structure than the sensitive ones. Very few interactions were necessary to connect all strains, implying that the network was 'small world'. The network was highly nested, having a core of highly interacting strains, with which the less antagonistic or highly sensitive interact. A probabilistic model was built, which captured most features of the network. Biological interpretation of the model implied a state in which many different antagonistic mechanisms were present, and most strains were resistant to them. Our work shows that strains of Pseudomonas from the water column at Cuatro Ciénegas have the potential to interact antagonistically with many closely related strains and that these interactions are usually not reciprocal. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  5. Actinobacteria phylogenomics, selective isolation from an iron oligotrophic environment and siderophore functional characterization, unveil new desferrioxamine traits.

    Science.gov (United States)

    Cruz-Morales, Pablo; Ramos-Aboites, Hilda E; Licona-Cassani, Cuauhtémoc; Selem-Mójica, Nelly; Mejía-Ponce, Paulina M; Souza-Saldívar, Valeria; Barona-Gómez, Francisco

    2017-09-01

    Desferrioxamines are hydroxamate siderophores widely conserved in both aquatic and soil-dwelling Actinobacteria. While the genetic and enzymatic bases of siderophore biosynthesis and their transport in model families of this phylum are well understood, evolutionary studies are lacking. Here, we perform a comprehensive desferrioxamine-centric (des genes) phylogenomic analysis, which includes the genomes of six novel strains isolated from an iron and phosphorous depleted oasis in the Chihuahuan desert of Mexico. Our analyses reveal previously unnoticed desferrioxamine evolutionary patterns, involving both biosynthetic and transport genes, likely to be related to desferrioxamines chemical diversity. The identified patterns were used to postulate experimentally testable hypotheses after phenotypic characterization, including profiling of siderophores production and growth stimulation of co-cultures under iron deficiency. Based in our results, we propose a novel des gene, which we term desG, as responsible for incorporation of phenylacetyl moieties during biosynthesis of previously reported arylated desferrioxamines. Moreover, a genomic-based classification of the siderophore-binding proteins responsible for specific and generalist siderophore assimilation is postulated. This report provides a much-needed evolutionary framework, with specific insights supported by experimental data, to direct the future ecological and functional analysis of desferrioxamines in the environment. © FEMS 2017.

  6. Food quality dominates the impact of food quantity on Daphnia life history : possible implications for re-oligotrophication

    NARCIS (Netherlands)

    Sarpe, Dirk; Domis, Lisette N. de Senerpont; Declerck, Steven A. J.; van Donk, Ellen; Ibelings, Bas W.

    2014-01-01

    The elemental composition of phytoplankton is highly variable compared to the relatively narrow stoichiometry of zooplankton grazers. Using a full factorial design, we tested the effects of alterations in algal elemental composition (i.e., food quality) combined with food quantity on the life histor

  7. Food quality dominates the impact of food quantity on Daphnia life history: possible implications for re-oligotrophication

    NARCIS (Netherlands)

    Sarpe, D.; Senerpont Domis, de L.N.; Declerck, S.A.J.; Donk, van E.; Ibelings, B.W.

    2014-01-01

    The elemental composition of phytoplankton is highly variable compared to the relatively narrow stoichiometry of zooplankton grazers. Using a full factorial design, we tested the effects of alterations in algal elemental composition (i.e., food quality) combined with food quantity on the life histor

  8. Novel 2,4-Dichlorophenoxyacetic Acid Degradation Genes from Oligotrophic Bradyrhizobium sp. Strain HW13 Isolated from a Pristine Environment

    Science.gov (United States)

    Kitagawa, Wataru; Takami, Sachiko; Miyauchi, Keisuke; Masai, Eiji; Kamagata, Yoichi; Tiedje, James M.; Fukuda, Masao

    2002-01-01

    The tfd genes of Ralstonia eutropha JMP134 are the only well-characterized set of genes responsible for 2,4-dichlorophenoxyacetic acid (2,4-D) degradation among 2,4-D-degrading bacteria. A new family of 2,4-D degradation genes, cadRABKC, was cloned and characterized from Bradyrhizobium sp. strain HW13, a strain that was isolated from a buried Hawaiian soil that has never experienced anthropogenic chemicals. The cadR gene was inferred to encode an AraC/XylS type of transcriptional regulator from its deduced amino acid sequence. The cadABC genes were predicted to encode 2,4-D oxygenase subunits from their deduced amino acid sequences that showed 46, 44, and 37% identities with the TftA and TftB subunits of 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) oxygenase of Burkholderia cepacia AC1100 and with a putative ferredoxin, ThcC, of Rhodococcus erythropolis NI86/21, respectively. They are thoroughly different from the 2,4-D dioxygenase gene, tfdA, of R. eutropha JMP134. The cadK gene was presumed to encode a 2,4-D transport protein from its deduced amino acid sequence that showed 60% identity with the 2,4-D transporter, TfdK, of strain JMP134. Sinorhizobium meliloti Rm1021 cells containing cadRABKC transformed several phenoxyacetic acids, including 2,4-D and 2,4,5-T, to corresponding phenol derivatives. Frameshift mutations indicated that each of the cadRABC genes was essential for 2,4-D conversion in strain Rm1021 but that cadK was not. Five 2,4-D degraders, including Bradyrhizobium and Sphingomonas strains, were found to have cadA gene homologs, suggesting that these 2,4-D degraders share 2,4-D degradation genes similar to those of strain HW13 cadABC. PMID:11751829

  9. Water Constituents and Water Depth Retrieval from Sentinel-2A—A First Evaluation in an Oligotrophic Lake

    Directory of Open Access Journals (Sweden)

    Katja Dörnhöfer

    2016-11-01

    Full Text Available Satellite remote sensing may assist in meeting the needs of lake monitoring. In this study, we aim to evaluate the potential of Sentinel-2 to assess and monitor water constituents and bottom characteristics of lakes at spatio-temporal synoptic scales. In a field campaign at Lake Starnberg, Germany, we collected validation data concurrently to a Sentinel-2A (S2-A overpass. We compared the results of three different atmospheric corrections, i.e., Sen2Cor, ACOLITE and MIP, with in situ reflectance measurements, whereof MIP performed best (r = 0.987, RMSE = 0.002 sr−1. Using the bio-optical modelling tool WASI-2D, we retrieved absorption by coloured dissolved organic matter (aCDOM(440, backscattering and concentration of suspended particulate matter (SPM in optically deep water; water depths, bottom substrates and aCDOM(440 were modelled in optically shallow water. In deep water, SPM and aCDOM(440 showed reasonable spatial patterns. Comparisons with in situ data (mean: 0.43 m−1 showed an underestimation of S2-A derived aCDOM(440 (mean: 0.14 m−1; S2-A backscattering of SPM was slightly higher than backscattering from in situ data (mean: 0.027 m−1 vs. 0.019 m−1. Chlorophyll-a concentrations (~1 mg·m−3 of the lake were too low for a retrieval. In shallow water, retrieved water depths exhibited a high correlation with echo sounding data (r = 0.95, residual standard deviation = 0.12 m up to 2.5 m (Secchi disk depth: 4.2 m, though water depths were slightly underestimated (RMSE = 0.56 m. In deeper water, Sentinel-2A bands were incapable of allowing a WASI-2D based separation of macrophytes and sediment which led to erroneous water depths. Overall, the results encourage further research on lakes with varying optical properties and trophic states with Sentinel-2A.

  10. A Mesophilic, Autotrophic, Ammonia-Oxidizing Archaeon of Thaumarchaeal Group I.1a Cultivated from a Deep Oligotrophic Soil Horizon

    NARCIS (Netherlands)

    Jung, M.Y.; Park, S.J.; Kim, S.J.; Kim, J.G.; Sinninghe Damsté, J.S.; Jeon, C.O.; Rhee, S.K.

    2014-01-01

    Soil nitrification plays an important role in the reduction of soil fertility and in nitrate enrichment of groundwater. Various ammonia- oxidizing archaea (AOA) are considered to be members of the pool of ammonia-oxidizing microorganisms in soil. This study reports the discovery of a chemolithoautot

  11. Chlorophyll-a Algorithms for Oligotrophic Oceans: A Novel Approach Based on Three-Band Reflectance Difference

    Science.gov (United States)

    Hu, Chuanmin; Lee, Zhongping; Franz, Bryan

    2011-01-01

    A new empirical algorithm is proposed to estimate surface chlorophyll-a concentrations (Chl) in the global ocean for Chl less than or equal to 0.25 milligrams per cubic meters (approximately 77% of the global ocean area). The algorithm is based on a color index (CI), defined as the difference between remote sensing reflectance (R(sub rs), sr(sup -1) in the green and a reference formed linearly between R(sub rs) in the blue and red. For low Chl waters, in situ data showed a tighter (and therefore better) relationship between CI and Chl than between traditional band-ratios and Chl, which was further validated using global data collected concurrently by ship-borne and SeaWiFS satellite instruments. Model simulations showed that for low Chl waters, compared with the band-ratio algorithm, the CI-based algorithm (CIA) was more tolerant to changes in chlorophyll-specific backscattering coefficient, and performed similarly for different relative contributions of non-phytoplankton absorption. Simulations using existing atmospheric correction approaches further demonstrated that the CIA was much less sensitive than band-ratio algorithms to various errors induced by instrument noise and imperfect atmospheric correction (including sun glint and whitecap corrections). Image and time-series analyses of SeaWiFS and MODIS/Aqua data also showed improved performance in terms of reduced image noise, more coherent spatial and temporal patterns, and consistency between the two sensors. The reduction in noise and other errors is particularly useful to improve the detection of various ocean features such as eddies. Preliminary tests over MERIS and CZCS data indicate that the new approach should be generally applicable to all existing and future ocean color instruments.

  12. Spatial patterns and light-driven variation of microbial population gene expression in surface waters of the oligotrophic open ocean.

    Science.gov (United States)

    Hewson, Ian; Poretsky, Rachel S; Tripp, H James; Montoya, Joseph P; Zehr, Jonathan P

    2010-07-01

    Because bacterioplankton production rates do not vary strongly across vast expanses of the ocean, it is unclear how variability in community structure corresponds with functional variability in the open ocean. We surveyed community transcript functional profiles at eight locations in the open ocean, in both the light and in the dark, using the genomic subsystems approach, to understand variability in gene expression patterns in surface waters. Metatranscriptomes from geographically distinct areas and collected during the day and night shared a large proportion of metabolic functional similarity (74%) at the finest metabolic resolution possible. The variability between metatranscriptomes could be explained by phylogenetic differences between libraries (Mantel test, P < 0.0001). Several key gene expression pathways, including Photosystem I, Photosystem II and ammonium uptake, demonstrated the most variability both geographically and between light and dark. Libraries were dominated by transcripts of the cyanobacterium Prochlorocococcus marinus, where most geographical and diel variability between metatranscriptomes reflected between-station differences in cyanobacterial phototrophic metabolism. Our results demonstrate that active genetic machinery in surface waters of the ocean is dominated by photosynthetic microorganisms and their site-to-site variability, while variability in the remainder of assemblages is dependent on local taxonomic composition.

  13. Chironomid communities as indicators of local and global changes in an oligotrophic high mountain lake (Enol Lake, Northwestern Spain

    Directory of Open Access Journals (Sweden)

    Pol Tarrats

    2016-12-01

    Full Text Available The benthos of the high mountain Enol Lake (Picos de Europa National Park, Spain was analyzed in order to understand the spatiotemporal factors and patterns controlling its current Chironomidae community. In total, more than 14,000 chironomid larvae were identified, belonging to 27 taxa. The results have pointed out the presence of 3 main chironomid assemblages in the lake: i littoral community, which is mainly controlled by temperature and oxygen seasonal changes, ii Chara-dominant community, which is mainly controlled by the presence and abundance of Characeae in the lake, and iii profundal community, which is affected by low oxygen levels caused by sediment and organic matter discharge to the lake due to human pressures in the lake basin. We provide valuable insights for the managers to understand the current ecological status of Enol Lake and to evaluate which measures should be implemented to preserve or improve it. Moreover, our results constitute an essential step forward to improve the interpretation of the past changes of the lake by means of the subfossil chironomid community. 

  14. Development Strategy for Effective Sampling to Detect Possible Nutrient Fluxes in Oligotrophic Coastal Reef Waters in the Caribbean

    Science.gov (United States)

    2009-01-01

    Amazon and Orinoco rivers. Non-detectability of nutrients in the upper 50m is a reflection of poor sampling frequency or the absence of a highly...global warming, sea level rise, drought , freshening, eutrophication and pollution - affect the susceptibility of coastal areas to natural...northeast Caribbean Sea.18 Also, the southern region of PR is intermittently influenced by plumes from two major rivers, the Orinoco and Amazon . Thus

  15. Cosmogenic 32P and 33P in the Atmosphere and Oligotrophic Ocean and Applications to the Study of Phosphorus Cycling

    Science.gov (United States)

    1993-02-01

    Woods Hc’e Oceanographic Institution Joint Program. 3 Reproduction in whole or in part is permitted for any purpos of the United States Government. This...al., 1961). Also it has been shown that zooplankton have the ability to store P as fat in the males and in the reproductive system in the females... sargassum . I Table 4.4. Specific activities of 3 2 P and 3 3 P and activity ratio 3 3 p/ 2 p in plankton tows and suspended particulate (> 3 pmo

  16. Relationship between bacteria and phytoplankton during the giant jellyfish Nemopilema nomurai bloom in an oligotrophic temperate marine ecosystem

    Institute of Scientific and Technical Information of China (English)

    ZENG Yang; HUANG Xuguang; HUANG Bangqin; MI Tiezhu

    2016-01-01

    Bacterial abundance, phytoplankton community structure and environmental parameters were investigated to study the relationships between bacteria and phytoplankton during giant jellyfish Nemopilema nomurai blooms in the central Yellow Sea during 2013. N. nomurai appeared in June, increased in August, reached a peak and began to degrade in September 2013. Results showed that phosphate was possible a key nutrient for both phytoplankton and bacteria in June, but it changed to nitrate in August and September. Phytoplankton composition significantly changed that pico-phytoplankton relative biomass significantly increased, whereas other size phytoplankton significantly decreased during jellyfish bloom. In June, a significantly positive correlation was observed between chlorophyll a concentration and bacterial abundance (r=0.67, P0.05, n=25), but the relationship (r=0.71, P<0.001, n=31) was rebuilt with jellyfish degradation in September. In August, small size phytoplankton occupied the mixed layer in offshore stations, while bacteria almost distributed evenly in vertical. Chlorophyll a concentration significantly increased from (0.42±0.056) μg/L in June to (0.74±0.174) μg/L in August , while bacterial abundance just slightly increased. Additionally, the negative net community production indicated that community respiration was not entirely determined by the local primary productivity in August. These results indicated that jellyfish blooms potentially affect coupling of phytoplankton and bacteria in marine ecosystems.

  17. Nutrient control of N2 fixation in the oligotrophic Mediterranean Sea and the impact of Saharan dust events

    Directory of Open Access Journals (Sweden)

    M. Pujo-Pay

    2011-09-01

    Full Text Available A better understanding of the factors controlling N2 fixation is a pre-requisite for improving our knowledge on the contribution of N2 fixation process in the nitrogen cycling. Trace-metal clean nutrient/dust addition bioassays (+P, +PFe, +dust were performed at three stations located in the western, central and eastern Mediterranean Sea, in summer 2008 as part of the BOUM cruise. The main goals were (1 to investigate the nutrient factor(s limiting N2 fixation (uptake of 15N2 and (2 to evaluate the potential impact of a Saharan dust event on this biological process during the stratification period. Initially, surface waters at the three stations were DIP-depleted (2 fixation (from 130 % to 430 %. The highest dust stimulation of N2 fixation was recorded at the station located in the eastern basin. The response of diazotrophic activity to nutrient additions was variable between the sampled stations suggesting a spatial variability of the factor controlling N2 fixation over the whole basin. At all stations, N2 fixation was not limited by Fe nor co-limited by P and Fe. At the western station, N2 fixation was DIP limited while at the eastern one, N2 fixation was first DIP limited, then was limited by one or several chemical element(s released by dust. Our results demonstrated that a Saharan dust input was able to relieve these successive on going limitations. Very interestingly, at the station located in the central basin, N2 fixation was not limited by the availability of P yet it was strongly stimulated by dust addition (x3.1. A chemical element or a combination of several, released by the added dust may have been responsible for the observed stimulations of N2 fixation. These results indicated that Saharan dust pulses to the surface Mediterranean waters, in addition to P and Fe, could be a source of chemical(s element(s that are necessary for metabolic processes and therefore influence rates of N2 fixation.

  18. Paleolimnological record as an indication of incipient eutrophication in an oligotrophic subtropical coastal lake in Southern Brazil.

    Science.gov (United States)

    Hennemann, Mariana Coutinho; Simonassi, José Carlos; Petrucio, Mauricio Mello

    2015-08-01

    Paleolimnology of lake sediments can be a powerful tool to assess various aspects of lake history and catchment change through elemental, isotopic and molecular analysis of the sedimented organic matter (OM). In this sense, the objective of the present study was to investigate the source, depositional history and preservation of OM in the sediments of two different sites in Peri Lake (southern Brazil) to better understand the nature and direction of environmental changes. Therefore, two sediment cores were sampled and analysed for total organic carbon (TOC), total nitrogen (TN) and total phosphorus (TP) concentrations and elemental ratios, and stable isotope ratios of C and N (δ(13)C and δ(15)N). Both cores showed similar general tendencies, with increasing amounts of OM (range 1-35%), TOC (2.55-258.40 mg g(-1)), TN (0.30-25.97 mg g(-1)) and TP (0.03-4.72 mg g(-1)) from the bottom toward the top more recent layers. TOC:TN ratios (range 8.1-14.7) showed a slight decrease in recent times and indicated a mixture of allochthonous and autochthonous contribution to the OM, with predominance of the last source. TN:TP (range 0.2-51.3) indicated a condition of potential limitation by P in general. Both δ(13)C (range -25.58 to -20.85) and δ(15)N (range 2.6 to 7.1) showed a decreasing pattern toward the top of the cores, in opposition to macronutrient concentration. Differences in the depth variation pattern between the two cores were associated to the marginal location of one of the cores. The results suggest that nutrients and primary production are increasing in the lake.

  19. Metabolic and molecular characterization of bacterial community associated to Patagonian Chilean oligotrophic-lakes of quaternary glacial origin.

    Science.gov (United States)

    Leon, Carla; Campos, Víctor; Urrutia, Roberto; Mondaca, María-Angélica

    2012-04-01

    The Patagonian Lakes have particular environmental conditions with or without intermittent disturbances. The study of the microorganisms present in aquatic ecosystems has increased notably because they can be used as micro-scale bioindicators of, among others, anthropogenic pollution and climatic change. The aim of the work was to compare the composition of the bacterial communities associated with sediments of three Patagonian Lakes with different geomorphologic patterns and disturbances. The lake sediments were characterized by molecular techniques, physiology profiles and physico-chemical analyses. The metabolic and physiological profiles of the microbial community demonstrated that non-impacted Tranquilo Lake is statistically different to impacted Bertrand and Plomo Lakes. Similar results were detected by DGGE profiles. FISH results demonstrated that betaproteobacteria showed the highest count in the Tranquilo Lake while gammaproteobacteria showed the highest counts in the Bertrand and Plomo Lakes, indicating that their sediments are highly dystrophic. The results demonstrate differences in the metabolic activity and structural and functional composition of bacterial communities of the studied lakes, which have different geomorphological patterns due to disturbances such as volcanic activity and the climatic change.

  20. NCBI nr-aa BLAST: CBRC-GGOR-01-1350 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GGOR-01-1350 ref|NP_419199.1| beta-glucanase [Caulobacter crescentus CB15] ref...|YP_002515759.1| glucan endo-1,3-beta-glucosidase [Caulobacter crescentus NA1000] gb|AAK22367.1| beta-glucanase [Caulobacter crescent...us CB15] gb|ACL93851.1| glucan endo-1,3-beta-glucosidase [Caulobacter crescentus NA1000] NP_419199.1 3.7 27% ...

  1. Activation and polar sequestration of PopA, a c-di-GMP effector protein involved in Caulobacter crescentus cell cycle control

    DEFF Research Database (Denmark)

    Ozaki, Shogo; Schalch-Moser, Annina; Zumthor, Ludwig

    2014-01-01

    transduction in PopA with the GGDEF domain adopting input function and the receiver domain serving as regulatory output. We show that the N-terminal receiver domain of PopA specifically interacts with RcdA, a component required for CtrA degradation. In contrast, the GGDEF domain serves to target Pop...

  2. Análise funcional das proteínas HrcA, GroES/GroEL e DnaK/DnaJ em Caulobacter crescentus

    OpenAIRE

    Michelle Fernanda Susin

    2005-01-01

    O operon groESL de C. crescentus apresenta dupla regulação. A indução deste operon por choque térmico é dependente do fator sigma de choque térmico σ32. A temperaturas fisiológicas, a expressão de groESL apresenta regulação temporal durante o ciclo celular da bactéria e o controle envolve a proteína repressora HrcA e o elemento CIRCE (controlling inverted repeat of chaperonin expression). Para estudar a atividade da proteína repressora in vitro, produzimos e purificamos de E. coli a Hrc...

  3. Characteristics of dimethylaminium and trimethylaminium in atmospheric particles ranging from supermicron to nanometer sizes over eutrophic marginal seas of China and oligotrophic open oceans.

    Science.gov (United States)

    Yu, Peiran; Hu, Qingjing; Li, Kai; Zhu, Yujiao; Liu, Xiaohuan; Gao, Huiwang; Yao, Xiaohong

    2016-12-01

    In this study, we characterized dimethylaminium (DMA(+)) and trimethylaminium (TMA(+)) in size-segregated atmospheric particles during three cruise campaigns in the marginal seas of China and one cruise campaign mainly in the northwest Pacific Ocean (NWPO). An 14-stage nano-MOUDI sampler was utilized for sampling atmospheric particles ranging from 18μm to 0.010μm. Among the four cruise campaigns, the highest concentrations of DMA(+) and TMA(+) in PM10 were observed over the South Yellow Sea (SYS) in August 2015, i.e., 0.76±0.12nmolm(-3) for DMA(+) (average value±standard deviation) and 0.93±0.13nmolm(-3) for TMA(+). The lowest values were observed over the NWPO in April 2015, i.e., 0.28±0.16nmolm(-3) for DMA(+) and 0.22±0.12nmolm(-3) for TMA(+). In general, size distributions of the two ions exhibited a bi-modal pattern, i.e., one mode at 0.01-0.1μm and the other at 0.1-1.8μm. The two ions' mode at 0.01-0.1μm was firstly observed. The mode was largely enhanced in samples collected over the SYS in August 2015, leading to high mole ratios of (DMA(+)+TMA(+))/NH4(+) in PM0.1 (0.4±0.8, median value±standard deviation) and the ions' concentrations in PM0.1 accounting for ~10% and ~40% of their corresponding concentrations in PM10. This implied that (DMA(+)+TMA(+)) likely played an important role in neutralizing acidic species in the smaller particles. Using SO4(2-), NO3(-) and NH4(+) as references, we confirm that the elevated concentrations of DMA(+) and TMA(+) in the 0.01-0.1μm size range were probably real signals rather than sampling artifacts.

  4. Plankton assembly in an ultra-oligotrophic Antarctic lake over the summer transition from the ice-cover to ice-free period: A size spectra approach

    Science.gov (United States)

    Rochera, Carlos; Quesada, Antonio; Toro, Manuel; Rico, Eugenio; Camacho, Antonio

    2017-03-01

    Lakes from the Antarctic maritime region experience climate change as a main stressor capable of modifying their plankton community structure and function, essentially because summer temperatures are commonly over the freezing point and the lake's ice cap thaws. This study was conducted in such seasonally ice-covered lake (Lake Limnopolar, Byers Peninsula, Livingston Is., Antarctica), which exhibits a microbial dominated pelagic food web. An important feature is also the occurrence of benthic mosses (Drepanocladus longifolius) covering the lake bottom. Plankton dynamics were investigated during the ice-thawing transition to the summer maximum. Both bacterioplankton and viral-like particles were higher near the lake's bottom, suggesting a benthic support. When the lake was under dim conditions because of the snow-and-ice cover, autotrophic picoplankters dominated at deep layers. The taxa-specific photopigments indicated dominance of picocyanobacteria among them when the light availability was lower. By contrast, larger and less edible phytoplankton dominated at the onset of the ice melting. The plankton size spectra were fitted to the continuous model of Pareto distribution. Spectra evolved similarly at two sampled depths, in surface and near the bottom, with slopes increasing until mid-January. However, slopes were less steep (i.e., size classes more uniformly distributed) at the bottom, thus denoting a more efficient utilization of resources. These findings suggest that microbial loop pathways in the lake are efficiently channelized during some periods to the metazoan production (mainly the copepod Boeckella poppei). Our results point to that trophic interactions may still occur in these lakes despite environmental harshness. This results of interest in a framework of increasing temperatures that may reduce the climatic restrictions and therefore stimulate biotic interactions.

  5. Incorporation of inorganic mercury (Hg{sup 2+}) in pelagic food webs of ultraoligotrophic and oligotrophic lakes: The role of different plankton size fractions and species assemblages

    Energy Technology Data Exchange (ETDEWEB)

    Soto Cárdenas, Carolina, E-mail: sotocardenascaro@gmail.com [Laboratorio de Fotobiología, Instituto de Investigaciones en Biodiversidad y Medioambiente (INIBIOMA, UNComahue-CONICET), Quintral 1250, 8400 San Carlos de Bariloche, Río Negro (Argentina); Diéguez, Maria C. [Laboratorio de Fotobiología, Instituto de Investigaciones en Biodiversidad y Medioambiente (INIBIOMA, UNComahue-CONICET), Quintral 1250, 8400 San Carlos de Bariloche, Río Negro (Argentina); Ribeiro Guevara, Sergio [Laboratorio de Análisis por Activación Neutrónica, CAB, CNEA, Av. Bustillo Km 9.5, 8400, San Carlos de Bariloche, Río Negro (Argentina); Marvin-DiPasquale, Mark [United States Geological Survey, 345 Middlefield Rd./MS 480, Menlo Park, CA 94025 (United States); Queimaliños, Claudia P. [Laboratorio de Fotobiología, Instituto de Investigaciones en Biodiversidad y Medioambiente (INIBIOMA, UNComahue-CONICET), Quintral 1250, 8400 San Carlos de Bariloche, Río Negro (Argentina)

    2014-10-01

    In lake food webs, pelagic basal organisms such as bacteria and phytoplankton incorporate mercury (Hg{sup 2+}) from the dissolved phase and pass the adsorbed and internalized Hg to higher trophic levels. This experimental investigation addresses the incorporation of dissolved Hg{sup 2+} by four plankton fractions (picoplankton: 0.2–2.7 μm; pico + nanoplankton: 0.2–20 μm; microplankton: 20–50 μm; and mesoplankton: 50–200 μm) obtained from four Andean Patagonian lakes, using the radioisotope {sup 197}Hg{sup 2+}. Species composition and abundance were determined in each plankton fraction. In addition, morphometric parameters such as surface and biovolume were calculated using standard geometric models. The incorporation of Hg{sup 2+} in each plankton fraction was analyzed through three concentration factors: BCF (bioconcentration factor) as a function of cell or individual abundance, SCF (surface concentration factor) and VCF (volume concentration factor) as functions of individual exposed surface and biovolume, respectively. Overall, this investigation showed that through adsorption and internalization, pico + nanoplankton play a central role leading the incorporation of Hg{sup 2+} in pelagic food webs of Andean lakes. Larger planktonic organisms included in the micro- and mesoplankton fractions incorporate Hg{sup 2+} by surface adsorption, although at a lesser extent. Mixotrophic bacterivorous organisms dominate the different plankton fractions of the lakes connecting trophic levels through microbial loops (e.g., bacteria–nanoflagellates–crustaceans; bacteria–ciliates–crustaceans; endosymbiotic algae–ciliates). These bacterivorous organisms, which incorporate Hg from the dissolved phase and through their prey, appear to explain the high incorporation of Hg{sup 2+} observed in all the plankton fractions. - Highlights: • Hg{sup 2+} incorporation in lake plankton fractions was studied using the isotope {sup 197}Hg{sup 2+}. • Hg{sup 2+} incorporation was assessed using three different bioconcentration factors. • Bioconcentration factors related with fraction cell abundance, surface and biovolume. • Picoplankton and nanoplankton lead the incorporation of Hg{sup 2+} in pelagic food webs. • Mixotrophic species appear to enhance Hg incorporation via bacteria feeding.

  6. Incorporation of inorganic mercury (Hg2+) in pelagic food webs of ultraoligotrophic and oligotrophic lakes: the role of different plankton size fractions and species assemblages

    Science.gov (United States)

    Soto Cárdenas, Carolina; Diéguez, Maria C.; Ribeiro Guevara, Sergio; Marvin-DiPasquale, Mark; Queimaliños, Claudia P.

    2014-01-01

    In lake food webs, pelagic basal organisms such as bacteria and phytoplankton incorporate mercury (Hg2+) from the dissolved phase and pass the adsorbed and internalized Hg to higher trophic levels. This experimental investigation addresses the incorporation of dissolved Hg2+ by four plankton fractions (picoplankton: 0.2–2.7 μm; pico + nanoplankton: 0.2–20 μm; microplankton: 20–50 μm; and mesoplankton: 50–200 μm) obtained from four Andean Patagonian lakes, using the radioisotope 197Hg2+. Species composition and abundance were determined in each plankton fraction. In addition, morphometric parameters such as surface and biovolume were calculated using standard geometric models. The incorporation of Hg2+ in each plankton fraction was analyzed through three concentration factors: BCF (bioconcentration factor) as a function of cell or individual abundance, SCF (surface concentration factor) and VCF (volume concentration factor) as functions of individual exposed surface and biovolume, respectively. Overall, this investigation showed that through adsorption and internalization, pico + nanoplankton play a central role leading the incorporation of Hg2+ in pelagic food webs of Andean lakes. Larger planktonic organisms included in the micro- and mesoplankton fractions incorporate Hg2+ by surface adsorption, although at a lesser extent. Mixotrophic bacterivorous organisms dominate the different plankton fractions of the lakes connecting trophic levels through microbial loops (e.g., bacteria–nanoflagellates–crustaceans; bacteria–ciliates–crustaceans; endosymbiotic algae–ciliates). These bacterivorous organisms, which incorporate Hg from the dissolved phase and through their prey, appear to explain the high incorporation of Hg2+ observed in all the plankton fractions.

  7. Spatial and temporal changes in the concentration of various phosphorus pools and their possible biogeochemical roles in the oligotrophic subtropical western North Pacific

    Science.gov (United States)

    Lee, Sangkyun; Hama, Takeo; Ishii, Masao; Saito, Shu; Yanagi, Katsumi

    2008-03-01

    Spatial and temporal variations in the concentration of phosphorus pools, including total phosphorus (TP), reactive phosphorus (RP), and nonreactive phosphorus (NP), were evaluated in subtropical regions (10-30°N) of the western North Pacific Ocean along 137°E through eight sampling periods from summer 2003 to spring 2005. RP was depleted at the water surface throughout our observation, varying less than 0.1 μM. The low concentration of RP was restricted to the surface mixing layer, and the concentration obviously increased concomitant with the decrease in water temperature. NP concentration was generally highest at the water surface and gradually decreased with increasing depth, but the depth and temporal variations were not definite compared with those of RP. NP was further divided into two fractions depending on its reactivity to specific ultraviolet (UV) irradiation. The concentration of UV labile organic phosphorus (UVL OP) was consistently low, being comparable with that of RP; their inventories from 0 to 50 m of RP and UVL OP fluctuated with in the range of 0.9-3.2 and 2.0-3.1 mmol m-2, respectively. As for incubation experiments in coastal waters using glucose-1-phosphate, which is fractionated into UVL OP, the UVL OP most likely has a short turnover time due to rapid utilization by microorganisms, indicating a significant role in the phosphorus cycle through the microbial food web. Inventory of the UV stable OP (UVS OP), on the other hand, varied nearly eightfold in the upper 50 m, and the temporal change in TP inventory was exclusively due to that of UVS OP.

  8. Short-term changes in particulate fluxes measured by drifting sediment traps during end summer oligotrophic regime in the NW Mediterranean Sea

    Directory of Open Access Journals (Sweden)

    J. C. Marty

    2009-05-01

    Full Text Available Short-term changes in the flux of particulate matter were determined in the central north western Mediterranean Sea (near DYFAMED site using drifting sediment traps at 200 m depth in the course of the DYNAPROC 2 cruise (14 September–17 October 2004. In this period of marked water column stratification, POC fluxes varied by an order of magnitude, in the range of 0.03–0.29 mgC m−2 h−1 over the month and showed very rapid and high variations. Particulate carbon export represented less than 5% of integrated primary production, suggesting that phytoplankton production was essentially sustained by internal recycling of organic matter and retained within the photic zone. While PON and POP fluxes paralleled one another, the elemental ratios POC/PON and POC/POP, varied widely over short-term periods. Values of these ratios generally higher than the conventional Redfield ratio, together with the very low chlorophyll a flux recorded in the traps (mean 0.017 μg m−2 h−1, and the high phaeopigment and acyl lipid hydrolysis metabolite concentrations of the settling material, indicated that the organic matter reaching 200 m depth was reworked (by grazing, fecal pellets production, degradation and that algal sinking, dominated by nano- and picoplankton, made a small contribution to the downward flux. Over time, the relative abundance of individual lipid classes in organic matter (OM changed from glycolipids-dominated to neutral (wax esters, triacylglycerols and phospholipids-dominated, suggesting ecosystem maturation as well as rapid and continual exchanges between dissolved, suspended and sinking pools. Our most striking result was documenting the rapid change in fluxes of the various measured parameters. In the situation encountered here, with dominant regenerated production, a decrease of fluxes was noticed during windy periods (possibly through reduction of grazing. But fluxes increased as soon as calm conditions settle.

  9. Degradation of pyrene and characterization of Saccharothrix sp. PYX-6 from the oligotrophic Tianchi Lake in Xinjiang Uygur Autonomous Region, China

    Institute of Scientific and Technical Information of China (English)

    HU Yuting; REN Fenghua; ZHOU Peijin; XIA Min; LIU Shuangjiang

    2003-01-01

    Bacterial strain PYX-6 that utilizes anthracene, phenanthrene, or pyrene for carbon and energy sources for growth was isolated from a non-polluted lake (Tianchi Lake) in Xinjiang Uygur Autonomons Region of China. Its morphology, physiological and biochemical properties, cell wall pattern and G+C mol% content of DNA molecules were characterized. The 16S rRNA gene of strain PYX-6 was sequenced and analyzed for similarities to related bacterial species. Results indicated that strain PYX-6 is a member of the Genus Saccharothrix, and the strain was named Saccharothrix sp. PYX-6. When pyrene was the sole carbon source in cultural medium, the strain PYX-6 assimilated pyrene for growth and 0.005% of yeast extract stimulated pyrene degradation and assimilation. The optimal pH of cultural medium and the optimal shaking frequency during cultivation were 6-8 and 200 r/m, respectively. It was found that the disappearance of pyrene in medium occurred before significant growth of strain PYX-6 took place. Phthalic acid, benzylacetic acid, and benzylpropenoic acid were detected as catabolic intermediates during pyrene degradation with mass spectroscopy and this result indicated that Saccharothrix sp. PYX-6 adopted a pathway that is different from the pathway of the previously reported pyrene-degrading Mycobacterium sp. PYR-1.

  10. Non-destructive methods for peat layer assessment in oligotrophic peat bogs: a case study from Poiana Ştampei, Romania

    Directory of Open Access Journals (Sweden)

    Iuliana F. Gheorghe

    2011-01-01

    Full Text Available Practices currently employed in the investigation and characterisation of peat deposits are destructive and may irremediable perturb peat bog development even in cases when exploitation is not carried out. We investigated the correlation between vegetation characteristics in the active area of Poiana Ştampei peat bog, Romania, and the underlying peat layer depth, aiming at establishing a non-destructive method of peat layer depth estimation. The presence of the Sphagneto-Eriophoretum vaginati association, dominated by Sphagnum fimbriatum, Eriophorum vaginatum, Andromeda polifolia, Vaccinium oxycoccos, V. myrtillus, V. vitis-idaea, Polytrichum commune, Picea excelsa, Pinus sylvestris and Betula verrucosa was found to predict the existence of the peat layer but not its depth. Out of the seven identified vegetation types, one type was associated with a very thin or no peat layer, one type was characterised by the presence of a thick (over 100 cm peat layer and five types indicated the presence of variable average depths of the peat layer. pH values correlated with peat layer depth only within the vegetation type associated with thick peat layers.

  11. Environment and phenology: CO2 net ecosystem exchange and CO2 flux partitioning at an acid and oligotrophic mire system in northern Sweden

    Science.gov (United States)

    Gažovič, Michal; Peichl, Matthias; Vermeij, Ilse; Limpens, Juul; Nilsson, Mats. B.

    2015-04-01

    Static chamber and environmental measurements in combination with vegetation indices (i.e. vascular green area (VGA) and the greenness chromatic color index (gcc) derived from digital camera images) were used to investigate effects of environment and phenology on the CO2 net ecosystem exchange (NEE) and CO2 flux partitioning at the Degerö Stormyr site in northern Sweden (64°11' 23.565" N, 19°33' 55.291 E) during two environmentally different years. Our measurement design included a control plot, a moss plot (where vascular plants were removed by clipping) and four heterotrophic respiration (RH) collars (where all green moss and vascular plant biomass were removed) to partition between soil heterotrophic and plant autotrophic (moss and vascular plants) respiration (RA), as well as between moss and vascular plant gross primary production (GPP). Environmental conditions, especially the shallow snow cover, peat soil frost and cold spring in 2014 caused delayed onset of spring green up, reduced soil respiration flux and reduced GPP of vascular plants. Soil temperature measured in 26 cm depth started to rise from spring temperatures of ~ 0.6 °C in 2013 and 0.15 °C in 2014 about 20 days earlier in 2013 compared to 2014. With earlier onset of the growing season and higher soil temperatures in 2013, heterotrophic soil respiration was higher in year 2013 than in year 2014. In 2013, RH dominated the total ecosystem respiration in all months but June and August. On contrary, autotrophic respiration dominated ecosystem respiration in all months of 2014. In both years, vascular plants and mosses were more or less equally contributing to autotrophic respiration. We measured higher GPP in year 2013 compared to year 2014. Also VGA and gcc were higher in spring and throughout the rest of 2013 compared to 2014. The onset of VGA was delayed by ~ 10 days in 2014. In general, total GPP was dominated by GPP of vascular plants in both years, although moss GPP had substantial contribution and exceeded GPP of vascular plants in June of 2013 and July and September of 2014, respectively. NEE was lower (i.e. greater net CO2 uptake) in 2013 compared to 2014. Our study highlights the importance of phenology in relation to ongoing environmental change, which in turn exerts a strong control on the CO2 exchange and its partitioning into component fluxes.

  12. Complexity of diatom response to Lateglacial and Holocene climate and environmental change in ancient, deep and oligotrophic Lake Ohrid (Macedonia and Albania)

    Science.gov (United States)

    Zhang, X. S.; Reed, J. M.; Lacey, J. H.; Francke, A.; Leng, M. J.; Levkov, Z.; Wagner, B.

    2016-03-01

    Lake Ohrid (Macedonia and Albania) is a rare example of a deep, ancient Mediterranean lake and is a key site for palaeoclimate research in the northeastern Mediterranean region. This study conducts the analysis of diatoms as a proxy for Lateglacial and Holocene climate and environmental change in Lake Ohrid at a higher resolution than in previous studies. While Lake Ohrid has the potential to be sensitive to water temperature change, the data demonstrate a highly complex diatom response, probably comprising a direct response to temperature-induced lake productivity in some phases and an indirect response to temperature-related lake stratification or mixing and epilimnetic nutrient availability in others. The data also demonstrate the possible influence of physical limnological (e.g. the influence of wind stress on stratification or mixing) and chemical processes (e.g. the influence of catchment dynamics on nutrient input) in mediating the complex response of diatoms. During the Lateglacial (ca. 12 300-11 800 cal yr BP), the low-diversity dominance of hypolimnetic Cyclotella fottii indicates low lake productivity, linked to low water temperature. Although the subsequent slight increase in small, epilimnetic C. minuscula during the earliest Holocene (ca. 11 800-10 600 cal yr BP) suggests climate warming and enhanced stratification, diatom concentration remains as low as during the Lateglacial, suggesting that water temperature increase was muted across this major transition. The early Holocene (ca. 10 600-8200 cal yr BP) is characterised by a sustained increase in epilimnetic taxa, with mesotrophic C. ocellata indicating high water-temperature-induced productivity between ca. 10 600-10 200 cal yr BP and between ca. 9500-8200 cal yr BP and with C. minuscula in response to low nutrient availability in the epilimnion between ca. 10 200-9500 cal yr BP. During the middle Holocene (ca. 8200-2600 cal yr BP), when sedimentological and geochemical proxies provide evidence for maximum Holocene water temperature, anomalously low C. ocellata abundance is probably a response to epilimnetic nutrient limitation, almost mimicking the Lateglacial flora apart from the occurrence of mesotrophic Stephanodiscus transylvanicus in the hypolimnion. During the late Holocene (ca. 2600 cal yr BP-present), high abundance and fluctuating composition of epilimnetic taxa are probably a response more to enhanced anthropogenic nutrient input, particularly nitrogen enrichment, than to climate. Overall, the data indicate that previous assumptions concerning the linearity of diatom response in this deep, ancient lake are invalid, and multi-proxy analysis is essential to improve understanding of palaeolimnological dynamics in future research on the long, Quaternary sequence.

  13. Coupled dynamics of iron and phosphorus in sediments of an oligotrophic coastal basin and the impact of anaerobic oxidation of methane

    NARCIS (Netherlands)

    Slomp, C.P.; Mort, H.P.; Jilbert, T.; Reed, D.C.; Gustafsson, B.G.; Wolthers, M.

    2013-01-01

    Studies of phosphorus (P) dynamics in surface sediments of lakes and coastal seas typically emphasize the role of coupled iron (Fe), sulfur (S) and P cycling for sediment P burial and release. Here, we show that anaerobic oxidation of methane (AOM) also may impact sediment P cycling in such systems.

  14. A comparative analysis of the cultured micromycetes in oligotrophic peatlands of natural biosphere reservations located in the northern and central parts of Russia

    Directory of Open Access Journals (Sweden)

    Elena N. Bilanenko

    2016-08-01

    Full Text Available The current study addresses the fungal diversity of the peatlands that vary geographically and geologically, in the central and northern parts of Russia. The central (Smolenskaya area, National Park «Smolenskoye Poozerie» and northern (White Sea Biological Station area peatlands have a different geological history, the former have a glacial origin, while the latter are of marine origin. Our aim was to investigate the fungal biota of these zones full of Sphagnum moss, and possibly reveal the species that are pertinent to these particular habitats. In both investigated areas, we found an overall poor species diversity with a high fraction of sterile mycelia. Many penicillia species were dominating in all peat samples – P. thomii, P. spinulosum, P. glabrum, P. funiculosum, P. aurantiogriseum (together with Geotrichum candidum, many of them are known to degrade Sphagnum. We were particularly interested in the dynamics of the species from the genus Oidiodendron that have been shown to be involved in primary Sphagnum degradation. The northern area contained a larger species diversity of Oidiodendron, as compared to the central zone. Interestingly, insect-associated species of the genus Tolypocladium and Beauveria, along with the psychrotolerant species of Geomyces and Tolypocladium were recovered only from the northern peatlands. Aspergillus spp. were found in peatlands of the central zone only. We link this result to the climatic features of the area, but also different invertebrate contents that may be utilized by fungi. We detected a tendency of an increasing diversity of Oidiodendron species, psychrotolerant and entomopathogenic fungal species as well as a proportion of sterile forms in the northern area.

  15. Bacteria, fungi and biokarst in Lechuguilla Cave, Carlsbad Caverns National Park, New Mexico

    Science.gov (United States)

    Cunningham, K. I.; Northup, D. E.; Pollastro, R. M.; Wright, W. G.; Larock, E. J.

    1995-02-01

    Lechuguilla Cave is a deep, extensive, gypsumand sulfur-bearing hypogenic cave in Carlsbad Caverns National Park, New Mexico, most of which (>90%) lies more than 300 m beneath the entrance. Located in the arid Guadalupe Mountains, Lechuguilla's remarkable state of preservation is partially due to the locally continuous Yates Formation siltstone that has effectively diverted most vadose water away from the cave. Allocthonous organic input to the cave is therefore very limited, but bacterial and fungal colonization is relatively extensive: (1) Aspergillus sp. fungi and unidentified bacteria are associated with iron-, manganese-, and sulfur-rich encrustations on calcitic folia near the suspected water table 466 m below the entrance; (2) 92 species of fungi in 19 genera have been identified throughout the cave in oligotrophic (nutrient-poor) “soils” and pools; (3) cave-air condensate contains unidentified microbes; (4) indigenous chemoheterotrophic Seliberius and Caulobacter bacteria are known from remote pool sites; and (5) at least four genera of heterotrophic bacteria with population densities near 5×105 colony-forming units (CFU) per gram are present in ceiling-bound deposits of supposedly abiogenic condensation-corrosion residues. Various lines of evidence suggest that autotrophic bacteria are present in the ceiling-bound residues and could act as primary producers in a unique subterranean microbial food chain. The suspected autotrophic bacteria are probably chemolithoautotrophic (CLA), utilizing trace iron, manganese, or sulfur in the limestone and dolomitic bedrock to mechanically (and possibly biochemically) erode the substrate to produce residual floor deposits. Because other major sources of organic matter have not been detected, we suggest that these CLA bacteria are providing requisite organic matter to the known heterotrophic bacteria and fungi in the residues. The cavewide bacterial and fungal distribution, the large volumes of corrosion residues

  16. Frequency of infection with A and B supergroup Wolbachia in insects and pests associated with mulberry and silkworm

    Indian Academy of Sciences (India)

    B M Prakash; H P Puttaraju

    2007-06-01

    Wolbachia is a ubiquitous, Gram-negative, vertically transmitted, alpha-proteobacterium that causes an array of reproductive abnormalities including cytoplasmic incompatibility, feminization of genetic males, parthenogenesis in a number of insect species, among others. Wolbachia is now being exploited as an agent for pest and vector control. Previous surveys indicated that it is commonly seen in 16–76% of arthropods. In this paper, using polymerase chain reaction assay based on specific amplification of the ftsZ-A and -B supergroup Wolbachia gene fragments, we found that 30% of insects and pests screened were positive for Wolbachia. Among them 66.7% harbour double Wolbachia infection, while 33.3% harbour single Wolbachia infection. These results indicate widespread infection with both double and single Wolbachia, and provide a wealth of information to exploit this endobacterium for the management of pests and vectors.

  17. Exploring mitochondrial evolution and metabolism organization principles by comparative analysis of metabolic networks.

    Science.gov (United States)

    Chang, Xiao; Wang, Zhuo; Hao, Pei; Li, Yuan-Yuan; Li, Yi-Xue

    2010-06-01

    The endosymbiotic theory proposed that mitochondrial genomes are derived from an alpha-proteobacterium-like endosymbiont, which was concluded from sequence analysis. We rebuilt the metabolic networks of mitochondria and 22 relative species, and studied the evolution of mitochondrial metabolism at the level of enzyme content and network topology. Our phylogenetic results based on network alignment and motif identification supported the endosymbiotic theory from the point of view of systems biology for the first time. It was found that the mitochondrial metabolic network were much more compact than the relative species, probably related to the higher efficiency of oxidative phosphorylation of the specialized organelle, and the network is highly clustered around the TCA cycle. Moreover, the mitochondrial metabolic network exhibited high functional specificity to the modules. This work provided insight to the understanding of mitochondria evolution, and the organization principle of mitochondrial metabolic network at the network level.

  18. Simultaneous nutrients and carbon removal from low-strength domestic wastewater with an immobilised-microorganism biological aerated filter.

    Science.gov (United States)

    Chen, Q; Qu, L; Tong, G; Ni, J

    2011-01-01

    To improve the efficiency of low-strength domestic wastewater treatment, an immobilised-microorganism biological aerated filter (I-BAF) was established for simultaneous carbon, nitrogen and phosphorus removal. The I-BAF performance was systematically evaluated under continuous and intermittent aeration modes. At the optimal condition with an intermittent aeration control schedule of 2 h on/1 h off, the maximum removal rates of COD, NH(4)(+)-N, TN and P were 82.54%, 94.83%, 51.85% and 61.49%, respectively, and the corresponding averaged effluents could meet the first class standards of China. Further analysis of PCR-DGGE profile revealed that members of the gamma and alpha proteobacterium bacterial groups were probably responsible for the nitrogen and phosphorus removal. The I-BAF system showed excellent performance in carbon and nutrients removal, which provided a cost-effective solution for the treatment of low-strength domestic wastewater.

  19. Wolbachia infections in natural Anopheles populations affect egg laying and negatively correlate with Plasmodium development

    Science.gov (United States)

    Shaw, W. Robert; Marcenac, Perrine; Childs, Lauren M.; Buckee, Caroline O.; Baldini, Francesco; Sawadogo, Simon P.; Dabiré, Roch K.; Diabaté, Abdoulaye; Catteruccia, Flaminia

    2016-01-01

    The maternally inherited alpha-proteobacterium Wolbachia has been proposed as a tool to block transmission of devastating mosquito-borne infectious diseases like dengue and malaria. Here we study the reproductive manipulations induced by a recently identified Wolbachia strain that stably infects natural mosquito populations of a major malaria vector, Anopheles coluzzii, in Burkina Faso. We determine that these infections significantly accelerate egg laying but do not induce cytoplasmic incompatibility or sex-ratio distortion, two parasitic reproductive phenotypes that facilitate the spread of other Wolbachia strains within insect hosts. Analysis of 221 blood-fed A. coluzzii females collected from houses shows a negative correlation between the presence of Plasmodium parasites and Wolbachia infection. A mathematical model incorporating these results predicts that infection with these endosymbionts may reduce malaria prevalence in human populations. These data suggest that Wolbachia may be an important player in malaria transmission dynamics in Sub-Saharan Africa. PMID:27243367

  20. Sex and the eukaryotic cell cycle is consistent with a viral ancestry for the eukaryotic nucleus.

    Science.gov (United States)

    Bell, Philip John Livingstone

    2006-11-07

    The origin of the eukaryotic cell cycle, including mitosis, meiosis, and sex are as yet unresolved aspects of the evolution of the eukaryotes. The wide phylogenetic distribution of both mitosis and meiosis suggest that these processes are integrally related to the origin of the earliest eukaryotic cells. According to the viral eukaryogenesis (VE) hypothesis, the eukaryotes are a composite of three phylogenetically unrelated organisms: a viral lysogen that evolved into the nucleus, an archaeal cell that evolved into the eukaryotic cytoplasm, and an alpha-proteobacterium that evolved into the mitochondria. In the extended VE hypothesis presented here, the eukaryotic cell cycle arises as a consequence of the derivation of the nucleus from a lysogenic DNA virus.

  1. Genetic Diversity Affects the Daily Transcriptional Oscillations of Marine Microbial Populations.

    Science.gov (United States)

    Shilova, Irina N; Robidart, Julie C; DeLong, Edward F; Zehr, Jonathan P

    2016-01-01

    Marine microbial communities are genetically diverse but have robust synchronized daily transcriptional patterns at the genus level that are similar across a wide variety of oceanic regions. We developed a microarray-inspired gene-centric approach to resolve transcription of closely-related but distinct strains/ecotypes in high-throughput sequence data. Applying this approach to the existing metatranscriptomics datasets collected from two different oceanic regions, we found unique and variable patterns of transcription by individual taxa within the abundant picocyanobacteria Prochlorococcus and Synechococcus, the alpha Proteobacterium Pelagibacter and the eukaryotic picophytoplankton Ostreococcus. The results demonstrate that marine microbial taxa respond differentially to variability in space and time in the ocean. These intra-genus individual transcriptional patterns underlie whole microbial community responses, and the approach developed here facilitates deeper insights into microbial population dynamics.

  2. Identifying the bacterial community on the surface of Intralox belting in a meat boning room by culture-dependent and culture-independent 16S rDNA sequence analysis.

    Science.gov (United States)

    Brightwell, Gale; Boerema, Jackie; Mills, John; Mowat, Eilidh; Pulford, David

    2006-05-25

    We examined the bacterial community present on an Intralox conveyor belt system in an operating lamb boning room by sequencing the 16S ribosomal DNA (rDNA) of bacteria extracted in the presence or absence of cultivation. RFLP patterns for 16S rDNA clone library and cultures were generated using HaeIII and MspI restriction endonucleases. 16S rDNA amplicons produced 8 distinct RFLP pattern groups. RFLP groups I-IV were represented in the clone library and RFLP groups I and V-VIII were represented amongst the cultured isolates. Partial DNA sequences from each RFLP group revealed that all group I, II and VIII representatives were Pseudomonas spp., group III were Sphingomonas spp., group IV clones were most similar to an uncultured alpha proteobacterium, group V was similar to a Serratia spp., group VI with an Alcaligenes spp., and group VII with Microbacterium spp. Sphingomonads were numerically dominant in the culture-independent clone library and along with the group IV alpha proteobacterium were not represented amongst the cultured isolates. Serratia, Alcaligenes and Microbacterium spp. were only represented with cultured isolates. Pseudomonads were detected by both culture-dependent (84% of isolates) and culture-independent (12.5% of clones) methods and their presence at high frequency does pose the risk of product spoilage if transferred onto meat stored under aerobic conditions. The detection of sphingomonads in large numbers by the culture-independent method demands further analysis because sphingomonads may represent a new source of meat spoilage that has not been previously recognised in the meat processing environment. The 16S rDNA collections generated by both methods were important at representing the diversity of the bacterial population associated with an Intralox conveyor belt system.

  3. NCBI nr-aa BLAST: CBRC-CFAM-07-0011 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CFAM-07-0011 ref|NP_421849.1| hypothetical protein CC_3055 [Caulobacter crescent...us CB15] gb|AAK25017.1| hypothetical protein CC_3055 [Caulobacter crescentus CB15] NP_421849.1 4e-06 28% ...

  4. NCBI nr-aa BLAST: CBRC-MLUC-01-0541 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MLUC-01-0541 ref|NP_421849.1| hypothetical protein CC_3055 [Caulobacter crescent...us CB15] gb|AAK25017.1| hypothetical protein CC_3055 [Caulobacter crescentus CB15] NP_421849.1 5e-05 27% ...

  5. NCBI nr-aa BLAST: CBRC-XTRO-01-0132 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0132 ref|NP_419922.1| hypothetical protein CC_1106 [Caulobacter crescent...us CB15] gb|AAK23090.1| conserved hypothetical protein [Caulobacter crescentus CB15] NP_419922.1 4e-19 26% ...

  6. NCBI nr-aa BLAST: CBRC-LAFR-01-3885 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-3885 ref|NP_422131.1| peptidase, M20/M25/M40 family [Caulobacter crescent...us CB15] gb|AAK25299.1| peptidase, M20/M25/M40 family [Caulobacter crescentus CB15] NP_422131.1 8.2 27% ...

  7. NCBI nr-aa BLAST: CBRC-TNIG-22-0088 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TNIG-22-0088 ref|ZP_01419722.1| Penicillin-binding protein 1C [Caulobacter sp. K31] gb|EAU12387.1| Peni...cillin-binding protein 1C [Caulobacter sp. K31] ZP_01419722.1 0.072 30% ...

  8. Nitrógeno: elemento limitante para el crecimiento fitoplanctónico en un lago oligotrófico tropical Nitrogen: The limiting nutrient for phytoplankton growth in an oligotrophic, tropical lake

    Directory of Open Access Journals (Sweden)

    Erik Ramos-Higuera

    2008-08-01

    Full Text Available El presente estudio tuvo como propósito evaluar si las concentraciones naturales de fósforo y nitrógeno limitaban la biomasa fitoplanctónica en el Lago Alchichica. Se aplicaron tratamientos de enriquecimiento con nitrógeno, fósforo y nitrógeno + fósforo durante las dos etapas características de la hidrodinámica del lago: estratificación y mezcla. Se dio seguimiento al potencial de crecimiento algal mediante la evaluación fluorométrica in vivo de los cambios diarios de la concentración de clorofila a expresados como porcentaje de cambio relativo con respecto al testigo (T. Durante la época de mezcla la respuesta al enriquecimiento fue limitada y la comunidad de fitoplancton no modificó su composición ni abundancia. Contrariamente, durante la época de estratificación la adición de nutrimentos favoreció el crecimiento del fitoplancton modificando su composición y abundancia. Los resultados muestran que el nitrógeno es el nutrimento que principalmente podría limitar el crecimiento algal en el lago Alchichica como ha sido encontrado en otros lagos tropicales, a diferencia de los lagos templados en donde se ha encontrado que el nutriente limitante es el fósforo. La adición de un solo nutrimento produjo sólo un breve incremento en el crecimiento en tanto el otro nutrimento se agotó. Sin embargo, cuando ambos nutrientes fueron añadidos, el crecimiento fue sostenido y se produjo una respuesta importante y detectable.The purpose of the present study was to evaluate if the available phosphorus and nitrogen concentrations were limiting the phytoplankton biomass in Lake Alchichica. Here we present a series of nutrient enrichment experiments (nitrogen, phosphorus, nitrogen + phosphorus conducted in the two characteristic stages of the lake's hydrodynamics: stratification and mixing. We followed up the algal growth potential through in vivo fluorometric analysis of chlorophyll a changes expressed as relative change percentage regarding the control. The nutrient enrichment experiment during the circulation period did not modify significantly the phytoplankton growth or the species. Opposite, during the stratification period, the nutrient addition promoted the phytoplankton growth significantly by modifying its composition and abundance. The results showed that nitrogen is the most likely nutrient limiting phytoplankton growth in Lake Alchichica as has been found in other tropical lakes, opposite to temperate lakes where phosphorus has been found to be the limiting nutrient. The enrichment of one nutrient without the other produced only a brief growth enhancement until depletion of the other nutrient occured. However, when both nitrogen and phosphorus were added, enhanced algal growth were sustained and produced substantial and detectable responses.

  9. Fiscal 1997 report on the survey of biological CO2 fixation using arid land and oligotrophic waters; 1997 nendo chosa hokokusho (kansochi, hin`eiyo kaiiki wo riyoshita seibutsuteki CO2 kotei ni kansuru chosa)

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-03-01

    This survey is aimed to investigate various measures to be taken for biological CO2 fixation, to synthetically study feasibilities of the measures from various aspects of CO2 fixation mechanism, scale, speed, and environmental effects and technical problems in case of introducing those, and to assess the measures quantitatively. In this fiscal year, a study was proceeded with of possibilities of carbon fixation by afforestation and that by fertilization into ocean. The paper defined significance of afforestation in arid land, and especially advantages in conducting researches in West Australia. Relationships were examined among afforestation, precipitation and topography. The result of the survey was described of water- and salt-transfer simulation methods. Studies of arid land were made in terms of photosynthetic speed, transpiration speed, soil characteristics, measuring methods for precipitation and vaporization amount, and the examples. Seven places of Leonora where water source and water quality were examined were selected, and the measuring results were described. The paper summed up the state of utilization of biomass energy obtained from forest and commented on a scenario on tree-planting. Finally, a possibility was stated of the carbon fixation by fermentation into ocean. 178 refs., 121 figs., 53 tabs.

  10. The Genome of the Obligately Intracellular Bacterium Ehrlichia canis Reveals Themes of Complex Membrane Structure and Immune Evasion Strategies

    Energy Technology Data Exchange (ETDEWEB)

    Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Doyle, C Kuyler [Center for Biodenfense and Emerging Infectious Diseases; Lykidis, A [U.S. Department of Energy, Joint Genome Institute; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Francino, M P [U.S. Department of Energy, Joint Genome Institute; Chain, Patrick S [ORNL; Shin, M [U.S. Department of Energy, Joint Genome Institute; Malfatti, Stephanie [Lawrence Livermore National Laboratory (LLNL); Larimer, Frank W [ORNL; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Detter, J C [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Richardson, P M [U.S. Department of Energy, Joint Genome Institute; Yu, X J [Center for Biodenfense and Emerging Infectious Diseases; Walker, D H [Center for Biodenfense and Emerging Infectious Diseases; McBride, J W [Center for Biodenfense and Emerging Infectious Diseases; Kyripides, N C [U.S. Department of Energy, Joint Genome Institute

    2006-01-01

    Ehrlichia canis, a small obligately intracellular, tick-transmitted, gram-negative, {alpha}-proteobacterium, is the primary etiologic agent of globally distributed canine monocytic ehrlichiosis. Complete genome sequencing revealed that the E. canis genome consists of a single circular chromosome of 1,315,030 bp predicted to encode 925 proteins, 40 stable RNA species, 17 putative pseudogenes, and a substantial proportion of noncoding sequence (27%). Interesting genome features include a large set of proteins with transmembrane helices and/or signal sequences and a unique serine-threonine bias associated with the potential for O glycosylation that was prominent in proteins associated with pathogen-host interactions. Furthermore, two paralogous protein families associated with immune evasion were identified, one of which contains poly(G-C) tracts, suggesting that they may play a role in phase variation and facilitation of persistent infections. Genes associated with pathogen-host interactions were identified, including a small group encoding proteins (n = 12) with tandem repeats and another group encoding proteins with eukaryote-like ankyrin domains (n = 7).

  11. The cobinamide amidohydrolase (cobyric acid-forming) CbiZ enzyme: a critical activity of the cobamide remodelling system of Rhodobacter sphaeroides.

    Science.gov (United States)

    Gray, Michael J; Escalante-Semerena, Jorge C

    2009-12-01

    The chemical structures of cobamides [cobalamin (Cbl)-like compounds] are the same, except for the lower ligand, which in adenosylcobalamin (AdoCbl) is 5,6-dimethylbenzimidazole, and in adenosylpseudocobalamin (AdopseudoCbl) is adenine. Why the lower ligand of cobamides varies and what the mechanism of lower ligand replacement is are long-standing questions in the field of B(12) biosynthesis. Work reported here uncovers the strategy used by the photosynthetic alpha-proteobacterium Rhodobacter sphaeroides to procure the cobamide it needs to grow on acetate as a carbon and energy source. On the basis of genetic and biochemical evidence we conclude that, in R. sphaeroides, the activity of the cobyric acid-producing amidohydrolase CbiZ enzyme is essential for the conversion of AdopseudoCbl into AdoCbl, the cobamide needed for the catabolism of acetate. The CbiZ enzyme uses AdopseudoCbl as a substrate, but not AdoCbl. Implications of these findings for cobamide remodelling in R. sphaeroides and in other CbiZ-containing microorganisms are discussed.

  12. A novel thermoalkalostable esterase from Acidicaldus sp. strain USBA-GBX-499 with enantioselectivity isolated from an acidic hot springs of Colombian Andes.

    Science.gov (United States)

    López, Gina; Chow, Jennifer; Bongen, Patrick; Lauinger, Benjamin; Pietruszka, Jörg; Streit, Wolfgang R; Baena, Sandra

    2014-10-01

    Several thermo- and mesoacidophilic bacterial strains that revealed high lipolytic activity were isolated from water samples derived from acidic hot springs in Los Nevados National Natural Park (Colombia). A novel lipolytic enzyme named 499EST was obtained from the thermoacidophilic alpha-Proteobacterium Acidicaldus USBA-GBX-499. The gene estA encoded a 313-amino-acid protein named 499EST. The deduced amino acid sequence showed the highest identity (58 %) with a putative α/β hydrolase from Acidiphilium sp. (ZP_08632277.1). Sequence alignments and phylogenetic analysis indicated that 499EST is a new member of the bacterial esterase/lipase family IV. The esterase reveals its optimum catalytic activity at 55 °C and pH 9.0. Kinetic studies showed that 499EST preferentially hydrolyzed middle-length acyl chains (C6-C8), especially p-nitrophenyl (p-NP) caproate (C6). Its thermostability and activity were strongly enhanced by adding 6 mM FeCl3. High stability in the presence of water-miscible solvents such as dimethyl sulfoxide and glycerol was observed. This enzyme also exhibits stability under harsh environmental conditions and enantioselectivity towards naproxen and ibuprofen esters, yielding the medically relevant (S)-enantiomers. In conclusion, according to our knowledge, 499EST is the first thermoalkalostable esterase derived from a Gram-negative thermoacidophilic bacterium.

  13. Detection of Alpha and Gamma-Proteobacteria in Amblyomma triste (Acari: Ixodidae) from Uruguay.

    Science.gov (United States)

    Venzal, José Manuel; Estrada-Peña, Agustín; Portillo, Aránzazu; Mangold, Atilio J; Castro, Oscar; de Souza, Carlos G; Félix, María L; Pérez-Martínez, Laura; Santibánez, Sonia; Oteo, José A

    2008-01-01

    Amblyomma triste is the most prevalent tick species reported in human tick bites in Uruguay and has been found to be infected with Rickettsia parkeri, but no other microorganisms have been reported from this tick. A sample of 254 adults of A. triste was collected by flagging on vegetation in suburban areas in southern Uruguay. Pools of five ticks were assembled and a screening for the DNA from the resulting 51 pools was realized by PCR assays using primers for amplifying a fragment of 16S rRNA gene for members of Anaplasmataceae. Seventeen pools were positive (33%) and the sequenciation of the gene fragment amplified revealed the presence of a putative new Alpha-Proteobacterium (denominated Atri-uru). The phylogenetic analysis showed that this microorganism is closely related to the symbiont of I. ricinus denominated 'Candidatus Midichloria mitochondrii' and other associated organisms. This rickettsial symbiont of ticks is included in a recent new clade proposed for the Alpha subclass of the Proteobacteria. The discovery of this bacterium in A. triste is the first evidence of this group of Rickettsiales detected in the Genus Amblyomma, and the first record in South America. Also, in two of 17 positive samples a Gamma-Proteobacterium related to Francisella-like organisms was detected.

  14. Global changes in gene expression in Sinorhizobium meliloti 1021 under microoxic and symbiotic conditions.

    Science.gov (United States)

    Becker, Anke; Bergès, Hélène; Krol, Elizaveta; Bruand, Claude; Rüberg, Silvia; Capela, Delphine; Lauber, Emmanuelle; Meilhoc, Eliane; Ampe, Frédéric; de Bruijn, Frans J; Fourment, Joëlle; Francez-Charlot, Anne; Kahn, Daniel; Küster, Helge; Liebe, Carine; Pühler, Alfred; Weidner, Stefan; Batut, Jacques

    2004-03-01

    Sinorhizobium meliloti is an alpha-proteobacterium that alternates between a free-living phase in bulk soil or in the rhizosphere of plants and a symbiotic phase within the host plant cells, where the bacteria ultimately differentiate into nitrogen-fixing organelle-like cells, called bacteroids. As a step toward understanding the physiology of S. meliloti in its free-living and symbiotic forms and the transition between the two, gene expression profiles were determined under two sets of biological conditions: growth under oxic versus microoxic conditions, and in free-living versus symbiotic state. Data acquisition was based on both macro- and microarrays. Transcriptome profiles highlighted a profound modification of gene expression during bacteroid differentiation, with 16% of genes being altered. The data are consistent with an overall slow down of bacteroid metabolism during adaptation to symbiotic life and acquisition of nitrogen fixation capability. A large number of genes of unknown function, including potential regulators, that may play a role in symbiosis were identified. Transcriptome profiling in response to oxygen limitation indicated that up to 5% of the genes were oxygen regulated. However, the microoxic and bacteroid transcriptomes only partially overlap, implying that oxygen contributes to a limited extent to the control of symbiotic gene expression.

  15. The viral eukaryogenesis hypothesis: a key role for viruses in the emergence of eukaryotes from a prokaryotic world environment.

    Science.gov (United States)

    Bell, Philip John Livingstone

    2009-10-01

    Understanding how the gulf between prokaryotic and eukaryotic cellular design arose is a major challenge. The viral eukaryogenesis (VE) hypothesis addresses the challenge of eukaryotic origins by suggesting the first eukaryotic cell was a multimember consortium consisting of a viral ancestor of the nucleus, an archaeal ancestor of the eukaryotic cytoplasm, and a bacterial ancestor of the mitochondria. Using only prokaryotes and their viruses, and invoking selective pressures observed in modern organisms, the VE hypothesis can explain the origins of the eukaryotic cell, sex, and meiosis. In the VE hypothesis, a cell wall-less archaeon and an alpha-prot