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Sample records for diazotrophic cyanobacterium important

  1. The genome of Cyanothece 51142, a unicellular diazotrophic cyanobacterium important in the marine nitrogen cycle

    Energy Technology Data Exchange (ETDEWEB)

    Welsh, Eric A.; Liberton, Michelle L.; Stockel, Jana; Loh, Thomas; Elvitigala, Thanura R.; Wang, Chunyan; Wollam, Aye; Fulton, Robert S.; Clifton, Sandra W.; Jacobs, Jon M.; Aurora, Rajeev; Ghosh, Bijoy K.; Sherman, Louis A.; Smith, Richard D.; Wilson, Richard K.; Pakrasi, Himadri B.

    2008-09-30

    Cyanobacteria are oxygenic photosynthetic bacteria that have significant roles in global biological carbon sequestration and oxygen production. They occupy a diverse range of habitats, from open ocean, to hot springs, deserts, and arctic waters. Cyanobacteria are known as the progenitors of the chloroplasts of plants and algae, and are the simplest known organisms to exhibit circadian behavior4. Cyanothece sp. ATCC 51142 is a unicellular marine cyanobacterium capable of N2-fixation, a process that is biochemically incompatible with oxygenic photosynthesis. To resolve this problem, Cyanothece performs photosynthesis during the day and nitrogen fixation at night, thus temporally separating these processes in the same cell. The genome of Cyanothece 51142 was completely sequenced and found to contain a unique arrangement of one large circular chromosome, four small plasmids, and one linear chromosome, the first report of such a linear element in a photosynthetic bacterium. Annotation of the Cyanothece genome was aided by the use of highthroughput proteomics data, enabling the reclassification of 25% of the proteins with no informative sequence homology. Phylogenetic analysis suggests that nitrogen fixation is an ancient process that arose early in evolution and has subsequently been lost in many cyanobacterial strains. In cyanobacterial cells, the circadian clock influences numerous processes, including carbohydrate synthesis, nitrogen fixation, photosynthesis, respiration, and the cell division cycle. During a diurnal period, Cyanothece cells actively accumulate and degrade different storage inclusion bodies for the products of photosynthesis and N2-fixation. This ability to utilize metabolic compartmentalization and energy storage makes Cyanothece an ideal system for bioenergy research, as well as studies of how a unicellular organism balances multiple, often incompatible, processes in the same cell.

  2. Dissolved organic nitrogen and carbon release by a marine unicellular diazotrophic cyanobacterium

    NARCIS (Netherlands)

    Benavides, M.; Agawin, N.S.R.; Aristegui, J.; Peene, J.; Stal, L.J.

    2013-01-01

    Dinitrogen (N-2) fixation rates may be underestimated when recently fixed N2 is released as dissolved organic nitrogen (DON). DON release (DONr) is substantial in the filamentous cyanobacterium Trichodesmium but has never been reported in unicellular diazotrophic cyanobacteria. We used axenic

  3. Dissolved organic nitrogen and carbon release by a marine unicellular diazotrophic cyanobacterium

    NARCIS (Netherlands)

    Benavides, M.; Agawin, N.S.R.; Aristegui, J.; Peene, J.; Stal, L.J.

    2013-01-01

    Dinitrogen (N2) fixation rates may be underestimated when recently fixed N2 is released as dissolved organic nitrogen (DON). DON release (DONr) is substantial in the filamentous cyanobacterium Trichodesmium but has never been reported in unicellular diazotrophic cyanobacteria. We used axenic

  4. Response of the unicellular diazotrophic cyanobacterium Crocosphaera watsonii to iron limitation.

    Directory of Open Access Journals (Sweden)

    Violaine Jacq

    Full Text Available Iron (Fe is widely suspected as a key controlling factor of N2 fixation due to the high Fe content of nitrogenase and photosynthetic enzymes complex, and to its low concentrations in oceanic surface seawaters. The influence of Fe limitation on the recently discovered unicellular diazotrophic cyanobacteria (UCYN is poorly understood despite their biogeochemical importance in the carbon and nitrogen cycles. To address this knowledge gap, we conducted culture experiments on Crocosphaera watsonii WH8501 growing under a range of dissolved Fe concentrations (from 3.3 to 403 nM. Overall, severe Fe limitation led to significant decreases in growth rate (2.6-fold, C, N and chlorophyll a contents per cell (up to 4.1-fold, N2 and CO2 fixation rates per cell (17- and 7-fold as well as biovolume (2.2-fold. We highlighted a two phased response depending on the degree of limitation: (i under a moderate Fe limitation, the biovolume of C. watsonii was strongly reduced, allowing the cells to keep sufficient energy to maintain an optimal growth, volume-normalized contents and N2 and CO2 fixation rates; (ii with increasing Fe deprivation, biovolume remained unchanged but the entire cell metabolism was affected, as shown by a strong decrease in the growth rate, volume-normalized contents and N2 and CO2 fixation rates. The half-saturation constant for growth of C. watsonii with respect to Fe is twice as low as that of the filamentous Trichodesmium indicating a better adaptation of C. watsonii to poor Fe environments than filamentous diazotrophs. The physiological response of C. watsonii to Fe limitation was different from that previously shown on the UCYN Cyanothece sp, suggesting potential differences in Fe requirements and/or Fe acquisition within the UCYN community. These results contribute to a better understanding of how Fe bioavailability can control the activity of UCYN and explain the biogeography of diverse N2 fixers in ocean.

  5. Transcriptional analysis of the unicellular, diazotrophic cyanobacterium Cyanothece sp. ATCC 51142 grown under short day/night cycles

    Energy Technology Data Exchange (ETDEWEB)

    Toepel, Jorg; McDermott, Jason E.; Summerfield, Tina; Sherman, Louis A.

    2009-06-01

    Cyanothece sp. strain ATCC 51142 is a unicellular, diazotrophic cyanobacterium that demonstrates extensive metabolic periodicities of photosynthesis, respiration and nitrogen fixation when grown under N2-fixing conditions. We have performed a global transcription analysis of this organism using 6 h light/dark cycles in order to determine the response of the cell to these conditions and to differentiate between diurnal and circadian regulated genes. In addition, we used a context-likelihood of relatedness (CLR) analysis with this data and those from two-day light/dark and light-dark plus continuous light experiments to better differentiate between diurnal and circadian regulated genes. Cyanothece sp. adapted in several ways to growth under short light/dark conditions. Nitrogen was fixed in every second dark period and only once in each 24 h period. Nitrogen fixation was strongly correlated to the energy status of the cells and glycogen breakdown and high respiration rates were necessary to provide appropriate energy and anoxic conditions for this process. We conclude that glycogen breakdown is a key regulatory step within these complex processes. Our results demonstrated that the main metabolic genes involved in photosynthesis, respiration, nitrogen fixation and central carbohydrate metabolism have strong (or total) circadian-regulated components. The short light/dark cycles enable us to identify transcriptional differences among the family of psbA genes, as well as the differing patterns of the hup genes, which follow the same pattern as nitrogenase genes, relative to the hox genes which displayed a diurnal, dark-dependent gene expression.

  6. Analysis of carbohydrate storage granules in the diazotrophic cyanobacterium Cyanothece sp. PCC 7822

    Energy Technology Data Exchange (ETDEWEB)

    Welkie, David G. [Purdue Univ., West Lafayette, IN (United States); Sherman, Debra M. [Purdue Univ., West Lafayette, IN (United States); Chrisler, William B. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Orr, Galya [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Sherman, Louis A. [Purdue Univ., West Lafayette, IN (United States)

    2013-10-19

    The unicellular diazotrophic cyanobacteria of the genus Cyanothece demonstrate oscillations in nitrogenase activity and H2 production when grown under 12h light-12h dark cycles. We established that Cyanothece sp. PCC 7822 allows for the construction of knock-out mutants and our objective was to improve the growth characteristics of this strain and to identify the nature of the intracellular storage granules. We report the physiological and morphological effects of reduction in nitrate and phosphate concentrations in BG-11 media on this strain. We developed a series of BG-11-derived growth media and monitored batch culture growth, nitrogenase activity and nitrogenase-mediated hydrogen production, culture synchronicity, and intracellular storage content. Reduction in NaNO3 and K2HPO4 concentrations from 17.6 and 0.23 mM to 4.41 and 0.06 mM, respectively, improved growth characteristics such as cell size and uniformity, and enhanced the rate of cell division. Cells grown in this low NP BG-11 were less complex, a parameter that related to the composition of the intracellular storage granules. Cells grown in low NP BG-11 had less polyphosphate, fewer polyhydroxybutyrate granules and many smaller granules became evident. Biochemical analysis and transmission electron microscopy using the histocytochemical PATO technique demonstrated that these small granules contained glycogen. The glycogen levels and the number of granules per cell correlated nicely with a 2.3 to 3.3-fold change from the minimum at L0 to the maximum at D0. The differences in granule morphology and enzymes between Cyanothece ATCC 51142 and Cyanothece PCC 7822 provide insights into the formation of large starch-like granules in some cyanobacteria.

  7. Analysis of the early heterocyst Cys-proteome in the multicellular cyanobacterium Nostoc punctiforme reveals novel insights into the division of labor within diazotrophic filaments.

    Science.gov (United States)

    Sandh, Gustaf; Ramström, Margareta; Stensjö, Karin

    2014-12-04

    In the filamentous cyanobacterium Nostoc punctiforme ATCC 29133, removal of combined nitrogen induces the differentiation of heterocysts, a cell-type specialized in N2 fixation. The differentiation involves genomic, structural and metabolic adaptations. In cyanobacteria, changes in the availability of carbon and nitrogen have also been linked to redox regulated posttranslational modifications of protein bound thiol groups. We have here employed a thiol targeting strategy to relatively quantify the putative redox proteome in heterocysts as compared to N2-fixing filaments, 24 hours after combined nitrogen depletion. The aim of the study was to expand the coverage of the cell-type specific proteome and metabolic landscape of heterocysts. Here we report the first cell-type specific proteome of newly formed heterocysts, compared to N2-fixing filaments, using the cysteine-specific selective ICAT methodology. The data set defined a good quantitative accuracy of the ICAT reagent in complex protein samples. The relative abundance levels of 511 proteins were determined and 74% showed a cell-type specific differential abundance. The majority of the identified proteins have not previously been quantified at the cell-type specific level. We have in addition analyzed the cell-type specific differential abundance of a large section of proteins quantified in both newly formed and steady-state diazotrophic cultures in N. punctiforme. The results describe a wide distribution of members of the putative redox regulated Cys-proteome in the central metabolism of both vegetative cells and heterocysts of N. punctiforme. The data set broadens our understanding of heterocysts and describes novel proteins involved in heterocyst physiology, including signaling and regulatory proteins as well as a large number of proteins with unknown function. Significant differences in cell-type specific abundance levels were present in the cell-type specific proteomes of newly formed diazotrophic filaments

  8. Genetically modified cyanobacterium Nostoc muscorum ...

    Indian Academy of Sciences (India)

    Madhu

    diazotrophic cultures of the cyanobacterium N. muscorum in quantities of 5 .... growth media and then used for estimation of their characteristics. Each reading is ..... 1224–1232. Christian J H B 1950 The influence of nutrition on the water rela-.

  9. Enhanced production of biomass, pigments and antioxidant capacity of a nutritionally important cyanobacterium Nostochopsis lobatus.

    Science.gov (United States)

    Pandey, Usha; Pandey, J

    2008-07-01

    A diazotrophic cyanobacterium Nostochopsis lobatus was evaluated for enhanced production of biomass, pigments and antioxidant capacity. N. lobatus showed potentially high antioxidant capacity (46.12 microM AEAC) with significant improvement under immobilized cell cultures (87.05 microM AEAC). When a mixture of P and Fe was supplemented, biomass, pigments, nutritive value and antioxidant capacity increased substantially at pH 7.8. When considered separately, P appeared to be a better supplement than Fe for the production of biomass, chlorophyll and carotenoids. However, for phycocyanin, phycoerythrin, nutritive value and antioxidant capacity, Fe appeared more effective than P. Our study indicates N. lobatus to be a promising bioresource for enhanced production of nutritionally rich biomass, pigments and antioxidants. The study also suggests that P and Fe are potentially effective supplements for scale-up production for commercial application.

  10. Decoupling of ammonium regulation and ntcA transcription in the diazotrophic marine cyanobacterium Trichodesmium sp. IMS101.

    Science.gov (United States)

    Post, Anton F; Rihtman, Branko; Wang, Qingfeng

    2012-03-01

    Nitrogen (N) physiology in the marine cyanobacterium Trichodesmium IMS101 was studied along with transcript accumulation of the N-regulatory gene ntcA and of two of its target genes: napA (nitrate assimilation) and nifH (N(2) fixation). N(2) fixation was impaired in the presence of nitrite, nitrate and urea. Strain IMS101 was capable of growth on these combined N sources at ammonium. Whereas ecologically relevant N concentrations (2-20 μM) suppressed growth and assimilation, much higher concentrations were required to affect transcript levels. Transcripts of nifH accumulated under nitrogen-fixing conditions; these transcript levels were maintained in the presence of nitrate (100 μM) and ammonium (20 μM). However, nifH transcript levels were below detection at ammonium concentrations >20 μM. napA mRNA was found at low levels in both N(2)-fixing and ammonium-utilizing filaments, and it accumulated in filaments grown with nitrate. The positive effect of nitrate on napA transcription was abolished by ammonium additions of >200 μM. This effect was restored upon addition of the glutamine synthetase inhibitor L-methionin-DL-sulfoximine. Surprisingly, ntcA transcript levels remained high in the presence of ammonium, even at elevated concentrations. These findings indicate that ammonium repression is decoupled from transcriptional activation of ntcA in Trichodesmium IMS101.

  11. Comparisons of diazotrophic communities in native and agricultural desert ecosystems reveal plants as important drivers in diversity.

    Science.gov (United States)

    Köberl, Martina; Erlacher, Armin; Ramadan, Elshahat M; El-Arabi, Tarek F; Müller, Henry; Bragina, Anastasia; Berg, Gabriele

    2016-02-01

    Diazotrophs provide the only biological source of fixed atmospheric nitrogen in the biosphere. Although they are the key player for plant-available nitrogen, less is known about their diversity and potential importance in arid ecosystems. We investigated the nitrogenase gene diversity in native and agricultural desert soil as well as within root-associated microbiota of medicinal plants grown in Egypt through the combination of nifH-specific qPCR, fingerprints, amplicon pyrosequencing and fluorescence in situ hybridization-confocal laser scanning microscopy. Although the diazotrophic microbiota were characterized by generally high abundances and diversity, statistically significant differences were found between both soils, the different microhabitats, and between the investigated plants (Matricaria chamomilla L., Calendula officinalis L. and Solanum distichum Schumach. and Thonn.). We observed a considerable community shift from desert to agriculturally used soil that demonstrated a higher abundance and diversity in the agro-ecosystem. The endorhiza was characterized by lower abundances and only a subset of species when compared to the rhizosphere. While the microbiomes of the Asteraceae were similar and dominated by potential root-nodulating rhizobia acquired primarily from soil, the perennial S. distichum generally formed associations with free-living nitrogen fixers. These results underline the importance of diazotrophs in desert ecosystems and additionally identify plants as important drivers in functional gene pool diversity. © FEMS 2015.

  12. A common transport system for methionine, L-methionine-DL-sulfoximine (MSX), and phosphinothricin (PPT) in the diazotrophic cyanobacterium Nostoc muscorum.

    Science.gov (United States)

    Singh, Arvind Kumar; Syiem, Mayashree B; Singh, Rajkumar S; Adhikari, Samrat; Rai, Amar Nath

    2008-05-01

    We present evidence, for the first time, of the occurrence of a transport system common for amino acid methionine, and methionine/glutamate analogues L-methionine-DL-sulfoximine (MSX) and phosphinothricin (PPT) in cyanobacterium Nostoc muscorum. Methionine, which is toxic to cyanobacterium, enhanced its nitrogenase activity at lower concentrations. The cyanobacterium showed a biphasic pattern of methionine uptake activity that was competitively inhibited by the amino acids alanine, isoleucine, leucine, phenylalanine, proline, valine, glutamine, and asparagine. The methionine/glutamate analogue-resistant N. muscorum strains (MSX-R and PPT-R strains) also showed methionine-resistant phenotype accompanied by a drastic decrease in 35S methionine uptake activity. Treatment of protein extracts from these mutant strains with MSX and PPT reduced biosynthetic glutamine synthetase (GS) activity only in vitro and not in vivo. This finding implicated that MSX- and PPT-R phenotypes may have arisen due to a defect in their MSX and PPT transport activity. The simultaneous decrease in methionine uptake activity and in vitro sensitivity toward MSX and PPT of GS protein in MSX- and PPT-R strains indicated that methionine, MSX, and PPT have a common transport system that is shared by other amino acids as well in N. muscorum. Such information can become useful for isolation of methionine-producing cyanobacterial strains.

  13. The Multiple Functions of Common Microbial Carbon Polymers, Glycogen and PHB, during Stress Responses in the Non-Diazotrophic Cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Damrow, Ramon; Maldener, Iris; Zilliges, Yvonne

    2016-01-01

    Classical microbial carbon polymers such as glycogen and polyhydroxybutyrate (PHB) have a crucial impact as both a sink and a reserve under macronutrient stress conditions. Most microbial species exclusively synthesize and degrade either glycogen or PHB. A few bacteria such as the phototrophic model organism Synechocystis sp. PCC 6803 surprisingly produce both physico-chemically different polymers under conditions of high C to N ratios. For the first time, the function and interrelation of both carbon polymers in non-diazotrophic cyanobacteria are analyzed in a comparative physiological study of single- and double-knockout mutants (ΔglgC; ΔphaC; ΔglgC/ΔphaC), respectively. Most of the observed phenotypes are explicitly related to the knockout of glycogen synthesis, highlighting the metabolic, energetic, and structural impact of this process whenever cells switch from an active, photosynthetic 'protein status' to a dormant 'glycogen status'. The carbon flux regulation into glycogen granules is apparently crucial for both phycobilisome degradation and thylakoid layer disassembly in the presence of light. In contrast, PHB synthesis is definitely not involved in this primary acclimation response. Moreover, the very weak interrelations between the two carbon-polymer syntheses indicate that the regulation and role of PHB synthesis in Synechocystis sp. PCC 6803 is different from glycogen synthesis.

  14. Lipopolysaccharides in diazotrophic bacteria.

    Science.gov (United States)

    Serrato, Rodrigo V

    2014-01-01

    Biological nitrogen fixation (BNF) is a process in which the atmospheric nitrogen (N2) is transformed into ammonia (NH3) by a select group of nitrogen-fixing organisms, or diazotrophic bacteria. In order to furnish the biologically useful nitrogen to plants, these bacteria must be in constant molecular communication with their host plants. Some of these molecular plant-microbe interactions are very specific, resulting in a symbiotic relationship between the diazotroph and the host. Others are found between associative diazotrophs and plants, resulting in plant infection and colonization of internal tissues. Independent of the type of ecological interaction, glycans, and glycoconjugates produced by these bacteria play an important role in the molecular communication prior and during colonization. Even though exopolysaccharides (EPS) and lipochitooligosaccharides (LCO) produced by diazotrophic bacteria and released onto the environment have their importance in the microbe-plant interaction, it is the lipopolysaccharides (LPS), anchored on the external membrane of these bacteria, that mediates the direct contact of the diazotroph with the host cells. These molecules are extremely variable among the several species of nitrogen fixing-bacteria, and there are evidences of the mechanisms of infection being closely related to their structure.

  15. Towards long-read metagenomics: complete assembly of three novel genomes from bacteria dependent on a diazotrophic cyanobacterium in a freshwater lake co-culture.

    Science.gov (United States)

    Driscoll, Connor B; Otten, Timothy G; Brown, Nathan M; Dreher, Theo W

    2017-01-01

    Here we report three complete bacterial genome assemblies from a PacBio shotgun metagenome of a co-culture from Upper Klamath Lake, OR. Genome annotations and culture conditions indicate these bacteria are dependent on carbon and nitrogen fixation from the cyanobacterium Aphanizomenon flos-aquae, whose genome was assembled to draft-quality . Due to their taxonomic novelty relative to previously sequenced bacteria, we have temporarily designated these bacteria as incertae sedis Hyphomonadaceae strain UKL13-1 (3,501,508 bp and 56.12% GC), incertae sedis Betaproteobacterium strain UKL13-2 (3,387,087 bp and 54.98% GC), and incertae sedis Bacteroidetes strain UKL13-3 (3,236,529 bp and 37.33% GC). Each genome consists of a single circular chromosome with no identified plasmids. When compared with binned Illumina assemblies of the same three genomes, there was ~7% discrepancy in total genome length. Gaps where Illumina assemblies broke were often due to repetitive elements. Within these missing sequences were essential genes and genes associated with a variety of functional categories. Annotated gene content reveals that both Proteobacteria are aerobic anoxygenic phototrophs, with Betaproteobacterium UKL13-2 potentially capable of phototrophic oxidation of sulfur compounds. Both proteobacterial genomes contain transporters suggesting they are scavenging fixed nitrogen from A. flos-aquae in the form of ammonium. Bacteroidetes UKL13-3 has few completely annotated biosynthetic pathways, and has a comparatively higher proportion of unannotated genes. The genomes were detected in only a few other freshwater metagenomes, suggesting that these bacteria are not ubiquitous in freshwater systems. Our results indicate that long-read sequencing is a viable method for sequencing dominant members from low-diversity microbial communities, and should be considered for environmental metagenomics when conditions meet these requirements.

  16. Grazing by Zooplankton on Diazotrophs in the Amazon River Plume and Western Tropical North Atlantic

    Science.gov (United States)

    Conroy, B.; Steinberg, D. K.; Song, B.; Foster, R.

    2016-02-01

    Organisms capable of fixing di-nitrogen (N2), known as diazotrophs, are important primary producers and a potentially significant source for new nitrogen entering the planktonic food web. However, limited evidence exists for zooplankton grazing on diazotrophs compared to other primary producers. In the western tropical North Atlantic Ocean (WTNA), the Amazon River plume creates a niche for symbiotic diatom-diazotroph associations (DDAs) which can form large blooms. In adjacent non-plume-influenced waters, the colonial cyanobacterium Trichodesmium is abundant. In order to reveal zooplankton-diazotroph grazing interactions and determine the fate of newly fixed nitrogen, gut contents of zooplankton captured in these two regions were compared based on quantitative PCR (qPCR) assay of nitrogenase genes (nifH), and their microbiomes compared using next generation sequencing (NGS) analysis of 16S rRNA genes. We sampled individual copepods from discrete depth intervals (0-25m and 25-50m) and in two size classes (0.5-1mm and 1-2mm) for analysis. A modified DNA extraction protocol was developed and 54 extracts were used as templates in nifH qPCR assays for the larger size fraction diazotrophs (>10µm): Trichodesmium, and Hemiaulus or Rhizosolenia (diatoms)-Richelia (diazotroph) associations. Copepod gut content nifH copies ranged from 1.6 to 13.6 copies individual-1 for the assay targeting the Hemiaulus-Richelia DDA and from 1.1 to 3.0 copies individual-1 for Trichodesmium. 16S NGS conducted on 35 extracts with an Ion Torrent PGM and mothur revealed that cyanobacteria sequences accounted for up to 20% of sequences per extract. Our results show that both DDAs and Trichodesmium are prey for zooplankton, and that new nitrogen moves through the food web via these grazing interactions. These interactions should be considered in future explorations of the global ocean nitrogen cycle.

  17. Occurrence of diazotrophic bacteria in Araucaria angustifolia

    OpenAIRE

    Neroni,Rafaela de Fátima; Cardoso,Elke Jurandy Bran Nogueira

    2007-01-01

    Araucaria angustifolia is an environmentally threatened tree and the whole biota of the Araucaria Forest should be investigated with the aim of its preservation. Diazotrophic bacteria are extremely important for the maintenance of ecosystems, but they have never been studied in Araucaria Forests. In this study, diazotrophic bacteria were isolated from Araucaria roots and soil, when grown in semi-specific, semi-solid media. The diazotrophic character of some recovered isolates could be confirm...

  18. Lipopolysaccharides in diazotrophic bacteria

    OpenAIRE

    Serrato, Rodrigo V.

    2014-01-01

    Biological nitrogen fixation (BNF) is a process in which the atmospheric nitrogen (N2) is transformed into ammonia (NH3) by a select group of nitrogen-fixing organisms, or diazotrophic bacteria. In order to furnish the biologically useful nitrogen to plants, these bacteria must be in constant molecular communication with their host plants. Some of these molecular plant-microbe interactions are very specific, resulting in a symbiotic relationship between the diazotroph and the host. Others are...

  19. The Green Berry Consortia of the Sippewissett Salt Marsh: Millimeter-Sized Aggregates of Diazotrophic Unicellular Cyanobacteria.

    Science.gov (United States)

    Wilbanks, Elizabeth G; Salman-Carvalho, Verena; Jaekel, Ulrike; Humphrey, Parris T; Eisen, Jonathan A; Buckley, Daniel H; Zinder, Stephen H

    2017-01-01

    Microbial interactions driving key biogeochemical fluxes often occur within multispecies consortia that form spatially heterogeneous microenvironments. Here, we describe the "green berry" consortia of the Sippewissett salt marsh (Falmouth, MA, United States): millimeter-sized aggregates dominated by an uncultured, diazotrophic unicellular cyanobacterium of the order Chroococcales (termed GB-CYN1). We show that GB-CYN1 is closely related to Crocosphaera watsonii (UCYN-B) and " Candidatus Atelocyanobacterium thalassa" (UCYN-A), two groups of unicellular diazotrophic cyanobacteria that play an important role in marine primary production. Other green berry consortium members include pennate diatoms and putative heterotrophic bacteria from the Alphaproteobacteria and Bacteroidetes . Tight coupling was observed between photosynthetic oxygen production and heterotrophic respiration. When illuminated, the green berries became supersaturated with oxygen. From the metagenome, we observed that GB-CYN1 encodes photosystem II genes and thus has the metabolic potential for oxygen production unlike UCYN-A. In darkness, respiratory activity rapidly depleted oxygen creating anoxia within the aggregates. Metagenomic data revealed a suite of nitrogen fixation genes encoded by GB-CYN1, and nitrogenase activity was confirmed at the whole-aggregate level by acetylene reduction assays. Metagenome reads homologous to marker genes for denitrification were observed and suggest that heterotrophic denitrifiers might co-occur in the green berries, although the physiology and activity of facultative anaerobes in these aggregates remains uncharacterized. Nitrogen fixation in the surface ocean was long thought to be driven by filamentous cyanobacterial aggregates, though recent work has demonstrated the importance of unicellular diazotrophic cyanobacteria (UCYN) from the order Chroococcales. The green berries serve as a useful contrast to studies of open ocean UCYN and may provide a tractable

  20. Biogeochemical tracers of the marine cyanobacterium Trichodesmium

    Science.gov (United States)

    Carpenter, Edward J.; Harvey, H. Rodger; Fry, Brian; Capone, Douglas G.

    1997-01-01

    We examined the utility of several biogeochemical tracers for following the fate of the planktonic diazotrophic cyanobacterium Trichodesmium in the sea. The presence of a (CIO) fatty acid previously reported was observed in a culture of Trichodesmium but was not found in natural samples. This cyanobacterium had high concentrations of C 14 and C 16 acids, with lesser amounts of several saturated and unsaturated C 18 fatty acids. This composition was similar to that of other marine cyanobacteria. The major hydrocarbon identified was the C 17n-alkane, which was present in all samples from the five stations examined. Sterols common to algae and copepods were observed in many samples along with hopanoids representative of bacteria, suggesting a varied community structure in colonies collected from different stations. We found no unique taxonomic marker of Trichodesmium among the sterols. Measurements of the σ 15N and σ 13C in Trichodesmium samples from the SW Sargasso and NW Caribbean Seas averaged -0.4960 (range from -0.7 to -0.25960) and -12.9%0 (range from -15.2 to -11.9960), respectively, thus confirming previous observations that this cyanobacterial diazotroph has both the lowest σ 15N and highest σ 13C of any marine phytoplankter observed to date. A culture of Trichodesmium grown under diazotrophic conditions had a σ 15N between -1.3 and -3.6960. Our results support the supposition that the relatively low σ 15N and high σ 13C values observed in suspended and sediment-trapped material from some tropical and subtropical seas result from substantial input of C and N by Trichodesmium.

  1. Diversity and Activity of Diazotrophs in Great Barrier Reef Surface Waters.

    Science.gov (United States)

    Messer, Lauren F; Brown, Mark V; Furnas, Miles J; Carney, Richard L; McKinnon, A D; Seymour, Justin R

    2017-01-01

    Discrepancies between bioavailable nitrogen (N) concentrations and phytoplankton growth rates in the oligotrophic waters of the Great Barrier Reef (GBR) suggest that undetermined N sources must play a significant role in supporting primary productivity. One such source could be biological dinitrogen (N 2 ) fixation through the activity of "diazotrophic" bacterioplankton. Here, we investigated N 2 fixation and diazotroph community composition over 10° S of latitude within GBR surface waters. Qualitative N 2 fixation rates were found to be variable across the GBR but were relatively high in coastal, inner and outer GBR waters, reaching 68 nmol L -1 d -1 . Diazotroph assemblages, identified by amplicon sequencing of the nifH gene, were dominated by the cyanobacterium Trichodesmium erythraeum , γ-proteobacteria from the Gamma A clade, and δ-proteobacterial phylotypes related to sulfate-reducing genera. However, diazotroph communities exhibited significant spatial heterogeneity, correlated with shifts in dissolved inorganic nutrient concentrations. Specifically, heterotrophic diazotrophs generally increased in relative abundance with increasing concentrations of phosphate and N, while Trichodesmium was proportionally more abundant when concentrations of these nutrients were low. This study provides the first in-depth characterization of diazotroph community composition and N 2 fixation dynamics within the oligotrophic, N-limited surface waters of the GBR. Our observations highlight the need to re-evaluate N cycling dynamics within oligotrophic coral reef systems, to include diverse N 2 fixing assemblages as a potentially significant source of dissolved N within the water column.

  2. Cellular and functional specificity among ferritin-like proteins in the multicellular cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Ekman, Martin; Sandh, Gustaf; Nenninger, Anja; Oliveira, Paulo; Stensjö, Karin

    2014-03-01

    Ferritin-like proteins constitute a remarkably heterogeneous protein family, including ferritins, bacterioferritins and Dps proteins. The genome of the filamentous heterocyst-forming cyanobacterium Nostoc punctiforme encodes five ferritin-like proteins. In the present paper, we report a multidimensional characterization of these proteins. Our phylogenetic and bioinformatics analyses suggest both structural and physiological differences among the ferritin-like proteins. The expression of these five genes responded differently to hydrogen peroxide treatment, with a significantly higher rise in transcript level for Npun_F3730 as compared with the other four genes. A specific role for Npun_F3730 in the cells tolerance against hydrogen peroxide was also supported by the inactivation of Npun_F3730, Npun_R5701 and Npun_R6212; among these, only the ΔNpun_F3730 strain showed an increased sensitivity to hydrogen peroxide compared with wild type. Analysis of promoter-GFP reporter fusions of the ferritin-like genes indicated that Npun_F3730 and Npun_R5701 were expressed in all cell types of a diazotrophic culture, while Npun_F6212 was expressed specifically in heterocysts. Our study provides the first comprehensive analysis combining functional differentiation and cellular specificity within this important group of proteins in a multicellular cyanobacterium. © 2013 John Wiley & Sons Ltd and Society for Applied Microbiology.

  3. Permanent Draft Genome of Strain ESFC-1: Ecological Genomics of a Newly Discovered Lineage of Filamentous Diazotrophic Cyanobacteria

    Science.gov (United States)

    Everroad, R. Craig; Stuart, Rhona K.; Bebout, Brad M.; Detweiler, Angela M.; Lee, Jackson Zan; Woebken, Dagmar; Bebout, Leslie E.; Pett-Ridge, Jennifer

    2016-01-01

    The nonheterocystous filamentous cyanobacterium, strain ESFC-1, is a recently described member of the order Oscillatoriales within the Cyanobacteria. ESFC-1 has been shown to be a major diazotroph in the intertidal microbial mat system at Elkhorn Slough, CA, USA. Based on phylogenetic analyses of the 16S RNA gene, ESFC-1 appears to belong to a unique, genus-level divergence; the draft genome sequence of this strain has now been determined. Here we report features of this genome as they relate to the ecological functions and capabilities of strain ESFC-1. The 5,632,035 bp genome sequence encodes 4914 protein-coding genes and 92 RNA genes. One striking feature of this cyanobacterium is the apparent lack of either uptake or bi-directional hydrogenases typically expected within a diazotroph. Additionally, a large genomic island is found that contains numerous low GC-content genes and genes related to extracellular polysaccharide production and cell wall synthesis and maintenance.

  4. Diversity and Structure of Diazotrophic Communities in Mangrove Rhizosphere, Revealed by High-Throughput Sequencing.

    Science.gov (United States)

    Zhang, Yanying; Yang, Qingsong; Ling, Juan; Van Nostrand, Joy D; Shi, Zhou; Zhou, Jizhong; Dong, Junde

    2017-01-01

    Diazotrophic communities make an essential contribution to the productivity through providing new nitrogen. However, knowledge of the roles that both mangrove tree species and geochemical parameters play in shaping mangove rhizosphere diazotrophic communities is still elusive. Here, a comprehensive examination of the diversity and structure of microbial communities in the rhizospheres of three mangrove species, Rhizophora apiculata , Avicennia marina , and Ceriops tagal , was undertaken using high - throughput sequencing of the 16S rRNA and nifH genes. Our results revealed a great diversity of both the total microbial composition and the diazotrophic composition specifically in the mangrove rhizosphere. Deltaproteobacteria and Gammaproteobacteria were both ubiquitous and dominant, comprising an average of 45.87 and 86.66% of total microbial and diazotrophic communities, respectively. Sulfate-reducing bacteria belonging to the Desulfobacteraceae and Desulfovibrionaceae were the dominant diazotrophs. Community statistical analyses suggested that both mangrove tree species and additional environmental variables played important roles in shaping total microbial and potential diazotroph communities in mangrove rhizospheres. In contrast to the total microbial community investigated by analysis of 16S rRNA gene sequences, most of the dominant diazotrophic groups identified by nifH gene sequences were significantly different among mangrove species. The dominant diazotrophs of the family Desulfobacteraceae were positively correlated with total phosphorus, but negatively correlated with the nitrogen to phosphorus ratio. The Pseudomonadaceae were positively correlated with the concentration of available potassium, suggesting that diazotrophs potentially play an important role in biogeochemical cycles, such as those of nitrogen, phosphorus, sulfur, and potassium, in the mangrove ecosystem.

  5. Diversity and Structure of Diazotrophic Communities in Mangrove Rhizosphere, Revealed by High-Throughput Sequencing

    Directory of Open Access Journals (Sweden)

    Yanying Zhang

    2017-10-01

    Full Text Available Diazotrophic communities make an essential contribution to the productivity through providing new nitrogen. However, knowledge of the roles that both mangrove tree species and geochemical parameters play in shaping mangove rhizosphere diazotrophic communities is still elusive. Here, a comprehensive examination of the diversity and structure of microbial communities in the rhizospheres of three mangrove species, Rhizophora apiculata, Avicennia marina, and Ceriops tagal, was undertaken using high-throughput sequencing of the 16S rRNA and nifH genes. Our results revealed a great diversity of both the total microbial composition and the diazotrophic composition specifically in the mangrove rhizosphere. Deltaproteobacteria and Gammaproteobacteria were both ubiquitous and dominant, comprising an average of 45.87 and 86.66% of total microbial and diazotrophic communities, respectively. Sulfate-reducing bacteria belonging to the Desulfobacteraceae and Desulfovibrionaceae were the dominant diazotrophs. Community statistical analyses suggested that both mangrove tree species and additional environmental variables played important roles in shaping total microbial and potential diazotroph communities in mangrove rhizospheres. In contrast to the total microbial community investigated by analysis of 16S rRNA gene sequences, most of the dominant diazotrophic groups identified by nifH gene sequences were significantly different among mangrove species. The dominant diazotrophs of the family Desulfobacteraceae were positively correlated with total phosphorus, but negatively correlated with the nitrogen to phosphorus ratio. The Pseudomonadaceae were positively correlated with the concentration of available potassium, suggesting that diazotrophs potentially play an important role in biogeochemical cycles, such as those of nitrogen, phosphorus, sulfur, and potassium, in the mangrove ecosystem.

  6. Diazotrophic Bacterial Community of Degraded Pastures

    OpenAIRE

    João Tiago Correia Oliveira; Everthon Fernandes Figueredo; Williane Patrícia da Silva Diniz; Lucianne Ferreira Paes de Oliveira; Pedro Avelino Maia de Andrade; Fernando Dini Andreote; Júlia Kuklinsky-Sobral; Danúbia Ramos de Lima; Fernando José Freire

    2017-01-01

    Pasture degradation can cause changes in diazotrophic bacterial communities. Thus, this study aimed to evaluate the culturable and total diazotrophic bacterial community, associated with regions of the rhizosphere and roots of Brachiaria decumbens Stapf. pastures in different stages of degradation. Samples of roots and rhizospheric soil were collected from slightly, partially, and highly degraded pastures. McCrady’s table was used to obtain the Most Probable Number (MPN) of bacteria per gram ...

  7. Diazotrophic Bacterial Community of Degraded Pastures

    Directory of Open Access Journals (Sweden)

    João Tiago Correia Oliveira

    2017-01-01

    Full Text Available Pasture degradation can cause changes in diazotrophic bacterial communities. Thus, this study aimed to evaluate the culturable and total diazotrophic bacterial community, associated with regions of the rhizosphere and roots of Brachiaria decumbens Stapf. pastures in different stages of degradation. Samples of roots and rhizospheric soil were collected from slightly, partially, and highly degraded pastures. McCrady’s table was used to obtain the Most Probable Number (MPN of bacteria per gram of sample, in order to determine population density and calculate the Shannon-Weaver diversity index. The diversity of total diazotrophic bacterial community was determined by the technique of Denaturing Gradient Gel Electrophoresis (DGGE of the nifH gene, while the diversity of the culturable diazotrophic bacteria was determined by the Polymerase Chain Reaction (BOX-PCR technique. The increase in the degradation stage of the B. decumbens Stapf. pasture did not reduce the population density of the cultivated diazotrophic bacterial community, suggesting that the degradation at any degree of severity was highly harmful to the bacteria. The structure of the total diazotrophic bacterial community associated with B. decumbens Stapf. was altered by the pasture degradation stage, suggesting a high adaptive capacity of the bacteria to altered environments.

  8. Isolation, characterization and localization of extracellular polymeric substances from the cyanobacterium Arthrospira platensis strain MMG-9

    NARCIS (Netherlands)

    Ahmed, M.; Moerdijk-Poortvliet, T.C.W.; Wijnholds, A.; Stal, L.J.; Hasnain, S.

    2014-01-01

    Arthrospira platensis is a cyanobacterium known for its nutritional value and secondary metabolites. Extracellular polymeric substances (EPS) are an important trait of most cyanobacteria, including A. platensis. Here, we extracted and analysed different fractions of EPS from a locally isolated

  9. Isolation, characterization and localization of extracellular polymeric substances from the cyanobacterium

    NARCIS (Netherlands)

    Ahmed, M.; Wijnholds, A.; Stal, L.J.; Hasnain, S.

    2014-01-01

    Arthrospira platensis is a cyanobacterium known for its nutritional value and secondary metabolites. Extracellular polymeric substances (EPS) are an important trait of most cyanobacteria, including A. platensis. Here, we extracted and analysed different fractions of EPS from a locally isolated

  10. Diazotroph Diversity in the Sea Ice, Melt Ponds, and Surface Waters of the Eurasian Basin of the Central Arctic Ocean.

    Science.gov (United States)

    Fernández-Méndez, Mar; Turk-Kubo, Kendra A; Buttigieg, Pier L; Rapp, Josephine Z; Krumpen, Thomas; Zehr, Jonathan P; Boetius, Antje

    2016-01-01

    The Eurasian basin of the Central Arctic Ocean is nitrogen limited, but little is known about the presence and role of nitrogen-fixing bacteria. Recent studies have indicated the occurrence of diazotrophs in Arctic coastal waters potentially of riverine origin. Here, we investigated the presence of diazotrophs in ice and surface waters of the Central Arctic Ocean in the summer of 2012. We identified diverse communities of putative diazotrophs through targeted analysis of the nifH gene, which encodes the iron protein of the nitrogenase enzyme. We amplified 529 nifH sequences from 26 samples of Arctic melt ponds, sea ice and surface waters. These sequences resolved into 43 clusters at 92% amino acid sequence identity, most of which were non-cyanobacterial phylotypes from sea ice and water samples. One cyanobacterial phylotype related to Nodularia sp. was retrieved from sea ice, suggesting that this important functional group is rare in the Central Arctic Ocean. The diazotrophic community in sea-ice environments appear distinct from other cold-adapted diazotrophic communities, such as those present in the coastal Canadian Arctic, the Arctic tundra and glacial Antarctic lakes. Molecular fingerprinting of nifH and the intergenic spacer region of the rRNA operon revealed differences between the communities from river-influenced Laptev Sea waters and those from ice-related environments pointing toward a marine origin for sea-ice diazotrophs. Our results provide the first record of diazotrophs in the Central Arctic and suggest that microbial nitrogen fixation may occur north of 77°N. To assess the significance of nitrogen fixation for the nitrogen budget of the Arctic Ocean and to identify the active nitrogen fixers, further biogeochemical and molecular biological studies are needed.

  11. Diazotroph diversity in the sea ice, melt ponds and surface waters of the Eurasian Basin of the Central Arctic Ocean

    Directory of Open Access Journals (Sweden)

    Mar Fernández-Méndez

    2016-11-01

    Full Text Available The Eurasian basin of the Central Arctic Ocean is nitrogen limited, but little is known about the presence and role of nitrogen-fixing bacteria. Recent studies have indicated the occurrence of diazotrophs in Arctic coastal waters potentially of riverine origin. Here, we investigated the presence of diazotrophs in ice and surface waters of the Central Arctic Ocean in the summer of 2012. We identified diverse communities of putative diazotrophs through targeted analysis of the nifH gene, which encodes the iron protein of the nitrogenase enzyme. We amplified 529 nifH sequences from 26 samples of Arctic melt ponds, sea ice and surface waters. These sequences resolved into 43 clusters at 92% amino acid sequence identity, most of which were non-cyanobacterial phylotypes from sea ice and water samples. One cyanobacterial phylotype related to Nodularia sp. was retrieved from sea ice, suggesting that this important functional group is rare in the Central Arctic Ocean. The diazotrophic community in sea-ice environments appear distinct from other cold-adapted diazotrophic communities, such as those present in the coastal Canadian Arctic, the Arctic tundra and glacial Antarctic lakes. Molecular fingerprinting of nifH and the intergenic spacer region of the rRNA operon revealed differences between the communities from river-influenced Laptev Sea waters and those from ice-related environments pointing towards a marine origin for sea-ice diazotrophs. Our results provide the first record of diazotrophs in the Central Arctic and suggest that microbial nitrogen fixation may occur north of 77ºN. To assess the significance of nitrogen fixation for the nitrogen budget of the Arctic Ocean and to identify the active nitrogen fixers, further biogeochemical and molecular biological studies are needed.

  12. Diel regulation of photosynthetic activity in the oceanic unicellular diazotrophic cyanobacterium Crocosphaera watsonii WH8501

    Czech Academy of Sciences Publication Activity Database

    Masuda, Takako; Bernát, Gábor; Bečková, Martina; Kotabová, Eva; Lawrenz, Evelyn; Lukeš, Martin; Komenda, Josef; Prášil, Ondřej

    2018-01-01

    Roč. 20, č. 2 (2018), s. 546-560 ISSN 1462-2912 R&D Projects: GA MŠk(CZ) LO1416; GA MŠk(CZ) ED2.1.00/19.0392; GA ČR GA16-15467S Institutional support: RVO:61388971 Keywords : NORTH PACIFIC-OCEAN * SYNECHOCYSTIS SP PCC-6803; * PHOTOSYSTEM-II Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 5.395, year: 2016

  13. Ultradian metabolic rhythm in the diazotrophic cyanobacterium Cyanothece sp. ATCC 51142

    Czech Academy of Sciences Publication Activity Database

    Červený, Jan; Sinětova, Maria A.; Valledor, Luis; Sherman, L. A.; Nedbal, Ladislav

    2013-01-01

    Roč. 110, č. 32 (2013), s. 13210-13215 ISSN 0027-8424 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0073; GA MŠk(CZ) EE2.3.20.0256; GA ČR GA206/09/1284 Institutional support: RVO:67179843 Keywords : cyanobacteria * diurnal * metabolism * oscillation Subject RIV: EH - Ecology, Behaviour Impact factor: 9.809, year: 2013

  14. Nitrogen signalling in plant interactions with associative and endophytic diazotrophic bacteria.

    Science.gov (United States)

    Carvalho, T L G; Balsemão-Pires, E; Saraiva, R M; Ferreira, P C G; Hemerly, A S

    2014-10-01

    Some beneficial plant-interacting bacteria can biologically fix N2 to plant-available ammonium. Biological nitrogen fixation (BNF) is an important source of nitrogen (N) input in agriculture and represents a promising substitute for chemical N fertilizers. Diazotrophic bacteria have the ability to develop different types of root associations with different plant species. Among the highest rates of BNF are those measured in legumes nodulated by endosymbionts, an already very well documented model of plant-diazotrophic bacterial association. However, it has also been shown that economically important crops, especially monocots, can obtain a substantial part of their N needs from BNF by interacting with associative and endophytic diazotrophic bacteria, that either live near the root surface or endophytically colonize intercellular spaces and vascular tissues of host plants. One of the best reported outcomes of this association is the promotion of plant growth by direct and indirect mechanisms. Besides fixing N, these bacteria can also produce plant growth hormones, and some species are reported to improve nutrient uptake and increase plant tolerance against biotic and abiotic stresses. Thus, this particular type of plant-bacteria association consists of a natural beneficial system to be explored; however, the regulatory mechanisms involved are still not clear. Plant N status might act as a key signal, regulating and integrating various metabolic processes that occur during association with diazotrophic bacteria. This review will focus on the recent progress in understanding plant association with associative and endophytic diazotrophic bacteria, particularly on the knowledge of the N networks involved in BNF and in the promotion of plant growth. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  15. Inoculation of sugarcane with diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Nivaldo Schultz

    2014-04-01

    Full Text Available The sugarcane industry, a strategic crop in Brazil, requires technological improvements in production efficiency to increase the crop energy balance. Among the various currently studied alternatives, inoculation with diazotrophic bacteria proved to be a technology with great potential. In this context, the efficiency of a mixture of bacterial inoculant was evaluated with regard to the agronomic performance and N nutrition of sugarcane. The experiment was carried out on an experimental field of Embrapa Agrobiologia, in Seropédica, Rio de Janeiro, using a randomized block, 2 × 3 factorial design (two varieties and three treatments with four replications, totaling 24 plots. The varieties RB867515 and RB72454 were tested in treatments consisting of: inoculation with diazotrophic bacteria, N-fertilized control with 120 kg ha-1 N and absolute control (no inoculation and no N fertilizer. The inoculum was composed of five strains of five diazotrophic species. The yield, dry matter accumulation, total N in the shoot dry matter and the contribution of N by biological fixation were evaluated, using the natural 15N abundance in non-inoculated sugarcane as reference. The bacterial inoculant increased the stalk yield of variety RB72454 similarly to fertilization with 120 kg ha-1 N in the harvests of plant-cane and first ratoon crops, however the contribution of biological N fixation was unchanged by inoculation, indicating that the benefits of the inoculant in sugarcane may have resulted from plant growth promotion.

  16. Basin scale variability of active diazotrophs and nitrogen fixation in the North Pacific, from the tropics to the subarctic Bering Sea

    Science.gov (United States)

    Shiozaki, Takuhei; Bombar, Deniz; Riemann, Lasse; Hashihama, Fuminori; Takeda, Shigenobu; Yamaguchi, Tamaha; Ehama, Makoto; Hamasaki, Koji; Furuya, Ken

    2017-06-01

    Nitrogen-fixing microorganisms (diazotrophs) provide biologically available nitrogen to plankton communities and thereby greatly influence the productivity in many marine regions. Various cyanobacterial groups have traditionally been considered the major oceanic diazotrophs, but later noncyanobacterial and presumably heterotrophic diazotrophs were also found to be widespread and potentially important in nitrogen fixation. However, the distribution and activity of different diazotroph groups is still poorly constrained for most oceanic ecosystems. Here we examined diazotroph community structure and activity along a 7500 km south-north transect between the central equatorial Pacific and the Bering Sea. Nitrogen fixation contributed up to 84% of new production in the upper waters of the subtropical gyre, where the diazotroph community included the gammaproteobacterium γ-24774A11 and highly active cyanobacterial phylotypes (>50% of total nifH transcript abundance). Nitrogen fixation was sometimes detectable down to 150 m depth and extended horizontally to the edge of the gyre at around 35°N. Nitrogen fixation was even detected far north on the Bering Sea shelf. In the Alaskan Coastal Waters on the Bering Sea shelf, low nitrate together with high dissolved iron concentrations seemed to foster diazotroph growth, including a prominent role of UCYN-A2, which was abundant near the surface (1.2×105 nifH gene copies L-1). Our study provides evidence for nitrogen fixation in the Bering Sea and suggests a clear contrast in the composition of diazotrophs between the tropical/subtropical gyre and the separate waters in the cold northern regions of the North Pacific.

  17. GENETIC VARIABILITY OF SUGARCANE-ASSOCIATED DIAZOTROPHIC BACTERIA CAPABLE OF INORGANIC PHOSPHATE SOLUBILIZING

    OpenAIRE

    Lira-Cadete, Luana; Barbosa de Farias, Andreza Raquel; de Souza Ramos, Andresa Priscila; da Costa, Diogo Paes; Freire, Fernando Jose; Kuklinsky-Sobral, Julia

    2013-01-01

    The sugarcane is a culture of great importance for the Brazilian agriculture. Every year this culture consumes great amounts of nitrogen and phosphate fertilizers. However, the use of plant growth-promoting bacteria can reduce the use of the chemical fertilizers, contributing to the economy and the environment conservation. So, the goal of this study was to select sugarcane-associated diazotrophic bacteria able to solubilize inorganic phosphate and to evaluate the genetic diversity of these b...

  18. Effects of long-term elevated CO2, warming, and prolonged drought on Pleurozium-associated diazotrophic activity and abundance

    Science.gov (United States)

    Dyrnum, Kristine; Priemé, Anders; Michelsen, Anders

    2014-05-01

    Nitrogen (N2) fixation is the primary natural influx of N to terrestrial ecosystems, and changes in N2 fixation may have consequences for primary productivity and thus ecosystem function. We studied the activity and abundance of diazotrophs associated with the feather moss Pleurozium schreberi in a temperate heathland, after seven years of global change manipulations, including elevated atmospheric CO2 (510 ppm), increased temperature (0.5-1.5 ° C), and prolonged pre-summer droughts (4-6 weeks /year). Acetylene reduction assay was carried out monthly to monitor N2 fixation rates throughout one year, while nif H copy abundance, serving as a diazotroph abundance estimate, was measured by quantitative polymerase chain reaction (q-PCR). Prolonged summer droughts significantly increased both N2 fixation and nif H copy abundance, contrasting previous studies that demonstrate a direct negative correlation between N2 fixation and water availability. A shift in the relative abundance of N2-fixing bacteria from the green, upper parts of the moss stem to the lower, brown parts was observed. This shift could make diazotrophs less sensitive to desiccation, enabling N2 fixation to be upheld for longer during drought and thus causing higher abundance. Increased temperature likewise had a positive effect on the diazotroph abundance, although this did not translate into increased activity. Possibly, warming protects diazotrophs during extreme cold events, while actual N2 fixation is limited by water, disregarding a rise in potential N2 fixation caused by higher abundance. Increased CO2 caused no significant diazotroph response. Our study showed that long-term increase in temperature and recurrent drought events cause higher diazotroph abundance in Pleurozium schreberi and thus enhance the potential N2 fixations rate. Furthermore, our results indicate that diazotrophs may alter colonization patterns and thereby actively remain in the moss fraction less likely affected by

  19. Temporal changes in the diazotrophic bacterial communities associated with Caribbean sponges Ircinia stroblina and Mycale laxissima

    Directory of Open Access Journals (Sweden)

    Fan eZhang

    2014-10-01

    Full Text Available Sponges that harbor microalgal or cyanobacterial symbionts may benefit from photosynthetically derived carbohydrates, which are rich in carbon but devoid of nitrogen, and may therefore encounter nitrogen limitation. Diazotrophic communities associated with two Caribbean sponges, Ircinia strobilina and Mycale laxissima were studied in a time series during which three individuals of each sponge were collected in four time points (5:00 AM, 12:00 noon, 5:00 PM, 10:00 PM. nifH genes were successfully amplified from the corresponding gDNA and cDNA pools and sequenced by high throughput 454 amplicon sequencing. In both sponges, over half the nifH transcripts were classified as from cyanobacteria and the remainder from heterotrophic bacteria. We found various groups of bacteria actively expressing the nifH gene during the entire day-night cycle, an indication that the nitrogen fixation potential was fully exploited by different nitrogen fixing bacteria groups associated with their hosts. This study showed for the first time the dynamic changes in the activity of the diazotrophic bacterial communities in marine sponges. Our study expands understanding of the diazotrophic groups that contribute to the fixed nitrogen pool in the benthic community. Sponge bacterial community-associated diazotrophy may have an important impact on the nitrogen biogeochemical cycle in the coral reef ecosystem.

  20. Temporal changes in the diazotrophic bacterial communities associated with Caribbean sponges Ircinia stroblina and Mycale laxissima.

    Science.gov (United States)

    Zhang, Fan; Vicente, Jan; Hill, Russell T

    2014-01-01

    Sponges that harbor microalgal or, cyanobacterial symbionts may benefit from photosynthetically derived carbohydrates, which are rich in carbon but devoid of nitrogen, and may therefore encounter nitrogen limitation. Diazotrophic communities associated with two Caribbean sponges, Ircinia strobilina and Mycale laxissima were studied in a time series during which three individuals of each sponge were collected in four time points (5:00 AM, 12:00 noon, 5:00 PM, 10:00 PM). nifH genes were successfully amplified from the corresponding gDNA and cDNA pools and sequenced by high throughput 454 amplicon sequencing. In both sponges, over half the nifH transcripts were classified as from cyanobacteria and the remainder from heterotrophic bacteria. We found various groups of bacteria actively expressing the nifH gene during the entire day-night cycle, an indication that the nitrogen fixation potential was fully exploited by different nitrogen fixing bacteria groups associated with their hosts. This study showed for the first time the dynamic changes in the activity of the diazotrophic bacterial communities in marine sponges. Our study expands understanding of the diazotrophic groups that contribute to the fixed nitrogen pool in the benthic community. Sponge bacterial community-associated diazotrophy may have an important impact on the nitrogen biogeochemical cycle in the coral reef ecosystem.

  1. Agrobacterium tumefaciens is a diazotrophic bacterium

    International Nuclear Information System (INIS)

    Kanvinde, L.; Sastry, G.R.K.

    1990-01-01

    This is the first report that Agrobacterium tumefaciens can fix nitrogen in a free-living condition as shown by its abilities to grown on nitrogen-free medium, reduce acetylene to ethylene, and incorporate 15 N supplied as 15 N 2 . As with most other well-characterized diazotrophic bacteria, the presence of NH 4 + in the medium and aerobic conditions repress nitrogen fixation by A. tumefaciens. The system requires molybdenum. No evidence for nodulation was found with pea, peanut, or soybean plants. Further understanding of the nitrogen-fixing ability of this bacterium, which has always been considered a pathogen, should cast new light on the evolution of a pathogenic versus symbiotic relationship

  2. Agrobacterium tumefaciens is a diazotrophic bacterium

    Energy Technology Data Exchange (ETDEWEB)

    Kanvinde, L.; Sastry, G.R.K. (Univ. of Leeds (England))

    1990-07-01

    This is the first report that Agrobacterium tumefaciens can fix nitrogen in a free-living condition as shown by its abilities to grown on nitrogen-free medium, reduce acetylene to ethylene, and incorporate {sup 15}N supplied as {sup 15}N{sub 2}. As with most other well-characterized diazotrophic bacteria, the presence of NH{sub 4}{sup +} in the medium and aerobic conditions repress nitrogen fixation by A. tumefaciens. The system requires molybdenum. No evidence for nodulation was found with pea, peanut, or soybean plants. Further understanding of the nitrogen-fixing ability of this bacterium, which has always been considered a pathogen, should cast new light on the evolution of a pathogenic versus symbiotic relationship.

  3. Differential distribution and abundance of diazotrophic bacterial communities across different soil niches using a gene-targeted clone library approach.

    Science.gov (United States)

    Yousuf, Basit; Kumar, Raghawendra; Mishra, Avinash; Jha, Bhavanath

    2014-11-01

    Diazotrophs are key players of the globally important biogeochemical nitrogen cycle, having a significant role in maintaining ecosystem sustainability. Saline soils are pristine and unexplored habitats representing intriguing ecosystems expected to harbour potential diazotrophs capable of adapting in extreme conditions, and these implicated organisms are largely obscure. Differential occurrence of diazotrophs was studied by the nifH gene-targeted clone library approach. Four nifH gene clone libraries were constructed from different soil niches, that is saline soils (low and high salinity; EC 3.8 and 7.1 ds m(-1) ), and agricultural and rhizosphere soil. Additionally, the abundance of diazotrophic community members was assessed using quantitative PCR. Results showed environment-dependent metabolic versatility and the presence of nitrogen-fixing bacteria affiliated with a range of taxa, encompassing members of the Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, Gammaproteobacteria, Cyanobacteria and Firmicutes. The analyses unveiled the dominance of Alphaproteobacteria and Gammaproteobacteria (Pseudomonas, Halorhodospira, Ectothiorhodospira, Bradyrhizobium, Agrobacterium, Amorphomonas) as nitrogen fixers in coastal-saline soil ecosystems, and Alphaproteobacteria and Betaproteobacteria (Bradyrhizobium, Azohydromonas, Azospirillum, Ideonella) in agricultural/rhizosphere ecosystems. The results revealed a repertoire of novel nitrogen-fixing bacterial guilds particularly in saline soil ecosystems. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  4. Characterization of Diazotrophs Containing Mo-Independent Nitrogenases, Isolated from Diverse Natural Environments

    Science.gov (United States)

    Molybdenum-independent nitrogenases were first described in the nitrogen-fixing bacterium Azotobacter vinelandii and have since been described in other diazotrophic bacteria. Previously, we reported the isolation of seven diazotrophs with Molybdenum-independent nitrogenases from aquatic environments...

  5. Oscillating behavior of carbohydrate granule formation and dinitrogen fixation in the cyanobacterium Cyanothece sp. strain ATCC 51142

    Science.gov (United States)

    Schneegurt, M. A.; Sherman, D. M.; Nayar, S.; Sherman, L. A.; Mitchell, C. A. (Principal Investigator)

    1994-01-01

    It has been shown that some aerobic, unicellular, diazotrophic cyanobacteria temporally separate photosynthetic O2 evolution and oxygen-sensitive N2 fixation. Cyanothece sp. ATCC strain 51142 is an aerobic, unicellular, diazotrophic cyanobacterium that fixes N2 during discrete periods of its cell cycle. When the bacteria are maintained under diurnal light-dark cycles, N2 fixation occurs in the dark. Similar cycling is observed in continuous light, implicating a circadian rhythm. Under N2-fixing conditions, large inclusion granules form between the thylakoid membranes. Maximum granulation, as observed by electron microscopy, occurs before the onset of N2 fixation, and the granules decrease in number during the period of N2 fixation. The granules can be purified from cell homogenates by differential centrifugation. Biochemical analyses of the granules indicate that these structures are primarily carbohydrate, with some protein. Further analyses of the carbohydrate have shown that it is a glucose polymer with some characteristics of glycogen. It is proposed that N2 fixation is driven by energy and reducing power stored in these inclusion granules. Cyanothece sp. strain ATCC 51142 represents an excellent experimental organism for the study of the protective mechanisms of nitrogenase, metabolic events in cyanobacteria under normal and stress conditions, the partitioning of resources between growth and storage, and biological rhythms.

  6. Nitrogen uptake dynamics of a persistent cyanobacterium ...

    African Journals Online (AJOL)

    Worldwide, persistent cyanobacterial blooms are becoming more frequent and are often associated with effects of global climate change. In June 2009, a widespread bloom of the unicellular cyanobacterium, Cyanothece sp., appeared in North Lake and False Bay of Lake St Lucia – a large (360 km2) estuarine lake system ...

  7. Growth Characteristics of an Estuarine Heterocystous Cyanobacterium

    NARCIS (Netherlands)

    Guimarães, P.; Yunes, J.S.; Cretoiu, M.S.; Stal, L.J.

    2017-01-01

    A new estuarine filamentous heterocystous cyanobacterium was isolated from intertidal sediment of the Lagoa dos Patos estuary (Brazil). The isolate may represent a new genus related to Cylindrospermopsis. While the latter is planktonic, contains gas vesicles, and is toxic, the newly isolated strain

  8. Photoacclimation of cultured strains of the cyanobacterium

    NARCIS (Netherlands)

    Bañares-España, E.; Kromkamp, J.C.; López-Rodas, V.; Costas, E.; Flores-Moya, A.

    2013-01-01

    The cyanobacterium Microcystis aeruginosa forms blooms that can consist of colonies. We have investigated how M.aeruginosa acclimatizes to changing light conditions such as can occur during blooms. Three different strains were exposed to two irradiance levels: lower (LL) and higher (HL) than the

  9. Bacterial N2-fixation in mangrove ecosystems: insights from a diazotroph-mangrove interaction.

    Science.gov (United States)

    Alfaro-Espinoza, Gabriela; Ullrich, Matthias S

    2015-01-01

    Mangrove forests are highly productive ecosystems but represent low nutrient environments. Nitrogen availability is one of the main factors limiting mangrove growth. Diazotrophs have been identified as key organisms that provide nitrogen to these environments. N2-fixation by such organisms was found to be higher in the mangrove roots than in surrounding rhizosphere. Moreover, previous studies showed that mangroves grew better in the presence of N2-fixers indicating a potentially mutualistic relationship. However, the molecular signals and mechanisms that govern these interactions are still poorly understood. Here we present novel insights in the interaction of a diazotroph with a mangrove species to improve our understanding of the molecular and ecophysiological relationship between these two organisms under controlled conditions. Our results showed that Marinobacterium mangrovicola is a versatile organism capable of competing with other organisms to survive for long periods in mangrove soils. N2-fixation by this bacterium was up-regulated in the presence of mangrove roots, indicating a possible beneficial interaction. The increase in N2-fixation was limited to cells of the exponential growth phase suggesting that N2-fixation differs over the bacterial growth cycle. Bacterial transformants harboring a transcriptional nifH::gusA fusion showed that M. mangrovicola successfully colonized mangrove roots and simultaneously conducted N2-fixation. The colonization process was stimulated by the lack of an external carbon source suggesting a possible mutualistic relationship. M. mangrovicola represents an interesting genetically accessible diazotroph, which colonize mangrove roots and exhibit higher N2-fixation in the presence of mangrove roots. Consequently, we propose this microorganism as a tool to study molecular interactions between N2-fixers and mangrove plants and to better understand how changes in the environment could impact these important and relatively unknown

  10. Technetium and diazotrophic organisms: toxicity, localization, transfer factors

    International Nuclear Information System (INIS)

    Vandecasteele, C.M.; Delmotte, A.; Roucoux, P.; Hove, C. van

    1982-01-01

    Three diazotrophic organisms, together with one leguminous organism in symbiosis with one of them, were cultivated in the presence of various quantities of technetium, of which the localization, transfer factors and toxicity were studied in relation to the age of the organisms and their type of metabolism. The paper discusses the biochemical aspects of the results. (author)

  11. Isolation of heterotrophic diazotrophic bacteria from estuarine surface waters.

    Science.gov (United States)

    Farnelid, Hanna; Harder, Jens; Bentzon-Tilia, Mikkel; Riemann, Lasse

    2014-10-01

    The wide distribution of diverse nitrogenase (nifH) genes affiliated with those of heterotrophic bacteria in marine and estuarine waters indicates ubiquity and an ecologically relevant role for heterotrophic N2 -fixers (diazotrophs) in aquatic nitrogen (N) cycling. However, the lack of cultivated representatives currently precludes an evaluation of their N2 -fixing capacity. In this study, microoxic or anoxic N-free media were inoculated with estuarine Baltic Sea surface water to select for N2 -fixers. After visible growth and isolation of single colonies on oxic plates or in anoxic agar tubes, nifH gene amplicons were obtained from 64 strains and nitrogenase activity, applying the acetylene reduction assay, was confirmed for 40 strains. Two strains, one Gammaproteobacterium affiliated with Pseudomonas and one Alphaproteobacterium affiliated with Rhodopseudomonas were shown to represent established members of the indigenous diazotrophic community in the Baltic Sea, with abundances of up to 7.9 × 10(4) and 4.7 × 10(4)  nifH copies l(-1) respectively. This study reports media for successful isolation of heterotrophic diazotrophs. The applied methodology and the obtained strains will facilitate future identification of factors controlling heterotrophic diazotrophic activity in aquatic environments, which is a prerequisite for understanding and evaluating their ecology and contribution to N cycling at local and regional scales. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  12. Database of diazotrophs in global ocean: abundance, biomass and nitrogen fixation rates

    Directory of Open Access Journals (Sweden)

    Y.-W. Luo

    2012-08-01

    Full Text Available Marine N2 fixing microorganisms, termed diazotrophs, are a key functional group in marine pelagic ecosystems. The biological fixation of dinitrogen (N2 to bioavailable nitrogen provides an important new source of nitrogen for pelagic marine ecosystems and influences primary productivity and organic matter export to the deep ocean. As one of a series of efforts to collect biomass and rates specific to different phytoplankton functional groups, we have constructed a database on diazotrophic organisms in the global pelagic upper ocean by compiling about 12 000 direct field measurements of cyanobacterial diazotroph abundances (based on microscopic cell counts or qPCR assays targeting the nifH genes and N2 fixation rates. Biomass conversion factors are estimated based on cell sizes to convert abundance data to diazotrophic biomass. The database is limited spatially, lacking large regions of the ocean especially in the Indian Ocean. The data are approximately log-normal distributed, and large variances exist in most sub-databases with non-zero values differing 5 to 8 orders of magnitude. Reporting the geometric mean and the range of one geometric standard error below and above the geometric mean, the pelagic N2 fixation rate in the global ocean is estimated to be 62 (52–73 Tg N yr−1 and the pelagic diazotrophic biomass in the global ocean is estimated to be 2.1 (1.4–3.1 Tg C from cell counts and to 89 (43–150 Tg C from nifH-based abundances. Reporting the arithmetic mean and one standard error instead, these three global estimates are 140 ± 9.2 Tg N yr−1, 18 ± 1.8 Tg C and 590 ± 70 Tg C, respectively. Uncertainties related to biomass conversion factors can change the estimate of geometric mean pelagic diazotrophic biomass in the global ocean by about ±70%. It was recently established that the most commonly applied method used to measure N2

  13. Effect of salinity on diazotrophic activity and microbial composition of phototrophic communities from Bitter-1 soda lake (Kulunda Steppe, Russia).

    Science.gov (United States)

    Namsaraev, Zorigto; Samylina, Olga; Sukhacheva, Marina; Borisenko, Gennadii; Sorokin, Dimitry Y; Tourova, Tatiana

    2018-04-16

    Bitter-1 is a shallow hypersaline soda lake in Kulunda Steppe (Altai region, Russia). During a study period between 2005 and 2016, the salinity in the littoral area of the lake fluctuated within the range from 85 to 400 g/L (in July of each year). Light-dependent nitrogen fixation occurred in this lake up to the salt-saturating conditions. The rates increased with a decrease in salinity, both under environmental conditions and in laboratory simulations. The salinities below 100 g/L were favorable for light-dependent nitrogen fixation, while the process was dramatically inhibited above 200 g/L salts. The analysis of nifH genes in environmental samples and in enrichment cultures of diazotrophic phototrophs suggested that anaerobic fermenting and sulfate-reducing bacteria could participate in the dark nitrogen fixation process up to soda-saturating conditions. However, we cannot exclude the possibility that haloalkaliphilic nonheterocystous cyanobacteria (Euhalothece sp. and Geitlerinema sp.) and anoxygenic purple sulfur bacteria (Ectothiorhodospira sp.) might also play a role in the process at light conditions. The heterocystous cyanobacterium Nodularia sp. develops at low salinity (below 80 g/L) that is not characteristic for Bitter-1 Lake and thus does not make a significant contribution to the nitrogen fixation in this lake.

  14. Attachment of associative diazotroph alcaligenes faecalis to rice roots

    International Nuclear Information System (INIS)

    Lin Min; Fang Xuanjun; You Chongbiao

    1993-01-01

    The process of attachment of diazotroph Alcaligenes faecalis to host plant rice was studied by using 15 N-labelled bacteria and Tn5-induced mutants. A three-step attachment mechanism of A. faecalis to rice root surface is proposed on the basis of experimental data. Adsorption is the first step. The number of adsorbed bacteria reaches maximal level after 3 h of inoculation, it consists 3.7% of the total number of bacteria inoculated. Adsorbed bacteria could be removed from rice root surface quantitatively by shaking in water. Therefore, the adsorption forces are weak. Anchoring is the second step. It begins only after 9h of inoculation and reaches a maximal level (21%) after 16 h. Anchored bacteria could not be removed by shaking. Colonization is the third step. After 20 h of inoculation. part of anchored bacteria colonizes on rice root surface tightly, and it can not be removed by vortex. At this time, the pectolytic activity of bacteria appears. Chemotaxis and exopolysaccharide (EPS) play important roles in the attachment of A. faecalis to rice root surface. EPS mutants (Exo - , Exo ++ ) showed less anchoring-capability in comparison with wild type of bacterium, but they remained the adsorption capability. While chemotaxis (Che - ) mutants are defective in adsorption, but not in anchoring. Che - , Exo - mutant lost both adsorption and anchoring capabilities. A. faecalis absorbed on all part of rice root, but the anchoring and colonization of bacteria were occurred mainly on root hairs, particularly on the joint area of main root and lateral root

  15. Enhancement of Chlorella vulgaris growth and bioremediation ability of aquarium wastewater using diazotrophs.

    Science.gov (United States)

    Ali, Sayeda Mohammed; Nasr, Hoda Shafeek; Abbas, Wafaa Tawfik

    2012-08-15

    Treatment of aquarium wastewater represents an important process to clean and recycle wastewater to be safely returned to the environment, used for cultivation or to minimize the multiple renewal of water. Chlorella vulgaris was an important freshwater microalgae which used in wastewater treatment, and increasing its potential of treatment can be achieved with existence of N2-fixing bacteria. Co-culturing of Chlorella vulgaris with the diazotrophs, Azospirillum brasilense or Azotobacter chroococcum in three different media; aquarium wastewater (AWW), sterile enriched natural aquarium wastewater (GPM) and synthetic wastewater media (SWW) were studied. Biomass yield of the microalgae was estimated by determination of chlorophylls (a and b), total carotenoid and the dry weight of C. vulgaris. Also determination of ammonia, nitrite, phosphate and nitrate in the culture were done. The presence of diazotrophs significantly increased the biomass of C. vulgaris by increasing its microalgae pigments (chlorophylls a and b, and total carotenoids). The highest pigments percentage was reported due to addition of A. brasilense to C. vulgaris (18.3-133.5%) compared to A. chroococcum (23.9-56.9%). As well as increased dry weight from 12 to 50%. There was also improved removal of nitrate, nitrite, ammonia and phosphate; where, the highest removal percentage was reported due to addition of A. chroococcum to C. vulgaris (0.0-52%) compared to A. brasilense (0.6-16.4%). A. brasilense and A. chroococcum can support C. vulgaris biomass production and bioremediation activity in the aquarium to minimize the periodical water renewal.

  16. Genome-wide identification of microRNA and siRNA responsive to endophytic beneficial diazotrophic bacteria in maize.

    Science.gov (United States)

    Thiebaut, Flávia; Rojas, Cristian A; Grativol, Clícia; Motta, Mariana Romeiro; Vieira, Tauan; Regulski, Michael; Martienssen, Robert A; Farinelli, Laurent; Hemerly, Adriana S; Ferreira, Paulo C G

    2014-09-06

    Small RNA (sRNA) has been described as a regulator of gene expression. In order to understand the role of maize sRNA (Zea mays-hybrid UENF 506-8) during association with endophytic nitrogen-fixing bacteria, we analyzed the sRNA regulated by its association with two diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. Deep sequencing analysis was done with RNA extracted from plants inoculated with H. seropedicae, allowing the identification of miRNA and siRNA. A total of 25 conserved miRNA families and 15 novel miRNAs were identified. A dynamic regulation in response to inoculation was also observed. A hypothetical model involving copper-miRNA is proposed, emphasizing the fact that the up-regulation of miR397, miR398, miR408 and miR528, which is followed by inhibition of their targets, can facilitate association with diazotrophic bacteria. Similar expression patterns were observed in samples inoculated with A. brasilense. Moreover, novel miRNA and siRNA were classified in the Transposable Elements (TE) database, and an enrichment of siRNA aligned with TE was observed in the inoculated samples. In addition, an increase in 24-nt siRNA mapping to genes was observed, which was correlated with an increase in methylation of the coding regions and a subsequent reduction in transcription. Our results show that maize has RNA-based silencing mechanisms that can trigger specific responses when plants interact with beneficial endophytic diazotrophic bacteria. Our findings suggest important roles for sRNA regulation in maize, and probably in other plants, during association with diazotrophic bacteria, emphasizing the up-regulation of Cu-miRNA.

  17. The cyanobacterium Mastigocladus fulfills the nitrogen demand of a terrestrial hot spring microbial mat.

    Science.gov (United States)

    Estrella Alcamán, María; Fernandez, Camila; Delgado, Antonio; Bergman, Birgitta; Díez, Beatriz

    2015-10-01

    Cyanobacteria from Subsection V (Stigonematales) are important components of microbial mats in non-acidic terrestrial hot springs. Despite their diazotrophic nature (N2 fixers), their impact on the nitrogen cycle in such extreme ecosystems remains unknown. Here, we surveyed the identity and activity of diazotrophic cyanobacteria in the neutral hot spring of Porcelana (Northern Patagonia, Chile) during 2009 and 2011-2013. We used 16S rRNA and the nifH gene to analyze the distribution and diversity of diazotrophic cyanobacteria. Our results demonstrate the dominance of the heterocystous genus Mastigocladus (Stigonematales) along the entire temperature gradient of the hot spring (69-38 °C). In situ nitrogenase activity (acetylene reduction), nitrogen fixation rates (cellular uptake of (15)N2) and nifH transcription levels in the microbial mats showed that nitrogen fixation and nifH mRNA expression were light-dependent. Nitrogen fixation activities were detected at temperatures ranging from 58 °C to 46 °C, with maximum daily rates of 600 nmol C2H4 cm(-2) per day and 94.1 nmol N cm(-2) per day. These activity patterns strongly suggest a heterocystous cyanobacterial origin and reveal a correlation between nitrogenase activity and nifH gene expression during diurnal cycles in thermal microbial mats. N and C fixation in the mats contributed ~3 g N m(-2) per year and 27 g C m(-2) per year, suggesting that these vital demands are fully met by the diazotrophic and photoautotrophic capacities of the cyanobacteria in the Porcelana hot spring.

  18. Performance of pineapple slips inoculated with diazotrophic phosphate-solubilizing bacteria and rock phosphate

    OpenAIRE

    Lílian Estrela Borges Baldotto; Marihus Altoé Baldotto; Fábio Lopes Olivares; Adriane Nunes de Souza

    2014-01-01

    Besides fixing N2, some diazotrophic bacteria or diazotrophs, also synthesize organic acids and are able to solubilize rock phosphates, increasing the availability of P for plants. The application of these bacteria to pineapple leaf axils in combination with rock phosphate could increase N and P availability for the crop, due to the bacterial activity of biological nitrogen fixation and phosphate solubilization. The objectives of this study were: (i) to select and characterize diazotrophs abl...

  19. Transfer of diazotroph-derived nitrogen towards non-diazotrophic planktonic communities: a comparative study between Trichodesmium erythraeum, Crocosphaera watsonii and Cyanothece sp.

    Science.gov (United States)

    Berthelot, Hugo; Bonnet, Sophie; Grosso, Olivier; Cornet, Véronique; Barani, Aude

    2016-07-01

    Biological dinitrogen (N2) fixation is the major source of new nitrogen (N) for the open ocean, and thus promotes marine productivity, in particular in the vast N-depleted regions of the surface ocean. Yet, the fate of the diazotroph-derived N (DDN) in marine ecosystems is poorly understood, and its transfer to auto- and heterotrophic surrounding plankton communities is rarely measured due to technical limitations. Moreover, the different diazotrophs involved in N2 fixation (Trichodesmium spp. vs. UCYN) exhibit distinct patterns of N2 fixation and inhabit different ecological niches, thus having potentially different fates in the marine food webs that remain to be explored. Here we used nanometer scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labelling and flow cytometry cell sorting to examine the DDN transfer to specific groups of natural phytoplankton and bacteria during artificially induced diazotroph blooms in New Caledonia (southwestern Pacific). The fate of the DDN was compared according to the three diazotrophs: the filamentous and colony-forming Trichodesmium erythraeum (IMS101), and the unicellular strains Crocosphaera watsonii WH8501 and Cyanothece ATCC51142. After 48 h, 7-17 % of the N2 fixed during the experiment was transferred to the dissolved pool and 6-12 % was transferred to non-diazotrophic plankton. The transfer was twice as high in the T. erythraeum bloom than in the C. watsonii and Cyanothece blooms, which shows that filamentous diazotrophs blooms are more efficient at promoting non-diazotrophic production in N-depleted areas. The amount of DDN released in the dissolved pool did not appear to be a good indicator of the DDN transfer efficiency towards the non-diazotrophic plankton. In contrast, the 15N-enrichment of the extracellular ammonium (NH4+) pool was a good indicator of the DDN transfer efficiency: it was significantly higher in the T. erythraeum than in unicellular diazotroph blooms, leading to a DDN

  20. Anthropogenic impact on diazotrophic diversity in the mangrove rhizosphere revealed by nifH pyrosequencing.

    Science.gov (United States)

    Jing, Hongmei; Xia, Xiaomin; Liu, Hongbin; Zhou, Zhi; Wu, Chen; Nagarajan, Sanjay

    2015-01-01

    Diazotrophs in the mangrove rhizosphere play a major role in providing new nitrogen to the mangrove ecosystem and their composition and activity are strongly influenced by anthropogenic activity and ecological conditions. In this study, the diversity of the diazotroph communities in the rhizosphere sediment of five tropical mangrove sites with different levels of pollution along the north and south coastline of Singapore were studied by pyrosequencing of the nifH gene. Bioinformatics analysis revealed that in all the studied locations, the diazotroph communities comprised mainly of members of the diazotrophic cluster I and cluster III. The detected cluster III diazotrophs, which were composed entirely of sulfate-reducing bacteria, were more abundant in the less polluted locations. The metabolic capacities of these diazotrophs indicate the potential for bioremediation and resiliency of the ecosystem to anthropogenic impact. In heavily polluted locations, the diazotrophic community structures were markedly different and the diversity of species was significantly reduced when compared with those in a pristine location. This, together with the increased abundance of Marinobacterium, which is a bioindicator of pollution, suggests that anthropogenic activity has a negative impact on the genetic diversity of diazotrophs in the mangrove rhizosphere.

  1. CyanOmics: an integrated database of omics for the model cyanobacterium Synechococcus sp. PCC 7002

    OpenAIRE

    Yang, Yaohua; Feng, Jie; Li, Tao; Ge, Feng; Zhao, Jindong

    2015-01-01

    Cyanobacteria are an important group of organisms that carry out oxygenic photosynthesis and play vital roles in both the carbon and nitrogen cycles of the Earth. The annotated genome of Synechococcus sp. PCC 7002, as an ideal model cyanobacterium, is available. A series of transcriptomic and proteomic studies of Synechococcus sp. PCC 7002 cells grown under different conditions have been reported. However, no database of such integrated omics studies has been constructed. Here we present Cyan...

  2. Genetic manipulation of a metabolic enzyme and a transcriptional regulator increasing succinate excretion from unicellular cyanobacterium

    Directory of Open Access Journals (Sweden)

    Takashi eOsanai

    2015-10-01

    Full Text Available Succinate is a building block compound that the U.S. Department of Energy has declared as important in biorefineries, and it is widely used as a commodity chemical. Here, we identified the two genes increasing succinate production of the unicellular cyanobacterium Synechocystis sp. PCC 6803. Succinate was excreted under dark, anaerobic conditions, and its production level increased by knocking out ackA, which encodes an acetate kinase, and by overexpressing sigE, which encodes an RNA polymerase sigma factor. Glycogen catabolism and organic acid biosynthesis were enhanced in the mutant lacking ackA and overexpressing sigE, leading to an increase in succinate production reaching 5 times of the wild-type levels. Our genetic and metabolomic analyses thus demonstrated the effect of genetic manipulation of a metabolic enzyme and a transcriptional regulator on succinate excretion from this cyanobacterium with the data based on metabolomic technique.

  3. Understanding predicted shifts in diazotroph biogeography using resource competition theory

    Directory of Open Access Journals (Sweden)

    S. Dutkiewicz

    2014-10-01

    Full Text Available We examine the sensitivity of the biogeography of nitrogen fixers to a warming climate and increased aeolian iron deposition in the context of a global earth system model. We employ concepts from the resource-ratio theory to provide a simplifying and transparent interpretation of the results. First we demonstrate that a set of clearly defined, easily diagnosed provinces are consistent with the theory. Using this framework we show that the regions most vulnerable to province shifts and changes in diazotroph biogeography are the equatorial and South Pacific, and central Atlantic. Warmer and dustier climates favor diazotrophs due to an increase in the ratio of supply rate of iron to fixed nitrogen. We suggest that the emergent provinces could be a standard diagnostic for global change models, allowing for rapid and transparent interpretation and comparison of model predictions and the underlying mechanisms. The analysis suggests that monitoring of real world province boundaries, indicated by transitions in surface nutrient concentrations, would provide a clear and easily interpreted indicator of ongoing global change.

  4. Nitrogenase (nifH gene expression in diazotrophic cyanobacteria in the Tropical North Atlantic in response to nutrient amendments

    Directory of Open Access Journals (Sweden)

    Kendra A Turk-Kubo

    2012-11-01

    Full Text Available The Tropical North Atlantic (TNAtl plays a critical role in the marine nitrogen cycle, as it supports high rates of biological nitrogen (N2 fixation, yet it is unclear whether this process is limited by the availability of iron (Fe, phosphate (P or is co-limited by both. In order to investigate the impact of nutrient limitation on the N2-fixing microorganisms (diazotrophs in the TNAtl, trace metal clean nutrient amendment experiments were conducted, and the expression of nitrogenase (nifH in cyanobacterial diazotrophs in response to the addition of Fe, P, or Fe+P was measured using quantitative PCR. To provide context, N2 fixation rates associated with the <10 μm community and diel nifH expression in natural cyanobacterial populations were measured. In the western TNAtl, nifH expression in Crocosphaera, Trichodesmium, and Richelia was stimulated by Fe and Fe+P additions, but not by P, implying that diazotrophs may be Fe-limited in this region. In the eastern TNAtl, nifH expression in unicellular cyanobacteria UCYN-A and Crocosphaera was stimulated by P, implying P-limitation. In equatorial waters, nifH expression in Trichodesmium was highest in Fe+P treatments, implying co-limitation in this region. Nutrient additions did not measurably stimulate N2 fixation rates in the <10 μm fraction in most of the experiments, even when upregulation of nifH expression was evident. These results demonstrate the utility of using gene expression to investigate the physiological state of natural populations of microorganisms, while underscoring the complexity of nutrient limitation on diazotrophy, and providing evidence that diazotroph populations are slow to respond to the addition of limiting nutrients and may be limited by different nutrients on basin-wide spatial scales. This has important implications for our current understanding of controls on N2 fixation in the TNAtl and may partially explain why it appears to be intermittently limited by Fe, P, or

  5. Specificity of salt marsh diazotrophs for vegetation zones and plant hosts

    Directory of Open Access Journals (Sweden)

    Debra Aline Davis

    2012-03-01

    Full Text Available Salt marshes located on the east coast of temperate North America are highly productive, typically nitrogen-limited, and support diverse assemblages of nitrogen fixing (diazotrophic bacteria. The distributions of these diazotrophs are strongly influenced by plant host and abiotic environmental parameters. Crab Haul Creek Basin, North Inlet, SC, USA is a tidally dominated marsh that displays discrete plant zones distributed along an elevation gradient from the tidal creek bank to the terrestrial forest. These zones are defined by gradients of abiotic environmental variables, particularly salinity and sulfide. DGGE fingerprinting and phylogenetic analyses of recovered sequences demonstrated that the distributions of some diazotrophs indicate plant host specificity and that diazotroph assemblages across the marsh gradient are heavily influenced by edaphic conditions. Broadly distributed diazotrophs capable of maintaining populations in all environmental conditions across the gradient are also present in these assemblages. Parsimony test results confirm that diazotroph assemblages in different plant zones are significantly (p<0.01 different across the marsh landscape. Results also indicated that diazotroph assemblages associated with different plant hosts growing in the same area of the marsh were structurally similar confirming the influence of edaphic parameters on these assemblages. Principal Component Analysis of DGGE gel banding patterns confirmed these results. This article reviews and analyzes data from North Inlet Estuary, addressing diazotroph assemblage structure and the influence of plant host and environmental conditions. New data demonstrate the heterogeneity of salt marsh rhizosphere microenvironments, and corroborate previous findings from different plant hosts growing at several locations within this estuary. These data support the hypothesis that the biogeography of microorganisms is non-random and is partially driven by

  6. Diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple plantlets during acclimatization

    OpenAIRE

    Silva,Aurilena de Aviz; Carvalho,Almy Junior Cordeiro de; Freitas,Flávia Paiva de; Pessanha,Patrícia Gomes de Oliveira; Santos,Paulo Cesar dos; Silva,Mírian Peixoto Soares da; Vasconcelos,Tábatha de Souza; Olivares,Fábio Lopes

    2016-01-01

    ABSTRACT: This study examines the effect of inoculation with diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple cv. 'Vitória' plantlets during the acclimatization period. The experiment was carried out in a greenhouse in Campos dos Goytacazes, in randomized blocks, using a 2x5x5 factorial scheme, with the factors being two types of inocula (absence or presence of a mixture of diazotrophic bacteria that contained Burkholderia sp. UENF 114111, Burkholder...

  7. Evidence for the functional significance of diazotroph community structure in soil.

    Science.gov (United States)

    Hsu, Shi-Fang; Buckley, Daniel H

    2009-01-01

    Microbial ecologists continue to seek a greater understanding of the factors that govern the ecological significance of microbial community structure. Changes in community structure have been shown to have functional significance for processes that are mediated by a narrow spectrum of organisms, such as nitrification and denitrification, but in some cases, functional redundancy in the community seems to buffer microbial ecosystem processes. The functional significance of microbial community structure is frequently obscured by environmental variation and is hard to detect in short-term experiments. We examine the functional significance of free-living diazotrophs in a replicated long-term tillage experiment in which extraneous variation is minimized and N-fixation rates can be related to soil characteristics and diazotroph community structure. Soil characteristics were found to be primarily impacted by tillage management, whereas N-fixation rates and diazotroph community structure were impacted by both biomass management practices and interactions between tillage and biomass management. The data suggest that the variation in diazotroph community structure has a greater impact on N-fixation rates than do soil characteristics at the site. N-fixation rates displayed a saturating response to increases in diazotroph community diversity. These results show that the changes in the community structure of free-living diazotrophs in soils can have ecological significance and suggest that this response is related to a change in community diversity.

  8. Impact of copper on the diazotroph abundance and community composition during swine manure composting.

    Science.gov (United States)

    Yin, Yanan; Gu, Jie; Wang, Xiaojuan; Zhang, Kaiyu; Hu, Ting; Ma, Jiyue; Wang, Qianzhi

    2018-05-01

    Biological nitrogen fixation is a major pathway in ecosystems. This study investigated the effects of adding Cu at different levels (0, 200, and 2000 mg kg -1 ) on the diazotroph community during swine manure composting. Quantitative PCR and high-throughput sequencing were used to analyze the abundances of diazotrophs and the community composition based on the nifH gene. The nifH gene copy number was relatively high in the early stage of composting and Cu had a significant inhibitory effect on the nifH copy number. Furthermore, Cu decreased the diversity of nifH and changed the microbial community structure in the early stage. The nifH genes from members of Firmicutes and Clostridium were most abundant. Co-occurrence ecological network analysis showed that the Cu treatments affected the co-occurrence patterns of diazotroph communities and reduced the associations between different diazotrophs. Interestingly, Cu may weaken symbiotic diazotrophic interactions and enhance the roles of free-living diazotrophs. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. Binding characteristics of copper and cadmium by cyanobacterium Spirulina platensis.

    Science.gov (United States)

    Fang, Linchuan; Zhou, Chen; Cai, Peng; Chen, Wenli; Rong, Xingmin; Dai, Ke; Liang, Wei; Gu, Ji-Dong; Huang, Qiaoyun

    2011-06-15

    Cyanobacteria are promising biosorbent for heavy metals in bioremediation. Although sequestration of metals by cyanobacteria is known, the actual mechanisms and ligands involved are not very well understood. The binding characteristics of Cu(II) and Cd(II) by the cyanobacterium Spirulina platensis were investigated using a combination of chemical modifications, batch adsorption experiments, Fourier transform infrared (FTIR) spectroscopy and X-ray absorption fine structure (XAFS) spectroscopy. A significant increase in Cu(II) and Cd(II) binding was observed in the range of pH 3.5-5.0. Dramatical decrease in adsorption of Cu(II) and Cd(II) was observed after methanol esterification of the nonliving cells demonstrating that carboxyl functional groups play an important role in the binding of metals by S. platensis. The desorption rate of Cu(II) and Cd(II) from S. platensis surface was 72.7-80.7% and 53.7-58.0% by EDTA and NH(4)NO(3), respectively, indicating that ion exchange and complexation are the dominating mechanisms for Cu(II) and Cd(II) adsorption. XAFS analysis provided further evidence on the inner-sphere complexation of Cu by carboxyl ligands and showed that Cu is complexed by two 5-membered chelate rings on S. platensis surface. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Growth Characteristics of an Estuarine Heterocystous Cyanobacterium

    Directory of Open Access Journals (Sweden)

    Pablo Guimarães

    2017-06-01

    Full Text Available A new estuarine filamentous heterocystous cyanobacterium was isolated from intertidal sediment of the Lagoa dos Patos estuary (Brazil. The isolate may represent a new genus related to Cylindrospermopsis. While the latter is planktonic, contains gas vesicles, and is toxic, the newly isolated strain is benthic and does not contain gas vesicles. It is not known whether the new strain is toxic. It grows equally well in freshwater, brackish and full salinity growth media, in the absence of inorganic or organic combined nitrogen, with a growth rate 0.6 d-1. Nitrogenase, the enzyme complex responsible for fixing dinitrogen, was most active during the initial growth phase and its activity was not different between the different salinities tested (freshwater, brackish, and full salinity seawater. Salinity shock also did not affect nitrogenase activity. The frequency of heterocysts was high, coinciding with high nitrogenase activity during the initial growth phase, but decreased subsequently. However, the frequency of heterocysts decreased considerably more at higher salinity, while no change in nitrogenase activity occurred, indicating a higher efficiency of dinitrogen fixation. Akinete frequency was low in the initial growth phase and higher in the late growth phase. Akinete frequency was much lower at high salinity, which might indicate better growth conditions or that akinete differentiation was under the same control as heterocyst differentiation. These trends have hitherto not been reported for heterocystous cyanobacteria but they seem to be well fitted for an estuarine life style.

  11. Diazotrophic diversity in the rhizosphere of two exotic weed plants, Prosopis juliflora and Parthenium hysterophorus.

    Science.gov (United States)

    Cibichakravarthy, B; Preetha, R; Sundaram, S P; Kumar, K; Balachandar, D

    2012-02-01

    This study is aimed at assessing culturable diazotrophic bacterial diversity in the rhizosphere of Prosopis juliflora and Parthenium hysterophorus, which grow profusely in nutritionally-poor soils and environmentally-stress conditions so as to identify some novel strains for bioinoculant technology. Diazotrophic isolates from Prosopis and Parthenium rhizosphere were characterized for nitrogenase activity by Acetylene Reduction Assay (ARA) and 16S rRNA gene sequencing. Further, the culture-independent quantitative PCR (qPCR) was performed to compare the abundance of diazotrophs in rhizosphere with bulk soils. The proportion of diazotrophs in total heterotrophs was higher in rhizosphere than bulk soils and 32 putative diazotrophs from rhizosphere of two plants were identified by nifH gene amplification. The ARA activity of the isolates ranged from 40 to 95 nmol ethylene h(-1) mg protein(-1). The 16S rRNA gene analysis identified the isolates to be members of alpha, beta and gamma Proteobacteria and firmicutes. The qPCR assay also confirmed that abundance of nif gene in rhizosphere of these two plants was 10-fold higher than bulk soil.

  12. Evidence for the endophytic colonization of Phaseolus vulgaris (common bean) roots by the diazotroph Herbaspirillum seropedicae.

    Science.gov (United States)

    Schmidt, M A; Souza, E M; Baura, V; Wassem, R; Yates, M G; Pedrosa, F O; Monteiro, R A

    2011-03-01

    Herbaspirillum seropedicae is an endophytic diazotrophic bacterium, which associates with important agricultural plants. In the present study, we have investigated the attachment to and internal colonization of Phaseolus vulgaris roots by the H. seropedicae wild-type strain SMR1 and by a strain of H. seropedicae expressing a red fluorescent protein (DsRed) to track the bacterium in the plant tissues. Two-day-old P. vulgaris roots were incubated at 30°C for 15 min with 6 x 10(8) CFU/mL H. seropedicae SMR1 or RAM4. Three days after inoculation, 4 x 10(4) cells of endophytic H. seropedicae SMR1 were recovered per gram of fresh root, and 9 days after inoculation the number of endophytes increased to 4 x 10(6) CFU/g. The identity of the recovered bacteria was confirmed by amplification and sequencing of the 16SrRNA gene. Furthermore, confocal microscopy of P. vulgaris roots inoculated with H. seropedicae RAM4 showed that the bacterial cells were attached to the root surface 15 min after inoculation; fluorescent bacteria were visible in the internal tissues after 24 h and were found in the central cylinder after 72 h, showing that H. seropedicae RAM4 is capable of colonizing the roots of the dicotyledon P. vulgaris. Determination of dry weight of common bean inoculated with H. seropedicae SMR1 suggested that this bacterium has a negative effect on the growth of P. vulgaris.

  13. Wastewater utilization for poly-β-hydroxybutyrate production by the cyanobacterium Aulosira fertilissima in a recirculatory aquaculture system.

    Science.gov (United States)

    Samantaray, Shilalipi; Nayak, Jitendra Kumar; Mallick, Nirupama

    2011-12-01

    Intensive aquaculture releases large quantities of nutrients into aquatic bodies, which can lead to eutrophication. The objective of this study was the development of a biological recirculatory wastewater treatment system with a diazotrophic cyanobacterium, Aulosira fertilissima, and simultaneous production of valuable product in the form of poly-β-hydroxybutyrate (PHB). To investigate this possible synergy, batch scale tests were conducted under a recirculatory aquaculture system in fiber-reinforced plastic tanks enhanced by several manageable parameters (e.g., sedimentation, inoculum size, depth, turbulence, and light intensity), an adequate combination of which showed better productivity. The dissolved-oxygen level increased in the range of 3.2 to 6.9 mg liter⁻¹ during the culture period. Nutrients such as ammonia, nitrite, and phosphate decreased to as low as zero within 15 days of incubation, indicating the system's bioremediation capability while yielding valuable cyanobacterial biomass for PHB production. Maximum PHB accumulation in A. fertilissima was found in sedimented fish pond discharge at 20-cm culture depth with stirring and an initial inoculum size of 80 mg dry cell weight (dcw) liter⁻¹. Under optimized conditions, the PHB yield was boosted to 92, 89, and 80 g m⁻², respectively for the summer, rainy, and winter seasons. Extrapolation of the result showed that a hectare of A. fertilissima cultivation in fish pond discharge would give an annual harvest of ∼17 tons dry biomass, consisting of 14 tons of PHB with material properties comparable to those of the bacterial polymer, with simultaneous treatment of 32,640 m³ water discharge.

  14. Wastewater Utilization for Poly-β-Hydroxybutyrate Production by the Cyanobacterium Aulosira fertilissima in a Recirculatory Aquaculture System▿

    Science.gov (United States)

    Samantaray, Shilalipi; Nayak, Jitendra Kumar; Mallick, Nirupama

    2011-01-01

    Intensive aquaculture releases large quantities of nutrients into aquatic bodies, which can lead to eutrophication. The objective of this study was the development of a biological recirculatory wastewater treatment system with a diazotrophic cyanobacterium, Aulosira fertilissima, and simultaneous production of valuable product in the form of poly-β-hydroxybutyrate (PHB). To investigate this possible synergy, batch scale tests were conducted under a recirculatory aquaculture system in fiber-reinforced plastic tanks enhanced by several manageable parameters (e.g., sedimentation, inoculum size, depth, turbulence, and light intensity), an adequate combination of which showed better productivity. The dissolved-oxygen level increased in the range of 3.2 to 6.9 mg liter−1 during the culture period. Nutrients such as ammonia, nitrite, and phosphate decreased to as low as zero within 15 days of incubation, indicating the system's bioremediation capability while yielding valuable cyanobacterial biomass for PHB production. Maximum PHB accumulation in A. fertilissima was found in sedimented fish pond discharge at 20-cm culture depth with stirring and an initial inoculum size of 80 mg dry cell weight (dcw) liter−1. Under optimized conditions, the PHB yield was boosted to 92, 89, and 80 g m−2, respectively for the summer, rainy, and winter seasons. Extrapolation of the result showed that a hectare of A. fertilissima cultivation in fish pond discharge would give an annual harvest of ∼17 tons dry biomass, consisting of 14 tons of PHB with material properties comparable to those of the bacterial polymer, with simultaneous treatment of 32,640 m3 water discharge. PMID:21984242

  15. Sheep manure vermicompost supplemented with a native diazotrophic bacteria and mycorrhizas for maize cultivation.

    Science.gov (United States)

    Gutiérrez-Miceli, F A; Moguel-Zamudio, B; Abud-Archila, M; Gutiérrez-Oliva, V F; Dendooven, L

    2008-10-01

    An orthogonal experimental design L9 (3(4)) with 10 repetitions was used to investigate the effect of Glomus claroideum (0, 1 or 2g(-1) plant), G. fasciculatum (0, 1 or 2g plant(-1)), native diazotrophic bacteria (0, 10(3) and 10(5) UFC ml(-1)) and sheep manure vermicompost (0%, 5% and 10% v/v) on maize plant growth, N and P in leaves and mycorrhization percent. Vermicompost explained most of the variation found for leaf number, wet weight, stem height, and diameter. Both mycorrhizas increased the plant wet weight but G. fasciculatum the most. Mycorrhization increased the P content, but not the N content. Mycorrhizal colonization increased when diazotrophic bacteria and vermicompost were added. It was found that weight of maize plants cultivated in peat moss amended with vermicompost increased when supplemented with G. fasciculatum and diazotrophic bacteria.

  16. Divergent Responses of the Diazotrophic Microbiome to Elevated CO2 in Two Rice Cultivars

    Directory of Open Access Journals (Sweden)

    Yongjie Yu

    2018-06-01

    Full Text Available The species-specific responses of plant growth to elevated atmospheric CO2 concentration (eCO2 could lead to N limitation and potentially influence the sustainability of ecosystem. Questions remain unanswered with regards to the response of soil N2-fixing community to eCO2 when developing high-yielding agroecosystem to dampen the future rate of increase in CO2 levels and associated climate warming. This study demonstrates the divergent eCO2 influences on the paddy diazotrophic community between weak- and strong-responsive rice cultivars. In response to eCO2, the diazotrophic abundance increased more for the strong-responsive cultivar treatments than for the weak-responsive ones. Only the strong-responsive cultivars decreased the alpha diversity and separated the composition of diazotrophic communities in response to eCO2. The topological indices of the ecological networks further highlighted the different co-occurrence patterns of the diazotrophic microbiome in rice cultivars under eCO2. Strong-responsive cultivars destabilized the diazotrophic community by complicating and centralizing the co-occurrence network as well as by shifting the hub species from Bradyrhizobium to Dechloromonas in response to eCO2. On the contrary, the network pattern of the weak-responsive cultivars was simplified and decentralized in response to eCO2, with the hub species shifting from Halorhodospira under aCO2 to Sideroxydans under eCO2. Collectively, the above information indicates that the strong-responsive cultivars could potentially undermine the belowground ecosystem from the diazotrophs perspective in response to eCO2. This information highlights that more attention should be paid to the stability of the belowground ecosystem when developing agricultural strategies to adapt prospective climatic scenarios by growing high-yielding crop cultivars under eCO2.

  17. Antagonistic interactions between filamentous heterotrophs and the cyanobacterium Nostoc muscorum

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    Wolf Sarah

    2011-09-01

    Full Text Available Abstract Background Little is known about interactions between filamentous heterotrophs and filamentous cyanobacteria. Here, interactions between the filamentous heterotrophic bacteria Fibrella aestuarina (strain BUZ 2 and Fibrisoma limi (BUZ 3 with an axenic strain of the autotrophic filamentous cyanobacterium Nostoc muscorum (SAG 25.82 were studied in mixed cultures under nutrient rich (carbon source present in medium and poor (carbon source absent in medium conditions. Findings F. aestuarina BUZ 2 significantly reduced the cyanobacterial population whereas F. limi BUZ 3 did not. Physical contact between heterotrophs and autotroph was observed and the cyanobacterial cells showed some level of damage and lysis. Therefore, either contact lysis or entrapment with production of extracellular compounds in close vicinity of host cells could be considered as potential modes of action. The supernatants from pure heterotrophic cultures did not have an effect on Nostoc cultures. However, supernatant from mixed cultures of BUZ 2 and Nostoc had a negative effect on cyanobacterial growth, indicating that the lytic compounds were only produced in the presence of Nostoc. The growth and survival of tested heterotrophs was enhanced by the presence of Nostoc or its metabolites, suggesting that the heterotrophs could utilize the autotrophs and its products as a nutrient source. However, the autotroph could withstand and out-compete the heterotrophs under nutrient poor conditions. Conclusions Our results suggest that the nutrients in cultivation media, which boost or reduce the number of heterotrophs, were the important factor influencing the outcome of the interplay between filamentous heterotrophs and autotrophs. For better understanding of these interactions, additional research is needed. In particular, it is necessary to elucidate the mode of action for lysis by heterotrophs, and the possible defense mechanisms of the autotrophs.

  18. Binding characteristics of copper and cadmium by cyanobacterium Spirulina platensis

    International Nuclear Information System (INIS)

    Fang Linchuan; Zhou Chen; Cai Peng; Chen Wenli; Rong Xingmin; Dai Ke; Liang Wei; Gu Jidong; Huang Qiaoyun

    2011-01-01

    Highlights: → The carboxyl groups play a vital role in the binding of Cu(II) and Cd(II) to S. platensis cells. → Ion exchange and complexation are the dominating mechanism for Cu(II) and Cd(II) adsorption. → XAFS analysis provided evidence for the inner-sphere complexation of Cu by carboxyl ligands and showed that Cu is complexed by two 5-membered chelate rings on S. platensis surface. - Abstract: Cyanobacteria are promising biosorbent for heavy metals in bioremediation. Although sequestration of metals by cyanobacteria is known, the actual mechanisms and ligands involved are not very well understood. The binding characteristics of Cu(II) and Cd(II) by the cyanobacterium Spirulina platensis were investigated using a combination of chemical modifications, batch adsorption experiments, Fourier transform infrared (FTIR) spectroscopy and X-ray absorption fine structure (XAFS) spectroscopy. A significant increase in Cu(II) and Cd(II) binding was observed in the range of pH 3.5-5.0. Dramatical decrease in adsorption of Cu(II) and Cd(II) was observed after methanol esterification of the nonliving cells demonstrating that carboxyl functional groups play an important role in the binding of metals by S. platensis. The desorption rate of Cu(II) and Cd(II) from S. platensis surface was 72.7-80.7% and 53.7-58.0% by EDTA and NH 4 NO 3 , respectively, indicating that ion exchange and complexation are the dominating mechanisms for Cu(II) and Cd(II) adsorption. XAFS analysis provided further evidence on the inner-sphere complexation of Cu by carboxyl ligands and showed that Cu is complexed by two 5-membered chelate rings on S. platensis surface.

  19. Binding characteristics of copper and cadmium by cyanobacterium Spirulina platensis

    Energy Technology Data Exchange (ETDEWEB)

    Fang Linchuan [State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070 (China); Zhou Chen; Cai Peng [Key Laboratory of Subtropical Agricultural Resources and Environment, Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070 (China); Chen Wenli [State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070 (China); Rong Xingmin; Dai Ke; Liang Wei [Key Laboratory of Subtropical Agricultural Resources and Environment, Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070 (China); Gu Jidong [Department of Ecology and Biodiversity, University of Hong Kong, Pokfulam Road, Hong Kong (Hong Kong); Huang Qiaoyun, E-mail: qyhuang@mail.hzau.edu.cn [State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070 (China); Key Laboratory of Subtropical Agricultural Resources and Environment, Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070 (China)

    2011-06-15

    Highlights: {yields} The carboxyl groups play a vital role in the binding of Cu(II) and Cd(II) to S. platensis cells. {yields} Ion exchange and complexation are the dominating mechanism for Cu(II) and Cd(II) adsorption. {yields} XAFS analysis provided evidence for the inner-sphere complexation of Cu by carboxyl ligands and showed that Cu is complexed by two 5-membered chelate rings on S. platensis surface. - Abstract: Cyanobacteria are promising biosorbent for heavy metals in bioremediation. Although sequestration of metals by cyanobacteria is known, the actual mechanisms and ligands involved are not very well understood. The binding characteristics of Cu(II) and Cd(II) by the cyanobacterium Spirulina platensis were investigated using a combination of chemical modifications, batch adsorption experiments, Fourier transform infrared (FTIR) spectroscopy and X-ray absorption fine structure (XAFS) spectroscopy. A significant increase in Cu(II) and Cd(II) binding was observed in the range of pH 3.5-5.0. Dramatical decrease in adsorption of Cu(II) and Cd(II) was observed after methanol esterification of the nonliving cells demonstrating that carboxyl functional groups play an important role in the binding of metals by S. platensis. The desorption rate of Cu(II) and Cd(II) from S. platensis surface was 72.7-80.7% and 53.7-58.0% by EDTA and NH{sub 4}NO{sub 3}, respectively, indicating that ion exchange and complexation are the dominating mechanisms for Cu(II) and Cd(II) adsorption. XAFS analysis provided further evidence on the inner-sphere complexation of Cu by carboxyl ligands and showed that Cu is complexed by two 5-membered chelate rings on S. platensis surface.

  20. Studies on the process of attachment of diazotroph alcaligenes faecalis and its Tn5 mutants to rice roots using 15N-labelling technique

    International Nuclear Information System (INIS)

    Fang Xuanjun; Lin Min; You Chongbiao

    1993-09-01

    By using 15 N-labelling technique and Tn5-induced mutants the attachment of associative diazotroph Alcaligenes faecalis to intact rice plants was examined in vitro. Three distinguished modes of attachment of Alcaligenes faecalis: adsorption, anchoring and colonization were proposed by using 15 N-labelling bacterial cells and Tn5-induced mutants. Che - mutants affected on adsorption, but not on anchoring. Exo - Che - mutant is defective in both adsorption and anchoring. Exo - or exo ++ mutants are only defective in anchoring. Effective colonization is benefit for establishment on the associative system. The data also indicated that EPS (exopolysaccharide) play rather important roles in the association between the host plant and bacteria

  1. Occurrence of diazotrophic bacteria in Araucaria angustifolia Ocorrência de bactérias diazotróficas em Araucaria angustifolia

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    Rafaela de Fátima Neroni

    2007-06-01

    Full Text Available Araucaria angustifolia is an environmentally threatened tree and the whole biota of the Araucaria Forest should be investigated with the aim of its preservation. Diazotrophic bacteria are extremely important for the maintenance of ecosystems, but they have never been studied in Araucaria Forests. In this study, diazotrophic bacteria were isolated from Araucaria roots and soil, when grown in semi-specific, semi-solid media. The diazotrophic character of some recovered isolates could be confirmed using the acetylene reduction assay. According to their 16S rRNA sequences, most of these isolates belong to the genus Burkholderia.Araucaria angustifolia é uma espécie arbórea ameaçada de extinção e toda a comunidade viva das Florestas de Araucária deve ser estudada com a finalidade de sua preservação. Bactérias diazotróficas apresentam grande importância para a manutenção de ecossistemas, mas sua ocorrência nunca foi pesquisada em araucária. Neste estudo foi demonstrada a associação de bactérias fixadoras de nitrogênio com A. angustifolia. Amostras de solo e raízes de araucária apresentaram a ocorrência de bactérias diazotróficas, quando inoculadas em meios semi-sólidos semi-específícos. O caráter diazotrófico de alguns isolados recuperados foi confirmado através da análise de redução do acetileno. De acordo com a análise das seqüências do 16S rRNA, estes isolados são na sua maioria pertencentes ao gênero Burkholderia.

  2. Assimilation of diazotrophic nitrogen into pelagic food webs.

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    Ryan J Woodland

    Full Text Available The fate of diazotrophic nitrogen (N(D fixed by planktonic cyanobacteria in pelagic food webs remains unresolved, particularly for toxic cyanophytes that are selectively avoided by most herbivorous zooplankton. Current theory suggests that N(D fixed during cyanobacterial blooms can enter planktonic food webs contemporaneously with peak bloom biomass via direct grazing of zooplankton on cyanobacteria or via the uptake of bioavailable N(D (exuded from viable cyanobacterial cells by palatable phytoplankton or microbial consortia. Alternatively, N(D can enter planktonic food webs post-bloom following the remineralization of bloom detritus. Although the relative contribution of these processes to planktonic nutrient cycles is unknown, we hypothesized that assimilation of bioavailable N(D (e.g., nitrate, ammonium by palatable phytoplankton and subsequent grazing by zooplankton (either during or after the cyanobacterial bloom would be the primary pathway by which N(D was incorporated into the planktonic food web. Instead, in situ stable isotope measurements and grazing experiments clearly documented that the assimilation of N(D by zooplankton outpaced assimilation by palatable phytoplankton during a bloom of toxic Nodularia spumigena Mertens. We identified two distinct temporal phases in the trophic transfer of N(D from N. spumigena to the plankton community. The first phase was a highly dynamic transfer of N(D to zooplankton with rates that covaried with bloom biomass while bypassing other phytoplankton taxa; a trophic transfer that we infer was routed through bloom-associated bacteria. The second phase was a slowly accelerating assimilation of the dissolved-N(D pool by phytoplankton that was decoupled from contemporaneous variability in N. spumigena concentrations. These findings provide empirical evidence that N(D can be assimilated and transferred rapidly throughout natural plankton communities and yield insights into the specific processes

  3. Assimilation of diazotrophic nitrogen into pelagic food webs.

    Science.gov (United States)

    Woodland, Ryan J; Holland, Daryl P; Beardall, John; Smith, Jonathan; Scicluna, Todd; Cook, Perran L M

    2013-01-01

    The fate of diazotrophic nitrogen (N(D)) fixed by planktonic cyanobacteria in pelagic food webs remains unresolved, particularly for toxic cyanophytes that are selectively avoided by most herbivorous zooplankton. Current theory suggests that N(D) fixed during cyanobacterial blooms can enter planktonic food webs contemporaneously with peak bloom biomass via direct grazing of zooplankton on cyanobacteria or via the uptake of bioavailable N(D) (exuded from viable cyanobacterial cells) by palatable phytoplankton or microbial consortia. Alternatively, N(D) can enter planktonic food webs post-bloom following the remineralization of bloom detritus. Although the relative contribution of these processes to planktonic nutrient cycles is unknown, we hypothesized that assimilation of bioavailable N(D) (e.g., nitrate, ammonium) by palatable phytoplankton and subsequent grazing by zooplankton (either during or after the cyanobacterial bloom) would be the primary pathway by which N(D) was incorporated into the planktonic food web. Instead, in situ stable isotope measurements and grazing experiments clearly documented that the assimilation of N(D) by zooplankton outpaced assimilation by palatable phytoplankton during a bloom of toxic Nodularia spumigena Mertens. We identified two distinct temporal phases in the trophic transfer of N(D) from N. spumigena to the plankton community. The first phase was a highly dynamic transfer of N(D) to zooplankton with rates that covaried with bloom biomass while bypassing other phytoplankton taxa; a trophic transfer that we infer was routed through bloom-associated bacteria. The second phase was a slowly accelerating assimilation of the dissolved-N(D) pool by phytoplankton that was decoupled from contemporaneous variability in N. spumigena concentrations. These findings provide empirical evidence that N(D) can be assimilated and transferred rapidly throughout natural plankton communities and yield insights into the specific processes underlying

  4. Colony formation in the cyanobacterium Microcystis.

    Science.gov (United States)

    Xiao, Man; Li, Ming; Reynolds, Colin S

    2018-02-22

    Morphological evolution from a unicellular to multicellular state provides greater opportunities for organisms to attain larger and more complex living forms. As the most common freshwater cyanobacterial genus, Microcystis is a unicellular microorganism, with high phenotypic plasticity, which forms colonies and blooms in lakes and reservoirs worldwide. We conducted a systematic review of field studies from the 1990s to 2017 where Microcystis was dominant. Microcystis was detected as the dominant genus in waterbodies from temperate to subtropical and tropical zones. Unicellular Microcystis spp. can be induced to form colonies by adjusting biotic and abiotic factors in laboratory. Colony formation by cell division has been induced by zooplankton filtrate, high Pb 2+ concentration, the presence of another cyanobacterium (Cylindrospermopsis raciborskii), heterotrophic bacteria, and by low temperature and light intensity. Colony formation by cell adhesion can be induced by zooplankton grazing, high Ca 2+ concentration, and microcystins. We hypothesise that single cells of all Microcystis morphospecies initially form colonies with a similar morphology to those found in the early spring. These colonies gradually change their morphology to that of M. ichthyoblabe, M. wesenbergii and M. aeruginosa with changing environmental conditions. Colony formation provides Microcystis with many ecological advantages, including adaption to varying light, sustained growth under poor nutrient supply, protection from chemical stressors and protection from grazing. These benefits represent passive tactics responding to environmental stress. Microcystis colonies form at the cost of decreased specific growth rates compared with a unicellular habit. Large colony size allows Microcystis to attain rapid floating velocities (maximum recorded for a single colony, ∼ 10.08 m h -1 ) that enable them to develop and maintain a large biomass near the surface of eutrophic lakes, where they may shade

  5. Performance of pineapple slips inoculated with diazotrophic phosphate-solubilizing bacteria and rock phosphate

    Directory of Open Access Journals (Sweden)

    Lílian Estrela Borges Baldotto

    2014-06-01

    Full Text Available Besides fixing N2, some diazotrophic bacteria or diazotrophs, also synthesize organic acids and are able to solubilize rock phosphates, increasing the availability of P for plants. The application of these bacteria to pineapple leaf axils in combination with rock phosphate could increase N and P availability for the crop, due to the bacterial activity of biological nitrogen fixation and phosphate solubilization. The objectives of this study were: (i to select and characterize diazotrophs able to solubilize phosphates in vitro and (ii evaluate the initial performance of the pineapple cultivars Imperial and Pérola in response to inoculation with selected bacteria in combination with rock phosphate. The experiments were conducted at Universidade Estadual do Norte Fluminense Darcy Ribeiro, in 2009. In the treatments with bacteria the leaf contents of N, P and K were higher than those of the controls, followed by an increase in plant growth. These results indicate that the combined application of diazotrophic phosphate-solubilizing bacteria Burkholderia together with Araxá rock phosphate can be used to improve the initial performance of pineapple slips.

  6. Phylogenetic changes in soil microbial and diazotrophic diversity with application of butachlor.

    Science.gov (United States)

    Yen, Jui-Hung; Wang, Yei-Shung; Hsu, Wey-Shin; Chen, Wen-Ching

    2013-01-01

    We investigated changes in population and taxonomic distribution of cultivable bacteria and diazotrophs with butachlor application in rice paddy soils. Population changes were measured by the traditional plate-count method, and taxonomic distribution was studied by 16S rDNA sequencing, then maximum parsimony phylogenic analysis with bootstrapping (1,000 replications). The bacterial population was higher after 39 than 7 days of rice cultivation, which indicated the augmentation of soil microbes by rice root exudates. The application of butachlor increased the diazotrophic population in both upper (0-3 cm) and lower (3-15 cm) layers of soils. Especially at day 39, the population of diazotrophs was 1.8 and 1.6 times that of the control in upper and lower layer soils, respectively. We found several bacterial strains only with butachlor application; examples are strains closest to Bacillus arsenicus, B. marisflavi, B. luciferensis, B. pumilus, and Pseudomonas alvei. Among diazotrophs, three strains closely related to Streptomyces sp. or Rhrizobium sp. were found only with butachlor application. The population of cultivable bacteria and the species composition were both changed with butachlor application, which explains in part the contribution of butachlor to augmenting soil nitrogen-fixing ability.

  7. The effects of nitrogen fixation and plant growth-promoting in rice-diazotroph association

    International Nuclear Information System (INIS)

    Lin Fan; Wang Lu

    1999-05-01

    This is a review of studies on applications of the genetic engineered ammonium-tolerant diazotroph as an inoculum with the effects of nitrogen-fixation, plant growth-promoting and yield-increasing on rice and some crops by using 15 N tracer in mini-plot and field experiments in resent years

  8. Characterization of Diazotrophs Containing Mo-Independent Nitrogenases, Isolated from Diverse Natural Environments▿

    Science.gov (United States)

    Betancourt, Doris A.; Loveless, Telisa M.; Brown, James W.; Bishop, Paul E.

    2008-01-01

    Molybdenum-independent nitrogenases were first described in the nitrogen-fixing bacterium Azotobacter vinelandii and have since been described in other diazotrophic bacteria. Previously, we reported the isolation of seven diazotrophs with Mo-independent nitrogenases from aquatic environments. In the present study, we extend these results to include diazotrophs isolated from wood chip mulch, soil, “paraffin dirt,” and sediments from mangrove swamps. Mo-deficient, N-free media under both aerobic and anaerobic conditions were used for the isolations. A total of 26 isolates were genetically and physiologically characterized. Their phylogenetic placement was determined using 16S rRNA gene sequence analysis. Most of the isolates are members of the gamma subdivision of the class Proteobacteria and appear to be specifically related to fluorescent pseudomonads and azotobacteria. Two other isolates, AN1 and LPF4, are closely related to Enterobacter spp. and Paenibacillus spp., respectively. PCR and/or Southern hybridization were used to detect the presence of nitrogenase genes in the isolates. PCR amplification of vnfG and anfG was used to detect the genetic potential for the expression of the vanadium-containing nitrogenase and the iron-only nitrogenase in the isolates. This study demonstrates that diazotrophs with Mo-independent nitrogenases can be readily isolated from diverse natural environments. PMID:18378646

  9. Seasonal variations in the diversity and abundance of diazotrophic communities across soils

    NARCIS (Netherlands)

    Pereira e Silva, Michele C.; Semenov, Alexander V.; van Elsas, Jan Dirk; Salles, Joana Falcao

    The nitrogen (N)-fixing community is a key functional community in soil, as it replenishes the pool of biologically available N that is lost to the atmosphere via anaerobic ammonium oxidation and denitrification. We characterized the structure and dynamic changes in diazotrophic communities, based

  10. [Dynamics of diazotrophic bacteria number in the root zone of wheat Vrn lines isogenic by genes].

    Science.gov (United States)

    Samoĭlov, A M; Zhmurko, V V

    2012-01-01

    The number of diazotrophic bacteria and nitrogenase activity in the root zone of isogenic monogene-dominant Vrn lines were measured in the field experiments throughout their vegetation from tillering to heading. The total number of diazotrophic bacteria and nitrogenase activity in the root zone of these lines during this period were increased irrespective of their genotypes. The above indices of the winter cultivar (Vrn loci bottom recessive) were lower than those of the spring lines--Vrn-A1, Vrn-B1 and Vrn-D1. Plants of Vrn-B1 line have the lowest indices among the spring lines with the exception of some indices. This line plants flowered later than those of Vrn-A1 and Vrn-D1 lines. We hypothesized the differences between plants of these lines as to nitrogen fixation activity and the number of diazotrophic bacteria are mediately determined by Vrn loci through their effects on metabolism intensity and assimilate reflux in the form of root exudates, therefore the total number of diazotrophic bacteria and nitrogenase activity increases.

  11. Dominance of unicellular cyanobacteria in the diazotrophic community in the Atlantic Ocean

    NARCIS (Netherlands)

    Agawin, N.S.R.; Benavides, M.; Busquets, A.; Ferriol, P.; Stal, L.J.; Aristegui, J.

    2014-01-01

    ABSTRACT: The horizontal and vertical distribution of representatives of diazotrophic unicellular cyanobacteria was investigated in the subtropical northeast Atlantic Ocean (28.87 to 42.00°N; 9.01 to 20.02°W). Samples from stations encompassing different water conditions (from oceanic oligotrophic

  12. Dominance of unicellular cyanobacteria in the diazotrophic community in the Atlantic Ocean

    NARCIS (Netherlands)

    Agawin, N.S.R.; Benavides, M.; Busquets, A.; Ferriol, P.; Stal, L.J.; Arístegui, J.

    2014-01-01

    The horizontal and vertical distribution of representatives of diazotrophic unicellular cyanobacteria was investigated in the subtropical northeast Atlantic Ocean (28.87 to 42.00°N; 9.01 to 20.02°W). Samples from stations encompassing different water conditions (from oceanic oligotrophic waters to

  13. Analogous nutrient limitations in unicellular diazotrophs and Prochlorococcus in the South Pacific Ocean.

    Science.gov (United States)

    Moisander, Pia H; Zhang, Ruifeng; Boyle, Edward A; Hewson, Ian; Montoya, Joseph P; Zehr, Jonathan P

    2012-04-01

    Growth limitation of phytoplankton and unicellular nitrogen (N(2)) fixers (diazotrophs) were investigated in the oligotrophic Western South Pacific Ocean. Based on change in abundances of nifH or 23S rRNA gene copies during nutrient-enrichment experiments, the factors limiting net growth of the unicellular diazotrophs UCYN-A (Group A), Crocosphaera watsonii, γ-Proteobacterium 24774A11, and the non-diazotrophic picocyanobacterium Prochlorococcus, varied within the region. At the westernmost stations, numbers were enhanced by organic carbon added as simple sugars, a combination of iron and an organic chelator, or iron added with phosphate. At stations nearest the equator, the nutrient-limiting growth was not apparent. Maximum net growth rates for UCYN-A, C. watsonii and γ-24774A11 were 0.19, 0.61 and 0.52 d(-1), respectively, which are the first known empirical growth rates reported for the uncultivated UCYN-A and the γ-24774A11. The addition of N enhanced total phytoplankton biomass up to 5-fold, and the non-N(2)-fixing Synechococcus was among the groups that responded favorably to N addition. Nitrogen was the major nutrient-limiting phytoplankton biomass in the Western South Pacific Ocean, while availability of organic carbon or iron and organic chelator appear to limit abundances of unicellular diazotrophs. Lack of phytoplankton response to nutrient additions in the Pacific warm pool waters suggests diazotroph growth in this area is controlled by different factors than in the higher latitudes, which may partially explain previously observed variability in community composition in the region.

  14. The Assimilation of Diazotroph-Derived Nitrogen by Scleractinian Corals Depends on Their Metabolic Status

    Directory of Open Access Journals (Sweden)

    Vanessa N. Bednarz

    2017-01-01

    Full Text Available Tropical corals are associated with a diverse community of dinitrogen (N2-fixing prokaryotes (diazotrophs providing the coral an additional source of bioavailable nitrogen (N in oligotrophic waters. The overall activity of these diazotrophs changes depending on the current environmental conditions, but to what extent it affects the assimilation of diazotroph-derived N (DDN by corals is still unknown. Here, in a series of 15N2 tracer experiments, we directly quantified DDN assimilation by scleractinian corals from the Red Sea exposed to different environmental conditions. We show that DDN assimilation strongly varied with the corals’ metabolic status or with phosphate availability in the water. The very autotrophic shallow-water (~5 m corals showed low or no DDN assimilation, which significantly increased under elevated phosphate availability (3 µM. Corals that depended more on heterotrophy (i.e., bleached and deep-water [~45 m] corals assimilated significantly more DDN, which contributed up to 15% of the corals’ N demand (compared to 1% in shallow corals. Furthermore, we demonstrate that a substantial part of the DDN assimilated by deep corals was likely obtained from heterotrophic feeding on fixed N compounds and/or diazotrophic cells in the mucus. Conversely, in shallow corals, the net release of mucus, rich in organic carbon compounds, likely enhanced diazotroph abundance and activity and thereby the release of fixed N to the pelagic and benthic reef community. Overall, our results suggest that DDN assimilation by corals varies according to the environmental conditions and is likely linked to the capacity of the coral to acquire nutrients from seawater.

  15. Enumeration and characterization of arsenic-tolerant diazotrophic bacteria in a long-term heavy-metal-contaminated soil

    OpenAIRE

    Oliveira, A.; Pampulha, M.E.; Neto, M.M.; Almeida, A.C.

    2009-01-01

    The abundance of arsenic-tolerant diazotrophic bacteria was compared in a long-term contaminated soil versus a non-contaminated one. In addition, the characterization of tolerant diazotrophic bacteria was carried out. Differences in the number of heterotrophic N2 fixers were found between soils. Contaminated soil showed a decrease in the microbial population size of about 80%, confirming the great sensitivity of this group of soil bacteria to metals. However, quantitat...

  16. Utilization of a terrestrial cyanobacterium, Nostoc sp. HK-01, for space habitation

    Science.gov (United States)

    Kimura, Shunta; Tomita-Yokotani, Kaori; Arai, Mayumi; Yamashita, Masamichi; Katoh, Hiroshi; Ajioka, Reiko; Inoue, Kotomi

    2016-07-01

    A terrestrial cyanobacterium, Nostoc sp. HK-01 (hereafter HK-01), has several useful abilities for space habitation; photosynthesis, nitrogen fixation, and space environmental tolerances to vacuum, UV, gamma-ray, heavy particle beam, low and high temperature. Space environmental tolerances are important for transportation to Mars. HK-01 can grow on Martian regolith simulant (MRS) in vitro. Furthermore, HK-01 is useful as food. HK-01 may be utilized as oxygen supply, soil formation and food material for bio-chemical circulation in closed bio-ecosystems, including space habitation such as Mars. HK-01 was adopted as a biological material for the "TANPOPO" mission (JAXA et al.,), because of their high environmental tolerances. The "TANPOPO" mission is performing the space exposure experiments on the Japan Experimental Module (JEM) of the International Space Station (ISS). The results of these experiments will show the ability of HK-01 to survive in space.

  17. Development and nitrate reductase activity of sugarcane inoculated with five diazotrophic strains.

    Science.gov (United States)

    Dos Santos, Silvana Gomes; da Silva Ribeiro, Flaviane; da Fonseca, Camila Sousa; Pereira, Willian; Santos, Leandro Azevedo; Reis, Veronica Massena

    2017-08-01

    Diazotrophs are able to stimulate plant growth. This study aimed at evaluating the effect of inoculation of five diazotrophic strains on growth promotion and nitrate reductase (NR, EC 1.7.1.1) activity in sugarcane. An experiment was carried out from three stages of cultivation: sprouting, tubes, and in hydroponics. On the first two stages, seven treatments were adopted: uninoculated control; mixed inoculation with five strains; and individual inoculation with Gluconacetobacter diazotrophicus (Gd), Herbaspirillum rubrisubalbicans (Hr), Herbaspirillum seropedicae (Hs), Nitrospirillum amazonense (Na), and Paraburkholderia tropica (Pt). The four treatments showing the best performance were transferred to the hydroponic system for analysis of NR activity. Hs, Pt, and the mixture of all strains led to the highest seedling biomass in tubes, followed by Hr. In hydroponics, the mixture and the strain Hr had the highest growth-promoting effect. NR activity was influenced by inoculation only under low N supply conditions, with positive effect of Hr, Pt, and the mixture.

  18. Characteristics of nitrogen fixation of mixed diazotrophs associated with rice plant

    International Nuclear Information System (INIS)

    Ling Fan; Wang Zhengfang; Wang Yaodong; Song Wei

    1997-01-01

    Characteristics of N 2 fixation of diazotrophs associated with rice plant in paddy soils in Nanjing was studied by 15 N tracing technique. The results showed that amount of N fixed by rice plant was 1.03 kg/666.7 m 2 and the rate of fixed N was 6.7%. The maximum N fixed was occurred during jointing-complete heading stage. The daily average amount of N fixed reached to 24.31 mg/m 2 ·day. The fixed N of jointing-complete heading stage was 40.9% of that whole rice growth stage. The amount of fixed N during jointing-maturing stage was over 70% of whole rice growth stage. The economic benefits for fertilizer saving was 13.3 kg/666.7 m 2 of ammonium sulphate. The yield of rice grain was increased by 4.14% after inoculation with the mixed diazotrophs

  19. Three New Malyngamides from the Marine Cyanobacterium Moorea producens

    Directory of Open Access Journals (Sweden)

    Kosuke Sueyoshi

    2017-11-01

    Full Text Available Three new compounds of the malyngamide series, 6,8-di-O-acetylmalyngamide 2 (1, 6-O-acetylmalyngamide 2 (2, and N-demethyl-isomalyngamide I (3, were isolated from the marine cyanobacterium Moorea producens. Their structures were determined by spectroscopic analysis and chemical derivatization and degradation. These compounds stimulated glucose uptake in cultured L6 myotubes. In particular, 6,8-di-O-acetylmalyngamide 2 (1 showed potent activity and activated adenosine monophosphate-activated protein kinase (AMPK.

  20. Associative diazotrophic bacteria in grass roots and soils from heavy metal contaminated sites.

    Science.gov (United States)

    Moreira, Fátima M S; Lange, Anderson; Klauberg-Filho, Osmar; Siqueira, José O; Nóbrega, Rafaela S A; Lima, Adriana S

    2008-12-01

    This work aimed to evaluate density of associative diazotrophic bacteria populations in soil and grass root samples from heavy metal contaminated sites, and to characterize isolates from these populations, both, phenotypically (Zinc, Cadmium and NaCl tolerance in vitro, and protein profiles) and genotypically (16S rDNA sequencing), as compared to type strains of known diazotrophic species. Densities were evaluated by using NFb, Fam and JNFb media, commonly used for enrichment cultures of diazotrophic bacteria. Bacterial densities found in soil and grass root samples from contaminated sites were similar to those reported for agricultural soils. Azospirillum spp. isolates from contaminated sites and type strains from non-contaminated sites varied substantially in their in vitro tolerance to Zn+2 and Cd+2, being Cd+2 more toxic than Zn+2. Among the most tolerant isolates (UFLA 1S, 1R, S181, S34 and S22), some (1R, S34 and S22) were more tolerant to heavy metals than rhizobia from tropical and temperate soils. The majority of the isolates tolerant to heavy metals were also tolerant to salt stress as indicated by their ability to grow in solid medium supplemented with 30 g L(-1) NaCl. Five isolates exhibited high dissimilarity in protein profiles, and the 16S rDNA sequence analysis of two of them revealed new sequences for Azospirillum.

  1. "Cold" Fixation: Reconciliation of Nitrogen Fixation Rates and Diazotroph Assemblages in the Arctic Ocean

    Science.gov (United States)

    Fong, A. A.; Waite, A.; Rost, B.; Richter, K. U.

    2016-02-01

    Measurements of biological nitrogen fixation are typically conducted in oligotrophic subtropical and tropical marine environments where concentrations of fixed inorganic nitrogen are low. To date, only a handful of nitrogen fixation studies have been conducted in high latitude marine environments, but further investigation is needed to resolve the distribution of cold ocean diazotrophic assemblages. Nitrogen fixation rates and nifH gene distributions were measured at seven stations from 5°E to 20°E, north of 81°N in the Arctic Ocean at the onset of summer 2015. Discrete water samples in ice-covered regions were collected from the sea surface to 200 m for 15N2-tracer additions and targeted nifH gene and transcript analyses. Previous work suggests that heterotrophic bacteria dominate diazotrophic communities in the Arctic Ocean. Therefore, additional nifH gene surveys of sinking particles were conducted to test for enrichment on organic matter-rich microenvironments. Together, these measurements aim to reconcile diazotrophic activity with microbial community composition, further elucidating how nitrogen fixers could impact current concepts in polar carbon and nutrient cycling.

  2. Strong shift in the diazotrophic endophytic bacterial community inhabiting rice (Oryza sativa) plants after flooding.

    Science.gov (United States)

    Ferrando, Lucía; Fernández Scavino, Ana

    2015-09-01

    Flooding impacts soil microbial communities, but its effect on endophytic communities has rarely been explored. This work addresses the effect of flooding on the abundance and diversity of endophytic diazotrophic communities on rice plants established in a greenhouse experiment. The nifH gene was significantly more abundant in roots after flooding, whereas the nifH gene copy numbers in leaves were unaffected and remained low. The PCA (principal component analysis) of T-RFLP (terminal restriction fragment length polymorphism) profiles indicated that root communities of replicate plots were more similar and diverse after flooding than before flooding. The nifH libraries obtained by cloning and 454 pyrosequencing consistently showed a remarkable shift in the diazotrophic community composition after flooding. Gammaproteobacteria (66-98%), mainly of the genus Stenotrophomonas, prevailed in roots before flooding, whereas Betaproteobacteria was the dominant class (26-34%) after flooding. A wide variety of aerotolerant and anaerobic diazotrophic bacteria (e.g. Dechloromonas, Rhodopseudomonas, Desulfovibrio, Geobacter, Chlorobium, Spirochaeta, Selenomonas and Dehalobacter) with diverse metabolic traits were retrieved from flooded rice roots. These findings suggest that endophytic communities could be significantly impacted by changes in plant-soil conditions derived from flooding during rice cropping. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  3. Diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple plantlets during acclimatization

    Directory of Open Access Journals (Sweden)

    Aurilena de Aviz Silva

    Full Text Available ABSTRACT: This study examines the effect of inoculation with diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple cv. 'Vitória' plantlets during the acclimatization period. The experiment was carried out in a greenhouse in Campos dos Goytacazes, in randomized blocks, using a 2x5x5 factorial scheme, with the factors being two types of inocula (absence or presence of a mixture of diazotrophic bacteria that contained Burkholderia sp. UENF 114111, Burkholderia silvatlantica UENF 11711, and Herbaspirillum seropedicae HRC 54, five levels of urea (0, 2, 5, 7, and 10g L-1, and five evaluation periods (30, 60, 90, 120, and 150 days following the planting of plantlets in trays, and with four replications of each treatment. An increase in the dried weight of shoots and roots of micropropagated pineapple cv. 'Vitória' plants depended on inoculation with diazotrophic bacteria, and plantlets acclimatization could be reduced by up to 57 days, depending on the concentration of urea fertilizer.

  4. Isolation and molecular identification of endophytic diazotrophs from seeds and stems of three cereal crops.

    Directory of Open Access Journals (Sweden)

    Huawei Liu

    Full Text Available Ten strains of endophytic diazotroph were isolated and identified from the plants collected from three different agricultural crop species, wheat, rice and maize, using the nitrogen-free selective isolation conditions. The nitrogen-fixing ability of endophytic diazotroph was verified by the nifH-PCR assay that showed positive nitrogen fixation ability. These identified strains were classified by 879F-RAPD and 16S rRNA sequence analysis. RAPD analyses revealed that the 10 strains were clustered into seven 879F-RAPD groups, suggesting a clonal origin. 16S rRNA sequencing analyses allowed the assignment of the 10 strains to known groups of nitrogen-fixing bacteria, including organisms from the genera Paenibacillus, Enterobacter, Klebsiella and Pantoea. These representative genus are not endophytic diazotrophs in the conventional sense. They may have obtained nitrogen fixation ability through lateral gene transfer, however, the evolutionary forces of lateral gene transfer are not well known. Molecular identification results from 16S rRNA analyses were also confirmed by morphological and biochemical data. The test strains SH6A and MZB showed positive effect on the growth of plants.

  5. Diurnal Regulation of Cellular Processes in the Cyanobacterium Synechocystis sp. Strain PCC 6803: Insights from Transcriptomic, Fluxomic, and Physiological Analyses

    Directory of Open Access Journals (Sweden)

    Rajib Saha

    2016-05-01

    Full Text Available Synechocystis sp. strain PCC 6803 is the most widely studied model cyanobacterium, with a well-developed omics level knowledgebase. Like the lifestyles of other cyanobacteria, that of Synechocystis PCC 6803 is tuned to diurnal changes in light intensity. In this study, we analyzed the expression patterns of all of the genes of this cyanobacterium over two consecutive diurnal periods. Using stringent criteria, we determined that the transcript levels of nearly 40% of the genes in Synechocystis PCC 6803 show robust diurnal oscillating behavior, with a majority of the transcripts being upregulated during the early light period. Such transcripts corresponded to a wide array of cellular processes, such as light harvesting, photosynthetic light and dark reactions, and central carbon metabolism. In contrast, transcripts of membrane transporters for transition metals involved in the photosynthetic electron transport chain (e.g., iron, manganese, and copper were significantly upregulated during the late dark period. Thus, the pattern of global gene expression led to the development of two distinct transcriptional networks of coregulated oscillatory genes. These networks help describe how Synechocystis PCC 6803 regulates its metabolism toward the end of the dark period in anticipation of efficient photosynthesis during the early light period. Furthermore, in silico flux prediction of important cellular processes and experimental measurements of cellular ATP, NADP(H, and glycogen levels showed how this diurnal behavior influences its metabolic characteristics. In particular, NADPH/NADP+ showed a strong correlation with the majority of the genes whose expression peaks in the light. We conclude that this ratio is a key endogenous determinant of the diurnal behavior of this cyanobacterium.

  6. Photosynthetic Versatility in the Genome of Geitlerinema sp. PCC 9228 (Formerly Oscillatoria limnetica 'Solar Lake'), a Model Anoxygenic Photosynthetic Cyanobacterium.

    Science.gov (United States)

    Grim, Sharon L; Dick, Gregory J

    2016-01-01

    Anoxygenic cyanobacteria that use sulfide as the electron donor for photosynthesis are a potentially influential but poorly constrained force on Earth's biogeochemistry. Their versatile metabolism may have boosted primary production and nitrogen cycling in euxinic coastal margins in the Proterozoic. In addition, they represent a biological mechanism for limiting the accumulation of atmospheric oxygen, especially before the Great Oxidation Event and in the low-oxygen conditions of the Proterozoic. In this study, we describe the draft genome sequence of Geitlerinema sp. PCC 9228, formerly Oscillatoria limnetica 'Solar Lake', a mat-forming diazotrophic cyanobacterium that can switch between oxygenic photosynthesis and sulfide-based anoxygenic photosynthesis (AP). Geitlerinema possesses three variants of psbA , which encodes protein D1, a core component of the photosystem II reaction center. Phylogenetic analyses indicate that one variant is closely affiliated with cyanobacterial psbA genes that code for a D1 protein used for oxygen-sensitive processes. Another version is phylogenetically similar to cyanobacterial psbA genes that encode D1 proteins used under microaerobic conditions, and the third variant may be cued to high light and/or elevated oxygen concentrations. Geitlerinema has the canonical gene for sulfide quinone reductase (SQR) used in cyanobacterial AP and a putative transcriptional regulatory gene in the same operon. Another operon with a second, distinct sqr and regulatory gene is present, and is phylogenetically related to sqr genes used for high sulfide concentrations. The genome has a comprehensive nif gene suite for nitrogen fixation, supporting previous observations of nitrogenase activity. Geitlerinema possesses a bidirectional hydrogenase rather than the uptake hydrogenase typically used by cyanobacteria in diazotrophy. Overall, the genome sequence of Geitlerinema sp. PCC 9228 highlights potential cyanobacterial strategies to cope with fluctuating

  7. Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae in the Brazilian Amazon

    Directory of Open Access Journals (Sweden)

    Paulo Ivan Fernandes Júnior

    2013-06-01

    Full Text Available The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg’s medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH+ bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fifty- one bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360bp nifH gene fragment. Among the 38 nifH+ isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus

  8. Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae) in the Brazilian Amazon.

    Science.gov (United States)

    Júnior, Paulo Ivan Fernandes; Pereira, Gilmara Maria Duarte; Perin, Liamara; da Silva, Luana Mesquita; Baraúna, Alexandre Cardoso; Alvess, Francilene Muniz; Passos, Samuel Ribeiro; Zilli, Jerri Edson

    2013-06-01

    The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg's medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fifty-one bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360 bp nifH gene fragment. Among the 38 nifH+ isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus. The growth

  9. Levels of daily light doses under changed day-night cycles regulate temporal segregation of photosynthesis and N2 Fixation in the cyanobacterium Trichodesmium erythraeum IMS101.

    Science.gov (United States)

    Cai, Xiaoni; Gao, Kunshan

    2015-01-01

    While the diazotrophic cyanobacterium Trichodesmium is known to display inverse diurnal performances of photosynthesis and N2 fixation, such a phenomenon has not been well documented under different day-night (L-D) cycles and different levels of light dose exposed to the cells. Here, we show differences in growth, N2 fixation and photosynthetic carbon fixation as well as photochemical performances of Trichodesmium IMS101 grown under 12L:12D, 8L:16D and 16L:8D L-D cycles at 70 μmol photons m-2 s-1 PAR (LL) and 350 μmol photons m-2 s-1 PAR (HL). The specific growth rate was the highest under LL and the lowest under HL under 16L:8D, and it increased under LL and decreased under HL with increased levels of daytime light doses exposed under the different light regimes, respectively. N2 fixation and photosynthetic carbon fixation were affected differentially by changes in the day-night regimes, with the former increasing directly under LL with increased daytime light doses and decreased under HL over growth-saturating light levels. Temporal segregation of N2 fixation from photosynthetic carbon fixation was evidenced under all day-night regimes, showing a time lag between the peak in N2 fixation and dip in carbon fixation. Elongation of light period led to higher N2 fixation rate under LL than under HL, while shortening the light exposure to 8 h delayed the N2 fixation peaking time (at the end of light period) and extended it to night period. Photosynthetic carbon fixation rates and transfer of light photons were always higher under HL than LL, regardless of the day-night cycles. Conclusively, diel performance of N2 fixation possesses functional plasticity, which was regulated by levels of light energy supplies either via changing light levels or length of light exposure.

  10. Quantification of diazotrophs bacteria isolated from cocoa soils (Theobroma cacao L., by the technique of Most Probable Number (MPN

    Directory of Open Access Journals (Sweden)

    Adriana Zulay Argüello Navarro

    2016-07-01

    Full Text Available The objective of this research was to quantify diazotrophic bacteria and compare physicochemically rhizospheric soils of three cocoa plantations (Theobroma cacao L. in Norte de Santander Department, Colombia; for which they were characterized, differing in cultivated area, agronomic management and crop age. From serial dilutions of the samples and using the technique of Most Probable Number (MPN, In semisolid culture media (NFb, JMV, LGI, JNFb, the diazotrophs were quantified, evaluating as positive the formation of a subsurface film in the medium contained in sealed vials; equal samples were sent to the Bioambiental laboratory (UNET for physicochemical analyzes. As a result, the evaluated samples showed deficiencies in the percentage of organic matter and elements such as Potassium, Phosphorus and Magnesium. Statistically highly significant differences in MPN were reported. The highest quantification of diazotrophs was reported in the Florilandia farm, which was characterized by drip irrigation. The highest quantification of diazotrophs was recorded in the media NFb and JMV, demonstrating a greater presence of the presumed genera Azospirillum sp. and Burkholderia sp. which are easily isolated from rhizospheric soils, unlike the genera Herbaspirillum sp. and Gluconacetobacter sp. which by their endophytic character tend to be less predominant in this type of samples. It is also concluded that the physicochemical characteristics of the soil, humidity and climatic relationships at the moment of sampling, condition the amount of root exudates and therefore are factors that conditioned the presence of diazotrophs in the samples.

  11. Influence of nitrogen fertilization on diazotrophic communities in the rhizosphere of the Jerusalem artichoke (Helianthus tuberosus L.).

    Science.gov (United States)

    Meng, Xianfa; Wang, Lin; Long, Xiaohua; Liu, Zhaopu; Zhang, Zhenhua; Zed, Rengel

    2012-06-01

    Diazotrophs in the soil may be influenced by plant factors as well as nitrogen (N) fertilization. In this study, we investigated potential diazotrophic communities in the rhizosphere of the Jerusalem artichoke (Helianthus tuberosus L.) supplied with differing amounts of N. The community structure of N(2)-fixing bacteria was profiled using the length heterogeneity polymerase chain reaction (LH-PCR) and terminal restriction fragment length polymorphism (T-RFLP) based on a variation in the nifH gene. Higher numbers of diazotrophs were detected by T-RFLP compared to LH-PCR. The lowest number of N(2)-fixing bacteria was observed in the rhizosphere soil with high N fertilization. T-RFLP was a better method than LH-PCR for profiling microbial diversity of diazotrophs using multidimensional scaling (MDS) and analysis of similarity (ANOSIM) of fingerprints as well as diversity measures. The supply of N fertilizer appeared to negatively influence the abundance of diazotrophs in the rhizophere of the Jerusalem artichoke. Copyright © 2012 Institut Pasteur. All rights reserved.

  12. Comparative study of endophytic and endophytic diazotrophic bacterial communities across rice landraces grown in the highlands of northern Thailand.

    Science.gov (United States)

    Rangjaroen, Chakrapong; Rerkasem, Benjavan; Teaumroong, Neung; Sungthong, Rungroch; Lumyong, Saisamorn

    2014-01-01

    Communities of bacterial endophytes within the rice landraces cultivated in the highlands of northern Thailand were studied using fingerprinting data of 16S rRNA and nifH genes profiling by polymerase chain reaction-denaturing gradient gel electrophoresis. The bacterial communities' richness, diversity index, evenness, and stability were varied depending on the plant tissues, stages of growth, and rice cultivars. These indices for the endophytic diazotrophic bacteria within the landrace rice Bue Wah Bo were significantly the lowest. The endophytic bacteria revealed greater diversity by cluster analysis with seven clusters compared to the endophytic diazotrophic bacteria (three clusters). Principal component analysis suggested that the endophytic bacteria showed that the community structures across the rice landraces had a higher stability than those of the endophytic diazotrophic bacteria. Uncultured bacteria were found dominantly in both bacterial communities, while higher generic varieties were observed in the endophytic diazotrophic bacterial community. These differences in bacterial communities might be influenced either by genetic variation in the rice landraces or the rice cultivation system, where the nitrogen input affects the endophytic diazotrophic bacterial community.

  13. Moessbauer study of cobalt and iron in the cyanobacterium (blue green alga)

    International Nuclear Information System (INIS)

    Ambe, Shizuko

    1990-01-01

    Moessbauer emission and absorption studies have been performed on cobalt and iron in the cyanobacterium (blue-green alga). The Moessbauer spectrum of the cyanobacterium cultivated with 57 Co is decomposed into two doublets. The parameters of the major doublet are in good agreement with those of cyanocobalamin (vitamin B 12 ) labeled with 57 Co. The other minor doublet has parameters close to those of Fe(II) coordinated with six nitrogen atoms. These suggest that cobalt is used for the biosynthesis of vitamin B 12 or its analogs in the cyanobacterium. The spectra of the cyanobacterium grown with 57 Fe show that iron is in the high-spin trivalent state and possibly in the form of ferritin, iron storage protein. (orig.)

  14. Identification of genes implicated in toxin production in the cyanobacterium Cylindrospermopsis raciborskii

    DEFF Research Database (Denmark)

    Schembri, Mark; Neilan, B.A.; Saint, C.P.

    2001-01-01

    Cylindrospermopsis raciborskii is a bloom-forming cyanobacterium found in both tropical and temperate climates which produces cylindrospermopsin, a potent hepatotoxic secondary metabolite. This organism is notorious for its association with a significant human poisoning incident on Palm Island...

  15. Evidence for the endophytic colonization of Phaseolus vulgaris(common bean roots by the diazotroph Herbaspirillum seropedicae

    Directory of Open Access Journals (Sweden)

    M.A. Schmidt

    2011-03-01

    Full Text Available Herbaspirillum seropedicae is an endophytic diazotrophic bacterium, which associates with important agricultural plants. In the present study, we have investigated the attachment to and internal colonization of Phaseolus vulgaris roots by the H. seropedicae wild-type strain SMR1 and by a strain of H. seropedicae expressing a red fluorescent protein (DsRed to track the bacterium in the plant tissues. Two-day-old P. vulgaris roots were incubated at 30°C for 15 min with 6 x 10(8 CFU/mL H. seropedicae SMR1 or RAM4. Three days after inoculation, 4 x 10(4 cells of endophytic H. seropedicae SMR1 were recovered per gram of fresh root, and 9 days after inoculation the number of endophytes increased to 4 x 10(6 CFU/g. The identity of the recovered bacteria was confirmed by amplification and sequencing of the 16SrRNA gene. Furthermore, confocal microscopy of P. vulgaris roots inoculated with H. seropedicae RAM4 showed that the bacterial cells were attached to the root surface 15 min after inoculation; fluorescent bacteria were visible in the internal tissues after 24 h and were found in the central cylinder after 72 h, showing that H. seropedicae RAM4 is capable of colonizing the roots of the dicotyledon P. vulgaris. Determination of dry weight of common bean inoculated with H. seropedicae SMR1 suggested that this bacterium has a negative effect on the growth of P. vulgaris.

  16. Evidence for paralytic shellfish poisons in the freshwater cyanobacterium Lyngbya wollei (Farlow ex Gomont) comb. nov.

    OpenAIRE

    Carmichael, W W; Evans, W R; Yin, Q Q; Bell, P; Moczydlowski, E

    1997-01-01

    Lyngbya wollei (Farlow ex Gomont) comb. nov., a perennial mat-forming filamentous cyanobacterium prevalent in lakes and reservoirs of the southeastern United States, was found to produce a potent, acutely lethal neurotoxin when tested in the mouse bioassay. Signs of poisoning were similar to those of paralytic shellfish poisoning. As part of the Tennessee Valley Authority master plan for Guntersville Reservoir, the mat-forming filamentous cyanobacterium L. wollei, a species that had recently ...

  17. Heterologous expression of an algal hydrogenase in a heterocystous cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Thorsten Heidorn; Peter Lindblad [Dept. of Physiological Botany, Uppsala University, Villavogen 6, SE-752 36 Uppsala, (Sweden)

    2006-07-01

    For the expression of an active algal [FeFe] hydrogenase in the heterocystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyanobacteria cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

  18. Heterologous expression of an algal hydrogenase in a heterocystous cyanobacterium

    International Nuclear Information System (INIS)

    Thorsten Heidorn; Peter Lindblad

    2006-01-01

    For the expression of an active algal [FeFe] hydrogenase in the heterocystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyanobacteria cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

  19. Ultraviolet radiation effects on pigmentation in the cyanobacterium ''Phormidium uncinatum''

    International Nuclear Information System (INIS)

    Donkor, V.A.; Haeder, D.P.

    1997-01-01

    The Baikal strain of the cyanobacterium Phormidium uncinatum was found to possess the photosynthetic pigments chlorophyll a, carotenoids, phycocyanin and allophycocyanin, while the Tuebingen strain of Phormidium contained, in addition to these, the biliprotein phycoerythrin. Sucrose gradient centrifugation of the pigment extracts resulted in a separation of the phycobiliproteins into several bands, which according to their absorption and fluorescence properties, were identified as monomers, trimers and hexamers. With increasing UV-B irradiation the heavier aggregates were broken down into smaller components. Photobleaching of these accessory pigments also occurred. FPLC gel filtration analyses of the pigments also showed loss of heavier aggregates of the phycobilins and bleaching of the pigments. SDS-polyacrylamide gel electrophoresis of the sucrose gradient and FPLC fractions indicated loss of the biliproteins with increasing UV-B irradiation. The loss of the β- were more rapid than that of the α- subunits. Increasing levels of ultraviolet irradiation is therefore deleterious to these organism. (author)

  20. Diversity of diazotrophic gut inhabitants of pikas (Ochotonidae) revealed by PCR-DGGE analysis.

    Science.gov (United States)

    Kizilova, A K; Kravchenko, I K

    2014-01-01

    Diazotrophic gut symbionts are considered to act as nitrogen providers for their hosts, as was shown for various termite species. Although the diet of lagomorphs, like pikas or rabbits, is very poor in nitrogen and energy, their fecal matter contains 30-40% of protein. Since our hypothesis was that pikas maintained a diazotrophic consortium in their gastrointestinal tract, we conducted the first investigation of microbial diversity in pika guts. We obtained gut samples from animals of several Ochotona species, O. hyperborea (Northern pika), O. mantchurica (Manchurian pika), and O. dauurica (Daurian pika), in order to retrieve and compare the nitrogen-fixing communities of different pika species. The age and gender of the animals were taken into consideration. We amplified 320-bp long fragments of the nifH gene using the DNA extracted directly from the colon and cecum samples of pika's gut, resolved them by DGGE, and performed phylogenetic reconstruction of 51 sequences obtained from excised bands. No significant difference was detected between the nitrogen-fixing gut inhabitants of different pika species. NifH sequences fell into two clusters. The first cluster contained the sequences affiliated with NifH Cluster I (Zehr et al., 2003) with similarity to Sphingomonas sp., Bradyrhizobium sp., and various uncultured bacteria from soil and rhizosphere. Sequences from the second group were related to Treponema sp., Fibrobacter succinogenes, and uncultured clones from the guts of various termites and belonged to NifH Cluster III. We suggest that diazotrophic organisms from the second cluster are genuine endosymbionts of pikas and provide nitrogen for further synthesis processes thus allowing these animals not to be short of protein.

  1. Genome sequence of Bradyrhizobium sp. LMTR 3, a diazotrophic symbiont of Lima bean (Phaseolus lunatus

    Directory of Open Access Journals (Sweden)

    Ernesto Ormeño-Orrillo

    2017-09-01

    Full Text Available Bradyrhizobium sp. LMTR 3 is a representative strain of one of the geno(species of diazotrophic symbionts associated with Lima bean (Phaseolus lunatus in Peru. Its 7.83 Mb genome was sequenced using the Illumina technology and found to encode a complete set of genes required for nodulation and nitrogen fixation, and additional genes putatively involved in root colonization. Its draft genome sequence and annotation have been deposited at GenBank under the accession number MAXC00000000.

  2. Seagrass (Zostera marina) Colonization Promotes the Accumulation of Diazotrophic Bacteria and Alters the Relative Abundances of Specific Bacterial Lineages Involved in Benthic Carbon and Sulfur Cycling.

    Science.gov (United States)

    Sun, Feifei; Zhang, Xiaoli; Zhang, Qianqian; Liu, Fanghua; Zhang, Jianping; Gong, Jun

    2015-10-01

    Seagrass colonization changes the chemistry and biogeochemical cycles mediated by microbes in coastal sediments. In this study, we molecularly characterized the diazotrophic assemblages and entire bacterial community in surface sediments of a Zostera marina-colonized coastal lagoon in northern China. Higher nitrogenase gene (nifH) copy numbers were detected in the sediments from the vegetated region than in the sediments from the unvegetated region nearby. The nifH phylotypes detected were mostly affiliated with the Geobacteraceae, Desulfobulbus, Desulfocapsa, and Pseudomonas. Redundancy analysis based on terminal restriction fragment length polymorphism analysis showed that the distribution of nifH genotypes was mostly shaped by the ratio of total organic carbon to total organic nitrogen, the concentration of cadmium in the sediments, and the pH of the overlying water. High-throughput sequencing and phylogenetic analyses of bacterial 16S rRNA genes also indicated the presence of Geobacteraceae and Desulfobulbaceae phylotypes in these samples. A comparison of these results with those of previous studies suggests the prevalence and predominance of iron(III)-reducing Geobacteraceae and sulfate-reducing Desulfobulbaceae diazotrophs in coastal sedimentary environments. Although the entire bacterial community structure was not significantly different between these two niches, Desulfococcus (Deltaproteobacteria) and Anaerolineae (Chloroflexi) presented with much higher proportions in the vegetated sediments, and Flavobacteriaceae (Bacteroidetes) occurred more frequently in the bare sediments. These data suggest that the high bioavailability of organic matter (indicated by relatively lower carbon-to-nitrogen ratios) and the less-reducing anaerobic condition in vegetated sediments may favor Desulfococcus and Anaerolineae lineages, which are potentially important populations in benthic carbon and sulfur cycling in the highly productive seagrass ecosystem. Copyright © 2015

  3. Characterization of plant growth-promoting traits of free-living diazotrophic bacteria and their inoculation effects on growth and nitrogen uptake of crop plants.

    Science.gov (United States)

    Islam, Md Rashedul; Madhaiyan, M; Deka Boruah, Hari P; Yim, Woojong; Lee, Gillseung; Saravanan, V S; Fu, Qingling; Hu, Hongqing; Sa, Tongmin

    2009-10-01

    The search for diverse plant growth-promoting (PGP) diazotrophic bacteria is gaining momentum as efforts are made to exploit them as biofertilizers for various economically important crops. In the present study, 17 diazotrophic strains belonging to eight different genera isolated from rice paddy fields were screened for multiple PGP traits and evaluated for their inoculation effects on canola and rice plants. All of the strains tested positive for 1- aminocyclopropane-1-carboxylate (ACC) deaminase activity and production of indole 3-acetic acid (IAA) and ammonia (NH3). Additionally, four of the strains were able to solubilize phosphorus (P), five tested positive for zinc (Zn) solubilization and sulfur (S) oxidation, and eight strains produced siderophores. Based on the presence of multiple PGP traits, 10 strains were selected for inoculation studies. Treatment with Herbaspirillum sp. RFNB26 resulted in maximum root length (54.3%), seedling vigor, and dry biomass in canola, whereas Paenibacillus sp. RFNB4 exhibited the lowest activity under gnotobiotic conditions. However, under pot culture conditions, Paenibacillus sp. RFNB4 significantly increased plant height and dry biomass production by 42.3% and 29.5%, respectively. Canola plants and rhizosphere soils inoculated with Bacillus sp. RFNB6 exhibited significantly higher nitrogenase activity. In greenhouse experiments, Serratia sp. RFNB18 increased rice plant height by 35.1%, Xanthomonas sp. RFNB24 enhanced biomass production by 84.6%, and rice rhizosphere soils inoculated with Herbaspirillum sp. RFNB26 exhibited the highest nitrogenase activity. Our findings indicate that most of the selected strains possess multiple PGP properties that significantly improve the growth parameters of the two plants when tested under controlled conditions.

  4. Seasonal resource conditions favor a summertime increase in North Pacific diatom-diazotroph associations.

    Science.gov (United States)

    Follett, Christopher L; Dutkiewicz, Stephanie; Karl, David M; Inomura, Keisuke; Follows, Michael J

    2018-02-15

    In the North Pacific Subtropical Gyre (NPSG), an annual pulse of sinking organic carbon is observed at 4000 m between July and August, driven by large diatoms found in association with nitrogen fixing, heterocystous, cyanobacteria: Diatom-Diazotroph Associations (DDAs). Here we ask what drives the bloom of DDAs and present a simplified trait-based model of subtropical phototroph populations driven by observed, monthly averaged, environmental characteristics. The ratio of resource supply rates favors nitrogen fixation year round. The relative fitness of DDA traits is most competitive in early summer when the mixed layer is shallow, solar irradiance is high, and phosphorus and iron are relatively abundant. Later in the season, as light intensity drops and phosphorus is depleted, the traits of small unicellular diazotrophs become more competitive. The competitive transition happens in August, at the time when the DDA export event occurs. This seasonal dynamic is maintained when embedded in a more complex, global-scale, ecological model, and provides predictions for the extent of the North Pacific DDA bloom. The model provides a parsimonious and testable hypothesis for the stimulation of DDA blooms.

  5. Spatial and temporal distribution of two diazotrophic bacteria in the Chesapeake Bay.

    Science.gov (United States)

    Short, Steven M; Jenkins, Bethany D; Zehr, Jonathan P

    2004-04-01

    The aim of this study was to initiate autecological studies on uncultivated natural populations of diazotrophic bacteria by examining the distribution of specific diazotrophs in the Chesapeake Bay. By use of quantitative PCR, the abundance of two nifH sequences (907h22 and 912h4) was quantified in water samples collected along a transect from the head to the mouth of the Chesapeake Bay during cruises in April and October 2001 and 2002. Standard curves for the quantitative PCR assays demonstrated that the relationship between gene copies and cycle threshold was linear and highly reproducible from 1 to 10(7) gene copies. The maximum number of 907h22 gene copies detected was approximately 140 ml(-1) and the maximum number of 912h4 gene copies detected was approximately 340 ml(-1). Sequence 912h4 was most abundant at the mouth of the Chesapeake Bay, and in general, its abundance increased with increasing salinity, with the highest abundances observed in April 2002. Overall, the 907h22 phylotype was most abundant at the mid-bay station. Additionally, 907h22 was most abundant in the April samples from the mid-bay and mouth of the Chesapeake Bay. Despite the fact that the Chesapeake Bay is rarely nitrogen limited, our results show that individual nitrogen-fixing bacteria have distinct nonrandom spatial and seasonal distributions in the Chesapeake Bay and are either distributed by specific physical processes or adapted to different environmental niches.

  6. Characterization and crop production efficiency of diazotrophic bacterial isolates from coastal saline soils.

    Science.gov (United States)

    Barua, Shilajit; Tripathi, Sudipta; Chakraborty, Ashis; Ghosh, Sagarmoy; Chakrabarti, Kalyan

    2012-01-20

    Use of eco-friendly area specific salt tolerant bioinoculants is better alternatives to chemical fertilizer for sustainable agriculture in coastal saline soils. We isolated diverse groups of diazotrophic bacteria from coastal saline soils of different forest and agricultural lands in the Sundarbans, West Bengal, India, to study their effect on crop productivity in saline soils. Phenotypic, biochemical and molecular identifications of the isolates were performed. The isolates produced indole acetic acid, phosphatase, and solubilized insoluble phosphates. Sequence analysis of 16S rDNA identified the SUND_BDU1 strain as Agrobacterium and the strains SUND_LM2, Can4 and Can6 belonging to the genus Bacillus. The ARA activity, dinitrogen fixation and presence of nifH genes indicated they were diazotrophs. Field trials with these strains as bioinoculants were carried out during 2007-2009, with rice during August-December followed by Lady's finger during April-June. Microplots, amended with FYM inoculated with four bioinoculants individually were compared against sole FYM (5 t ha(-1)) and a sole chemical fertilizer (60:30:30 kg ha(-1) NPK) treated plot. The strain Can6 was by far the best performer in respect of yield attributes and productivity of studied crops. Copyright © 2011 Elsevier GmbH. All rights reserved.

  7. Isolation and characterization of plant growth promoting endophytic diazotrophic bacteria from Korean rice cultivars.

    Science.gov (United States)

    Ji, Sang Hye; Gururani, Mayank Anand; Chun, Se-Chul

    2014-01-20

    We have isolated 576 endophytic bacteria from the leaves, stems, and roots of 10 rice cultivars and identified 12 of them as diazotrophic bacteria using a specific primer set of nif gene. Through 16S rDNA sequence analysis, nifH genes were confirmed in the two species of Penibacillus, three species of Microbacterium, three Bacillus species, and four species of Klebsiella. Rice seeds treated with these plant growth-promoting bacteria (PGPB) showed improved plant growth, increased height and dry weight and antagonistic effects against fungal pathogens. In addition, auxin and siderophore producing ability, and phosphate solubilizing activity were studied for the possible mechanisms of plant growth promotion. Among 12 isolates tested, 10 strains have shown higher auxin producing activity, 6 isolates were confirmed as strains with high siderophore producing activity while 4 isolates turned out to have high phosphate-solubilizing activity. These results strongly suggest that the endophytic diazotrophic bacteria characterized in this study could be successfully used to promote plant growth and inducing fungal resistance in plants. Copyright © 2013 Elsevier GmbH. All rights reserved.

  8. Quantification and characterization of putative diazotrophic bacteria from forage palm under saline water irrigation

    Directory of Open Access Journals (Sweden)

    Gabiane dos Reis Antunes

    2017-09-01

    Full Text Available The aim of this study was to evaluate the density and phenotypical diversity of diazotrophic endophytic bacteria from the forage palm irrigated with different saline water depths. Opuntia stricta (IPA-200016 received five depths of saline water (L1: 80%. ETo; L2: 60%.ETo; L3: 40%; ETo; L4: 20%; ETo and, L5: 0% ETo, where ETo is the reference evapotranspiration. The roots were collected in the field, disinfected, grounded and serial diluted from 10-1 to 10-4. The total concentration of diazotrophic bacteria was determined by the most probable number method (MPN and the isolated bacteria were characterized phenotipically. The concentration of bacteria found in forage palm roots ranged from 0.36 x 104 to 109.89 104 cells per gram of root, with highest occurrence on the 60 and 80% ETo. In the dendrogram of similarity it was possible to observe the formation of 24 phenotypic groups with 100% similarity. All bacteria presented similarity superior to 40%. Among these groups, 14 are rare groups, formed by only a single bacterial isolate. In the Semi-Arid conditions, the forage palm that receives the highest amount of saline water, presents a higher density of putative nitrogen-fixing endophytic bacteria with high phenotypic diversity.

  9. Diazotrophic potential among bacterial communities associated with wild and cultivated Agave species.

    Science.gov (United States)

    Desgarennes, Damaris; Garrido, Etzel; Torres-Gomez, Miryam J; Peña-Cabriales, Juan J; Partida-Martinez, Laila P

    2014-12-01

    Agaves are major biotic resources in arid and semi-arid ecosystems. Despite their ecological, economical and cultural relevance, many aspects of the microbial communities associated with agaves are still unknown. Here, we investigated the bacterial communities associated with two Agave species by 16S rRNA- Denaturing gradient gel electrophoresis fingerprinting and sequencing. We also evaluated the effects of biotic and abiotic factors in the structure of the bacterial communities. In parallel, we isolated and characterized diazotrophic bacteria associated with agaves, as Agave soils are characterized by their low nitrogen content. Our results demonstrate that in Agave, the structure of prokaryotic assemblages was mostly influenced by the community group, where the soil, episphere, and endosphere were clearly distinct. Proteobacteria (γ and α), Actinobacteria, and Acidobacteria were the dominant phyla. Bacterial communities in the episphere of agaves were mainly influenced by the host species, whereas in the endosphere were affected by the season. Fifteen bacterial taxa were common and abundant in the endosphere of both Agave species during the dry season. Notably, some of the confirmed diazotrophic strains belonged to this group, suggesting a possible beneficial role in planta. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  10. Toxicity of herbicides used in the sugarcane crop to diazotrophic bacterium Herbaspirillum seropedicae

    Directory of Open Access Journals (Sweden)

    Sergio de Oliveira Procópio

    2014-10-01

    Full Text Available The objective of this work was to identify herbicides used in the sugarcane crop that affects neither the growth, the development, of nor the process of biological nitrogen fixation (BNF by the diazotrophic bacterium Herbaspirillum seropedicae. Eighteen herbicides (paraquat, ametryne, tebuthiuron, amicarbazone, diuron, metribuzin, [hexazinone + diuron], [hexazinone + clomazone], clomazone, isoxaflutole, sulfentrazone, oxyfluorfen, imazapic, imazapyr, [trifloxysulfuron sodium + ametryne], glyphosate, MSMA e 2,4-D were tested in their respective commercial doses regarding their impact on the growth of the bacteria in liquid medium DIGs. For this, we determined the duration of lag phase, generation time and maximum cell density of H. seropedicae, calculated from optical density data obtained at regular intervals during the incubation of cultures for 33 h at 32oC. We also evaluated the impact of herbicides on nitrogenase activity of H. seropedicae grown in semi-solid N-free JNFb medium. The effects of herbicides on the growth variables and the ARA were compared with the untreated control by Dunnett test. A completely randomized design was used. The herbicides paraquat, imazapyr, ametryne, glyphosate and oxyfluorfen inhibited the growth of H. seropedicae in vitro. Ametryne, oxyfluorfen and glyphosate caused a small reduction in the duration of the lag phase of diazotrophic bacteria H. seropedicae. Oxyfluorfen, ametryne and imazapyr resulted in increased the generation time by H. seropedicae. Glyphosate promoted drastic reduction in biological nitrogen fixation in vitro by H. seropedicae. The other tested herbicides did not affect the growth or the same BNF by H. seropedicae.

  11. Genome Sequence and Composition of a Tolyporphin-Producing Cyanobacterium-Microbial Community.

    Science.gov (United States)

    Hughes, Rebecca-Ayme; Zhang, Yunlong; Zhang, Ran; Williams, Philip G; Lindsey, Jonathan S; Miller, Eric S

    2017-10-01

    The cyanobacterial culture HT-58-2 was originally described as a strain of Tolypothrix nodosa with the ability to produce tolyporphins, which comprise a family of distinct tetrapyrrole macrocycles with reported efflux pump inhibition properties. Upon reviving the culture from what was thought to be a nonextant collection, studies of culture conditions, strain characterization, phylogeny, and genomics have been undertaken. Here, HT-58-2 was shown by 16S rRNA analysis to closely align with Brasilonema strains and not with Tolypothrix isolates. Light, fluorescence, and scanning electron microscopy revealed cyanobacterium filaments that are decorated with attached bacteria and associated with free bacteria. Metagenomic surveys of HT-58-2 cultures revealed a diversity of bacteria dominated by Erythrobacteraceae , 97% of which are Porphyrobacter species. A dimethyl sulfoxide washing procedure was found to yield enriched cyanobacterial DNA (presumably by removing community bacteria) and sequence data sufficient for genome assembly. The finished, closed HT-58-2Cyano genome consists of 7.85 Mbp (42.6% G+C) and contains 6,581 genes. All genes for biosynthesis of tetrapyrroles (e.g., heme, chlorophyll a , and phycocyanobilin) and almost all for cobalamin were identified dispersed throughout the chromosome. Among the 6,177 protein-encoding genes, coding sequences (CDSs) for all but two of the eight enzymes for conversion of glutamic acid to protoporphyrinogen IX also were found within one major gene cluster. The cluster also includes 10 putative genes (and one hypothetical gene) encoding proteins with domains for a glycosyltransferase, two cytochrome P450 enzymes, and a flavin adenine dinucleotide (FAD)-binding protein. The composition of the gene cluster suggests a possible role in tolyporphin biosynthesis. IMPORTANCE A worldwide search more than 25 years ago for cyanobacterial natural products with anticancer activity identified a culture (HT-58-2) from Micronesia that

  12. Effect of salinity on nitrogenase activity and composition of the active diazotrophic community in intertidal microbial mats

    NARCIS (Netherlands)

    Severin, I.; Confurius-Guns, V.; Stal, L.J.

    2012-01-01

    Microbial mats are often found in intertidal areas experiencing a large range of salinities. This study investigated the effect of changing salinities on nitrogenase activity and on the composition of the active diazotrophic community ( transcript libraries) of three types of microbial mats situated

  13. Isolation and Screening of Bacteria for Their Diazotrophic Potential and Their Influence on Growth Promotion of Maize Seedlings in Greenhouses.

    Science.gov (United States)

    Kifle, Medhin H; Laing, Mark D

    2015-01-01

    Poor soil fertility is one of the major constraints for crop production. Nitrogen is the most limiting nutrient for increasing crop productivity. Therefore, there is a need to identify diazotrophic inoculants as an alternative or supplement to N-fertilizers for sustainable agriculture. In the current study, a number of free-living diazotrophic bacteria were isolated from soils collected from maize rhizosphere and from leaves and roots of maize within the KwaZulu-Natal Province, Republic of South Africa. Ninety-two isolates were selected for further screening because they were able to grow on N-free media containing different carbon sources. Isolates that were very slow to grow on N-free media were discarded. The isolates were screened in vitro for diazotrophic potential tests for ammonia production and acetylene reduction. Ethylene (C2H4) production was quantified and ranged from 4 to 73 nmoles of C2H4h(-1) culture(-1). The top 20 isolates were re-screened on maize seedlings, and eight isolates significantly (P = 0.001) enhanced some growth parameters of maize above the un-inoculated control. Isolates that showed significant effect on at least two growth parameters were identified at species or genera level. In conclusion, selected diazotrophic isolates may be potentially beneficial but they should be tested more in greenhouse and field conditions with maize to confirm their potential for application as biofertilizers.

  14. Isolation and screening of bacteria for their diazotrophic potential and their influence on growth promotion of maize seedlings in greenhouses

    Directory of Open Access Journals (Sweden)

    Medhin Hadish Kifle

    2016-01-01

    Full Text Available Poor soil fertility is one of the major constraints for crop production. Nitrogen is the most limiting nutrient for increasing crop productivity. Therefore, there is a need to identify diazotrophic inoculants as an alternative or supplement to N-fertilizers for sustainable agriculture. In the current study, a number of free-living diazotrophic bacteria were isolated from soils collected from maize rhizosphere and from leaves and roots of maize within the KwaZulu-Natal Province, Republic of South Africa. Ninety two isolates were selected for further screening because they were able to grow on N-free media containing different carbon sources. Isolates that were very slow to grow on N-free media were discarded. The isolates were screened in vitro for diazotrophic potential tests for ammonia production and acetylene reduction. Ethylene (C2H4 production was quantified and ranged from 4 to 73 nmoles of C2H4 h-1 culture-1. The top twenty isolates were re-screened on maize seedlings, and eight isolates significantly (P=0.001 enhanced some growth parameters of maize above the un-inoculated control. Isolates that showed significant effect on at least two growth parameters were identified at species or genera level. In conclusion, selected diazotrophic isolates may be potentially beneficial but they should be tested more in greenhouse and field conditions with maize to confirm their potential for application as biofertilizers.

  15. Distribution and drivers of symbiotic and free-living diazotrophic cyanobacteria in the western tropical South Pacific

    Science.gov (United States)

    Stenegren, Marcus; Caputo, Andrea; Berg, Carlo; Bonnet, Sophie; Foster, Rachel A.

    2018-03-01

    The abundance and distribution of cyanobacterial diazotrophs were quantified in two regions (Melanesian archipelago, MA; and subtropical gyre, SG) of the western tropical South Pacific using nifH quantitative polymerase chain reaction (qPCR) assays. UCYN-A1 and A2 host populations were quantified using 18S rRNA qPCR assays including one newly developed assay. All phylotypes were detected in the upper photic zone (0-50 m), with higher abundances in the MA region. Trichodesmium and UCYN-B dominated and ranged from 2.18 × 102 to 9.41 × 106 and 1.10 × 102 to 2.78 × 106 nifH copies L-1, respectively. Het-1 (symbiont of Rhizosolenia diatoms) was the next most abundant (1.40 × 101-1.74 × 105 nifH copies L-1) and co-occurred with het-2 and het-3. UCYN-A1 and A2 were the least abundant diazotrophs and were below detection (bd) in 63 and 79, respectively, of 120 samples. In addition, in up to 39 % of samples in which UCYN-A1 and A2 were detected, their respective hosts were bd. Pairwise comparisons of the nifH abundances and various environmental parameters supported two groups: a deep-dwelling group (45 m) comprised of UCYN-A1 and A2 and a surface group (0-15 m) comprised of Trichodesmium, het-1 and het-2. Temperature and photosynthetically active radiation were positively correlated with the surface group, while UCYN-A1 and A2 were positively correlated with depth, salinity, and oxygen. Similarly, in a meta-analysis of 11 external datasets, all diazotrophs, except UCYN-A were correlated with temperature. Combined, our results indicate that conditions favoring the UCYN-A symbiosis differ from those of diatom diazotroph associations and free-living cyanobacterial diazotrophs.

  16. CRISPR/Cas9 mediated targeted mutagenesis of the fast growing cyanobacterium Synechococcus elongatus UTEX 2973.

    Science.gov (United States)

    Wendt, Kristen E; Ungerer, Justin; Cobb, Ryan E; Zhao, Huimin; Pakrasi, Himadri B

    2016-06-23

    As autotrophic prokaryotes, cyanobacteria are ideal chassis organisms for sustainable production of various useful compounds. The newly characterized cyanobacterium Synechococcus elongatus UTEX 2973 is a promising candidate for serving as a microbial cell factory because of its unusually rapid growth rate. Here, we seek to develop a genetic toolkit that enables extensive genomic engineering of Synechococcus 2973 by implementing a CRISPR/Cas9 editing system. We targeted the nblA gene because of its important role in biological response to nitrogen deprivation conditions. First, we determined that the Streptococcus pyogenes Cas9 enzyme is toxic in cyanobacteria, and conjugational transfer of stable, replicating constructs containing the cas9 gene resulted in lethality. However, after switching to a vector that permitted transient expression of the cas9 gene, we achieved markerless editing in 100 % of cyanobacterial exconjugants after the first patch. Moreover, we could readily cure the organisms of antibiotic resistance, resulting in a markerless deletion strain. High expression levels of the Cas9 protein in Synechococcus 2973 appear to be toxic and result in cell death. However, introduction of a CRISPR/Cas9 genome editing system on a plasmid backbone that leads to transient cas9 expression allowed for efficient markerless genome editing in a wild type genetic background.

  17. Advances in the Function and Regulation of Hydrogenase in the Cyanobacterium Synechocystis PCC6803

    Science.gov (United States)

    Cassier-Chauvat, Corinne; Veaudor, Théo; Chauvat, Franck

    2014-01-01

    In order to use cyanobacteria for the biological production of hydrogen, it is important to thoroughly study the function and the regulation of the hydrogen-production machine in order to better understand its role in the global cell metabolism and identify bottlenecks limiting H2 production. Most of the recent advances in our understanding of the bidirectional [Ni-Fe] hydrogenase (Hox) came from investigations performed in the widely-used model cyanobacterium Synechocystis PCC6803 where Hox is the sole enzyme capable of combining electrons with protons to produce H2 under specific conditions. Recent findings suggested that the Hox enzyme can receive electrons from not only NAD(P)H as usually shown, but also, or even preferentially, from ferredoxin. Furthermore, plasmid-encoded functions and glutathionylation (the formation of a mixed-disulfide between the cysteines residues of a protein and the cysteine residue of glutathione) are proposed as possible new players in the function and regulation of hydrogen production. PMID:25365180

  18. CyanOmics: an integrated database of omics for the model cyanobacterium Synechococcus sp. PCC 7002.

    Science.gov (United States)

    Yang, Yaohua; Feng, Jie; Li, Tao; Ge, Feng; Zhao, Jindong

    2015-01-01

    Cyanobacteria are an important group of organisms that carry out oxygenic photosynthesis and play vital roles in both the carbon and nitrogen cycles of the Earth. The annotated genome of Synechococcus sp. PCC 7002, as an ideal model cyanobacterium, is available. A series of transcriptomic and proteomic studies of Synechococcus sp. PCC 7002 cells grown under different conditions have been reported. However, no database of such integrated omics studies has been constructed. Here we present CyanOmics, a database based on the results of Synechococcus sp. PCC 7002 omics studies. CyanOmics comprises one genomic dataset, 29 transcriptomic datasets and one proteomic dataset and should prove useful for systematic and comprehensive analysis of all those data. Powerful browsing and searching tools are integrated to help users directly access information of interest with enhanced visualization of the analytical results. Furthermore, Blast is included for sequence-based similarity searching and Cluster 3.0, as well as the R hclust function is provided for cluster analyses, to increase CyanOmics's usefulness. To the best of our knowledge, it is the first integrated omics analysis database for cyanobacteria. This database should further understanding of the transcriptional patterns, and proteomic profiling of Synechococcus sp. PCC 7002 and other cyanobacteria. Additionally, the entire database framework is applicable to any sequenced prokaryotic genome and could be applied to other integrated omics analysis projects. Database URL: http://lag.ihb.ac.cn/cyanomics. © The Author(s) 2015. Published by Oxford University Press.

  19. Enhanced ferrihydrite dissolution by a unicellular, planktonic cyanobacterium: a biological contribution to particulate iron bioavailability.

    Science.gov (United States)

    Kranzler, Chana; Kessler, Nivi; Keren, Nir; Shaked, Yeala

    2016-12-01

    Iron (Fe) bioavailability, as determined by its sources, sinks, solubility and speciation, places severe environmental constraints on microorganisms in aquatic environments. Cyanobacteria are a widespread group of aquatic, photosynthetic microorganisms with especially high iron requirements. While iron exists predominantly in particulate form, little is known about its bioavailability to cyanobacteria. Some cyanobacteria secrete iron solubilizing ligands called siderophores, yet many environmentally relevant strains do not have this ability. This work explores the bioavailability of amorphous synthetic Fe-oxides (ferrihydrite) to the non-siderophore producing, unicellular cyanobacterium, Synechocystis sp PCC 6803. Iron uptake assays with 55 ferrihydrite established dissolution as a critical prerequisite for iron transport. Dissolution assays with the iron binding ligand, desferrioxamine B, demonstrated that Synechocystis 6803 enhances ferrihydrite dissolution, exerting siderophore-independent biological influence on ferrihydrite bioavailability. Dissolution mechanisms were studied using a range of experimental conditions; both cell-particle physical proximity and cellular electron flow were shown to be important determinants of bio-dissolution by Synechocystis 6803. Finally, the effects of ferrihydrite stability on bio-dissolution rates and cell physiology were measured, integrating biological and chemical aspects of ferrihydrite bioavailability. Collectively, these findings demonstrate that Synechocystis 6803 actively dissolves ferrihydrite, highlighting a significant biological component to mineral phase iron bioavailability in aquatic environments. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  20. In silico differential display of defense-related expressed sequence tags from sugarcane tissues infected with diazotrophic endophytes

    Directory of Open Access Journals (Sweden)

    Lambais Marcio R.

    2001-01-01

    Full Text Available The expression patterns of 277 sugarcane expressed sequence tags (EST-contigs encoding putative defense-related (DR proteins were evaluated using the Sugarcane EST database. The DR proteins evaluated included chitinases, beta-1,3-glucanases, phenylalanine ammonia-lyases, chalcone synthases, chalcone isomerases, isoflavone reductases, hydroxyproline-rich glycoproteins, proline-rich glycoproteins, peroxidases, catalases, superoxide dismutases, WRKY-like transcription factors and proteins involved in cell death control. Putative sugarcane WRKY proteins were compared and their phylogenetic relationships determined. A hierarchical clustering approach was used to identify DR ESTs with similar expression profiles in representative cDNA libraries. To identify DR ESTs differentially expressed in sugarcane tissues infected with Gluconacetobacter diazotrophicus or Herbaspirillum rubrisubalbicans, 179 putative DR EST-contigs expressed in non-infected tissues (leaves and roots and/or infected tissues were selected and arrayed by similarity of their expression profiles. Changes in the expression levels of 124 putative DR EST-contigs, expressed in non-infected tissues, were evaluated in infected tissues. Approximately 42% of these EST-contigs showed no expression in infected tissues, whereas 15% and 3% showed more than 2-fold suppression in tissues infected with G. diazotrophicus or H. rubrisubalbicans, respectively. Approximately 14 and 8% of the DR EST-contigs evaluated showed more than 2-fold induction in tissues infected with G. diazotrophicus or H. rubrisubalbicans, respectively. The differential expression of clusters of DR genes may be important in the establishment of a compatible interaction between sugarcane and diazotrophic endophytes. It is suggested that the hierarchical clustering approach can be used on a genome-wide scale to identify genes likely involved in controlling plant-microorganism interactions.

  1. Two New Lyngbyatoxin Derivatives from the Cyanobacterium, Moorea producens

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    Weina Jiang

    2014-12-01

    Full Text Available The toxin-producing cyanobacterium, Moorea producens, is a known causative organism of food poisoning and seaweed dermatitis (also known as “swimmer’s itch”. Two new toxic compounds were isolated and structurally elucidated from an ethyl acetate extract of M. producens collected from Hawaii. Analyses of HR-ESI-MS and NMR spectroscopies, as well as optical rotations and CD spectra indicated two new lyngbyatoxin derivatives, 2-oxo-3(R-hydroxy-lyngbyatoxin A (1 and 2-oxo-3(R-hydroxy-13-N-desmethyl-lyngbyatoxin A (2. The cytotoxicity and lethal activities of 1 and 2 were approximately 10- to 150-times less potent than lyngbyatoxin A. Additionally, the binding activities of 1 and 2 possessed 10,000-times lower affinity for the protein kinase Cδ (PKCδ-C1B peptide when compared to lyngbyatoxin A. These findings suggest that these new lyngbyatoxin derivatives may mediate their acute toxicities through a non-PKC activation pathway.

  2. Nitrogen-fixing cyanobacterium with a high phycoerythrin content.

    Science.gov (United States)

    Rodriguez, H; Rivas, J; Guerrero, M G; Losada, M

    1989-03-01

    The elemental and molecular composition, pigment content, and productivity of a phycoerythrin-rich nitrogen-fixing cyanobacterium-an Anabaena strain isolated from the coastal lagoon Albufera de Valencia, Spain-has been investigated. When compared with other heterocystous species, this strain exhibits similar chlorophyll a, carotene, and total phycobiliprotein contents but differs remarkably in the relative proportion of specific phycobiliproteins; the content of C-phycoerythrin amounts to 8.3% (versus about 1% in the other species) of cell dry weight. Absorption and fluorescence spectra of intact phycobilisomes isolated from this Anabaena sp. corroborate the marked contribution of phycoerythrin as an antenna pigment, a circumstance that is unusual for cyanobacteria capable of fixing N(2). The pigment content of cells is affected by variations in irradiance and cell density, these adaptive changes being more patent for C-phycoerythrin than for phycocyanins. The Anabaena strain is clumpy and capable of rapid flocculation. It exhibits outdoor productivities higher than 20 g (dry weight) m day during summer.

  3. Non-symbiotic bacterial diazotrophs from of agricultural crops of San Carlos. Córdoba, Colombia

    Directory of Open Access Journals (Sweden)

    Cecilia Lara Mantilla

    2007-07-01

    Full Text Available Diazotrophic Azotobacter sp and Azospirrillum sp bacteria were isolated from the rhizosphere of plantain, (Musa paradisiaca, corn, wheat and fallow areas (uncultivated, but covered by fallen leaves in the San Carlos region (in the Sinú valley in the Córdoba department, Colombia. Microbial populations were identified by biochemical tests; physiological characters were examined using Gram reaction in different mediums: Burk’s, Ashby and Jensen’s (Azotobacter sp and Burk’s, NFB and Congo-red medium (Azospirillum sp. The study was aimed at determining ion ammonium production from Azotobacter sp and Azospirrillum sp isolates; ion ammonium was quantified according to Berthelot’s colorimetric technique (phenol-hypochlorite. This method was modified and standardised in line with using a Perkin-Elmer Lamba 11 UV-Vis spectrometer. As a result of this study, 14 isolates have been shown to be ammonium-producers at concentrations ranging from 0.9 mg/l to 5.2 mg/l; A16PG (5.1545 mg/l and A26M1P (5.1743 mg/l yielded the highest ammonium concentrations for Azotobacter sp and A5M1G (4.6741 mg/l for Azospirrillum sp. Biological N2 fixation (BNF by associative diazotrophic bacteria has contributed towards increasing harvest yield, thereby reducing the need for nitrogenised fertilisers and the emission of greenhouse gases (such as N2O and obtaining economic and environmental benefits for farming.

  4. Nitrogen fixation dynamics of two diazotrophic communities in Mono Lake, California

    Science.gov (United States)

    Oremland, R.S.

    1990-01-01

    Two types of diazotrophic microbial communities were found in the littoral zone of alkaline hypersaline Mono Lake, California. One consisted of anaerobic bacteria inhabiting the flocculent surface layers of sediments. Nitrogen fixation (acetylene reduction) by flocculent surface layers occurred under anaerobic conditions, was not stimulated by light or by additions of organic substrates, and was inhibited by O2, nitrate, and ammonia. The second community consisted of a ball-shaped association of a filamentous chlorophyte (Ctenocladus circinnatus) with diazotrophic, nonheterocystous cyanobacteria, as well as anaerobic bacteria (Ctenocladus balls). Nitrogen fixation by Ctenocladus balls was usually, but not always, stimulated by light. Rates of anaerobic dark fixation equaled those in the light under air. Fixation in the light was stimulated by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and by propanil [N-(3,4-dichlorophenyl)propanamide]. 3-(3,4-Dichlorophenyl)-1,1-dimethyl urea-elicited nitrogenase activity was inhibited by ammonia (96%) and nitrate (65%). Fixation was greatest when Ctenocladus balls were incubated anaerobically in the light with sulfide. Dark anaerobic fixation was not stimulated by organic substrates in short-term (4-h) incubations, but was in long-term (67-h) ones. Areal estimates of benthic N2 fixation were measured seasonally, using chambers. Highest rates (~29.3 ??mol of C2H4 m-2 h-1) occurred under normal diel regimens of light and dark. These estimates indicate that benthic N2 fixation has the potential to be a significant nitrogen source in Mono Lake.

  5. Bacterial and diazotrophic diversities of endophytes in Dendrobium catenatum determined through barcoded pyrosequencing.

    Science.gov (United States)

    Li, Ou; Xiao, Rong; Sun, Lihua; Guan, Chenglin; Kong, Dedong; Hu, Xiufang

    2017-01-01

    As an epiphyte orchid, Dendrobium catenatum relies on microorganisms for requisite nutrients. Metagenome pyrosequencing based on 16S rRNA and nifH genes was used to characterize the bacterial and diazotrophic communities associated with D. catenatum collected from 5 districts in China. Based on Meta-16S rRNA sequencing, 22 bacterial phyla and 699 genera were identified, distributed as 125 genera from 8 phyla and 319 genera from 10 phyla shared by all the planting bases and all the tissues, respectively. The predominant Proteobacteria varied from 71.81% (GZ) to 96.08% (YN), and Delftia (10.39-38.42%), Burkholderia (2.71-15.98%), Escherichia/Shigella (4.90-25.12%), Pseudomonas (2.68-30.72%) and Sphingomonas (1.83-2.05%) dominated in four planting bases. Pseudomonas (17.94-22.06%), Escherichia/Shigella (6.59-11.59%), Delftia (9.65-22.14%) and Burkholderia (3.12-11.05%) dominated in all the tissues. According to Meta-nifH sequencing, 4 phyla and 45 genera were identified, while 17 genera and 24 genera from 4 phyla were shared by all the planting bases and all the tissues, respectively. Burkholderia and Bradyrhizobium were the most popular in the planting bases, followed by Methylovirgula and Mesorhizobium. Mesorhizobium was the most popular in different tissues, followed by Beijerinckia, Xanthobacter, and Burkholderia. Among the genera, 39 were completely overlapped with the results based on the 16S rRNA gene. In conclusion, abundant bacteria and diazotrophs were identified in common in different tissues of D. catenatum from five planting bases, which might play a great role in the supply of nutrients such as nitrogen. The exact abundance of phylum and genus on the different tissues from different planting bases need deeper sequencing with more samples.

  6. Effect of diazotrophic bacteria as phosphate solubilizing and indolic compound producers on maize plants

    Directory of Open Access Journals (Sweden)

    Mónica Del Pilar López Ortega

    2013-07-01

    Full Text Available Phosphorus is limiting for growth of maize plants, and because of that use of fertilizers like Rock Phosphate has been proposed. However, direct use of Rock Phosphate is not recommended because of its low availability, so it is necessary to improve it. In this study, a group of diazotrophic bacteria were evaluated as phosphate-solubilizing bacteria, for their production of indolic compounds and for their effects on growth of maize plants. Strains of the genera Azosporillum, Azotobacter, Rhizobium and Klebsiella, were quantitatively evaluated for solubilization of Ca3(PO42 and rock phosphate as a single source of phosphorous in SRS culture media. Additionally, the phosphatase enzyme activity was quantified at pH 5.0, 7.0 and 8.0 using p-nitrophenyl phosphate, and production of indolic compound was determined by colorimetric quantification. The effect of inoculation of bacteria on maize was determined in a completely randomized greenhouse experiment where root and shoot dry weights and phosphorus content were assessed. Results showed that strain C50 produced 107.2 mg .L-1 of available-P after 12 days of fermentation, and AC10 strain had the highest phosphatase activity at pH 8 with 12.7 mg of p-nitrophenol mL .h-1. All strains synthetized indolic compounds, and strain AV5 strain produced the most at 63.03 µg .mL-1. These diazotrophic bacteria increased plant biomass up to 39 % and accumulation of phosphorus by 10%. Hence, use of diazotrphic phosphate-solubilizing bacteria may represent an alternative technology for fertilization systems in maize plants.

  7. Bacterial and diazotrophic diversities of endophytes in Dendrobium catenatum determined through barcoded pyrosequencing.

    Directory of Open Access Journals (Sweden)

    Ou Li

    Full Text Available As an epiphyte orchid, Dendrobium catenatum relies on microorganisms for requisite nutrients. Metagenome pyrosequencing based on 16S rRNA and nifH genes was used to characterize the bacterial and diazotrophic communities associated with D. catenatum collected from 5 districts in China. Based on Meta-16S rRNA sequencing, 22 bacterial phyla and 699 genera were identified, distributed as 125 genera from 8 phyla and 319 genera from 10 phyla shared by all the planting bases and all the tissues, respectively. The predominant Proteobacteria varied from 71.81% (GZ to 96.08% (YN, and Delftia (10.39-38.42%, Burkholderia (2.71-15.98%, Escherichia/Shigella (4.90-25.12%, Pseudomonas (2.68-30.72% and Sphingomonas (1.83-2.05% dominated in four planting bases. Pseudomonas (17.94-22.06%, Escherichia/Shigella (6.59-11.59%, Delftia (9.65-22.14% and Burkholderia (3.12-11.05% dominated in all the tissues. According to Meta-nifH sequencing, 4 phyla and 45 genera were identified, while 17 genera and 24 genera from 4 phyla were shared by all the planting bases and all the tissues, respectively. Burkholderia and Bradyrhizobium were the most popular in the planting bases, followed by Methylovirgula and Mesorhizobium. Mesorhizobium was the most popular in different tissues, followed by Beijerinckia, Xanthobacter, and Burkholderia. Among the genera, 39 were completely overlapped with the results based on the 16S rRNA gene. In conclusion, abundant bacteria and diazotrophs were identified in common in different tissues of D. catenatum from five planting bases, which might play a great role in the supply of nutrients such as nitrogen. The exact abundance of phylum and genus on the different tissues from different planting bases need deeper sequencing with more samples.

  8. Bacterial and diazotrophic diversities of endophytes in Dendrobium catenatum determined through barcoded pyrosequencing

    Science.gov (United States)

    Li, Ou; Sun, Lihua; Guan, Chenglin; Kong, Dedong

    2017-01-01

    As an epiphyte orchid, Dendrobium catenatum relies on microorganisms for requisite nutrients. Metagenome pyrosequencing based on 16S rRNA and nifH genes was used to characterize the bacterial and diazotrophic communities associated with D. catenatum collected from 5 districts in China. Based on Meta-16S rRNA sequencing, 22 bacterial phyla and 699 genera were identified, distributed as 125 genera from 8 phyla and 319 genera from 10 phyla shared by all the planting bases and all the tissues, respectively. The predominant Proteobacteria varied from 71.81% (GZ) to 96.08% (YN), and Delftia (10.39–38.42%), Burkholderia (2.71–15.98%), Escherichia/Shigella (4.90–25.12%), Pseudomonas (2.68–30.72%) and Sphingomonas (1.83–2.05%) dominated in four planting bases. Pseudomonas (17.94–22.06%), Escherichia/Shigella (6.59–11.59%), Delftia (9.65–22.14%) and Burkholderia (3.12–11.05%) dominated in all the tissues. According to Meta-nifH sequencing, 4 phyla and 45 genera were identified, while 17 genera and 24 genera from 4 phyla were shared by all the planting bases and all the tissues, respectively. Burkholderia and Bradyrhizobium were the most popular in the planting bases, followed by Methylovirgula and Mesorhizobium. Mesorhizobium was the most popular in different tissues, followed by Beijerinckia, Xanthobacter, and Burkholderia. Among the genera, 39 were completely overlapped with the results based on the 16S rRNA gene. In conclusion, abundant bacteria and diazotrophs were identified in common in different tissues of D. catenatum from five planting bases, which might play a great role in the supply of nutrients such as nitrogen. The exact abundance of phylum and genus on the different tissues from different planting bases need deeper sequencing with more samples. PMID:28931073

  9. The Cyanobacterium Cylindrospermopsis raciborskii (CYRF-01 Responds to Environmental Stresses with Increased Vesiculation Detected at Single-Cell Resolution

    Directory of Open Access Journals (Sweden)

    Victor Zarantonello

    2018-02-01

    Full Text Available Secretion of membrane-limited vesicles, collectively termed extracellular vesicles (EVs, is an important biological process of both eukaryotic and prokaryotic cells. This process has been observed in bacteria, but remains to be better characterized at high resolution in cyanobacteria. In the present work, we address the release of EVs by Cylindrospermopsis raciborskii (CYRF-01, a filamentous bloom-forming cyanobacterium, exposed to environmental stressors. First, non-axenic cultures of C. raciborskii (CYRF-01 were exposed to ultraviolet radiation (UVA + UVB over a 6 h period, which is known to induce structural damage to this species. Second, C. raciborskii was co-cultured in interaction with another cyanobacterium species, Microcystis aeruginosa (MIRF-01, over a 24 h period. After the incubation times, cell density and viability were analyzed, and samples were processed for transmission electron microscopy (TEM. Our ultrastructural analyses revealed that C. raciborskii constitutively releases EVs from the outer membrane during its normal growth and amplifies such ability in response to environmental stressors. Both situations induced significant formation of outer membrane vesicles (OMVs by C. raciborskii compared to control cells. Quantitative TEM revealed an increase of 48% (UV and 60% (interaction in the OMV numbers compared to control groups. Considering all groups, the OMVs ranged in size from 20 to 300 nm in diameter, with most OMVs showing diameters between 20 and 140 nm. Additionally, we detected that OMV formation is accompanied by phosphatidylserine exposure, a molecular event also observed in EV-secreting eukaryotic cells. Altogether, we identified for the first time that C. raciborskii has the competence to secrete OMVs and that under different stress situations the genesis of these vesicles is increased. The amplified ability of cyanobacteria to release OMVs may be associated with adaptive responses to changes in environmental

  10. Temperature and irradiance influences on cadmium and zinc uptake and toxicity in a freshwater cyanobacterium, Microcystis aeruginosa

    International Nuclear Information System (INIS)

    Zeng Jin; Wang Wenxiong

    2011-01-01

    Highlights: → This study is the first to study the influences of temperature and light irradiance, two critical factors for the occurrence of cyanobacterial blooms, on metal uptake, subcellular distribution, and toxicity in a freshwater cyanobacterium commonly blooming in eutrophic lakes. → With increasing metal exposure, both cellular growth rate and photosynthesis became more sensitive to metal toxicity under elevated irradiance and temperature, primarily as a result of increased uptake and accumulation. → Cd in the metal rich granule faction increased under Cd exposure, suggesting that MRG may partially detoxify Cd in the cyanobacterial cells. → This study implies that temperature and irradiance may influence the chemical cycling of metals during cyanobacterial blooming in eutrophic freshwater ecosystems. - Abstract: Temperature and light irradiance are important factors affecting the occurrence of cyanobacterial blooms. In this study, we examined the influences of different temperatures (15, 24, and 30 ° C ) and irradiances (18, 32, and 55 μmol photons m -2 s -1 ) on the uptake and toxicity of cadmium (Cd) and zinc (Zn) in a freshwater cyanobacterium Microcystis aeruginosa. The subcellular distribution of Cd and Zn was analyzed. Enhanced growth rates were observed for the cyanobacterial cells incubated at higher temperature or irradiance conditions with lower metal concentrations. With increasing ambient Cd or Zn concentrations, both cellular growth rate and photosynthesis were significantly inhibited at elevated irradiance conditions. The observed increase in Cd and Zn toxicity might be attributed to the enhanced metal uptake and accumulation in Microcystis. Based on the intracellular Cd concentration, the 50% inhibition concentration (IC 50 ) values were higher at the higher temperature or irradiance treatment. The subcellular distribution demonstrated that Cd in the metal rich granule (MRG) faction increased with elevated [Cd 2+ ] concentration

  11. Arsenic biotransformation by a cyanobacterium Nostoc sp. PCC 7120.

    Science.gov (United States)

    Xue, Xi-Mei; Yan, Yu; Xiong, Chan; Raber, Georg; Francesconi, Kevin; Pan, Ting; Ye, Jun; Zhu, Yong-Guan

    2017-09-01

    Nostoc sp. PCC 7120 (Nostoc), a typical filamentous cyanobacterium ubiquitous in aquatic system, is recognized as a model organism to study prokaryotic cell differentiation and nitrogen fixation. In this study, Nostoc cells incubated with arsenite (As(III)) for two weeks were extracted with dichloromethane/methanol (DCM/MeOH) and the extract was partitioned between water and DCM. Arsenic species in aqueous and DCM layers were determined using high performance liquid chromatography - inductively coupled plasma mass spectrometer/electrospray tandem mass spectrometry (HPLC-ICPMS/ESIMSMS). In addition to inorganic arsenic (iAs), the aqueous layer also contained monomethylarsonate (MAs(V)), dimethylarsinate (DMAs(V)), and the two arsenosugars, namely a glycerol arsenosugar (Oxo-Gly) and a phosphate arsenosugar (Oxo-PO4). Two major arsenosugar phospholipids (AsSugPL982 and AsSugPL984) were detected in DCM fraction. Arsenic in the growth medium was also investigated by HPLC/ICPMS and shown to be present mainly as the inorganic forms As(III) and As(V) accounting for 29%-38% and 29%-57% of the total arsenic respectively. The total arsenic of methylated arsenic, arsenosugars, and arsenosugar phospholipids in Nostoc cells with increasing As(III) exposure were not markedly different, indicating that the transformation to organoarsenic in Nostoc was not dependent on As(III) concentration in the medium. Our results provide new insights into the role of cyanobacteria in the biogeochemical cycling of arsenic. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Impacts of diurnal variation of ultraviolet-B and photosynthetically active radiation on phycobiliproteins of the hot-spring cyanobacterium Nostoc sp. strain HKAR-2.

    Science.gov (United States)

    Kannaujiya, Vinod K; Sinha, Rajeshwar P

    2017-01-01

    The effects of diurnal variation of photosynthetically active radiation (PAR; 400-700 nm) and ultraviolet-B (UV-B; 280-315 nm) radiation on phycobiliproteins (PBPs) and photosynthetic pigments (PP) have been studied in the hot-spring cyanobacterium Nostoc sp. strain HKAR-2. The variations in PBPs and PP were monitored by alternating light and dark under PAR, UV-B, and PAR + UV-B radiations over a period of 25 h. There was a decline in the amount of Chl a and PBPs during light periods of UV-B and PAR + UV-B and an increase during dark periods showing a circadian rhythm by destruction and resynthesis of pigment-protein complex. However, a marked induction in carotenoids was recorded during light periods of the same radiations. Moreover, the ratio of Chl a/PE and Chl a/PC was increased in dark periods showing the resynthesis of bleached Chl a. The wavelength shift in emission fluorescence of PBPs toward shorter wavelengths further indicated the bleaching and destruction of PBPs during light periods. Oxidative damage upon exposure to PAR, UV-B, and PAR + UV-B was alleviated by induction of antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). The studied cyanobacterium exhibits a significant increase in the activities of SOD, CAT, and APX upon exposure to UV-B and PAR + UV-B radiations. The results indicate that pigment-protein composition of Nostoc sp. stain HKAR-2 was significantly altered during diurnal variation of light/radiation, which might play an important role in optimization for their productivity in a particular cyanobacterium.

  13. Photosynthetic versatility in the genome of Geitlerinema sp. PCC 9228 (formerly Oscillatoria limnetica ‘Solar Lake’, a model anoxygenic photosynthetic cyanobacterium

    Directory of Open Access Journals (Sweden)

    Sharon L Grim

    2016-10-01

    Full Text Available Anoxygenic cyanobacteria that use sulfide as the electron donor for photosynthesis are a potentially influential but poorly constrained force on Earth’s biogeochemistry. Their versatile metabolism may have boosted primary production and nitrogen cycling in euxinic coastal margins in the Proterozoic. In addition, they represent a biological mechanism for limiting the accumulation of atmospheric oxygen, especially before the Great Oxidation Event and in the low-oxygen conditions of the Proterozoic. In this study, we describe the draft genome sequence of Geitlerinema sp. PCC 9228, formerly Oscillatoria limnetica ‘Solar Lake’, a mat-forming diazotrophic cyanobacterium that can switch between oxygenic photosynthesis and sulfide-based anoxygenic photosynthesis. Geitlerinema possesses three variants of psbA, which encodes protein D1, a core component of the photosystem II reaction center. Phylogenetic analyses indicate that one variant is closely affiliated with cyanobacterial psbA genes that code for a D1 protein used for oxygen-sensitive processes. Another version is phylogenetically similar to cyanobacterial psbA genes that encode D1 proteins used under microaerobic conditions, and the third variant may be cued to high light and/or elevated oxygen concentrations. Geitlerinema has the canonical gene for sulfide quinone reductase (SQR used in cyanobacterial anoxygenic photosynthesis and a putative transcriptional regulatory gene in the same operon. Another operon with a second, distinct sqr and regulatory gene is present, and is phylogenetically related to sqr genes used for high sulfide concentrations. The genome has a comprehensive nif gene suite for nitrogen fixation, supporting previous observations of nitrogenase activity. Geitlerinema possesses a bidirectional hydrogenase rather than the uptake hydrogenase typically used by cyanobacteria in diazotrophy. Overall, the genome sequence of Geitlerinema sp. PCC 9228 highlights potential

  14. Photosynthetic Versatility in the Genome of Geitlerinema sp. PCC 9228 (Formerly Oscillatoria limnetica ‘Solar Lake’), a Model Anoxygenic Photosynthetic Cyanobacterium

    Science.gov (United States)

    Grim, Sharon L.; Dick, Gregory J.

    2016-01-01

    Anoxygenic cyanobacteria that use sulfide as the electron donor for photosynthesis are a potentially influential but poorly constrained force on Earth’s biogeochemistry. Their versatile metabolism may have boosted primary production and nitrogen cycling in euxinic coastal margins in the Proterozoic. In addition, they represent a biological mechanism for limiting the accumulation of atmospheric oxygen, especially before the Great Oxidation Event and in the low-oxygen conditions of the Proterozoic. In this study, we describe the draft genome sequence of Geitlerinema sp. PCC 9228, formerly Oscillatoria limnetica ‘Solar Lake’, a mat-forming diazotrophic cyanobacterium that can switch between oxygenic photosynthesis and sulfide-based anoxygenic photosynthesis (AP). Geitlerinema possesses three variants of psbA, which encodes protein D1, a core component of the photosystem II reaction center. Phylogenetic analyses indicate that one variant is closely affiliated with cyanobacterial psbA genes that code for a D1 protein used for oxygen-sensitive processes. Another version is phylogenetically similar to cyanobacterial psbA genes that encode D1 proteins used under microaerobic conditions, and the third variant may be cued to high light and/or elevated oxygen concentrations. Geitlerinema has the canonical gene for sulfide quinone reductase (SQR) used in cyanobacterial AP and a putative transcriptional regulatory gene in the same operon. Another operon with a second, distinct sqr and regulatory gene is present, and is phylogenetically related to sqr genes used for high sulfide concentrations. The genome has a comprehensive nif gene suite for nitrogen fixation, supporting previous observations of nitrogenase activity. Geitlerinema possesses a bidirectional hydrogenase rather than the uptake hydrogenase typically used by cyanobacteria in diazotrophy. Overall, the genome sequence of Geitlerinema sp. PCC 9228 highlights potential cyanobacterial strategies to cope with

  15. Biofilm comprising phototrophic, diazotrophic, and hydrocarbon-utilizing bacteria: a promising consortium in the bioremediation of aquatic hydrocarbon pollutants.

    Science.gov (United States)

    Al-Bader, Dhia; Kansour, Mayada K; Rayan, Rehab; Radwan, Samir S

    2013-05-01

    Biofilms harboring simultaneously anoxygenic and oxygenic phototrophic bacteria, diazotrophic bacteria, and hydrocarbon-utilizing bacteria were established on glass slides suspended in pristine and oily seawater. Via denaturing gradient gel electrophoresis analysis on PCR-amplified rRNA gene sequence fragments from the extracted DNA from biofilms, followed by band amplification, biofilm composition was determined. The biofilms contained anoxygenic phototrophs belonging to alphaproteobacteria; pico- and filamentous cyanobacteria (oxygenic phototrophs); two species of the diazotroph Azospirillum; and two hydrocarbon-utilizing gammaproteobacterial genera, Cycloclasticus and Oleibacter. The coexistence of all these microbial taxa with different physiologies in the biofilm makes the whole community nutritionally self-sufficient and adequately aerated, a condition quite suitable for the microbial biodegradation of aquatic pollutant hydrocarbons.

  16. Phosphoproteome of the cyanobacterium Synechocystis sp. PCC 6803 and its dynamics during nitrogen starvation.

    Directory of Open Access Journals (Sweden)

    Philipp eSpät

    2015-03-01

    Full Text Available Cyanobacteria have shaped the earth’s biosphere as the first oxygenic photoautotrophs and still play an important role in many ecosystems. The ability to adapt to changing environmental conditions is an essential characteristic in order to ensure survival. To this end, numerous studies have shown that bacteria use protein post-translational modifications such as Ser/Thr/Tyr phosphorylation in cell signalling, adaptation and regulation. Nevertheless, our knowledge of cyanobacterial phosphoproteomes and their dynamic response to environmental stimuli is relatively limited. In this study, we applied gel-free methods and high accuracy mass spectrometry towards the unbiased detection of Ser/Thr/Tyr phosphorylation events in the model cyanobacterium Synechocystis sp. PCC 6803. We could identify over 300 phosphorylation events in cultures grown on nitrate as exclusive nitrogen source. Chemical dimethylation labelling was applied to investigate proteome and phosphoproteome dynamics during nitrogen starvation. Our dataset describes the most comprehensive (phosphoproteome of Synechocystis to date, identifying 2,382 proteins and 183 phosphorylation events and quantifying 2,111 proteins and 148 phosphorylation events during nitrogen starvation. Global protein phosphorylation levels were increased in response to nitrogen depletion after 24 hours. Among the proteins with increased phosphorylation, the PII signalling protein showed the highest fold-change, serving as positive control. Other proteins with increased phosphorylation levels comprised functions in photosynthesis and in carbon and nitrogen metabolism. This study reveals dynamics of Synechocystis phosphoproteome in response to environmental stimuli and suggests an important role of protein Ser/Thr/Tyr phosphorylation in fundamental mechanisms of homeostatic control in cyanobacteria.

  17. Hydrogen sulfide can inhibit and enhance oxygenic photosynthesis in a cyanobacterium from sulfidic springs

    NARCIS (Netherlands)

    Klatt, Judith M.; Haas, Sebastian; Yilmaz, Pelin; de Beer, Dirk; Polerecky, Lubos

    We used microsensors to investigate the combinatory effect of hydrogen sulfide (H2S) and light on oxygenic photosynthesis in biofilms formed by a cyanobacterium from sulfidic springs. We found that photosynthesis was both positively and negatively affected by H2S: (i) H2S accelerated the recovery of

  18. Genome Sequence of the Thermophilic Cyanobacterium Thermosynechococcus sp. Strain NK55a.

    Energy Technology Data Exchange (ETDEWEB)

    Stolyar, Sergey; Liu, Zhenfeng; Thiel, Vera; Tomsho, Lynn P.; Pinel, Nicolas; Nelson, William C.; Lindemann, Stephen R.; Romine, Margaret F.; Haruta, Shin; Schuster, Stephan C.; Bryant, Donald A.; Fredrickson, Jim K.

    2014-01-02

    The genome of the unicellular cyanobacterium, Thermosynechococcus sp. strain NK55a, isolated from Nakabusa hot spring, comprises a single, circular, 2.5-Mb chromosome. The genome is predicted to encode 2358 protein coding genes, including genes for all typical cyanobacterial photosynthetic and metabolic functions. No genes encoding hydrogenases or nitrogenase were identified.

  19. Sterol Compositions of the Filamentous Nitrogen-Fixing Terrestrial Cyanobacterium Scytonema sp

    Czech Academy of Sciences Publication Activity Database

    Řezanka, Tomáš; Dembitsky, V. M.; Go, J. V.; Dor, I.; Prell, Aleš; Hanuš, L.

    2003-01-01

    Roč. 48, č. 3 (2003), s. 357-360 ISSN 0015-5632 Institutional research plan: CEZ:AV0Z5020903 Keywords : nitrogen-fixing * cyanobacterium * scytonema Subject RIV: EE - Microbiology, Virology Impact factor: 0.857, year: 2003

  20. Highly plastic genome of Microcystis aeruginosa PCC 7806, a ubiquitous toxic freshwater cyanobacterium

    NARCIS (Netherlands)

    Frangeul, L.; Quillardet, P.; Castets, A.M.; Humbert, J.F.; Matthijs, H.C.P.; Cortez, D.; Tolonen, A.; Zhang, C.C.; Gribaldo, S.; Kehr, J.C.; Zilliges, Y.; Ziemert, N.; Becker, S.; Talla, E.; Latifi, A.; Billault, A.; Lepelletier, A.; Dittmann, E.; Bouchier, C.; Tandeau de Marsac, N.

    2008-01-01

    Background The colonial cyanobacterium Microcystis proliferates in a wide range of freshwater ecosystems and is exposed to changing environmental factors during its life cycle. Microcystis blooms are often toxic, potentially fatal to animals and humans, and may cause environmental problems. There

  1. Nostoc PCC7524, a cyanobacterium which contains five sequence-specific deoxyribonucleases

    NARCIS (Netherlands)

    Reaston, J.; Duybesteyn, M.G.C.; Waard, Adrian de

    1982-01-01

    Five nucleotide sequence-specific deoxyribonucleases present in cell-free extracts of the filamentous cyanobacterium Nostoc PCC7524 have been purified and characterized. One of these enzymes, designated Nsp(7524)I cleaves at a new kind of nucleotide sequence i.e. 5'-PuCATG λ Py-3'. The other four

  2. Horizontal transfer of the nitrogen fixation gene cluster in the cyanobacterium Microcoleus chthonoplastes

    NARCIS (Netherlands)

    Bolhuis, H.; Severin, I.; Confurius-Guns, V.; Wollenzien, U.I.A.; Stal, L.J.

    2010-01-01

    The filamentous, non-heterocystous cyanobacterium Microcoleus chthonoplastes is a cosmopolitan organism, known to build microbial mats in a variety of different environments. Although most of these cyanobacterial mats are known for their capacity to fix dinitrogen, M. chthonoplastes has not been

  3. Competition and facilitation between the marine nitrogen-fixing cyanobacterium Cyanothece and its associated bacterial community

    NARCIS (Netherlands)

    Brauer, Verena S; Stomp, Maayke; Bouvier, Thierry; Fouilland, Eric; Leboulanger, Christophe; Confurius-Guns, Veronique; Weissing, Franz J; Stal, Lucas J; Huisman, Jef

    2015-01-01

    N2-fixing cyanobacteria represent a major source of new nitrogen and carbon for marine microbial communities, but little is known about their ecological interactions with associated microbiota. In this study we investigated the interactions between the unicellular N2-fixing cyanobacterium Cyanothece

  4. Effect of Selected Plant Extracts and D- and L-Lysine on the Cyanobacterium Microcystis aeruginosa

    NARCIS (Netherlands)

    Lurling, M.F.L.L.W.; Oosterhout, J.F.X.

    2014-01-01

    We tested extracts from Fructus mume, Salvia miltiorrhiza and Moringa oleifera as well as L-lysine and D-Lysine as curative measures to rapidly suppress the cyanobacterium Microcystis aeruginosa NIVA-CYA 43. We tested these compounds under similar conditions to facilitate comparisons. We

  5. Comparative study of endophytic and endophytic diazotrophic bacterial communities across rice landraces grown in the highlands of northern Thailand

    OpenAIRE

    Rangjaroen, C.; Rerkasem, B.; Teaumroong, N.; Sungthong, R.; Lumyong, S.

    2014-01-01

    Communities of bacterial endophytes within the rice landraces cultivated in the highlands of northern Thailand were studied using fingerprinting data of 16S rRNA and nifH genes profiling by polymerase chain reaction-denaturing gradient gel electrophoresis. The bacterial communities' richness, diversity index, evenness, and stability were varied depending on the plant tissues, stages of growth, and rice cultivars. These indices for the endophytic diazotrophic bacteria within the landrace rice ...

  6. Screening and selection of most potent diazotrophic cyanobacterial isolate exhibiting natural tolerance to rice field herbicides for exploitation as biofertilizer.

    Science.gov (United States)

    Singh, Surendra; Datta, Pallavi

    2006-01-01

    Periodic applications of heavy dosages of herbicides in modern rice-agriculture are a necessary evil for obtaining high crop productivity. Such herbicides are not only detrimental to weeds but biofertilizer strains of diazotrophic cyanobacteria also. It is therefore, essential to screen and select such biofertilizer strains of diazotrophic cyanobacteria exhibiting natural tolerance to common rice-field herbicides that can be further improved by mutational techniques to make biofertilizer technology a viable one. Therefore, efforts have been made to screen five dominant diazotrophic cyanobacterial forms e.g. filamentous heterocystous Nostoc punctiforme , Nostoc calcicola , Anabaena variabilis and unicellular Gloeocapsa sp. and Aphanocapsa sp. along with standard laboratory strain Nostoc muscorum ISU against increasing concentrations (0-100 mg l(-1) of four commercial grade common rice-field herbicides i.e. Arozin, Butachlor, Alachlor and 2,4-D under diazotrophic growth conditions. The lethal and IGC(50) concentrations for all four herbicides tested were found highest for A. variabilis as compared to other test cyanobacteria. The lowest reduction in chlorophyll a content, photosynthetic oxygen evolution, and N(2)-fixation was found in A. variabilis as compared to other rice field isolates and standard laboratory strain N. muscorum ISU. On the basis of prolong survival potential and lowest reductions in vital metabolic activities tested at IGC(50) concentration of four herbicides, it is concluded that A. variabilis is the most potent and promising cyanobacterial isolate as compared with other forms. This could be further improved by mutational techniques for exploitation as most potential and viable biofertilizer strain.

  7. Elevated CO2 enhances nitrogen fixation and growth in the marine cyanobacterium Trichodesmium

    Czech Academy of Sciences Publication Activity Database

    Levitan, O.; Rosenberg, G.; Šetlík, Ivan; Šetlíková, Eva; Grígel, Juraj; Klepetář, Jiří; Prášil, Ondřej; Berman-Frank, I.

    2007-01-01

    Roč. 13, č. 2 (2007), s. 531-538 ISSN 1354-1013 R&D Projects: GA MŠk 1P05ME824; GA ČR GA206/05/0335; GA MŽP SL/1/6/04 Institutional research plan: CEZ:AV0Z50200510 Keywords : ccm * cyanobacteria * diazotrophs Subject RIV: EE - Microbiology, Virology Impact factor: 4.786, year: 2007

  8. Characterization of plant-growth promoting diazotrophic bacteria isolated from field grown Chinese cabbage under different fertilization conditions.

    Science.gov (United States)

    Yim, Woo-Jong; Poonguzhali, Selvaraj; Madhaiyan, Munusamy; Palaniappan, Pitchai; Siddikee, M A; Sa, Tongmin

    2009-04-01

    Diazotrophic bacteria isolated from the rhizosphere of Chinese cabbage were assessed for other plant growth promoting characteristics viz., production of IAA, ethylene, ACC deaminase, phosphate solubilization, and gnotobiotic root elongation. Their effect on inoculation to Chinese cabbage was also observed under growth chamber conditions. A total of 19 strains that showed higher nitrogenase activity identified by 16S rRNA gene sequence analysis were found to be the members of the genera Pseudomonas and Agrobacterium belonging to alpha- and gamma-Proteobacteria groups. These strains were also efficient in producing IAA and ACC deaminase though they produced low levels of ethylene and no phosphate solubilization. In addition, inoculation of selected diazotrophic bacterial strains significantly increased seedling length, dry weight, and total nitrogen when compared to uninoculated control. The colonization of crop plants by diazotrophic bacteria can be affected by many biotic and abiotic factors, and further studies are oriented towards investigating the factors that could influence the establishment of a selected bacterial community.

  9. Arsenic Demethylation by a C·As Lyase in Cyanobacterium Nostoc sp. PCC 7120.

    Science.gov (United States)

    Yan, Yu; Ye, Jun; Xue, Xi-Mei; Zhu, Yong-Guan

    2015-12-15

    Arsenic, a ubiquitous toxic substance, exists mainly as inorganic forms in the environment. It is perceived that organoarsenicals can be demethylated and degraded into inorganic arsenic by microorganisms. Few studies have focused on the mechanism of arsenic demethylation in bacteria. Here, we investigated arsenic demethylation in a typical freshwater cyanobacterium Nostoc sp. PCC 7120. This bacterium was able to demethylate monomethylarsenite [MAs(III)] rapidly to arsenite [As(III)] and also had the ability to demethylate monomethylarsenate [MAs(V)] to As(III). The NsarsI encoding a C·As lyase responsible for MAs(III) demethylation was cloned from Nostoc sp. PCC 7120 and heterologously expressed in an As-hypersensitive strain Escherichia coli AW3110 (ΔarsRBC). Expression of NsarsI was shown to confer MAs(III) resistance through arsenic demethylation. The purified NsArsI was further identified and functionally characterized in vitro. NsArsI existed mainly as the trimeric state, and the kinetic data were well-fit to the Hill equation with K0.5 = 7.55 ± 0.33 μM for MAs(III), Vmax = 0.79 ± 0.02 μM min(-1), and h = 2.7. Both of the NsArsI truncated derivatives lacking the C-terminal 10 residues (ArsI10) or 23 residues (ArsI23) had a reduced ability of MAs(III) demethylation. These results provide new insights for understanding the important role of cyanobacteria in arsenic biogeochemical cycling in the environment.

  10. Enhanced biohydrogen production by the N{sub 2}-fixing cyanobacterium Anabaena siamensis strain TISTR 8012

    Energy Technology Data Exchange (ETDEWEB)

    Khetkorn, Wanthanee [Program of Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330 (Thailand); Laboratory of Cyanobacterial Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phayathai Road, Bangkok, 10330 (Thailand); Department of Photochemistry and Molecular Science, Uppsala University, Box 523, SE-75120, Uppsala (Sweden); Lindblad, Peter [Department of Photochemistry and Molecular Science, Uppsala University, Box 523, SE-75120, Uppsala (Sweden); Incharoensakdi, Aran [Laboratory of Cyanobacterial Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phayathai Road, Bangkok, 10330 (Thailand)

    2010-12-15

    The efficiency of hydrogen production depends on several factors. We focused on external conditions leading to enhanced hydrogen production when using the N{sub 2}-fixing cyanobacterium Anabaena siamensis TISTR 8012, a novel strain isolated from a rice paddy field in Thailand. In this study, we controlled key factors affecting hydrogen production such as cell age, light intensity, time of light incubation and source of carbon. Our results showed an enhanced hydrogen production when cells, at log phase, were adapted under N{sub 2}-fixing condition using 0.5% fructose as carbon source and a continuous illumination of 200 {mu}E m{sup -2} s{sup -1} for 12 h under anaerobic incubation. The maximum hydrogen production rate was 32 {mu}mol H{sub 2} mg chl a{sup -1} h{sup -1}. This rate was higher than that observed in the model organisms Anabaena PCC 7120, Nostoc punctiforme ATCC 29133 and Synechocystis PCC 6803. This higher production was likely caused by a higher nitrogenase activity since we observed an upregulation of nifD. The production did not increase after 12 h which was probably due to an increased activity of the uptake hydrogenase as evidenced by an increased hupL transcript level. Interestingly, a proper adjustment of light conditions such as intensity and duration is important to minimize both the photodamage of the cells and the uptake hydrogenase activity. Our results indicate that A. siamensis TISTR 8012 has a high potential for hydrogen production with the ability to utilize sugars as substrate to produce hydrogen. (author)

  11. Composition and functional property of photosynthetic pigments under circadian rhythm in the cyanobacterium Spirulina platensis.

    Science.gov (United States)

    Kumar, Deepak; Kannaujiya, Vinod K; Richa; Pathak, Jainendra; Sundaram, Shanthy; Sinha, Rajeshwar P

    2018-05-01

    Circadian rhythm is an important endogenous biological signal for sustainable growth and development of cyanobacteria in natural ecosystems. Circadian effects of photosynthetically active radiation (PAR), ultraviolet-A (UV-A) and ultraviolet-B (UV-B) radiations on pigment composition have been studied in the cyanobacterium Spirulina platensis under light (L)/dark (D) oscillation with a combination of 4/20, 8/16, 12/12, 16/8, 20/4 and 24/24 h time duration. Circadian exposure of PAR + UV-A (PA) and PAR + UV-A + UV-B (PAB) showed more than twofold decline in Chl a, total protein and phycocyanin (PC) in light phase and significant recovery was achieved in dark phase. The fluorescence emission wavelength of PC was shifted towards lower wavelengths in the light phase of PAB in comparison to P and PA whereas the same wavelength was retrieved in the dark phase. The production of free radicals was accelerated twofold in the light phase (24 h L) whereas the same was retrieved to the level of control during the dark phase. Oxidatively induced damage was alleviated by antioxidative enzymes such as catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and ascorbate peroxidase (APX) in the light phase (0-24-h L) whereas the dark phase showed significant inhibition of the same enzymes. Similar characteristic inhibition of free radicals and recovery of PC was observed inside cellular filament after circadian rhythm of 24/24 h (L/D). Circadian exposure of P, PA and PAB significantly altered the synthesis and recovery of pigments that could be crucial for optimization and sustainable production of photosynthetic products for human welfare.

  12. Dependence of the cyanobacterium Prochlorococcus on hydrogen peroxide scavenging microbes for growth at the ocean's surface.

    Directory of Open Access Journals (Sweden)

    J Jeffrey Morris

    2011-02-01

    Full Text Available The phytoplankton community in the oligotrophic open ocean is numerically dominated by the cyanobacterium Prochlorococcus, accounting for approximately half of all photosynthesis. In the illuminated euphotic zone where Prochlorococcus grows, reactive oxygen species are continuously generated via photochemical reactions with dissolved organic matter. However, Prochlorococcus genomes lack catalase and additional protective mechanisms common in other aerobes, and this genus is highly susceptible to oxidative damage from hydrogen peroxide (HOOH. In this study we showed that the extant microbial community plays a vital, previously unrecognized role in cross-protecting Prochlorococcus from oxidative damage in the surface mixed layer of the oligotrophic ocean. Microbes are the primary HOOH sink in marine systems, and in the absence of the microbial community, surface waters in the Atlantic and Pacific Ocean accumulated HOOH to concentrations that were lethal for Prochlorococcus cultures. In laboratory experiments with the marine heterotroph Alteromonas sp., serving as a proxy for the natural community of HOOH-degrading microbes, bacterial depletion of HOOH from the extracellular milieu prevented oxidative damage to the cell envelope and photosystems of co-cultured Prochlorococcus, and facilitated the growth of Prochlorococcus at ecologically-relevant cell concentrations. Curiously, the more recently evolved lineages of Prochlorococcus that exploit the surface mixed layer niche were also the most sensitive to HOOH. The genomic streamlining of these evolved lineages during adaptation to the high-light exposed upper euphotic zone thus appears to be coincident with an acquired dependency on the extant HOOH-consuming community. These results underscore the importance of (indirect biotic interactions in establishing niche boundaries, and highlight the impacts that community-level responses to stress may have in the ecological and evolutionary outcomes for co

  13. Associative diazotrophic bacteria in grass roots and soils from heavy metal contaminated sites

    Directory of Open Access Journals (Sweden)

    Fátima M.S. Moreira

    2008-12-01

    Full Text Available This work aimed to evaluate density of associative diazotrophic bacteria populations in soil and grass root samples from heavy metal contaminated sites, and to characterize isolates from these populations, both, phenotypically (Zinc, Cadmium and NaCl tolerance in vitro, and protein profiles and genotypically (16S rDNA sequencing, as compared to type strains of known diazotrophic species. Densities were evaluated by using NFb, Fam and JNFb media, commonly used for enrichment cultures of diazotrophic bacteria. Bacterial densities found in soil and grass root samples from contaminated sites were similar to those reported for agricultural soils. Azospirillum spp. isolates from contaminated sites and type strains from non-contaminated sites varied substantially in their in vitro tolerance to Zn+2 and Cd+2, being Cd+2 more toxic than Zn+2. Among the most tolerant isolates (UFLA 1S, 1R, S181, S34 and S22, some (1R, S34 and S22 were more tolerant to heavy metals than rhizobia from tropical and temperate soils. The majority of the isolates tolerant to heavy metals were also tolerant to salt stress as indicated by their ability to grow in solid medium supplemented with 30 g L-1 NaCl. Five isolates exhibited high dissimilarity in protein profiles, and the 16S rDNA sequence analysis of two of them revealed new sequences for Azospirillum.Objetivou-se avaliar a densidade de populações de bactérias diazotróficas associativas em amostras de solos e de raízes de gramíneas oriundas de sítios contaminados com metais pesados, e caracterizar isolados destas populações através da análise fenotípica (tolerância aos metais pesados zinco e cádmio e à NaCl in vitro, perfis protéicos, e genotípica (seqüenciamento de 16S rDNA, comparados às estirpes tipo das mesmas espécies. As densidades foram avaliadas nos meios NFb, Fam e LGI, comumente utilizados para culturas de enriquecimento de populações de bactérias diazotróficas associativas. As densidades

  14. Dried Colony in Cyanobacterium, Nostoc sp. HK-01 — Several high Space Environment Tolerances for ``Tanpopo'' Mission

    Science.gov (United States)

    Tomita-Yokotani, K.; Kimura, S.; Kimura, Y.; Igarashi, Y.; Ajioka, R.; Sato, S.; Katoh, H.; Baba, K.

    2013-11-01

    A cyanobacterium, Nostoc sp. HK-01, has high several space environmental tolerance. Nostoc sp HK-01 would have high contribution for the “Tanpopo” mission in Japan Experimental Module of the International Space Station.

  15. Genome Sequence and Composition of a Tolyporphin-Producing Cyanobacterium-Microbial Community

    Energy Technology Data Exchange (ETDEWEB)

    Hughes, Rebecca-Ayme; Zhang, Yunlong; Zhang, Ran; Williams, Philip G.; Lindsey, Jonathan S.; Miller, Eric S.; Nojiri, Hideaki

    2017-07-28

    domains for a glycosyltransferase, two cytochrome P450 enzymes, and a flavin adenine dinucleotide (FAD)-binding protein. The composition of the gene cluster suggests a possible role in tolyporphin biosynthesis.

    IMPORTANCEA worldwide search more than 25 years ago for cyanobacterial natural products with anticancer activity identified a culture (HT-58-2) from Micronesia that produces tolyporphins. Tolyporphins are tetrapyrroles, like chlorophylls, but have several profound structural differences that reside outside the bounds of known biosynthetic pathways. To begin probing the biosynthetic origin and biological function of tolyporphins, our research has focused on studying the cyanobacterial strain, about which almost nothing has been previously reported. We find that the HT-58-2 culture is composed of the cyanobacterium and a community of associated bacteria, complicating the question of which organisms make tolyporphins. Elucidation of the cyanobacterial genome revealed an intriguing gene cluster that contains tetrapyrrole biosynthesis genes and a collection of unknown genes, suggesting that the cluster may be responsible for tolyporphin production. Knowledge of the genome and the gene cluster sharply focuses research to identify related cyanobacterial producers of tolyporphins and delineate the tolyporphin biosynthetic pathway.

  16. A high constitutive catalase activity confers resistance to methyl viologen-promoted oxidative stress in a mutant of the cyanobacterium Nostoc punctiforme ATCC 29133.

    Science.gov (United States)

    Moirangthem, Lakshmipyari Devi; Bhattacharya, Sudeshna; Stensjö, Karin; Lindblad, Peter; Bhattacharya, Jyotirmoy

    2014-04-01

    A spontaneous methyl viologen (MV)-resistant mutant of the nitrogen-fixing cyanobacterium Nostoc punctiforme ATCC 29133 was isolated and the major enzymatic antioxidants involved in combating MV-induced oxidative stress were evaluated. The mutant displayed a high constitutive catalase activity as a consequence of which, the intracellular level of reactive oxygen species in the mutant was lower than the wild type (N. punctiforme) in the presence of MV. The superoxide dismutase (SOD) activity that consisted of a SodA (manganese-SOD) and a SodB (iron-SOD) was not suppressed in the mutant following MV treatment. The mutant was, however, characterised by a lower peroxidase activity compared with its wild type, and its improved tolerance to externally added H₂O₂ could only be attributed to enhanced catalase activity. Furthermore, MV-induced toxic effects on the wild type such as (1) loss of photosynthetic performance assessed as maximal quantum yield of photosystem II, (2) nitrogenase inactivation, and (3) filament fragmentation and cell lysis were not observed in the mutant. These findings highlight the importance of catalase in preventing MV-promoted oxidative damage and cell death in the cyanobacterium N. punctiforme. Such oxidative stress resistant mutants of cyanobacteria are likely to be a better source of biofertilisers, as they can grow and fix nitrogen in an unhindered manner in agricultural fields that are often contaminated with the herbicide MV, also commonly known as paraquat.

  17. Characterization of Function of the GlgA2 Glycogen/Starch Synthase in Cyanobacterium sp. Clg1 Highlights Convergent Evolution of Glycogen Metabolism into Starch Granule Aggregation.

    Science.gov (United States)

    Kadouche, Derifa; Ducatez, Mathieu; Cenci, Ugo; Tirtiaux, Catherine; Suzuki, Eiji; Nakamura, Yasunori; Putaux, Jean-Luc; Terrasson, Amandine Durand; Diaz-Troya, Sandra; Florencio, Francisco Javier; Arias, Maria Cecilia; Striebeck, Alexander; Palcic, Monica; Ball, Steven G; Colleoni, Christophe

    2016-07-01

    At variance with the starch-accumulating plants and most of the glycogen-accumulating cyanobacteria, Cyanobacterium sp. CLg1 synthesizes both glycogen and starch. We now report the selection of a starchless mutant of this cyanobacterium that retains wild-type amounts of glycogen. Unlike other mutants of this type found in plants and cyanobacteria, this mutant proved to be selectively defective for one of the two types of glycogen/starch synthase: GlgA2. This enzyme is phylogenetically related to the previously reported SSIII/SSIV starch synthase that is thought to be involved in starch granule seeding in plants. This suggests that, in addition to the selective polysaccharide debranching demonstrated to be responsible for starch rather than glycogen synthesis, the nature and properties of the elongation enzyme define a novel determinant of starch versus glycogen accumulation. We show that the phylogenies of GlgA2 and of 16S ribosomal RNA display significant congruence. This suggests that this enzyme evolved together with cyanobacteria when they diversified over 2 billion years ago. However, cyanobacteria can be ruled out as direct progenitors of the SSIII/SSIV ancestral gene found in Archaeplastida. Hence, both cyanobacteria and plants recruited similar enzymes independently to perform analogous tasks, further emphasizing the importance of convergent evolution in the appearance of starch from a preexisting glycogen metabolism network. © 2016 American Society of Plant Biologists. All Rights Reserved.

  18. Diversity analysis of diazotrophic bacteria associated with the roots of tea (Camellia sinensis (L.) O. Kuntze).

    Science.gov (United States)

    Gulati, Arvind; Sood, Swati; Rahi, Praveen; Thakur, Rishu; Chauhan, Sunita; Chawla, Isha

    2011-06-01

    The diversity elucidation by amplified ribosomal DNA restriction analysis and 16S rDNA sequencing of 96 associative diazotrophs, isolated from the feeder roots of tea on enriched nitrogen-free semisolid media, revealed the predominance of Gram-positive over Gram-negative bacteria within the Kangra valley in Himachal Pradesh, India. The Gram-positive bacteria observed belong to two taxonomic groupings; Firmicutes, including the genera Bacillus and Paenibacillus; and Actinobacteria, represented by the genus Microbacterium. The Gram-negative bacteria included alpha-Proteobacteria genera Brevundimonas, Rhizobium, and Mesorhizobium; gamma-Proteobacteria genera Pseudomonas and Stenotrophomonas; and beta-Proteobacteria genera Azospira, Burkholderia, Delftia, Herbaspirillum and Ralstonia. The low level of similarity of two isolates, with the type strains Paenibacillus xinjiangensis and Mesorhizobium albiziae, suggests the possibility of raising species novum. The bacterial strains of different phylogenetic groups exhibited distinct carbon-source utilization patterns and fatty acid methyl ester profiles. The strains differed in their nitrogenase activities with relatively high activity seen in the Gramnegative strains exhibiting the highest similarity to Azospira oryzae, Delftia lacustris and Herbaspirillum huttiense.

  19. Association with an ammonium-excreting bacterium allows diazotrophic culture of oil-rich eukaryotic microalgae.

    Science.gov (United States)

    Ortiz-Marquez, Juan Cesar Federico; Do Nascimento, Mauro; Dublan, Maria de Los Angeles; Curatti, Leonardo

    2012-04-01

    Concerns regarding the depletion of the world's reserves of oil and global climate change have promoted an intensification of research and development toward the production of biofuels and other alternative sources of energy during the last years. There is currently much interest in developing the technology for third-generation biofuels from microalgal biomass mainly because of its potential for high yields and reduced land use changes in comparison with biofuels derived from plant feedstocks. Regardless of the nature of the feedstock, the use of fertilizers, especially nitrogen, entails a potential economic and environmental drawback for the sustainability of biofuel production. In this work, we have studied the possibility of nitrogen biofertilization by diazotrophic bacteria applied to cultured microalgae as a promising feedstock for next-generation biofuels. We have obtained an Azotobacter vinelandii mutant strain that accumulates several times more ammonium in culture medium than wild-type cells. The ammonium excreted by the mutant cells is bioavailable to promote the growth of nondiazotrophic microalgae. Moreover, this synthetic symbiosis was able to produce an oil-rich microalgal biomass using both carbon and nitrogen from the air. This work provides a proof of concept that artificial symbiosis may be considered an alternative strategy for the low-N-intensive cultivation of microalgae for the sustainable production of next-generation biofuels and other bioproducts.

  20. Endophytic colonization and in planta nitrogen fixation by a diazotrophic Serratia sp. in rice.

    Science.gov (United States)

    Sandhiya, G S; Sugitha, T C K; Balachandar, D; Kumar, K

    2005-09-01

    Nitrogen fixing endophytic Serratia sp. was isolated from rice and characterized. Re-colonization ability of Serratia sp. in the rice seedlings as endophyte was studied under laboratory condition. For detecting the re-colonization potential in the rice seedlings, Serratia sp. was marked with reporter genes (egfp and Kmr) using transposon mutagenesis. The conjugants were screened for re-colonization ability and presence of nif genes using PCR. Further, the influence of flavonoids and growth hormones on the endophytic colonization and in planta nitrogen fixation of Serratia was also investigated. The flavonoids, quercetin (3 microg/ml) and diadzein (2 microg/ml) significantly increased the re-colonization ability of the endophytic Serratia, whereas the growth hormones like IAA and NAA (5 microg/ml) reduced the endophytic colonization ability of Serratia sp. Similarly, the in planta nitrogen fixation by Serratia sp. in rice was significantly increased due to flavonoids. The inoculation of endophytic diazotrophs increased the plant biomass and biochemical constituents.

  1. Genome of Herbaspirillum seropedicae strain SmR1, a specialized diazotrophic endophyte of tropical grasses.

    Science.gov (United States)

    Pedrosa, Fábio O; Monteiro, Rose Adele; Wassem, Roseli; Cruz, Leonardo M; Ayub, Ricardo A; Colauto, Nelson B; Fernandez, Maria Aparecida; Fungaro, Maria Helena P; Grisard, Edmundo C; Hungria, Mariangela; Madeira, Humberto M F; Nodari, Rubens O; Osaku, Clarice A; Petzl-Erler, Maria Luiza; Terenzi, Hernán; Vieira, Luiz G E; Steffens, Maria Berenice R; Weiss, Vinicius A; Pereira, Luiz F P; Almeida, Marina I M; Alves, Lysangela R; Marin, Anelis; Araujo, Luiza Maria; Balsanelli, Eduardo; Baura, Valter A; Chubatsu, Leda S; Faoro, Helisson; Favetti, Augusto; Friedermann, Geraldo; Glienke, Chirlei; Karp, Susan; Kava-Cordeiro, Vanessa; Raittz, Roberto T; Ramos, Humberto J O; Ribeiro, Enilze Maria S F; Rigo, Liu Un; Rocha, Saul N; Schwab, Stefan; Silva, Anilda G; Souza, Eliel M; Tadra-Sfeir, Michelle Z; Torres, Rodrigo A; Dabul, Audrei N G; Soares, Maria Albertina M; Gasques, Luciano S; Gimenes, Ciela C T; Valle, Juliana S; Ciferri, Ricardo R; Correa, Luiz C; Murace, Norma K; Pamphile, João A; Patussi, Eliana Valéria; Prioli, Alberto J; Prioli, Sonia Maria A; Rocha, Carmem Lúcia M S C; Arantes, Olívia Márcia N; Furlaneto, Márcia Cristina; Godoy, Leandro P; Oliveira, Carlos E C; Satori, Daniele; Vilas-Boas, Laurival A; Watanabe, Maria Angélica E; Dambros, Bibiana Paula; Guerra, Miguel P; Mathioni, Sandra Marisa; Santos, Karine Louise; Steindel, Mario; Vernal, Javier; Barcellos, Fernando G; Campo, Rubens J; Chueire, Ligia Maria O; Nicolás, Marisa Fabiana; Pereira-Ferrari, Lilian; Silva, José L da Conceição; Gioppo, Nereida M R; Margarido, Vladimir P; Menck-Soares, Maria Amélia; Pinto, Fabiana Gisele S; Simão, Rita de Cássia G; Takahashi, Elizabete K; Yates, Marshall G; Souza, Emanuel M

    2011-05-01

    The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme--GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species.

  2. Genome of Herbaspirillum seropedicae strain SmR1, a specialized diazotrophic endophyte of tropical grasses.

    Directory of Open Access Journals (Sweden)

    Fábio O Pedrosa

    2011-05-01

    Full Text Available The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme--GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species.

  3. INOCULATION OF DIAZOTROPHIC BACTERIA AND NITROGEN FERTILIZATION IN TOPDRESSING IN IRRIGATED CORN

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    VANESSA ZIRONDI LONGHINI

    2016-01-01

    Full Text Available Corn is a nitrogen-intensive crop, and the use of management practices such as inoculation of the seed with diazotrophic bacteria, which can maximize crop productivity and reduce the need of nitrogen fertilizers, may result in lower production costs. The present study aimed to evaluate the effect of inoculation of corn seed with Azospirillum brasilense and controlled addition of nitrogen to topdressing on the nutrition, production components, and productivity of crop grain. The experimental design was a randomized block design, with four replications in a 2 × 5 factorial scheme. The treatments consisted of inoculation or not of corn seed with A. brasilense (at 100 mL per 25 kg of seed and five nitrogen (N levels in topdressing (0, 30, 60, 90, and 120 kg N ha-1 from urea [45% N] were applied when the corn was in the phenological growth stage V6. Foliar macronutrients, foliar chlorophyll index (FCI, production components, and yield of corn grain were valuated. Inoculation of corn seeds with A. brasilense increased plant height and grain yield. Fertilization in topdressing, with N levels up to 120 kg ha-1, linearly increased the foliar nutrients and productivity of corn cultivated in the spring/summer in the low-altitude Cerrado region of Brazil.

  4. A new UV-A/B protecting pigment in the terrestrial cyanobacterium Nostoc commune

    International Nuclear Information System (INIS)

    Scherer, S.; Chen, T.W.; Boeger, P.

    1988-01-01

    A new ultraviolet (UV)-A/B absorbing pigment with maxima at 312 and 330 nanometers from the cosmopolitan terrestrial cyanobacterium Nostoc commune is described. The pigment is found in high amounts (up to 10% of dry weight) in colonies grown under solar UV radiation but only in low concentrations in laboratory cultures illuminated by artificial light without UV. Its experimental induction by UV as well as its capacity to efficiently protect Nostoc against UV radiation is reported

  5. A new endonuclease recognizing the deoxynucleotide sequence CCNNGG from the cyanobacterium Synechocystis 6701.

    Science.gov (United States)

    Calléja, F; Tandeau de Marsac, N; Coursin, T; van Ormondt, H; de Waard, A

    1985-09-25

    A new sequence-specific endonuclease from the cyanobacterium Synechocystis species PCC 6701 has been purified and characterized. This enzyme, SecI, is unique in recognizing the nucleotide sequence: 5' -CCNNGG-3' 3' -GGNNCC-5' and cleaves it at the position indicated by the symbol. Two other restriction endonucleases, SecII and SecIII, found in this organism are isoschizomers of MspI and MstII, respectively.

  6. Detection of Bioactive Exometabolites Produced by the Filamentous Marine Cyanobacterium Geitlerinema sp.

    OpenAIRE

    Caicedo, Nelson H.; Kumirska, Jolanta; Neumann, Jennifer; Stolte, Stefan; Thöming, Jorg

    2011-01-01

    Marine cyanobacteria are noted for their ability to excrete metabolites with biotic properties. This paper focuses on such exometabolites obtained from the culture of the marine filamentous cyanobacterium Geitlerinema sp. strain, their purification and subsequent analyses. By this means the recoveries of the active compounds, a prerequisite for properly determining their concentration, are quantified here for the first time. We demonstrate a new procedure using Amberlite XAD-1180 resin in com...

  7. Dynamics of soil diazotrophic community structure, diversity, and functioning during the cropping period of cotton (Gossypium hirsutum).

    Science.gov (United States)

    Rai, Sandhya; Singh, Dileep Kumar; Annapurna, Kannepalli

    2015-01-01

    The soil sampled at different growth stages along the cropping period of cotton were analyzed using various molecular tools: restriction fragment length polymorphism (RFLP), terminal restriction length polymorphism (T-RFLP), and cloning-sequencing. The cluster analysis of the diazotrophic community structure of early sampled soil (0, 15, and 30 days) was found to be more closely related to each other than the later sampled one. Phylogenetic and diversity analysis of sequences obtained from the first (0 Day; C0) and last soil sample (180 day; C180) confirmed the data. The phylogenetic analysis revealed that C0 was having more unique sequences than C180 (presence of γ-Proteobacteria exclusively in C0). A relatively higher richness of diazotrophic community sequences was observed in C0 (S(ACE) : 30.76; S(Chao1) : 20.94) than C180 (S(ACE) : 18.00; S(Chao1) : 18.00) while the evenness component of Shannon diversity index increased from C0 (0.97) to C180 (1.15). The impact of routine agricultural activities was more evident based on diazotrophic activity (measured by acetylene reduction assay) than its structure and diversity. The nitrogenase activity of C0 (1264.85 ± 35.7 ηmol of ethylene production g(-1) dry soil h(-1) ) was statistically higher when compared to all other values (p structure/diversity and N2 fixation rates. Thus, considerable functional redundancy of nifH was concluded to be existing at the experimental site. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Single and mixed formulations of inoculants with diazotrophic bacteria, under different nitrogen rates and on the paddy rice crop

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    Paula Bianchet

    2013-12-01

    Full Text Available The use of diazotrophic bacteria as a biological input for the production of paddy rice can reduce nitrogen fertilizer applications and contribute to plant development. The use of mixed inoculants’ formulations can increase the efficiency of nitrogen fixation biological process. The objective of this study was to evaluate the effect of single and mixed formulations of inoculants with diazotrophic bacteria on the initial growth of paddy rice plants under different levels of N. The experiment was set in a greenhouse. Treatments consisted of four types of inoculation (no inoculation, inoculation with the isolated AI UDESC 27, inoculation with the isolated FE UDESC 22, and inoculation with the mixed formulation of isolated AI UDESC UDESC 27 and FE UDESC 22; and two levels of mineral nitrogen (30 and 60 mg kg-1 of N. The cultivar used was Epagri 109, which presents late maturity (over 140 days and high yield potential. Treatments were arranged in a factorial design (4 x 2 with five replicates. The experimental design was completely randomized. Inoculation with diazotrophic bacteria reduced by 18% and 26% shoot and root dry matter of rice plants, respectively. Plants also presented lower root area and volume when they were inoculated. There was no significant effect of inoculation and nitrogen rates on the number of leaves and tillers produced per plant or shoot nitrogen accumulation. The results showed that the isolated used in this work were not effective to stimulate shoot and root growth of cv Epagri 109, regardless of formulation type and rate of N.

  9. Estimation of Nitrogenase Enzyme Activities and Plant Growth of Legume and Non-legume Inoculated with Diazotrophic Bacteria

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    Salwani S.

    2012-06-01

    Full Text Available Biological Nitrogen Fixation (BNF process benefits the agriculture sector especially for reducing cost of nitrogenfertilizer. In the process, the diazotrophs convert N2 into ammonia (NH3 which is useable by plants. The BNF process iscatalysed by nitrogenase enzyme that involved protons and electrons together with evolution of H2 therefore, theassessment of N2 fixation is also available via H2 production and electron allocation analysis. Thus, the aims of thisexperiment were to estimate the nitrogenase enzyme activities and observe the influence of diazothrophs on growth oflegume (soybean and non legume (rice plants. Host plants were inoculated with respective inocula; Bradyrhizobiumjaponicum (strain 532C for soybean while Azospirillum brasilense (Sp7 and locally isolated diazotroph (isolate 5 forrice. At harvest, the plants were observed for plant growth parameters, H2 evolution, N2 fixation and electron allocationcoefficient (EAC values. The experiment recorded N2 fixation activities of inoculated soybean plants at 141.2 μmol N2 h-1g-1 dry weight nodule, and the evolution of H2 at 144.4 μmol H2 h-1 g-1 dry weight nodule. The electron allocationcoefficient (EAC of soybean was recorded at 0.982. For inoculated rice plants, none of the observations was successfully recorded. However, results for chlorophyll contents and plant dry weight of both plants inoculated with respective inocula were similar to the control treatments supplied with full nitrogen fertilization (+N. The experiment clearly showed that inoculation of diazotrophic bacteria could enhance growth of the host plants similar to plants treated with nitrogenous fertilizer due to efficient N2 fixation process

  10. Effects of the inoculation of Burkholderia vietnamensis and related endophytic diazotrophic bacteria on grain yield of rice.

    Science.gov (United States)

    Govindarajan, Munusamy; Balandreau, Jacques; Kwon, Soon-Wo; Weon, Hang-Yeon; Lakshminarasimhan, Cunthipuram

    2008-01-01

    During a survey of endophytic diazotrophic bacteria associated with different rice varieties in Tamilnadu, some "endophytes" were obtained. Thirteen bacterial isolates from surface-sterilized roots and shoots were obtained in pure culture, which produced indole acetic acid (IAA) and reduced acetylene to ethylene. Polymerase chain reaction (PCR) amplification confirmed the presence of nif-H gene in all the isolates. Morphological, biochemical, and molecular characteristics indicated that all of them belonged to the genus Burkholderia One of them, MGK3, was consistently more active in reducing acetylene, and 16S rDNA sequences of isolate MGK3 confirmed its identification as Burkholderia vietnamiensis. Colonization of rice root was confirmed by strain MGK3 marked with gusA gene. The inoculated roots showed a blue color, which was most intense at the points of lateral root emergence and at the root tip. Transverse sections of roots, 15 days after inoculation, revealed beta-glucuronidase (GUS) activity within many of the cortical intercellular spaces next to the stele and within the aerenchyma. Nitrogen fixation was quantified by using (15)N isotope dilution method with two different cultivars grown in pot and field experiments. Higher nitrogen fixation was observed in variety Ponni than in ADT-43, where nearly 42% (field) and 40% (pot) of the nitrogen was derived from the atmosphere (% Ndfa). Isolate MGK3 was used to inoculate rice seedlings in a comparison with four other diazotrophs, viz., Gluconacetobacter diazotrophicus LMG7603, Herbaspirillum seropedicae LMG6513, Azospirillum lipoferum 4B LMG4348, and B. vietnamiensis LMG10929. They were used to conduct two pot and four field inoculation experiments. MGK3 alone, and combined with other diazotrophs, performed best under both pot and field conditions: combined inoculation produced yield increases between 9.5 and 23.6%, while MGK3 alone increased yield by 5.6 to 12.16% over the uninoculated control treatment.

  11. Selection of diazotrophic bacteria isolated from wastewater treatment plant sludge at a poultry slaughterhouse for their effect on maize plants

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    Jorge Avelino Rodriguez Lozada

    Full Text Available ABSTRACT The economic and environmental costs of nitrogen fertilization have intensified the search for technologies that reduce mineral fertilization, for example atmospheric nitrogen-fixing (diazotrophic bacteria inoculation. In this context, the present study addressed the isolation and quantification of diazotrophic bacteria in the sludge from treated wastewater of a poultry slaughterhouse; a description of the bacteria, based on cell and colony morphology; and an assessment of growth and N content of maize plants in response to inoculation. Sixteen morphotypes of bacteria were isolated in six N-free culture media (JMV, JMVL, NFb, JNFb, LGI, and LGI-P. The bacteria stained gram-positive, with 10 rod- and six coccoid-shaped isolates. To evaluate the potential of bacteria to promote plant growth, maize seeds were inoculated. The experiment consisted of 17 treatments (control plus 16 bacterial isolates and was carried out in a completely randomized design with six replicates. The experimental units consisted of one pot containing two maize plants in a greenhouse. Forty-five days after planting, the variables plant height, leaf number, stem diameter, root and shoot fresh and dry weight, and N content were measured. The highest values were obtained with isolate UFV L-162, which produced 0.68 g total dry matter per plant and increased N content to 22.14 mg/plant, representing increments of 74 and 133%, respectively, compared with the control. Diazotrophs inhabit sludge from treated wastewater of poultry slaughterhouses and can potentially be used to stimulate plant development and enrich inoculants.

  12. Rates of Dinitrogen Fixation and the Abundance of Diazotrophs in North American Coastal Waters Between Cape Hatteras and Georges Bank

    Science.gov (United States)

    Mulholland, M.R.; Bernhardt, P. W.; Blanco-Garcia, J. L.; Mannino, A.; Hyde, K.; Mondragon, E.; Turk, K.; Moisander, P. H.; Zehr, J. P.

    2012-01-01

    We coupled dinitrogen (N2) fixation rate estimates with molecular biological methods to determine the activity and abundance of diazotrophs in coastal waters along the temperate North American Mid-Atlantic continental shelf during multiple seasons and cruises. Volumetric rates of N2 fixation were as high as 49.8 nmol N L(sup -1) d(sup -1) and areal rates as high as 837.9 micromol N m(sup -2) d(sup -1) in our study area. Our results suggest that N2 fixation occurs at high rates in coastal shelf waters that were previously thought to be unimportant sites of N2 fixation and so were excluded from calculations of pelagic marine N2 fixation. Unicellular N2-fixing group A cyanobacteria were the most abundant diazotrophs in the Atlantic coastal waters and their abundance was comparable to, or higher than, that measured in oceanic regimes where they were discovered. High rates of N2 fixation and the high abundance of diazotrophs along the North American Mid-Atlantic continental shelf highlight the need to revise marine N budgets to include coastal N2 fixation. Integrating areal rates of N2 fixation over the continental shelf area between Cape Hatteras and Nova Scotia, the estimated N2 fixation in this temperate shelf system is about 0.02 Tmol N yr(sup -1), the amount previously calculated for the entire North Atlantic continental shelf. Additional studies should provide spatially, temporally, and seasonally resolved rate estimates from coastal systems to better constrain N inputs via N2 fixation from the neritic zone.

  13. Effects of the Atrazine herbicide over the diazotrophic microorganisms associated with corn in a Saldana soil (Tolima)

    International Nuclear Information System (INIS)

    Ordonez, Alba R; Lozano de Yunda, A; Fuentes, C

    1999-01-01

    In this study, it was evaluated, at greenhouse in soil pots, the effect of the herbicide atrazine in different dose (corresponding to o, 0.75, 1.5 and 3.0 kg of soil hectare) over time (15, 30, 45, 60 and 90 days after the application) on the diazotrophic micro-organisms associated with rhizosphere soil and root of a corn cultivation in a clay loam soil of Saldana - Tolima. It was determined the population of diazotrophic microorganisms in soil by the most probable number technique; also it was observed the characteristic growth in the specific cultivation semisolid Nfb-malic acid medium and was evaluated the nitrogenase activity by means of the acetylene reduction technique. Of the same form, it was observed the characteristic growth and it was evaluated the nitrogenase activity of direct fragment sowings of root in the Nfb-malic acid medium. The 60.8% of soil isolates presented characteristic cultural growth in the form of whitish pellicle under the surface of the medium. in the root 83.3% showed this type of growth. The population of diazotrophic microorganisms of soil did not present significant differences (α = 0.05) between treatments neither over time the nitrogenase activity of soil dilutions demonstrated that there is no a significant effect (α = 0.05) of the treatments (dose) but if there is significant effect of the time. The highest values were presented to 45 days of application of the herbicide, with the application of the highest dose. the nitrogenase activity in the root to what is long of the time was different from rhizosphere soil. The treatment without application of atrazine presented greater values to what is long of the time, however, also in this case were not presented significant differences (α = 0.05) between the treatments (dose) neither to what is long of the time. For so much, as a conclusion, they were not presented effects on the population and the nitrogenase activity of the diazotrophic microorganisms until 90 days of after the

  14. Transcriptional patterns identify resource controls on the diazotroph Trichodesmium in the Atlantic and Pacific oceans.

    Science.gov (United States)

    Rouco, Mónica; Frischkorn, Kyle R; Haley, Sheean T; Alexander, Harriet; Dyhrman, Sonya T

    2018-02-28

    The N 2 -fixing cyanobacterium Trichodesmium is intensely studied because of the control this organism exerts over the cycling of carbon and nitrogen in the low nutrient ocean gyres. Although iron (Fe) and phosphorus (P) bioavailability are thought to be major drivers of Trichodesmium distributions and activities, identifying resource controls on Trichodesmium is challenging, as Fe and P are often organically complexed and their bioavailability to a single species in a mixed community is difficult to constrain. Further, Fe and P geochemistries are linked through the activities of metalloenzymes, such as the alkaline phosphatases (APs) PhoX and PhoA, which are used by microbes to access dissolved organic P (DOP). Here we identified significant correlations between Trichodesmium-specific transcriptional patterns in the North Atlantic (NASG) and North Pacific Subtropical Gyres (NPSG) and patterns in Fe and P biogeochemistry, with the relative enrichment of Fe stress markers in the NPSG, and P stress markers in the NASG. We also observed the differential enrichment of Fe-requiring PhoX transcripts in the NASG and Fe-insensitive PhoA transcripts in the NPSG, suggesting that metalloenzyme switching may be used to mitigate Fe limitation of DOP metabolism in Trichodesmium. This trait may underpin Trichodesmium success across disparate ecosystems.

  15. Metabolic engineering of a diazotrophic bacterium improves ammonium release and biofertilization of plants and microalgae.

    Science.gov (United States)

    Ambrosio, Rafael; Ortiz-Marquez, Juan Cesar Federico; Curatti, Leonardo

    2017-03-01

    The biological nitrogen fixation carried out by some Bacteria and Archaea is one of the most attractive alternatives to synthetic nitrogen fertilizers. However, with the exception of the symbiotic rhizobia-legumes system, progress towards a more extensive realization of this goal has been slow. In this study we manipulated the endogenous regulation of both nitrogen fixation and assimilation in the aerobic bacterium Azotobacter vinelandii. Substituting an exogenously inducible promoter for the native promoter of glutamine synthetase produced conditional lethal mutant strains unable to grow diazotrophically in the absence of the inducer. This mutant phenotype could be reverted in a double mutant strain bearing a deletion in the nifL gene that resulted in constitutive expression of nif genes and increased production of ammonium. Under GS non-inducing conditions both the single and the double mutant strains consistently released very high levels of ammonium (>20mM) into the growth medium. The double mutant strain grew and excreted high levels of ammonium under a wider range of concentrations of the inducer than the single mutant strain. Induced mutant cells could be loaded with glutamine synthetase at different levels, which resulted in different patterns of extracellular ammonium accumulation afterwards. Inoculation of the engineered bacteria into a microalgal culture in the absence of sources of C and N other than N 2 and CO 2 from the air, resulted in a strong proliferation of microalgae that was suppressed upon addition of the inducer. Both single and double mutant strains also promoted growth of cucumber plants in the absence of added N-fertilizer, while this property was only marginal in the parental strain. This study provides a simple synthetic genetic circuit that might inspire engineering of optimized inoculants that efficiently channel N 2 from the air into crops. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All

  16. DRIVERS OF THE DYNAMICS OF DIAZOTROPHS AND DENITRIFIERS IN NORTH SEA BOTTOM WATERS AND SEDIMENTS

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    Lucas eStal

    2015-07-01

    Full Text Available The fixation of dinitrogen (N2 and denitrification are two opposite processes in the nitrogen cycle. The former transfers atmospheric dinitrogen gas into bound nitrogen in the biosphere, while the latter returns this bound nitrogen back to atmospheric dinitrogen. It is unclear whether or not these processes are intimately connected in any microbial ecosystem or that they are spatially and/or temporally separated. Here, we measured seafloor nitrogen fixation and denitrification as well as pelagic nitrogen fixation by using the stable isotope technique. Alongside, we measured the diversity, abundance, and activity of nitrogen-fixing and denitrifying microorganisms at three stations in the southern North Sea. Nitrogen fixation ranged from undetectable to 2.4 nmol N L-1 d-1 and from undetectable to 8.2 nmol N g-1 d-1 in the water column and seafloor, respectively. The highest rates were measured in August at Doggersbank, both for the water column and for the seafloor. Denitrification ranged from 1.7 to 208.8 µmol m-2 d-1 and the highest rates were measured in May at the Oyster Grounds. DNA sequence analysis showed sequences of nifH, a structural gene for nitrogenase, related to sequences from anaerobic sulfur/iron reducers and sulfate reducers. Sequences of the structural gene for nitrite reductase, nirS, were related to environmental clones from marine sediments. Quantitative polymerase chain reaction (qPCR data revealed the highest abundance of nifH and nirS genes at the Oyster Grounds. Quantitative reverse transcription polymerase chain reaction (qRT-PCR data revealed the highest nifH expression at Doggersbank and the highest nirS expression at the Oyster Grounds. The distribution of the diazotrophic and denitrifying communities seems to be subject to different selecting factors, leading to spatial and temporal separation of nitrogen fixation and denitrification. These selecting factors include temperature, organic matter availability, and

  17. Seawater cultivation of freshwater cyanobacterium Synechocystis sp. PCC 6803 drastically alters amino acid composition and glycogen metabolism

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    Hiroko eIijima

    2015-04-01

    Full Text Available Water use assessment is important for bioproduction using cyanobacteria. For eco-friendly reasons, seawater should preferably be used for cyanobacteria cultivation instead of freshwater. In this study, we demonstrated that the freshwater unicellular cyanobacterium Synechocystis sp. PCC 6803 could be grown in a medium based on seawater. The Synechocystis wild-type strain grew well in an artificial seawater (ASW medium supplemented with nitrogen and phosphorus sources. The addition of HEPES buffer improved cell growth overall, although the growth in ASW medium was inferior to that in the synthetic BG-11 medium. The levels of proteins involved in sugar metabolism changed depending on the culture conditions. The biosynthesis of several amino acids including aspartate, glutamine, glycine, proline, ornithine, and lysine, was highly up-regulated by cultivation in ASW. Two types of natural seawater (NSW were also made available for the cultivation of Synechocystis cells, with supplementation of both nitrogen and phosphorus sources. These results revealed the potential use of seawater for the cultivation of freshwater cyanobacteria, which would help to reduce freshwater consumption during biorefinery using cyanobacteria.

  18. Azolla along a phosphorus gradient: biphasic growth response linked to diazotroph traits and phosphorus-induced iron chlorosis.

    Science.gov (United States)

    Temmink, Ralph J M; Harpenslager, Sarah F; Smolders, Alfons J P; van Dijk, Gijs; Peters, Roy C J H; Lamers, Leon P M; van Kempen, Monique M L

    2018-03-13

    Azolla spp., a water fern often used for phytoremediation, is a strong phosphorus (P) accumulator due to its high growth rate and N 2 fixing symbionts (diazotrophs). It is known that plant growth is stimulated by P, but the nature of the interactive response of both symbionts along a P gradient, and related changes in growth-limiting factors, are unclear. We determined growth, and N and P sequestration rates of Azolla filiculoides in N-free water at different P concentrations. The growth response appeared to be biphasic and highest at levels ≥10 P µmol l -1 . Diazotrophic N sequestration increased upon P addition, and rates were three times higher at high P than at low P. At 10 µmol P l -1 , N sequestration rates reached its maximum and A. filiculoides growth became saturated. Due to luxury consumption, P sequestration rates increased until 50 µmol P l -1 . At higher P concentrations (≥50 µmol l -1 ), however, chlorosis occurred that seems to be caused by iron- (Fe-), and not by N-deficiency. We demonstrate that traits of the complete symbiosis in relation to P and Fe availability determine plant performance, stressing the role of nutrient stoichiometry. The results are discussed regarding Azolla's potential use in a bio-based economy.

  19. Use of endophytic diazotrophic bacteria as a vector to express the cry3A gene from Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Salles Joana Falcão

    2000-01-01

    Full Text Available The goal of this study was to evaluate the potential of endophytic diazotrophic bacteria as a vector to express a cry gene from Bacillus thuringiensis, envisaging the control of pests that attack sugarcane plants. The endophytic nitrogen-fixing bacteria Gluconacetobacter diazotrophicus strain BR11281 and Herbaspirillum seropedicae strain BR11335 were used as models. The cry3A gene was transferred by conjugation using a suicide plasmid and the recombinant strains were selected by their ability to fix nitrogen in semi-solid N-free medium. The presence of the cry gene was detected by Southern-blot using an internal fragment of 1.0 kb as a probe. The production of delta-endotoxin by the recombinant H. seropedicae strain was detected by dot blot while for G. diazotrophicus the Western-blot technique was used. In both cases, a specific antibody raised against the B. thuringiensis toxin was applied. The delta-endotoxin production showed by the G. diazotrophicus recombinant strain was dependent on the nitrogen fixing conditions since the cry3A gene was fused to a nif promoter. In the case of H. seropedicae the delta-endotoxin expression was not affected by the promoter (rhi used. These results suggest that endophytic diazotrophic bacteria can be used as vectors to express entomopathogenic genes envisaging control of sugarcane pests.

  20. Macroalgal blooms favor heterotrophic diazotrophic bacteria in nitrogen-rich and phosphorus-limited coastal surface waters in the Yellow Sea

    Science.gov (United States)

    Zhang, Xiaoli; Song, Yanjing; Liu, Dongyan; Keesing, John K.; Gong, Jun

    2015-09-01

    Macroalgal blooms may lead to dramatic changes in physicochemical variables and biogeochemical cycling in affected waters. However, little is known about the effects of macroalgal blooms on marine bacteria, especially those functioning in nutrient cycles. We measured environmental factors and investigated bacterial diazotrophs in two niches, surface waters that were covered (CC) and non-covered (CF) with massive macroalgal canopies of Ulva prolifera, in the Yellow Sea in the summer of 2011 using real-time PCR and clone library analysis of nifH genes. We found that heterotrophic diazotrophs (Gammaproteobacteria) dominated the communities and were mostly represented by Vibrio-related phylotypes in both CC and CF. Desulfovibrio-related phylotypes were only detected in CC. There were significant differences in community composition in these two environments (p diazotrophic abundance and community composition and that vibrios and Desulfovibrio-related heterotrophic diazotrophs adapt well to the (N-rich but P-limited) environment during blooming. Potential ecological and microbiological mechanisms behind this scenario are discussed.

  1. Genome-derived insights into the biology of the hepatotoxic bloom-forming cyanobacterium Anabaena sp. strain 90

    Directory of Open Access Journals (Sweden)

    Wang Hao

    2012-11-01

    Full Text Available Abstract Background Cyanobacteria can form massive toxic blooms in fresh and brackish bodies of water and are frequently responsible for the poisoning of animals and pose a health risk for humans. Anabaena is a genus of filamentous diazotrophic cyanobacteria commonly implicated as a toxin producer in blooms in aquatic ecosystems throughout the world. The biology of bloom-forming cyanobacteria is poorly understood at the genome level. Results Here, we report the complete sequence and comprehensive annotation of the bloom-forming Anabaena sp. strain 90 genome. It comprises two circular chromosomes and three plasmids with a total size of 5.3 Mb, encoding a total of 4,738 genes. The genome is replete with mobile genetic elements. Detailed manual annotation demonstrated that almost 5% of the gene repertoire consists of pseudogenes. A further 5% of the genome is dedicated to the synthesis of small peptides that are the products of both ribosomal and nonribosomal biosynthetic pathways. Inactivation of the hassallidin (an antifungal cyclic peptide biosynthetic gene cluster through a deletion event and a natural mutation of the buoyancy-permitting gvpG gas vesicle gene were documented. The genome contains a large number of genes encoding restriction-modification systems. Two novel excision elements were found in the nifH gene that is required for nitrogen fixation. Conclusions Genome analysis demonstrated that this strain invests heavily in the production of bioactive compounds and restriction-modification systems. This well-annotated genome provides a platform for future studies on the ecology and biology of these important bloom-forming cyanobacteria.

  2. Visualizing Single Cell Biology: Nanosims Studies of Carbon and Nitrogen Metabolism in Diazotrophic Cyanobacteria

    Science.gov (United States)

    Pett-Ridge, J.; Finzi, J. A.; Capone, D. G.; Popa, R.; Nealson, K. H.; Ng, W.; Spormann, A. M.; Hutcheon, I. D.; Weber, P. K.

    2007-12-01

    Filamentous nitrogen fixing (diazotrophic) cyanobacteria are key players in global nutrient cycling, but the relationship between CO2- and N2-fixation and intercellular exchange of these elements remains poorly understood in many genera. These bacteria are faced with the challenge of isolating regions of N-fixation (O2 inhibited) and photosynthetic (O2 producing) activity. We used isotope labeling in conjunction with a high-resolution isotope and elemental mapping technique (NanoSIMS) to quantitatively describe 13C and 15N uptake and transport in two aquatic cyanobacteria grown on NaH13CO3 and 15N2. The technical challenges of tracing isotopes within individual bacteria can be overcome with high resolution Secondary Ion Mass Spectrometry (NanoSIMS). In NanoSIMS analysis, samples are sputtered with an energetic primary beam (Cs+, O-) liberating secondary ions that are separated by the mass spectrometer and detected in a suite of electron multipliers. Five isotopic species may be analyzed concurrently with spatial resolution as fine as 50nm. A high sensitivity isotope ratio 'map' can then be generated for the analyzed area. Using sequentially harvested cyanobacteria in conjunction with enriched H13CO3 and 15N2 incubations, we measured temporal enrichment patterns that evolve over the course of a day's growth and suggest tightly regulated changes in fixation kinetics. With a combination of TEM, SEM and NanoSIMS analyses, we also mapped the distribution of C, N and Mo (a critical nitrogenase co-factor) isotopes in intact cells. Our results suggest that NanoSIMS mapping of metal enzyme co-factors may be a powerful method of identifying physiological and morphological characteristics within individual bacterial cells, and could be used to provide a 3-dimensional context for more traditional analyses such as immunogold labeling. Finally, we resolved patterns of isotope enrichment at multiple spatial scales: sub-cellular variation, cell-cell differences along filaments

  3. Unraveling the molecular mechanisms of nitrogenase conformational protection against oxygen in diazotrophic bacteria.

    Science.gov (United States)

    Lery, Letícia M S; Bitar, Mainá; Costa, Mauricio G S; Rössle, Shaila C S; Bisch, Paulo M

    2010-12-22

    represents a large step towards the understanding of the conformational protection mechanism of nitrogenase against oxygen. In addition, this is the first study regarding the structural complementarities of FeSII-nitrogenase interactions in diazotrophic bacteria. The combination of bioinformatic tools for genome analysis, comparative protein modeling, docking calculations and molecular dynamics provided a powerful strategy for the elucidation of molecular mechanisms and structural features of FeSII-nitrogenase interaction.

  4. Genome erosion in a nitrogen-fixing vertically transmitted endosymbiotic multicellular cyanobacterium.

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    Liang Ran

    Full Text Available BACKGROUND: An ancient cyanobacterial incorporation into a eukaryotic organism led to the evolution of plastids (chloroplasts and subsequently to the origin of the plant kingdom. The underlying mechanism and the identities of the partners in this monophyletic event remain elusive. METHODOLOGY/PRINCIPAL FINDINGS: To shed light on this evolutionary process, we sequenced the genome of a cyanobacterium residing extracellularly in an endosymbiosis with a plant, the water-fern Azolla filiculoides Lam. This symbiosis was selected as it has characters which make it unique among extant cyanobacterial plant symbioses: the cyanobacterium lacks autonomous growth and is vertically transmitted between plant generations. Our results reveal features of evolutionary significance. The genome is in an eroding state, evidenced by a large proportion of pseudogenes (31.2% and a high frequency of transposable elements (approximately 600 scattered throughout the genome. Pseudogenization is found in genes such as the replication initiator dnaA and DNA repair genes, considered essential to free-living cyanobacteria. For some functional categories of genes pseudogenes are more prevalent than functional genes. Loss of function is apparent even within the 'core' gene categories of bacteria, such as genes involved in glycolysis and nutrient uptake. In contrast, serving as a critical source of nitrogen for the host, genes related to metabolic processes such as cell differentiation and nitrogen-fixation are well preserved. CONCLUSIONS/SIGNIFICANCE: This is the first finding of genome degradation in a plant symbiont and phenotypically complex cyanobacterium and one of only a few extracellular endosymbionts described showing signs of reductive genome evolution. Our findings suggest an ongoing selective streamlining of this cyanobacterial genome which has resulted in an organism devoted to nitrogen fixation and devoid of autonomous growth. The cyanobacterial symbiont of Azolla

  5. A Key Marine Diazotroph in a Changing Ocean: The Interacting Effects of Temperature, CO2 and Light on the Growth of Trichodesmium erythraeum IMS101.

    Directory of Open Access Journals (Sweden)

    Tobias G Boatman

    Full Text Available Trichodesmium is a globally important marine diazotroph that accounts for approximately 60 - 80% of marine biological N2 fixation and as such plays a key role in marine N and C cycles. We undertook a comprehensive assessment of how the growth rate of Trichodesmium erythraeum IMS101 was directly affected by the combined interactions of temperature, pCO2 and light intensity. Our key findings were: low pCO2 affected the lower temperature tolerance limit (Tmin but had no effect on the optimum temperature (Topt at which growth was maximal or the maximum temperature tolerance limit (Tmax; low pCO2 had a greater effect on the thermal niche width than low-light; the effect of pCO2 on growth rate was more pronounced at suboptimal temperatures than at supraoptimal temperatures; temperature and light had a stronger effect on the photosynthetic efficiency (Fv/Fm than did CO2; and at Topt, the maximum growth rate increased with increasing CO2, but the initial slope of the growth-irradiance curve was not affected by CO2. In the context of environmental change, our results suggest that the (i nutrient replete growth rate of Trichodesmium IMS101 would have been severely limited by low pCO2 at the last glacial maximum (LGM, (ii future increases in pCO2 will increase growth rates in areas where temperature ranges between Tmin to Topt, but will have negligible effect at temperatures between Topt and Tmax, (iii areal increase of warm surface waters (> 18°C has allowed the geographic range to increase significantly from the LGM to present and that the range will continue to expand to higher latitudes with continued warming, but (iv continued global warming may exclude Trichodesmium spp. from some tropical regions by 2100 where temperature exceeds Topt.

  6. Differential Responses of Dinitrogen Fixation, Diazotrophic Cyanobacteria and Ammonia Oxidation Reveal a Potential Warming-Induced Imbalance of the N-Cycle in Biological Soil Crusts.

    Directory of Open Access Journals (Sweden)

    Xiaobing Zhou

    Full Text Available N2 fixation and ammonia oxidation (AO are the two most important processes in the nitrogen (N cycle of biological soil crusts (BSCs. We studied the short-term response of acetylene reduction assay (ARA rates, an indicator of potential N2 fixation, and AO rates to temperature (T, -5°C to 35°C in BSC of different successional stages along the BSC ecological succession and geographic origin (hot Chihuahuan and cooler Great Basin deserts. ARA in all BSCs increased with T until saturation occurred between 15 and 20°C, and declined at 30-35°C. Culture studies using cyanobacteria isolated from these crusts indicated that the saturating effect was traceable to their inability to grow well diazotrophically within the high temperature range. Below saturation, temperature response was exponential, with Q10 significantly different in the two areas (~ 5 for Great Basin BSCs; 2-3 for Chihuahuan BSCs, but similar between the two successional stages. However, in contrast to ARA, AO showed a steady increase to 30-35°C in Great Basin, and Chihuhuan BSCs showed no inhibition at any tested temperature. The T response of AO also differed significantly between Great Basin (Q10 of 4.5-4.8 and Chihuahuan (Q10 of 2.4-2.6 BSCs, but not between successional stages. Response of ARA rates to T did not differ from that of AO in either desert. Thus, while both processes scaled to T in unison until 20°C, they separated to an increasing degree at higher temperature. As future warming is likely to occur in the regions where BSCs are often the dominant living cover, this predicted decoupling is expected to result in higher proportion of nitrates in soil relative to ammonium. As nitrate is more easily lost as leachate or to be reduced to gaseous forms, this could mean a depletion of soil N over large landscapes globally.

  7. Macrolactone Nuiapolide, Isolated from a Hawaiian Marine Cyanobacterium, Exhibits Anti-Chemotactic Activity.

    Science.gov (United States)

    Mori, Shogo; Williams, Howard; Cagle, Davey; Karanovich, Kristopher; Horgen, F David; Smith, Roger; Watanabe, Coran M H

    2015-10-09

    A new bioactive macrolactone, nuiapolide (1) was identified from a marine cyanobacterium collected off the coast of Niihau, near Lehua Rock. The natural product exhibits anti-chemotactic activity at concentrations as low as 1.3 μM against Jurkat cells, cancerous T lymphocytes, and induces a G2/M phase cell cycle shift. Structural characterization of the natural product revealed the compound to be a 40-membered macrolactone with nine hydroxyl functional groups and a rare tert-butyl carbinol residue.

  8. Macrolactone Nuiapolide, Isolated from a Hawaiian Marine Cyanobacterium, Exhibits Anti-Chemotactic Activity

    OpenAIRE

    Mori, Shogo; Williams, Howard; Cagle, Davey; Karanovich, Kristopher; Horgen, F. David; Smith, Roger; Watanabe, Coran M. H.

    2015-01-01

    A new bioactive macrolactone, nuiapolide (1) was identified from a marine cyanobacterium collected off the coast of Niihau, near Lehua Rock. The natural product exhibits anti-chemotactic activity at concentrations as low as 1.3 μM against Jurkat cells, cancerous T lymphocytes, and induces a G2/M phase cell cycle shift. Structural characterization of the natural product revealed the compound to be a 40-membered macrolactone with nine hydroxyl functional groups and a rare tert-butyl carbinol re...

  9. Macrolactone Nuiapolide, Isolated from a Hawaiian Marine Cyanobacterium, Exhibits Anti-Chemotactic Activity

    Directory of Open Access Journals (Sweden)

    Shogo Mori

    2015-10-01

    Full Text Available A new bioactive macrolactone, nuiapolide (1 was identified from a marine cyanobacterium collected off the coast of Niihau, near Lehua Rock. The natural product exhibits anti-chemotactic activity at concentrations as low as 1.3 μM against Jurkat cells, cancerous T lymphocytes, and induces a G2/M phase cell cycle shift. Structural characterization of the natural product revealed the compound to be a 40-membered macrolactone with nine hydroxyl functional groups and a rare tert-butyl carbinol residue.

  10. The Effects of the Toxic Cyanobacterium Limnothrix (Strain AC0243 on Bufo marinus Larvae

    Directory of Open Access Journals (Sweden)

    Olivia Daniels

    2014-03-01

    Full Text Available Limnothrix (strain AC0243 is a cyanobacterium, which has only recently been identified as toxin producing. Under laboratory conditions, Bufo marinus larvae were exposed to 100,000 cells mL−1 of Limnothrix (strain AC0243 live cultures for seven days. Histological examinations were conducted post mortem and revealed damage to the notochord, eyes, brain, liver, kidney, pancreas, gastrointestinal tract, and heart. The histopathological results highlight the toxicological impact of this strain, particularly during developmental stages. Toxicological similarities to β-N-Methylamino-L-alanine are discussed.

  11. The Effects of the Toxic Cyanobacterium Limnothrix (Strain AC0243) on Bufo marinus Larvae

    Science.gov (United States)

    Daniels, Olivia; Fabbro, Larelle; Makiela, Sandrine

    2014-01-01

    Limnothrix (strain AC0243) is a cyanobacterium, which has only recently been identified as toxin producing. Under laboratory conditions, Bufo marinus larvae were exposed to 100,000 cells mL−1 of Limnothrix (strain AC0243) live cultures for seven days. Histological examinations were conducted post mortem and revealed damage to the notochord, eyes, brain, liver, kidney, pancreas, gastrointestinal tract, and heart. The histopathological results highlight the toxicological impact of this strain, particularly during developmental stages. Toxicological similarities to β-N-Methylamino-l-alanine are discussed. PMID:24662524

  12. Acetylome analysis reveals the involvement of lysine acetylation in photosynthesis and carbon metabolism in the model cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Mo, Ran; Yang, Mingkun; Chen, Zhuo; Cheng, Zhongyi; Yi, Xingling; Li, Chongyang; He, Chenliu; Xiong, Qian; Chen, Hui; Wang, Qiang; Ge, Feng

    2015-02-06

    Cyanobacteria are the oldest known life form inhabiting Earth and the only prokaryotes capable of performing oxygenic photosynthesis. Synechocystis sp. PCC 6803 (Synechocystis) is a model cyanobacterium used extensively in research on photosynthesis and environmental adaptation. Posttranslational protein modification by lysine acetylation plays a critical regulatory role in both eukaryotes and prokaryotes; however, its extent and function in cyanobacteria remain unexplored. Herein, we performed a global acetylome analysis on Synechocystis through peptide prefractionation, antibody enrichment, and high accuracy LC-MS/MS analysis; identified 776 acetylation sites on 513 acetylated proteins; and functionally categorized them into an interaction map showing their involvement in various biological processes. Consistent with previous reports, a large fraction of the acetylation sites are present on proteins involved in cellular metabolism. Interestingly, for the first time, many proteins involved in photosynthesis, including the subunits of phycocyanin (CpcA, CpcB, CpcC, and CpcG) and allophycocyanin (ApcA, ApcB, ApcD, ApcE, and ApcF), were found to be lysine acetylated, suggesting that lysine acetylation may play regulatory roles in the photosynthesis process. Six identified acetylated proteins associated with photosynthesis and carbon metabolism were further validated by immunoprecipitation and Western blotting. Our data provide the first global survey of lysine acetylation in cyanobacteria and reveal previously unappreciated roles of lysine acetylation in the regulation of photosynthesis. The provided data set may serve as an important resource for the functional analysis of lysine acetylation in cyanobacteria and facilitate the elucidation of the entire metabolic networks and photosynthesis process in this model cyanobacterium.

  13. Analysis of diversity of diazotrophic bacteria associated with the rhizosphere of a tropical Arbor, Melastoma malabathricum L.

    Science.gov (United States)

    Sato, Atsuya; Watanabe, Toshihiro; Unno, Yusuke; Purnomo, Erry; Osaki, Mitsuru; Shinano, Takuro

    2009-01-01

    The diversity of diazotrophic bacteria in the rhizosphere of Melastoma malabathricum L. was investigated by cloning-sequencing of the nifH gene directly amplified from DNA extracted from soil. Samples were obtained from the rhizosphere and bulk soil of M. malabathricum growing in three different soil types (acid sulfate, peat and sandy clay soils) located very close to each other in south Kalimantan, Indonesia. Six clone libraries were constructed, generated from bulk and rhizosphere soil samples, and 300 nifH clones were produced, then assembled into 29 operational taxonomic units (OTUs) based on percent identity values. Our results suggested that nifH gene diversity is mainly dependent on soil properties, and did not differ remarkably between the rhizosphere and bulk soil of M. malabathricum except in acid sulfate soil. In acid sulfate soil, as the Shannon diversity index was lower in rhizosphere than in bulk soil, it is suggested that particular bacterial species might accumulate in the rhizosphere.

  14. A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry

    Directory of Open Access Journals (Sweden)

    L.P.S. Alves

    Full Text Available The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i cell permeabilization, ii Nile red staining, and iii analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99 compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.

  15. A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry.

    Science.gov (United States)

    Alves, L P S; Almeida, A T; Cruz, L M; Pedrosa, F O; de Souza, E M; Chubatsu, L S; Müller-Santos, M; Valdameri, G

    2017-01-16

    The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.

  16. A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry

    Science.gov (United States)

    Alves, L.P.S.; Almeida, A.T.; Cruz, L.M.; Pedrosa, F.O.; de Souza, E.M.; Chubatsu, L.S.; Müller-Santos, M.; Valdameri, G.

    2017-01-01

    The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots. PMID:28099582

  17. Comparative genomics reveals diversified CRISPR-Cas systems of globally distributed Microcystis aeruginosa, a freshwater bloom-forming cyanobacterium

    Directory of Open Access Journals (Sweden)

    Chen eYang

    2015-05-01

    Full Text Available Microcystis aeruginosa is one of the most common and dominant bloom-forming cyanobacteria in freshwater lakes around the world. Microcystis cells can produce toxic secondary metabolites, such as microcystins, which are harmful to human health. Two M. aeruginosa strains were isolated from two highly eutrophic lakes in China and their genomes were sequenced. Comparative genomic analysis was performed with the 12 other available M. aeruginosa genomes and closely related unicellular cyanobacterium. Each genome of M. aeruginosa containing at least one clustered regularly interspaced short palindromic repeat (CRISPR locus and total 71 loci were identified, suggesting it is ubiquitous in M. aeruginosa genomes. In addition to the previously reported subtype I-D cas gene sets, three CAS subtypes I-A, III-A and III-B were identified and characterized in this study. Seven types of CRISPR direct repeat have close association with CAS subtype, confirming that different and specific secondary structures of CRISPR repeats are important for the recognition, binding and process of corresponding cas gene sets. Homology search of the CRISPR spacer sequences provides a history of not only resistance to bacteriophages and plasmids known to be associated with M. aeruginosa, but also the ability to target much more exogenous genetic material in the natural environment. These adaptive and heritable defense mechanisms play a vital role in keeping genomic stability and self-maintenance by restriction of horizontal gene transfer. Maintaining genomic stability and modulating genomic plasticity are both important evolutionary strategies for M. aeruginosa in adaptation and survival in various habitats.

  18. First Evidence of Palytoxin and 42-Hydroxy-palytoxin in the Marine Cyanobacterium Trichodesmium

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    Dominique Laurent

    2011-03-01

    Full Text Available Marine pelagic diazotrophic cyanobacteria of the genus Trichodesmium (Oscillatoriales are widespread throughout the tropics and subtropics, and are particularly common in the waters of New Caledonia. Blooms of Trichodesmium are suspected to be a potential source of toxins in the ciguatera food chain and were previously reported to contain several types of paralyzing toxins. The toxicity of water-soluble extracts of Trichodesmium spp. were analyzed by mouse bioassay and Neuroblastoma assay and their toxic compounds characterized using liquid chromatography coupled with tandem mass spectrometry techniques. Here, we report the first identification of palytoxin and one of its derivatives, 42-hydroxy-palytoxin, in field samples of Trichodesmium collected in the New Caledonian lagoon. The possible role played by Trichodesmium blooms in the development of clupeotoxism, this human intoxication following the ingestion of plankton-eating fish and classically associated with Ostreopsis blooms, is also discussed.

  19. A Nostoc punctiforme sugar transporter necessary to establish a Cyanobacterium-plant symbiosis.

    Science.gov (United States)

    Ekman, Martin; Picossi, Silvia; Campbell, Elsie L; Meeks, John C; Flores, Enrique

    2013-04-01

    In cyanobacteria-plant symbioses, the symbiotic nitrogen-fixing cyanobacterium has low photosynthetic activity and is supplemented by sugars provided by the plant partner. Which sugars and cyanobacterial sugar uptake mechanism(s) are involved in the symbiosis, however, is unknown. Mutants of the symbiotically competent, facultatively heterotrophic cyanobacterium Nostoc punctiforme were constructed bearing a neomycin resistance gene cassette replacing genes in a putative sugar transport gene cluster. Results of transport activity assays using (14)C-labeled fructose and glucose and tests of heterotrophic growth with these sugars enabled the identification of an ATP-binding cassette-type transporter for fructose (Frt), a major facilitator permease for glucose (GlcP), and a porin needed for the optimal uptake of both fructose and glucose. Analysis of green fluorescent protein fluorescence in strains of N. punctiforme bearing frt::gfp fusions showed high expression in vegetative cells and akinetes, variable expression in hormogonia, and no expression in heterocysts. The symbiotic efficiency of N. punctiforme sugar transport mutants was investigated by testing their ability to infect a nonvascular plant partner, the hornwort Anthoceros punctatus. Strains that were specifically unable to transport glucose did not infect the plant. These results imply a role for GlcP in establishing symbiosis under the conditions used in this work.

  20. Sorption and desorption studies of chromium(VI) from nonviable cyanobacterium Nostoc muscorum biomass

    International Nuclear Information System (INIS)

    Gupta, V.K.; Rastogi, A.

    2008-01-01

    This communication presents results pertaining to the sorptive and desorptive studies carried out on chromium(VI) removal onto nonviable freshwater cyanobacterium (Nostoc muscorum) biomass. Influence of varying the conditions for removal of chromium(VI), such as the pH of aqueous solution, the dosage of biosorbent, the contact time with the biosorbent, the temperature for the removal of chromium, the effect of light metal ions and the adsorption-desorption studies were investigated. Sorption interaction of chromium on to cyanobacterial species obeyed both the first and the second-order rate equation and the experimental data showed good fit with both the Langmuir and freundlich adsorption isotherm models. The maximum adsorption capacity was 22.92 mg/g at 25 o C and pH 3.0. The adsorption process was endothermic and the values of thermodynamic parameters of the process were calculated. Various properties of the cyanobacterium, as adsorbent, explored in the characterization part were chemical composition of the adsorbent, surface area calculation by BET method and surface functionality by FTIR. Sorption-desorption of chromium into inorganic solutions and distilled water were observed and this indicated the biosorbent could be regenerated using 0.1 M HNO 3 and EDTA with upto 80% recovery. The biosorbents were reused in five biosorption-desorption cycles without a significant loss in biosorption capacity. Thus, this study demonstrated that the cyanobacterial biomass N. muscorum could be used as an efficient biosorbent for the treatment of chromium(VI) bearing wastewater

  1. Growth of Cyanobacterium aponinum influenced by increasing salt concentrations and temperature.

    Science.gov (United States)

    Winckelmann, Dominik; Bleeke, Franziska; Bergmann, Peter; Klöck, Gerd

    2015-06-01

    The increasing requirement of food neutral biofuels demands the detection of alternative sources. The use of non-arable land and waste water streams is widely discussed in this regard. A Cyanobacterium was isolated on the area of a possible algae production side near a water treatment plant in the arid desert region al-Wusta. It was identified as Cyanobacterium aponinum PB1 and is a possible lipid source. To determine its suitability of a production process using this organism, a set of laboratory experiments were performed. Its growth behavior was examined in regard to high temperatures and increasing NaCl concentrations. A productivity of 0.1 g L -1 per day was measured at an alga density below 0.75 g L -1 . C. aponinum PB1 showed no sign of altered growth behavior in media containing 70 g L -1 NaCl or less. Detection of a negative effect of NaCl on the growth using Pulse-Amplitude-Modulation chlorophyll fluorescence analysis was not more sensitive than optical density measurement.

  2. Unique thylakoid membrane architecture of a unicellular N2-fixing cyanobacterium revealed by electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Liberton, Michelle; Austin II, Jotham R; Berg, R. Howard; Pakrasi, Himadri B

    2011-04-01

    Cyanobacteria, descendants of the endosymbiont that gave rise to modern-day chloroplasts, are vital contributors to global biological energy conversion processes. A thorough understanding of the physiology of cyanobacteria requires detailed knowledge of these organisms at the level of cellular architecture and organization. In these prokaryotes, the large membrane protein complexes of the photosynthetic and respiratory electron transport chains function in the intracellular thylakoid membranes. Like plants, the architecture of the thylakoid membranes in cyanobacteria has direct impact on cellular bioenergetics, protein transport, and molecular trafficking. However, whole-cell thylakoid organization in cyanobacteria is not well understood. Here we present, by using electron tomography, an in-depth analysis of the architecture of the thylakoid membranes in a unicellular cyanobacterium, Cyanothece sp. ATCC 51142. Based on the results of three-dimensional tomographic reconstructions of near-entire cells, we determined that the thylakoids in Cyanothece 51142 form a dense and complex network that extends throughout the entire cell. This thylakoid membrane network is formed from the branching and splitting of membranes and encloses a single lumenal space. The entire thylakoid network spirals as a peripheral ring of membranes around the cell, an organization that has not previously been described in a cyanobacterium. Within the thylakoid membrane network are areas of quasi-helical arrangement with similarities to the thylakoid membrane system in chloroplasts. This cyanobacterial thylakoid arrangement is an efficient means of packing a large volume of membranes in the cell while optimizing intracellular transport and trafficking.

  3. Unique Thylakoid Membrane Architecture of a Unicellular N2-Fixing Cyanobacterium Revealed by Electron Tomography

    Energy Technology Data Exchange (ETDEWEB)

    Liberton, Michelle L.; Austin, Jotham R.; Berg, R. H.; Pakrasi, Himadri B.

    2011-04-01

    Cyanobacteria, descendants of the endosymbiont that gave rise to modern-day chloroplasts, are vital contributors to global biological energy conversion processes. A thorough understanding of the physiology of cyanobacteria requires detailed knowledge of these organisms at the level of cellular architecture and organization. In these prokaryotes, the large membrane protein complexes of the photosynthetic and respiratory electron transport chains function in the intracellular thylakoid membranes. Like plants, the architecture of the thylakoid membranes in cyanobacteria has direct impact on cellular bioenergetics, protein transport, and molecular trafficking. However, whole-cell thylakoid organization in cyanobacteria is not well understood. Here we present, by using electron tomography, an in-depth analysis of the architecture of the thylakoid membranes in a unicellular cyanobacterium, Cyanothece sp. ATCC 51142. Based on the results of three-dimensional tomographic reconstructions of near-entire cells, we determined that the thylakoids in Cyanothece 51142 form a dense and complex network that extends throughout the entire cell. This thylakoid membrane network is formed from the branching and splitting of membranes and encloses a single lumenal space. The entire thylakoid network spirals as a peripheral ring of membranes around the cell, an organization that has not previously been described in a cyanobacterium. Within the thylakoid membrane network are areas of quasi-helical arrangement with similarities to the thylakoid membrane system in chloroplasts. This cyanobacterial thylakoid arrangement is an efficient means of packing a large volume of membranes in the cell while optimizing intracellular transport and trafficking.

  4. Combined Effects of CO2 and Light on the N-2-Fixing Cyanobacterium Trichodesmium IMS101: A Mechanistic View

    Czech Academy of Sciences Publication Activity Database

    Levitan, O.; Kranz, S. A.; Spungin, D.; Prášil, Ondřej; Rost, B.; Berman-Frank, I.

    2010-01-01

    Roč. 154, č. 1 (2010), s. 346-356 ISSN 0032-0889 R&D Projects: GA ČR GA206/08/1683 Institutional research plan: CEZ:AV0Z50200510 Keywords : cyanobacterium Trichodesmium * ocean * photosystem I Subject RIV: EE - Microbiology, Virology Impact factor: 6.451, year: 2010

  5. Combined effect of CO2 and light on the N2-fixing cyanobacterium Trichodesmium IMS101: Physiological responses 1[OA

    Czech Academy of Sciences Publication Activity Database

    Kranz, S. A.; Levitan, O.; Richter, K.-U.; Prášil, Ondřej; Berman-Frank, I.; Rost, B.

    2010-01-01

    Roč. 154, č. 1 (2010), s. 334-345 ISSN 0032-0889 R&D Projects: GA ČR GA206/08/1683 Institutional research plan: CEZ:AV0Z50200510 Keywords : Trichodesmium IMS101 * cyanobacterium * CO2 Subject RIV: EE - Microbiology, Virology Impact factor: 6.451, year: 2010

  6. Mono-, di- and trimeric PS I reaction center complexes isolated from the thermophilic cyanobacterium Synechococcus sp. Size, shape and activity

    NARCIS (Netherlands)

    Rögner, M.; Mühlenhoff, U.; Boekema, E.J.; Witt, H.T.

    1990-01-01

    Photosystem I preparations from the cyanobacterium Synechococcus sp. were treated with high concentrations of Tris and octyl glucoside at alkaline pH and elevated temperature. A sucrose density gradient yielded three pigment-protein complexes; these were further purified on a HPLC anion-exchange

  7. Two-Step Separation of Nostotrebin 6 from Cultivated Soil Cyanobacterium (Nostoc sp.) by High Performance Countercurrent Chromatography

    Czech Academy of Sciences Publication Activity Database

    Cheel, José; Kučerová, P.; Garrard, I.; Ignatova, S.; Hrouzek, Pavel; Kopecký, Jiří

    2014-01-01

    Roč. 19, č. 4 (2014), s. 8773-8787 ISSN 1420-3049 R&D Projects: GA MŠk ED2.1.00/03.0110; GA MŠk EE2.3.30.0059 Institutional support: RVO:61388971 Keywords : nostotrebin 6 * cyanobacterium * Nostoc * HPLC separation Subject RIV: EE - Microbiology, Virology Impact factor: 2.416, year: 2014

  8. Backbone dynamics of reduced plastocyanin from the cyanobacterium Anabaena variabilis: Regions involved in electron transfer have enhanced mobility

    DEFF Research Database (Denmark)

    Ma, L.X.; Hass, M.A.S.; Vierick, N.

    2003-01-01

    The dynamics of the backbone of the electron-transfer protein plastocyanin from the cyanobacterium Anabaena variabilis were determined from the N-15 and C-13(alpha) R-1 and R-2) relaxation rates and steady-state [H-1]-N-15 and [H-1]-C-13 nuclear Overhauser effects (NOEs) using the model...

  9. Pulsed nitrogen supply induces dynamic changes in the amino acid composition and microcystin production of the harmful cyanobacterium Planktothrix agardhii

    NARCIS (Netherlands)

    van de Waal, D.B.; Ferreruela, G.; Tonk, L.; van Donk, E.; Huisman, J.; Visser, P.M.; Matthijs, H.C.P.

    2010-01-01

    Planktothrix agardhii is a widespread harmful cyanobacterium of eutrophic waters, and can produce the hepatotoxins [Asp3]microcystin-LR and [Asp3]microcystin-RR. These two microcystin variants differ in their first variable amino acid position, which is occupied by either leucine (L) or arginine

  10. Pulsed nitrogen supply induces dynamic changes in the amino acid compositionand microcystin production of the harmful cyanobacterium Planktothrix agardhii

    NARCIS (Netherlands)

    Van de Waal, D.B.; Ferreruela, G.; Tonk, L.; Van Donk, E.; Huisman, J.; Visser, P.M.; Matthijs, H.C.P.

    2010-01-01

    Planktothrix agardhii is a widespread harmful cyanobacterium of eutrophic waters, and can produce the hepatotoxins [Asp3]microcystin-LR and [Asp3]microcystin-RR. These two microcystin variants differ in their first variable amino acid position, which is occupied by either leucine (L) or arginine

  11. Evaluation of the cosmetic potential of the Cyanobacterium Spirulina platensis

    OpenAIRE

    Novak, Alessandra Cristine

    2013-01-01

    Abstract: Toiletry, Perfumery and Cosmetics Sector (TPCS) occupies a prominent place in modern society. The search for beauty and wellness, combined with increased life expectancy of the population, causes the intensive search to products that improve appearance, hygiene and health. Thus, the development of cosmetics is very stimulated. Another important feature is the continuously pressure from consumers and companies to this development of new and innovative products, raising competitivenes...

  12. LexA Binds to Transcription Regulatory Site of Cell Division Gene ftsZ in Toxic Cyanobacterium Microcystis aeruginosa.

    Science.gov (United States)

    Honda, Takashi; Morimoto, Daichi; Sako, Yoshihiko; Yoshida, Takashi

    2018-05-17

    Previously, we showed that DNA replication and cell division in toxic cyanobacterium Microcystis aeruginosa are coordinated by transcriptional regulation of cell division gene ftsZ and that an unknown protein specifically bound upstream of ftsZ (BpFz; DNA-binding protein to an upstream site of ftsZ) during successful DNA replication and cell division. Here, we purified BpFz from M. aeruginosa strain NIES-298 using DNA-affinity chromatography and gel-slicing combined with gel electrophoresis mobility shift assay (EMSA). The N-terminal amino acid sequence of BpFz was identified as TNLESLTQ, which was identical to that of transcription repressor LexA from NIES-843. EMSA analysis using mutant probes showed that the sequence GTACTAN 3 GTGTTC was important in LexA binding. Comparison of the upstream regions of lexA in the genomes of closely related cyanobacteria suggested that the sequence TASTRNNNNTGTWC could be a putative LexA recognition sequence (LexA box). Searches for TASTRNNNNTGTWC as a transcriptional regulatory site (TRS) in the genome of M. aeruginosa NIES-843 showed that it was present in genes involved in cell division, photosynthesis, and extracellular polysaccharide biosynthesis. Considering that BpFz binds to the TRS of ftsZ during normal cell division, LexA may function as a transcriptional activator of genes related to cell reproduction in M. aeruginosa, including ftsZ. This may be an example of informality in the control of bacterial cell division.

  13. Salinity tolerance of Picochlorum atomus and the use of salinity for contamination control by the freshwater cyanobacterium Pseudanabaena limnetica.

    Directory of Open Access Journals (Sweden)

    Nicolas von Alvensleben

    Full Text Available Microalgae are ideal candidates for waste-gas and -water remediation. However, salinity often varies between different sites. A cosmopolitan microalga with large salinity tolerance and consistent biochemical profiles would be ideal for standardised cultivation across various remediation sites. The aims of this study were to determine the effects of salinity on Picochlorum atomus growth, biomass productivity, nutrient uptake and biochemical profiles. To determine if target end-products could be manipulated, the effects of 4-day nutrient limitation were also determined. Culture salinity had no effect on growth, biomass productivity, phosphate, nitrate and total nitrogen uptake at 2, 8, 18, 28 and 36 ppt. 11 ppt, however, initiated a significantly higher total nitrogen uptake. While salinity had only minor effects on biochemical composition, nutrient depletion was a major driver for changes in biomass quality, leading to significant increases in total lipid, fatty acid and carbohydrate quantities. Fatty acid composition was also significantly affected by nutrient depletion, with an increased proportion of saturated and mono-unsaturated fatty acids. Having established that P. atomus is a euryhaline microalga, the effects of culture salinity on the development of the freshwater cyanobacterial contaminant Pseudanabaena limnetica were determined. Salinity at 28 and 36 ppt significantly inhibited establishment of P. limnetica in P. atomus cultures. In conclusion, P. atomus can be deployed for bioremediation at sites with highly variable salinities without effects on end-product potential. Nutrient status critically affected biochemical profiles--an important consideration for end-product development by microalgal industries. 28 and 36 ppt slow the establishment of the freshwater cyanobacterium P. limnetica, allowing for harvest of low contaminant containing biomass.

  14. Response of biological soil crust diazotrophs to season, altered summer precipitation and year-round increased temperature in an arid grassland of the Colorado Plateau, USA

    Directory of Open Access Journals (Sweden)

    Chris M Yeager

    2012-10-01

    Full Text Available Biological soil crusts (biocrusts, which supply significant amounts of fixed nitrogen into terrestrial ecosystems worldwide (~33 Tg y-1, are likely to respond to changes in temperature and precipitation associated with climate change. Using nifH gene-based surveys, we explored variation in the diazotrophic community of biocrusts of the Colorado Plateau, USA in response to season (autumn vs. spring, as well as field manipulations that increased the frequency of small-volume precipitation events and year-round soil temperature. Abundance of nifH genes in biocrusts ranged from 3x106 – 1x108 g-1 soil, and nifH from heterocystous cyanobacteria closely related to Scytonema hyalinum, Spirirestis rafaelensis, and Nostoc commune comprised > 98% of the total. Although there was no apparent seasonal effect on total nifH gene abundance in the biocrusts, T-RFLP analysis revealed a strong seasonal pattern in nifH composition. Spirirestis nifH abundance was estimated to oscillate 1 to >2 orders of magnitude between autumn (low and spring (high. A year-round increase of soil temperature (2 − 3 °C had little effect on the diazotroph community structure over 2 years. Altered summer precipitation had little impact on diazotroph community structure over the first 1.5 years of the study, when natural background patterns across years and seasons superseded any treatment effects. However, after the second summer of treatments, nifH abundance was 2.6 fold lower in biocrusts receiving altered precipitation. Heterocystous cyanobacteria were apparently more resilient to altered precipitation than other cyanobacteria. The results demonstrate that diazotrophic community composition of biocrusts in this semi-arid grassland undergoes strong seasonal shifts and that the abundance of its dominant members decreased in response to more frequent, small-volume precipitation events.

  15. Response of biological soil crust diazotrophs to season, altered summer precipitation, and year-round increased temperature in an arid grassland of the Colorado Plateau, USA

    Science.gov (United States)

    Yeager, Chris M.; Kuske, Cheryl R.; Carney, Travis D.; Johnson, Shannon L.; Ticknor, Lawrence O.; Belnap, Jayne

    2012-01-01

    Biological soil crusts (biocrusts), which supply significant amounts of fixed nitrogen into terrestrial ecosystems worldwide (~33Tg y-1), are likely to respond to changes in temperature and precipitation associated with climate change. Using nifH gene-based surveys, we explored variation in the diazotrophic community of biocrusts of the Colorado Plateau, USA in response to season (autumn vs. spring), as well as field manipulations that increased the frequency of small volume precipitation events and year-round soil temperature. Abundance of nifH genes in biocrusts ranged from 3×106 to 1×8 g-1 soil, and nifH from heterocystous cyanobacteria closely related to Scytonema hyalinum, Spirirestis rafaelensis, and Nostoc commune comprised >98% of the total. Although there was no apparent seasonal effect on total nifH gene abundance in the biocrusts, T-RFLP analysis revealed a strong seasonal pattern in nifH composition. Spirirestis nifH abundance was estimated to oscillate 1 to >2 orders of magnitude between autumn (low) and spring (high). A year-round increase of soil temperature (2–3°C) had little effect on the diazotroph community structure over 2 years. Altered summer precipitation had little impact on diazotroph community structure over the first 1.5years of the study, when natural background patterns across years and seasons superseded any treatment effects. However, after the second summer of treatments, nifH abundance was 2.6-fold lower in biocrusts receiving altered precipitation. Heterocystous cyanobacteria were apparently more resilient to altered precipitation than other cyanobacteria. The results demonstrate that diazotrophic community composition of biocrusts in this semi-arid grassland undergoes strong seasonal shifts and that the abundance of its dominant members decreased in response to more frequent, small volume precipitation events.

  16. On the Relationship Between Hydrogen Saturation in the Tropical Atlantic Ocean and Nitrogen Fixation by the Symbiotic Diazotroph UCYN-A

    Science.gov (United States)

    Moore, R. M.; Grefe, I.; Zorz, J.; Shan, S.; Thompson, K.; Ratten, J.; LaRoche, J.

    2018-04-01

    Dissolved hydrogen measurements were made at high resolution in surface waters along a tropical north Atlantic transect between Guadeloupe and Cape Verde in 2015 (Meteor 116). Parallel water samples acquired to assess the relative abundance of the nifH gene from several types of diazotrophs, indicated that Trichodesmium and UCYN-A were dominant in this region. We show that a high degree of correlation exists between the hydrogen saturations and UCYN-A nifH abundance, and a weak correlation with Trichodesmium. The findings suggest that nitrogen fixation by UCYN-A is a major contributor to hydrogen supersaturations in this region of the ocean. The ratio of hydrogen released to nitrogen fixed has not been determined for this symbiont, but the indications are that it may be high in comparison with the small number of diazotrophs for which the ratio has been measured in laboratory cultures. We speculate that this would be consistent with the diazotroph being an exosymbiont on its haptophyte host. Our high resolution measurements of hydrogen concentrations are capable of illustrating the time and space scales of inferred activity of diazotrophs in near real-time in a way that cannot be achieved by biological sampling and rate measurements requiring incubations with 15N2. Direct measurement of high resolution spatial variability would be relatively challenging through collection and analysis of biological samples by qPCR, and extremely challenging by 15N-uptake techniques, neither of which methods yields real-time data. Nonetheless, determination of fixation rates still firmly depends on the established procedure of incubations in the presence of 15N2.

  17. Biochemical studies of amylase, lipase and protease in Callosobruchus maculatus (Coleoptera: Chrysomelidae) populations fed with Vigna unguiculata grain cultivated with diazotrophic bacteria strains.

    Science.gov (United States)

    Silva, L B; Torres, É B; Nóbrega, R A S; Lopes, G N; Vogado, R F; Pavan, B E; Fernandes-Junior, P I

    2017-12-01

    The objective of this study was to evaluate the enzymatic activity of homogenates of insects fed on grain of cowpea, Vigna unguiculata (L.), cultivars grown with different nitrogen sources. For the experiment we used aliquots of the homogenate of 100 unsexed adult insects, emerged from 10 g of grain obtained from four cowpea cultivars: 'BRS Acauã', 'BRS Carijó', 'BRS Pujante', and 'BRS Tapaihum' grown under different regimes of nitrogen sources: mineral fertilizer, inoculation with strains of diazotrophs (BR 3267, BR 3262, BR 3299; INPA 03-11B, 03-84 UFLA, as well as the control (with soil nitrogen). The parameters evaluated were enzymatic activities of insect protease, amylase and lipase and the starch content of the grains. There were differences in the enzymatic activity of amylase, lipase and protease of insect homogenate according to the food source. A lower activity of the enzyme amylase from C. maculatus homogenate was observed when insects were fed grain of the cultivar BRS Carijó. A lower activity of lipase enzyme from C. maculatus homogenate was observed when the insects fed on grain from the interaction of the cultivar Tapaihum inoculated with BR 3262 diazotrophs. The lowest proteolytic activity was observed in homogenate of insects fed on interaction of 'BRS Carijó' inoculated with BR 3262 diazotrophs. Starch content correlated positively with the amylase activity of C. maculatus homogenate. The cultivar BRS Carijó had a different behavior from the other cultivars, according to the cluster analysis.

  18. Characterization of Function of the GlgA2 Glycogen/Starch Synthase in Cyanobacterium sp. Clg1 Highlights Convergent Evolution of Glycogen Metabolism into Starch Granule Aggregation1

    Science.gov (United States)

    Kadouche, Derifa; Arias, Maria Cecilia

    2016-01-01

    At variance with the starch-accumulating plants and most of the glycogen-accumulating cyanobacteria, Cyanobacterium sp. CLg1 synthesizes both glycogen and starch. We now report the selection of a starchless mutant of this cyanobacterium that retains wild-type amounts of glycogen. Unlike other mutants of this type found in plants and cyanobacteria, this mutant proved to be selectively defective for one of the two types of glycogen/starch synthase: GlgA2. This enzyme is phylogenetically related to the previously reported SSIII/SSIV starch synthase that is thought to be involved in starch granule seeding in plants. This suggests that, in addition to the selective polysaccharide debranching demonstrated to be responsible for starch rather than glycogen synthesis, the nature and properties of the elongation enzyme define a novel determinant of starch versus glycogen accumulation. We show that the phylogenies of GlgA2 and of 16S ribosomal RNA display significant congruence. This suggests that this enzyme evolved together with cyanobacteria when they diversified over 2 billion years ago. However, cyanobacteria can be ruled out as direct progenitors of the SSIII/SSIV ancestral gene found in Archaeplastida. Hence, both cyanobacteria and plants recruited similar enzymes independently to perform analogous tasks, further emphasizing the importance of convergent evolution in the appearance of starch from a preexisting glycogen metabolism network. PMID:27208262

  19. BMAA Inhibits Nitrogen Fixation in the Cyanobacterium Nostoc sp. PCC 7120

    Directory of Open Access Journals (Sweden)

    Birgitta Bergman

    2013-08-01

    Full Text Available Cyanobacteria produce a range of secondary metabolites, one being the neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA, proposed to be a causative agent of human neurodegeneration. As for most cyanotoxins, the function of BMAA in cyanobacteria is unknown. Here, we examined the effects of BMAA on the physiology of the filamentous nitrogen-fixing cyanobacterium Nostoc sp. PCC 7120. Our data show that exogenously applied BMAA rapidly inhibits nitrogenase activity (acetylene reduction assay, even at micromolar concentrations, and that the inhibition was considerably more severe than that induced by combined nitrogen sources and most other amino acids. BMAA also caused growth arrest and massive cellular glycogen accumulation, as observed by electron microscopy. With nitrogen fixation being a process highly sensitive to oxygen species we propose that the BMAA effects found here may be related to the production of reactive oxygen species, as reported for other organisms.

  20. Inhibition of motility in the cyanobacterium, Phormidium uncinatum, by solar and monochromatic UV irradiation

    International Nuclear Information System (INIS)

    Häder, D.P.; Watanabe, M.; Furuya, M.

    1986-01-01

    The effect of solar radiation and monochromatic UV radiation on the motility of the filamentous cyanobacterium Phormidium uncinatum was determined. Solar radiation (mid-day, in midsummer at a location near Lisboa, Portugal) was found to impair motility within about 30 min. This effect is neither a result of a temperature increase nor of visible light. The spectral sensitivity determined using the Okazaki Largé Spectrograph shows the maximal effectiveness of radiation of ≤300 nm. The short time requirement for the response and the lack of any photoreactivation of motility argues against DNA being the UV target. Investigations using reagents diagnostic of superoxide free radicals and singlet oxygen failed to confirm the involvement of photodynamic effects as the molecular mechanism causing UV inhibition of motility

  1. Molecular cloning of a recA-like gene from the cyanobacterium Anabaena variabilis

    International Nuclear Information System (INIS)

    Owttrim, G.W.; Coleman, J.R.

    1987-01-01

    A recA-like gene isolated from the cyanobacterium Anabaena variabilis was cloned and partially characterized. When introduced into Escherichia coli recA mutants, the 7.5-kilobase-pair plasmid-borne DNA insert restored resistance to methyl methanesulfonate and UV irradiation, as well as recombination proficiency when measured by Hfr-mediated conjugation. The cyanobacterial recA gene restored spontaneous but not mitomycin C-induced prophage production. Restriction analysis and subcloning yielded a 1.5-kilobase-pair Sau3A fragment which also restored methylmethane sulfonate resistance and coded for a 38- to 40-kilodalton polypeptide when expressed in an in vitro transcription-translation system

  2. The complex effects of ocean acidification on the prominent N2-fixing cyanobacterium Trichodesmium.

    Science.gov (United States)

    Hong, Haizheng; Shen, Rong; Zhang, Futing; Wen, Zuozhu; Chang, Siwei; Lin, Wenfang; Kranz, Sven A; Luo, Ya-Wei; Kao, Shuh-Ji; Morel, François M M; Shi, Dalin

    2017-05-05

    Acidification of seawater caused by anthropogenic carbon dioxide (CO 2 ) is anticipated to influence the growth of dinitrogen (N 2 )-fixing phytoplankton, which contribute a large fraction of primary production in the tropical and subtropical ocean. We found that growth and N 2 -fixation of the ubiquitous cyanobacterium Trichodesmium decreased under acidified conditions, notwithstanding a beneficial effect of high CO 2 Acidification resulted in low cytosolic pH and reduced N 2 -fixation rates despite elevated nitrogenase concentrations. Low cytosolic pH required increased proton pumping across the thylakoid membrane and elevated adenosine triphosphate production. These requirements were not satisfied under field or experimental iron-limiting conditions, which greatly amplified the negative effect of acidification. Copyright © 2017, American Association for the Advancement of Science.

  3. Microbial interactions with the cyanobacterium Microcystis aeruginosa and their dependence on temperature

    DEFF Research Database (Denmark)

    Dziallas, Claudia; Grossart, Hans-Peter

    2012-01-01

    and their associated community often masked this temperature effect. Both macro- and microenvironment of active cyanobacterial strains were characterized by high pH and oxygen values creating a unique habitat that potentially affects microbial diversity and function. For example, archaea including ‘anaerobic......Associated heterotrophic bacteria alter the microenvironment of cyanobacteria and potentially influence cyanobacterial development. Therefore, we studied interactions of the unicellular freshwater cyanobacterium Microcystis aeruginosa with heterotrophic bacteria. The associated bacterial community...... was greatly driven by temperature as seen by DNA Wngerprinting. However, the associated microbes also closely interacted with the cyanobacteria indicating changing ecological consequence of the associated bacterial community with temperature. Whereas concentration of dissolved organic carbon in cyanobacterial...

  4. Sacrolide A, a new antimicrobial and cytotoxic oxylipin macrolide from the edible cyanobacterium Aphanothece sacrum

    Directory of Open Access Journals (Sweden)

    Naoya Oku

    2014-08-01

    Full Text Available Macroscopic gelatinous colonies of freshwater cyanobacterium Aphanothece sacrum, a luxury ingredient for Japanese cuisine, were found to contain a new oxylipin-derived macrolide, sacrolide A (1, as an antimicrobial component. The configuration of two chiral centers in 1 was determined by a combination of chiral anisotropy analysis and conformational analysis of different ring-opened derivatives. Compound 1 inhibited the growth of some species of Gram-positive bacteria, yeast Saccharomyces cerevisiae and fungus Penicillium chrysogenum, and was also cytotoxic to 3Y1 rat fibroblasts. Concern about potential food intoxication caused by accidental massive ingestion of A. sacrum was dispelled by the absence of 1 in commercial products. A manual procedure for degrading 1 in raw colonies was also developed, enabling a convenient on-site detoxification at restaurants or for personal consumption.

  5. Space-environmental tolerances in a cyanobacterium, Nostoc sp. HK-01

    Science.gov (United States)

    Tomita-Yokotani, Kaori; Yokobori, Shin-ichi; Kimura, Shunta; Sato, Seigo; Katoh, Hiroshi; Ajioka, Reiko; Yamagishi, Akihiko; Inoue, Kotomi

    2016-07-01

    We have been investigating the tolerances to space-environments of a cyanobacterium, Nostoc sp. HK-01 (hereafter referred to as HK-01). Dry colonies of HK-01 had high tolerance to dry conditions, but more detailed information about tolerance to high-temperature, UV, gamma-ray and heavy particle beams were not deeply investigated. The obtained dry colonies of HK-01 after exposure to each of the conditions described above were investigated. In all of the tested colonies of HK-01 after exposure, all or some of the cells in the colonies were alive. One of the purposes of space agriculture is growing plants on Mars. In the early stages, of our research, cyanobacteria are introduced on Mars to promote the oxidation of the atmosphere and the formation of soil from Mars's regolith. HK-01 will contribute to each of these factors in the future.

  6. BMAA Inhibits Nitrogen Fixation in the Cyanobacterium Nostoc sp. PCC 7120

    Science.gov (United States)

    Berntzon, Lotta; Erasmie, Sven; Celepli, Narin; Eriksson, Johan; Rasmussen, Ulla; Bergman, Birgitta

    2013-01-01

    Cyanobacteria produce a range of secondary metabolites, one being the neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA), proposed to be a causative agent of human neurodegeneration. As for most cyanotoxins, the function of BMAA in cyanobacteria is unknown. Here, we examined the effects of BMAA on the physiology of the filamentous nitrogen-fixing cyanobacterium Nostoc sp. PCC 7120. Our data show that exogenously applied BMAA rapidly inhibits nitrogenase activity (acetylene reduction assay), even at micromolar concentrations, and that the inhibition was considerably more severe than that induced by combined nitrogen sources and most other amino acids. BMAA also caused growth arrest and massive cellular glycogen accumulation, as observed by electron microscopy. With nitrogen fixation being a process highly sensitive to oxygen species we propose that the BMAA effects found here may be related to the production of reactive oxygen species, as reported for other organisms. PMID:23966039

  7. Cadmium-mediated resistance to metals and antibiotics in a cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Singh, S.P.; Pandey, A.K.

    1982-01-01

    Cadmium-resistant strains of the cyanobacterium Nostoc calcicola were isolated through the step-wise transfer of the organism to higher levels of the metal. One of the Cd-resistant strains (CDsup(r)-10) showed cross-resistance to antibiotics like neomycin (1 ..mu..g/ml), chloramphenicol (3 ..mu..g/ml) but not to streptomycin. The Cd-resistant strain also tolerated elevated levels of metals such as zinc 20 ppm) and mercury (1 ppm). The stability of the metal-resistance required the presence of Cd/sup 2 +/ ions in the growth medium. It is suggested that metal resistance may also be determined by gene(s) on the antibiotic resistance plasmids in cyanobacteria.

  8. Composition and occurrence of lipid droplets in the cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Peramuna, Anantha; Summers, Michael L

    2014-12-01

    Inclusions of neutral lipids termed lipid droplets (LDs) located throughout the cell were identified in the cyanobacterium Nostoc punctiforme by staining with lipophylic fluorescent dyes. LDs increased in number upon entry into stationary phase and addition of exogenous fructose indicating a role for carbon storage, whereas high-light stress did not increase LD numbers. LD accumulation increased when nitrate was used as the nitrogen source during exponential growth as compared to added ammonia or nitrogen-fixing conditions. Analysis of isolated LDs revealed enrichment of triacylglycerol (TAG), α-tocopherol, and C17 alkanes. LD TAG from exponential phase growth contained mainly saturated C16 and C18 fatty acids, whereas stationary phase LD TAG had additional unsaturated fatty acids characteristic of whole cells. This is the first characterization of cyanobacterial LD composition and conditions leading to their production. Based upon their abnormally large size and atypical location, these structures represent a novel sub-organelle in cyanobacteria.

  9. Phycobiliprotein accumulation in cyanobacterium Nostoc linckia and modification of antioxidant activity

    Directory of Open Access Journals (Sweden)

    Ana VALUTA

    2015-01-01

    Full Text Available The article deals with iron(III coordination compounds with Schiff bases as ligands and their impact on phycobiliprotein accumulation by cyanobacterium Nostoc linckia. Stimulatory effect depends on the applied dose and in case of three compounds, the concentration 20 mg/L was determined as one with moderate intensity. Lower concentrations resulted in an increase of the phycobiliprotein synthesis. There was found a significant positive correlation between phycobiliprotein content and ABTS (2.2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid assay values displayed by aqueous extracts from Nostoc linckia biomass cultivated in nutrient medium with these coordination compounds. Hence, it is possible to modify the antioxidant activity of Nostoc biomass by applying low concentrations of chemical stimuli.

  10. Heterologous expression of an algal hydrogenase in a hetero-cystous cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Thorsten Heidorn; Peter Lindblad [Dept. of Physiological Botany, Uppsala University, V illavagen 6, SE-752 36 Uppsala, (Sweden)

    2006-07-01

    For the expression of an active algal [FeFe] hydrogenase in the hetero-cystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyano-bacterial cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

  11. Heterologous expression of an algal hydrogenase in a hetero-cystous cyanobacterium

    International Nuclear Information System (INIS)

    Thorsten Heidorn; Peter Lindblad

    2006-01-01

    For the expression of an active algal [FeFe] hydrogenase in the hetero-cystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyano-bacterial cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

  12. Jizanpeptins, Cyanobacterial Protease Inhibitors from a Symploca sp. Cyanobacterium Collected in the Red Sea.

    Science.gov (United States)

    Gallegos, David A; Saurí, Josep; Cohen, Ryan D; Wan, Xuemei; Videau, Patrick; Vallota-Eastman, Alec O; Shaala, Lamiaa A; Youssef, Diaa T A; Williamson, R Thomas; Martin, Gary E; Philmus, Benjamin; Sikora, Aleksandra E; Ishmael, Jane E; McPhail, Kerry L

    2018-05-29

    Jizanpeptins A-E (1-5) are micropeptin depsipeptides isolated from a Red Sea specimen of a Symploca sp. cyanobacterium. The planar structures of the jizanpeptins were established using NMR spectroscopy and mass spectrometry and contain 3-amino-6-hydroxy-2-piperidone (Ahp) as one of eight residues in a typical micropeptin motif, as well as a side chain terminal glyceric acid sulfate moiety. The absolute configurations of the jizanpeptins were assigned using a combination of Marfey's methodology and chiral-phase HPLC analysis of hydrolysis products compared to commercial and synthesized standards. Jizanpeptins A-E showed specific inhibition of the serine protease trypsin (IC 50 = 72 nM to 1 μM) compared to chymotrypsin (IC 50 = 1.4 to >10 μM) in vitro and were not overtly cytotoxic to HeLa cervical or NCI-H460 lung cancer cell lines at micromolar concentrations.

  13. Genetic Basis for Geosmin Production by the Water Bloom-Forming Cyanobacterium, Anabaena ucrainica

    Directory of Open Access Journals (Sweden)

    Zhongjie Wang

    2014-12-01

    Full Text Available Geosmin is a common, musty-smelling sesquiterpene, principally produced by cyanobacteria. Anabaena ucrainica (Schhorb. Watanabe, a water bloom-forming cyanobacterium, is the geosmin producer responsible for odor problems in Dianchi and Erhai lakes in China. In this study, the geosmin synthase gene (geo of A. ucrainica and its flanking regions were identified and cloned by polymerase chain reaction (PCR and genome walking. The geo gene was found to be located in a transcription unit with two cyclic nucleotide-binding protein genes (cnb. The two cnb genes were highly similar and were predicted members of the cyclic adenosine monophosphate (cAMP receptor protein/fumarate nitrate reductase regulator (Crp–Fnr family. Phylogenetic and evolutionary analyses implied that the evolution of the geosmin genes involved a horizontal gene transfer process in cyanobacteria. These genes showed a close relationship to 2-methylisoborneol genes in origin and evolution.

  14. Detection of bioactive exometabolites produced by the filamentous marine cyanobacterium Geitlerinema sp.

    Science.gov (United States)

    Caicedo, Nelson H; Kumirska, Jolanta; Neumann, Jennifer; Stolte, Stefan; Thöming, Jorg

    2012-08-01

    Marine cyanobacteria are noted for their ability to excrete metabolites with biotic properties. This paper focuses on such exometabolites obtained from the culture of the marine filamentous cyanobacterium Geitlerinema sp. strain, their purification and subsequent analyses. By this means the recoveries of the active compounds, a prerequisite for properly determining their concentration, are quantified here for the first time. We demonstrate a new procedure using Amberlite XAD-1180 resin in combination with the eluent isopropanol for extraction of the culture media and gas chromatography as simplified chemical analysis. This procedure reduced necessary bacteria cultivation time (from 150 to 21 days) at low volumes of culture media (300 mL) required for identification of two selected bioactive compounds: 4,4'-dihydroxybiphenyl and harmane.

  15. Isolation and Purification of Heterotetrameric Catalase from a Desiccation Tolerant Cyanobacterium Lyngbya arboricola

    Directory of Open Access Journals (Sweden)

    Kapoor, Shivali

    2013-02-01

    Full Text Available The desiccation tolerant cyanobacterium Lyngbya arboricola, isolated from bark surfaces of Mangifera indica, possessed up to four stable isoforms of catalase in addition to other antioxidative enzymes, for several years under a dry state. Purification of the two most persistent isoforms of catalase (Cat has been undertaken by employing acetone precipitation, ethanol: chloroform treatment, gel filtration and ion exchange chromatography. The two isoforms of catalase remained almost unchanged on varying matric and osmotic hydration levels of mats of the cyanobacterium. The purification procedures resulted in a 1.3 % yield of purified single isoform (0.22 mg mL-1 protein with 709 Units mg-1 specific activity and a purity index of 0.83. Five millimolar of dithiothreitol (DTT was observed to be pertinent in maintaining the optimum redox state of the enzyme. The purification procedures additionally facilitated the simultaneous elimination and procurement of phycoerythrins (PE and mycosporine-like amino acids (MAA. Each purified isoform gave a single band (~45kDa upon SDS-PAGE and denaturing urea isoelectric focusing (IEF depicted the presence of 2 subunits each of CatA and CatB. The monoisotopic mass and pI value of CatA and CatB as revealed by LC-MS analysis and internal amino acid sequencing was 78.96, 5.89 and 80.77, 5.92, respectively, showing resemblance with CatA of Erysiphe graminis subs. hordei and CatB of Ajellomyces capsulata. The heterotetrameric monofunctional catalase (~320 kDa, due to its stability in the form of resistance to ethanol: chloroform, its thermoalkaliphilic nature and the presence of innumerable hydrophobic amino acid residues (~40%, thus exhibited its potential for biotechnological applications.

  16. Cellular responses and bioremoval of nonylphenol by the bloom-forming cyanobacterium Planktothrix agardhii 1113

    Science.gov (United States)

    Medvedeva, Nadezda; Zaytseva, Tatyana; Kuzikova, Irina

    2017-07-01

    Nonylphenol (NP) is extensively used in agricultural, industrial and household applications. Moreover, NP is the major breakdown product of the nonionic surfactants, nonylphenol ethoxylates (NPEOs), the most widely used group of surfactants. Nonylphenol is persistent in the environment, highly toxic to aquatic organisms and is a potential endocrine disruptor. NP and NPEOs have been identified as priority hazardous substances under the Environmental Quality Standards Directive 2013/39/EU and are referred to in the list of substances of particular risk to the Baltic Sea. The toxicity of NP to the bloom-forming cyanobacterium Planktothrix agardhii 1113 isolated from the eastern Gulf of Finland, Baltic Sea and the bioremoval of NP by P. agardhii were studied. NP in concentrations > 0.4 mg L- 1 suppressed cyanobacterial growth. The median effective concentration of NP for P. agardhii after 4 days of treatment (EC50) was 1.5 mg L- 1. The removal of NP from the culture medium was primarily due to abiotic processes and biodegradation by the cyanobacterium rather than sorption by the cells. NP significantly increased the photosynthetic pigments, extracellular proteins and soluble exopolysaccharides content. The cyanobacterial growth inhibition was accompanied by the increased synthesis of microcystin dm-RR and of the odorous metabolites, geosmin and 2-methylisoborneol (MIB), by P. agardhii 1113. NP also notably increased the microcystin released into the environment. Increased levels of extracellular proteins, soluble exopolysaccharides, microcystins and odorous metabolites may affect the microbial loop in aquatic ecosystems. An increased level of malondialdehyde (MDA) was indicative of the formation of free radicals in P. agardhii under NP stress, whereas increased levels of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and proline indicated the occurrence of a scavenging mechanism.

  17. Sorption and desorption studies of chromium(VI) from nonviable cyanobacterium Nostoc muscorum biomass

    Energy Technology Data Exchange (ETDEWEB)

    Gupta, V.K. [Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247667 (India)], E-mail: vinodfcy@iitr.ernet.in; Rastogi, A. [Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247667 (India)

    2008-06-15

    This communication presents results pertaining to the sorptive and desorptive studies carried out on chromium(VI) removal onto nonviable freshwater cyanobacterium (Nostoc muscorum) biomass. Influence of varying the conditions for removal of chromium(VI), such as the pH of aqueous solution, the dosage of biosorbent, the contact time with the biosorbent, the temperature for the removal of chromium, the effect of light metal ions and the adsorption-desorption studies were investigated. Sorption interaction of chromium on to cyanobacterial species obeyed both the first and the second-order rate equation and the experimental data showed good fit with both the Langmuir and freundlich adsorption isotherm models. The maximum adsorption capacity was 22.92 mg/g at 25 {sup o}C and pH 3.0. The adsorption process was endothermic and the values of thermodynamic parameters of the process were calculated. Various properties of the cyanobacterium, as adsorbent, explored in the characterization part were chemical composition of the adsorbent, surface area calculation by BET method and surface functionality by FTIR. Sorption-desorption of chromium into inorganic solutions and distilled water were observed and this indicated the biosorbent could be regenerated using 0.1 M HNO{sub 3} and EDTA with upto 80% recovery. The biosorbents were reused in five biosorption-desorption cycles without a significant loss in biosorption capacity. Thus, this study demonstrated that the cyanobacterial biomass N. muscorum could be used as an efficient biosorbent for the treatment of chromium(VI) bearing wastewater.

  18. Effect of UV-B on enzymes of nitrogen metabolism in the cyanobacterium Nostoc calcicola

    International Nuclear Information System (INIS)

    Kumar, A.; Sinha, R.P.; Häder, D. P.

    1996-01-01

    The effects of ultraviolet-B (UV-B; 280–315 nm) irradiation on nitrogenase and nitrate reductase (NR) activity have been studied in the filamentous and heterocystous N 2 -fixing cyanobacterium Nostoc calcicola. Exposure of cultures to UV-B (5W/m 2 ) for as little as 30 min caused complete inactivation of nitrogenase activity whereas nitrate reductase activity was stimulated twofold in comparison to one exposed to fluorescent white light. GS activity was also inhibited by UV-B treatment, but there was no total loss of activity even after 4 h. NR activity showed a gradual stimulation up to 4 h and thereafter it became constant. Stimulation was also obtained in reductant deficient cultures (12 h incubation in the dark) suggesting independence of NR of PS-II under UV-B. NR activity was also unaffected in the presence of DCMU, a known inhibitor of PS-II. However, both O 2 evolution and 14 CO 2 uptake were completely abolished following 30 min of UV-B treatment. Addition of the protein synthesis inhibitor chloramphenicol (25 μg/mL) to cultures did not show any inhibitory effect on NR activity. SDS-PAGE analysis of UV-B treated cultures elicited gradual loss of protein bands with increasing duration of exposure. Our findings suggest that UV-B irradiance has differential effects on the enzymes of the nitrogen metabolism in the cyanobacterium Nostoc calcicola. Further studies are needed to reveal the exact mechanism involved in the stimulation of NR activity by UV-B. Whether UV-B has a direct effect on NO 2 − accumulation in the cells needs detailed investigation. (author)

  19. Temporal Gene Expression of the Cyanobacterium Arthrospira in Response to Gamma Rays

    Science.gov (United States)

    Badri, Hanène; Monsieurs, Pieter; Coninx, Ilse; Nauts, Robin; Wattiez, Ruddy; Leys, Natalie

    2015-01-01

    The edible cyanobacterium Arthrospira is resistant to ionising radiation. The cellular mechanisms underlying this radiation resistance are, however, still largely unknown. Therefore, additional molecular analysis was performed to investigate how these cells can escape from, protect against, or repair the radiation damage. Arthrospira cells were shortly exposed to different doses of 60Co gamma rays and the dynamic response was investigated by monitoring its gene expression and cell physiology at different time points after irradiation. The results revealed a fast switch from an active growth state to a kind of 'survival modus' during which the cells put photosynthesis, carbon and nitrogen assimilation on hold and activate pathways for cellular protection, detoxification, and repair. The higher the radiation dose, the more pronounced this global emergency response is expressed. Genes repressed during early response, suggested a reduction of photosystem II and I activity and reduced tricarboxylic acid (TCA) and Calvin-Benson-Bassham (CBB) cycles, combined with an activation of the pentose phosphate pathway (PPP). For reactive oxygen species detoxification and restoration of the redox balance in Arthrospira cells, the results suggested a powerful contribution of the antioxidant molecule glutathione. The repair mechanisms of Arthrospira cells that were immediately switched on, involve mainly proteases for damaged protein removal, single strand DNA repair and restriction modification systems, while recA was not induced. Additionally, the exposed cells showed significant increased expression of arh genes, coding for a novel group of protein of unknown function, also seen in our previous irradiation studies. This observation confirms our hypothesis that arh genes are key elements in radiation resistance of Arthrospira, requiring further investigation. This study provides new insights into phasic response and the cellular pathways involved in the radiation resistance of

  20. Temporal Gene Expression of the Cyanobacterium Arthrospira in Response to Gamma Rays.

    Directory of Open Access Journals (Sweden)

    Hanène Badri

    Full Text Available The edible cyanobacterium Arthrospira is resistant to ionising radiation. The cellular mechanisms underlying this radiation resistance are, however, still largely unknown. Therefore, additional molecular analysis was performed to investigate how these cells can escape from, protect against, or repair the radiation damage. Arthrospira cells were shortly exposed to different doses of 60Co gamma rays and the dynamic response was investigated by monitoring its gene expression and cell physiology at different time points after irradiation. The results revealed a fast switch from an active growth state to a kind of 'survival modus' during which the cells put photosynthesis, carbon and nitrogen assimilation on hold and activate pathways for cellular protection, detoxification, and repair. The higher the radiation dose, the more pronounced this global emergency response is expressed. Genes repressed during early response, suggested a reduction of photosystem II and I activity and reduced tricarboxylic acid (TCA and Calvin-Benson-Bassham (CBB cycles, combined with an activation of the pentose phosphate pathway (PPP. For reactive oxygen species detoxification and restoration of the redox balance in Arthrospira cells, the results suggested a powerful contribution of the antioxidant molecule glutathione. The repair mechanisms of Arthrospira cells that were immediately switched on, involve mainly proteases for damaged protein removal, single strand DNA repair and restriction modification systems, while recA was not induced. Additionally, the exposed cells showed significant increased expression of arh genes, coding for a novel group of protein of unknown function, also seen in our previous irradiation studies. This observation confirms our hypothesis that arh genes are key elements in radiation resistance of Arthrospira, requiring further investigation. This study provides new insights into phasic response and the cellular pathways involved in the radiation

  1. Radiation characteristics and effective optical properties of dumbbell-shaped cyanobacterium Synechocystis sp

    International Nuclear Information System (INIS)

    Heng, Ri-Liang; Pilon, Laurent

    2016-01-01

    This study presents experimental measurements of the radiation characteristics of unicellular freshwater cyanobacterium Synechocystis sp. during their exponential growth in F medium. Their scattering phase function at 633 nm average spectral absorption and scattering cross-sections between 400 and 750 nm were measured. In addition, an inverse method was used for retrieving the spectral effective complex index of refraction of overlapping or touching bispheres and quadspheres from their absorption and scattering cross-sections. The inverse method combines a genetic algorithm and a forward model based on Lorenz–Mie theory, treating bispheres and quadspheres as projected area and volume-equivalent coated spheres. The inverse method was successfully validated with numerically predicted average absorption and scattering cross-sections of suspensions consisting of bispheres and quadspheres, with realistic size distributions, using the T-matrix method. It was able to retrieve the monomers' complex index of refraction with size parameter up to 11, relative refraction index less than 1.3, and absorption index less than 0.1. Then, the inverse method was applied to retrieve the effective spectral complex index of refraction of Synechocystis sp. approximated as randomly oriented aggregates consisting of two overlapping homogeneous spheres. Both the measured absorption cross-section and the retrieved absorption index featured peaks at 435 and 676 nm corresponding to chlorophyll a, a peak at 625 nm corresponding to phycocyanin, and a shoulder around 485 nm corresponding to carotenoids. These results can be used to optimize and control light transfer in photobioreactors. The inverse method and the equivalent coated sphere model could be applied to other optically soft particles of similar morphologies. - Highlights: • Radiation characteristics of Synechocystis sp. were measured during exponential growth. • This unicellular freshwater cyanobacterium features an interesting

  2. First report of neurotoxic effect of the cyanobacterium Cylindrospermopsis raciborskii on the motility of trematode metacercariae.

    Science.gov (United States)

    Lopes, K C; Ferrão-Filho, A S; Santos, E G N; Santos, C P

    2018-03-01

    Cylindrospermopsis raciborskii (Woloszynska) is a photosynthetic cyanobacterium that can produce cytotoxic (cylindrospermopsin) and neurotoxic cyanotoxins (saxitoxins). In Brazil the strains of C. raciborskii are reported to produce only saxitoxins (STX) and their effect on fish parasites has not been tested to date. The fish Poecilia vivipara Bloch and Schneider is a common host for the trematode Pygidiopsis macrostomum Travassos off the coast of Rio de Janeiro, and this fish-parasite interaction is a model for behavioural and ecotoxicological studies. The aim of this work was to evaluate the motility of metacercariae of P. macrostomum from P. vivipara exposed to 40 mg l-1 and 400 mg l-1 of crude lyophilized extract of the cyanobacterium C. raciborskii (CYRF-01) for 48 h. The fish were separated into groups of ten individuals and, after exposure, five fish from each group were dissected for counting and checking the motility of metacercariae. The other five fish were dissected after 48 h in clean water. The detection and quantification of STX in the solutions of cyanobacteria, and the gills and guts of fish, were performed by an enzyme-linked immunosorbent assay. The crude extract of C. raciborskii caused temporary paralysis in metacercariae of P. macrostomum after exposure of fish to both concentrations, and the motility recovered after the fish were kept for 48 h in clean water. STX was detected in the guts and gills of all fish analysed, suggesting that this toxin is involved in the paralysis of metacercariae. This is the first report on the action of neurotoxins in metacercariae of fish.

  3. Differential responses of dinitrogen fixation, diazotrophic cyanobacteria and ammonia oxidation reveal a potential warming-induced imbalance of the N-cycle in biological soil crusts

    Science.gov (United States)

    Zhou, Xiaobing; Smith, Hilda J.; Giraldo Silva, Ana; Belnap, Jayne; Garcia-Pichel, Ferran

    2017-01-01

    N2 fixation and ammonia oxidation (AO) are the two most important processes in the nitrogen (N) cycle of biological soil crusts (BSCs). We studied the short-term response of acetylene reduction assay (ARA) rates, an indicator of potential N2 fixation, and AO rates to temperature (T, -5°C to 35°C) in BSC of different successional stages along the BSC ecological succession and geographic origin (hot Chihuahuan and cooler Great Basin deserts). ARA in all BSCs increased with T until saturation occurred between 15 and 20°C, and declined at 30–35°C. Culture studies using cyanobacteria isolated from these crusts indicated that the saturating effect was traceable to their inability to grow well diazotrophically within the high temperature range. Below saturation, temperature response was exponential, with Q10 significantly different in the two areas (~ 5 for Great Basin BSCs; 2–3 for Chihuahuan BSCs), but similar between the two successional stages. However, in contrast to ARA, AO showed a steady increase to 30–35°C in Great Basin, and Chihuhuan BSCs showed no inhibition at any tested temperature. The T response of AO also differed significantly between Great Basin (Q10 of 4.5–4.8) and Chihuahuan (Q10 of 2.4–2.6) BSCs, but not between successional stages. Response of ARA rates to T did not differ from that of AO in either desert. Thus, while both processes scaled to T in unison until 20°C, they separated to an increasing degree at higher temperature. As future warming is likely to occur in the regions where BSCs are often the dominant living cover, this predicted decoupling is expected to result in higher proportion of nitrates in soil relative to ammonium. As nitrate is more easily lost as leachate or to be reduced to gaseous forms, this could mean a depletion of soil N over large landscapes globally.

  4. Piecewise Structural Equation Model (SEM) Disentangles the Environmental Conditions Favoring Diatom Diazotroph Associations (DDAs) in the Western Tropical North Atlantic (WTNA).

    Science.gov (United States)

    Stenegren, Marcus; Berg, Carlo; Padilla, Cory C; David, Stefan-Sebastian; Montoya, Joseph P; Yager, Patricia L; Foster, Rachel A

    2017-01-01

    Diatom diazotroph associations (DDAs) are important components in the world's oceans, especially in the western tropical north Atlantic (WTNA), where blooms have a significant impact on carbon and nitrogen cycling. However, drivers of their abundances and distribution patterns remain unknown. Here, we examined abundance and distribution patterns for two DDA populations in relation to the Amazon River (AR) plume in the WTNA. Quantitative PCR assays, targeting two DDAs (het-1 and het-2) by their symbiont's nifH gene, served as input in a piecewise structural equation model (SEM). Collections were made during high (spring 2010) and low (fall 2011) flow discharges of the AR. The distributions of dissolved nutrients, chlorophyll- a , and DDAs showed coherent patterns indicative of areas influenced by the AR. A symbiotic Hemiaulus hauckii-Richelia (het-2) bloom (>10 6 cells L -1 ) occurred during higher discharge of the AR and was coincident with mesohaline to oceanic (30-35) sea surface salinities (SSS), and regions devoid of dissolved inorganic nitrogen (DIN), low concentrations of both DIP (>0.1 μmol L -1 ) and Si (>1.0 μmol L -1 ). The Richelia (het-1) associated with Rhizosolenia was only present in 2010 and at lower densities (10-1.76 × 10 5 nifH copies L -1 ) than het-2 and limited to regions of oceanic SSS (>36). The het-2 symbiont detected in 2011 was associated with H. membranaceus (>10 3 nifH copies L -1 ) and were restricted to regions with mesohaline SSS (31.8-34.3), immeasurable DIN, moderate DIP (0.1-0.60 μmol L -1 ) and higher Si (4.19-22.1 μmol L -1 ). The piecewise SEM identified a profound direct negative effect of turbidity on the het-2 abundance in spring 2010, while DIP and water turbidity had a more positive influence in fall 2011, corroborating our observations of DDAs at subsurface maximas. We also found a striking difference in the influence of salinity on DDA symbionts suggesting a niche differentiation and preferences in oceanic and

  5. Piecewise Structural Equation Model (SEM Disentangles the Environmental Conditions Favoring Diatom Diazotroph Associations (DDAs in the Western Tropical North Atlantic (WTNA

    Directory of Open Access Journals (Sweden)

    Marcus Stenegren

    2017-05-01

    Full Text Available Diatom diazotroph associations (DDAs are important components in the world’s oceans, especially in the western tropical north Atlantic (WTNA, where blooms have a significant impact on carbon and nitrogen cycling. However, drivers of their abundances and distribution patterns remain unknown. Here, we examined abundance and distribution patterns for two DDA populations in relation to the Amazon River (AR plume in the WTNA. Quantitative PCR assays, targeting two DDAs (het-1 and het-2 by their symbiont’s nifH gene, served as input in a piecewise structural equation model (SEM. Collections were made during high (spring 2010 and low (fall 2011 flow discharges of the AR. The distributions of dissolved nutrients, chlorophyll-a, and DDAs showed coherent patterns indicative of areas influenced by the AR. A symbiotic Hemiaulus hauckii-Richelia (het-2 bloom (>106 cells L-1 occurred during higher discharge of the AR and was coincident with mesohaline to oceanic (30–35 sea surface salinities (SSS, and regions devoid of dissolved inorganic nitrogen (DIN, low concentrations of both DIP (>0.1 μmol L-1 and Si (>1.0 μmol L-1. The Richelia (het-1 associated with Rhizosolenia was only present in 2010 and at lower densities (10-1.76 × 105nifH copies L-1 than het-2 and limited to regions of oceanic SSS (>36. The het-2 symbiont detected in 2011 was associated with H. membranaceus (>103nifH copies L-1 and were restricted to regions with mesohaline SSS (31.8–34.3, immeasurable DIN, moderate DIP (0.1–0.60 μmol L-1 and higher Si (4.19–22.1 μmol L-1. The piecewise SEM identified a profound direct negative effect of turbidity on the het-2 abundance in spring 2010, while DIP and water turbidity had a more positive influence in fall 2011, corroborating our observations of DDAs at subsurface maximas. We also found a striking difference in the influence of salinity on DDA symbionts suggesting a niche differentiation and preferences in oceanic and mesohaline

  6. Draft Genome Sequence of Leptolyngbya sp. KIOST-1, a Filamentous Cyanobacterium with Biotechnological Potential for Alimentary Purposes.

    Science.gov (United States)

    Kim, Ji Hyung; Kang, Do-Hyung

    2016-09-15

    Here, we report the draft genome of cyanobacterium Leptolyngbya sp. KIOST-1 isolated from a microalgal culture pond in South Korea. The genome consists of 13 contigs containing 6,320,172 bp, and a total of 5,327 coding sequences were predicted. This genomic information will allow further exploitation of its biotechnological potential for alimentary purposes. Copyright © 2016 Kim and Kang.

  7. Resistance to the photosystem II herbicide diuron is dominant to sensitivity in the cyanobacterium Synechococcus sp. PCC7942

    OpenAIRE

    Brusslan, Judy; Haselkorn, Robert

    1989-01-01

    The transformable cyanobacterium, Synechococcus sp. PCC7942, was used to study the genetics of resistance to the herbicide diuron. In wild-type cells, diuron binds to one of the core proteins, called D1, of photosystem II reaction centres. This binding prevents the transfer of electrons from QA, the primary quinone acceptor, to QB, which is necessary to create the charge separation that drives ATP synthesis. A single amino acid substitution in the D1 protein reduces diuron binding and confers...

  8. Identification of the n-1 fatty acid as an antibacterial constituent from the edible freshwater cyanobacterium Nostoc verrucosum.

    Science.gov (United States)

    Oku, Naoya; Yonejima, Kohsuke; Sugawa, Takao; Igarashi, Yasuhiro

    2014-01-01

    The cyanobacterium Nostoc verrucosum occurs in cool, clear streams and its gelatinous colonies, called "ashitsuki," have been eaten in ancient Japan. Its ethanolic extract was found to inhibit the growth of Gram-positive bacteria and activity-guided fractionation yielded an unusual n-1 fatty acid, (9Z,12Z)-9,12,15-hexadecatrienoic acid (1), as one of the active principles. It inhibited the growth of Staphylococcus aureus at MIC 64 μg/mL.

  9. Discovery of Rare and Highly Toxic Microcystins from Lichen-Associated Cyanobacterium Nostoc sp. Strain IO-102-I

    OpenAIRE

    Oksanen, Ilona; Jokela, Jouni; Fewer, David P.; Wahlsten, Matti; Rikkinen, Jouko; Sivonen, Kaarina

    2004-01-01

    The production of hepatotoxic cyclic heptapeptides, microcystins, is almost exclusively reported from planktonic cyanobacteria. Here we show that a terrestrial cyanobacterium Nostoc sp. strain IO-102-I isolated from a lichen association produces six different microcystins. Microcystins were identified with liquid chromatography-UV mass spectrometry by their retention times, UV spectra, mass fragmentation, and comparison to microcystins from the aquatic Nostoc sp. strain 152. The dominant micr...

  10. Significance of diazotrophic plant growth-promoting Herbaspirillum sp. GW103 on phytoextraction of Pband Zn by Zea mays L.

    Science.gov (United States)

    Praburaman, Loganathan; Park, Sung-Hee; Cho, Min; Lee, Kui-Jae; Ko, Jeong-Ae; Han, Sang-Sub; Lee, Sang-Hyun; Kamala-Kannan, Seralathan; Oh, Byung-Taek

    2017-01-01

    Microbe-assisted phytoremediation has been considered a promising measure for the remediation of heavy metal-polluted soil. The aim of this study was to assess the effect of diazotrophic plant growth-promoting Herbaspirillum sp. GW103 on growth and lead (Pb) and zinc (Zn) accumulation in Zea mays L. The strain GW103 exhibited plant growth-promoting traits such as indole-3-acetic acid, siderophores, and 1-aminocyclopropane-1-carboxylic deaminase. Treatment of Z. mays L. plants with GW103 significantly increased 19, 31, and 52% of plant biomass and 10, 50, and 126% of chlorophyll a contents in Pb, Zn, and Pb + Zn-amended soils, respectively. Similarly, the strain GW103 significantly increased Pb and Zn accumulation in shoots and roots of Z. mays L., which were 77 and 25% in Pb-amended soil, 42 and 73% in Zn-amended soil, and 27 and 84% in Pb + Zn-amended soil. Furthermore, addition of GW103 increased 8, 12, and 7% of total protein content, catalase, and superoxide dismutase levels, respectively, in Z. mays L. plants. The results pointed out that isolate GW103 could potentially reduce the phytotoxicity of metals and increase Pb and Zn accumulation in Z. mays L. plant.

  11. Analysis of a diverse assemblage of diazotrophic bacteria from Spartina alterniflora using DGGE and clone library screening.

    Science.gov (United States)

    Lovell, Charles R; Decker, Peter V; Bagwell, Christopher E; Thompson, Shelly; Matsui, George Y

    2008-05-01

    Methods to assess the diversity of the diazotroph assemblage in the rhizosphere of the salt marsh cordgrass, Spartina alterniflora were examined. The effectiveness of nifH PCR-denaturing gradient gel electrophoresis (DGGE) was compared to that of nifH clone library analysis. Seventeen DGGE gel bands were sequenced and yielded 58 nonidentical nifH sequences from a total of 67 sequences determined. A clone library constructed using the GC-clamp nifH primers that were employed in the PCR-DGGE (designated the GC-Library) yielded 83 nonidentical sequences from a total of 257 nifH sequences. A second library constructed using an alternate set of nifH primers (N-Library) yielded 83 nonidentical sequences from a total of 138 nifH sequences. Rarefaction curves for the libraries did not reach saturation, although the GC-Library curve was substantially dampened and appeared to be closer to saturation than the N-Library curve. Phylogenetic analyses showed that DGGE gel band sequencing recovered nifH sequences that were frequently sampled in the GC-Library, as well as sequences that were infrequently sampled, and provided a species composition assessment that was robust, efficient, and relatively inexpensive to obtain. Further, the DGGE method permits a large number of samples to be examined for differences in banding patterns, after which bands of interest can be sampled for sequence determination.

  12. Potential for hydrogen-oxidizing chemolithoautotrophic and diazotrophic populations to initiate biofilm formation in oligotrophic, deep terrestrial subsurface waters.

    Science.gov (United States)

    Wu, Xiaofen; Pedersen, Karsten; Edlund, Johanna; Eriksson, Lena; Åström, Mats; Andersson, Anders F; Bertilsson, Stefan; Dopson, Mark

    2017-03-23

    Deep terrestrial biosphere waters are separated from the light-driven surface by the time required to percolate to the subsurface. Despite biofilms being the dominant form of microbial life in many natural environments, they have received little attention in the oligotrophic and anaerobic waters found in deep bedrock fractures. This study is the first to use community DNA sequencing to describe biofilm formation under in situ conditions in the deep terrestrial biosphere. In this study, flow cells were attached to boreholes containing either "modern marine" or "old saline" waters of different origin and degree of isolation from the light-driven surface of the earth. Using 16S rRNA gene sequencing, we showed that planktonic and attached populations were dissimilar while gene frequencies in the metagenomes suggested that hydrogen-fed, carbon dioxide- and nitrogen-fixing populations were responsible for biofilm formation across the two aquifers. Metagenome analyses further suggested that only a subset of the populations were able to attach and produce an extracellular polysaccharide matrix. Initial biofilm formation is thus likely to be mediated by a few bacterial populations which were similar to Epsilonproteobacteria, Deltaproteobacteria, Betaproteobacteria, Verrucomicrobia, and unclassified bacteria. Populations potentially capable of attaching to a surface and to produce extracellular polysaccharide matrix for attachment were identified in the terrestrial deep biosphere. Our results suggest that the biofilm populations were taxonomically distinct from the planktonic community and were enriched in populations with a chemolithoautotrophic and diazotrophic metabolism coupling hydrogen oxidation to energy conservation under oligotrophic conditions.

  13. Most hydrocarbonoclastic bacteria in the total environment are diazotrophic, which highlights their value in the bioremediation of hydrocarbon contaminants.

    Science.gov (United States)

    Dashti, Narjes; Ali, Nedaa; Eliyas, Mohamed; Khanafer, Majida; Sorkhoh, Naser A; Radwan, Samir S

    2015-01-01

    Eighty-two out of the 100 hydrocarbonoclastic bacterial species that have been already isolated from oil-contaminated Kuwaiti sites, characterized by 16S rRNA nucleotide sequencing, and preserved in our private culture collection, grew successfully in a mineral medium free of any nitrogenous compounds with oil vapor as the sole carbon source. Fifteen out of these 82 species were selected for further study based on the predominance of most of the isolates in their specific sites. All of these species tested positive for nitrogenase using the acetylene reduction reaction. They belonged to the genera Agrobacterium, Sphingomonas, and Pseudomonas from oily desert soil and Nesiotobacter, Nitratireductor, Acinetobacter, Alcanivorax, Arthrobacter, Marinobacter, Pseudoalteromonas, Vibrio, Diatzia, Mycobacterium, and Microbacterium from the Arabian/Persian Gulf water body. A PCR-DGGE-based sequencing analysis of nifH genes revealed the common occurrence of the corresponding genes among all the strains tested. The tested species also grew well and consumed crude oil effectively in NaNO3 -containing medium with and without nitrogen gas in the top space. On the other hand, these bacteria only grew and consumed crude oil in the NaNO3 -free medium when the top space gas contained nitrogen. We concluded that most hydrocarbonoclastic bacteria are diazotrophic, which allows for their wide distribution in the total environment. Therefore, these bacteria are useful for the cost-effective, environmentally friendly bioremediation of hydrocarbon contaminants.

  14. Oil-bioremediation potential of two hydrocarbonoclastic, diazotrophic Marinobacter strains from hypersaline areas along the Arabian Gulf coasts.

    Science.gov (United States)

    Al-Mailem, D M; Eliyas, M; Radwan, S S

    2013-05-01

    Two halophilic, hydrocarbonoclastics bacteria, Marinobacter sedimentarum and M. flavimaris, with diazotrophic potential occured in hypersaline waters and soils in southern and northern coasts of Kuwait. Their numbers were in the magnitude of 10(3) colony forming units g(-1). The ambient salinity in the hypersaline environments was between 3.2 and 3.5 M NaCl. The partial 16S rRNA gene sequences of the two strains showed, respectively, 99 and 100% similarities to the sequences in the GenBank. The two strains failed to grow in the absence of NaCl, exhibited best growth and hydrocarbon biodegradation in the presence of 1 to 1.5 M NaCl, and still grew and maintained their hydrocarbonoclastic activity at salinities up to 5 M NaCl. Both species utilized Tween 80, a wide range of individual aliphatic hydrocarbons (C9-C40) and the aromatics benzene, biphenyl, phenanthrene, anthracene and naphthalene as sole sources of carbon and energy. Experimental evidence was provided for their nitrogen-fixation potential. The two halophilic Marinobacter strains successfully mineralized crude oil in nutrient media as well as in hypersaline soil and water microcosms without the use of any nitrogen fertilizers.

  15. Structural Interaction Between GFP-Labeled Diazotrophic Endophytic Bacterium Herbaspirillum seropedicae RAM10 and Pineapple Plantlets 'VitóRia'.

    Science.gov (United States)

    Estrela Borges Baldotto, Lílian; Lopes Olivares, Fábio; Bressan-Smith, Ricardo

    2011-01-01

    The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets 'Vitória' were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae.

  16. Structural interaction between GFP-labeled diazotrophic endophytic bacterium Herbaspirillum seropedicae RAM10 and pineapple plantlets 'Vitória'

    Directory of Open Access Journals (Sweden)

    Lílian Estrela Borges Baldotto

    2011-03-01

    Full Text Available The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets 'Vitória' were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae.

  17. Short Communication: Effects of temperature on growth, pigment composition and protein content of an Antarctic Cyanobacterium Nostoc commune

    Directory of Open Access Journals (Sweden)

    RANJANA TRIPATHI

    2012-11-01

    Full Text Available Tripathi R, Dhuldhaj UP, Singh S. 2012. Short Communication: Effects of temperature on growth, pigment composition and protein content of an Antarctic Cyanobacterium Nostoc commune. Nusantara Bioscience 4: 134-137. Effect of temperature variation on biomass accumulation, pigment composition and protein content were studied for the cyanobacterium Nostoc commune, isolated from Antarctica. Results confirmed the psychrotrophic behavior (optimum growth temperature 25◦C of the cyanobacterium. Low temperature increased the duration of lag phase and exponential growth phase. Maximum increase in biomass was recorded on 24th day at 25◦C and on 12th day at 50C. The downshift from 25 to 5◦C had almost negligible effect on chl a content. Maximal protein content was recorded for cultures growing at 50C on 12th day. The carotenoids/chl a ratio was maximum (2.48 at 50C on 9th day. It remained almost constant for cultures growing at 5 and 350C. There was an induction in protein synthesis following downshift in temperature from 25 to 5◦C.

  18. Programmed cell death in the marine cyanobacterium Trichodesmium mediates carbon and nitrogen export

    Science.gov (United States)

    Bar-Zeev, Edo; Avishay, Itamar; Bidle, Kay D; Berman-Frank, Ilana

    2013-01-01

    The extent of carbon (C) and nitrogen (N) export to the deep ocean depends upon the efficacy of the biological pump that transports primary production to depth, thereby preventing its recycling in the upper photic zone. The dinitrogen-fixing (diazotrophic) Trichodesmium spp. contributes significantly to oceanic C and N cycling by forming extensive blooms in nutrient-poor tropical and subtropical regions. These massive blooms generally collapse several days after forming, but the cellular mechanism responsible, along with the magnitude of associated C and N export processes, are as yet unknown. Here, we used a custom-made, 2-m high water column to simulate a natural bloom and to specifically test and quantify whether the programmed cell death (PCD) of Trichodesmium mechanistically regulates increased vertical flux of C and N. Our findings demonstrate that extremely rapid development and abrupt, PCD-induced demise (within 2–3 days) of Trichodesmium blooms lead to greatly elevated excretions of transparent exopolymers and a massive downward pulse of particulate organic matter. Our results mechanistically link autocatalytic PCD and bloom collapse to quantitative C and N export fluxes, suggesting that PCD may have an impact on the biological pump efficiency in the oceans. PMID:23887173

  19. Molecular exploration of the highly radiation resistant cyanobacterium Arthrospira sp. PCC 8005

    Science.gov (United States)

    Badri, Hanène; Leys, Natalie; Wattiez, Ruddy

    Arthrospira (Spirulina) is a photosynthetic cyanobacterium able to use sunlight to release oxygen from water and remove carbon dioxide and nitrate from water. In addition, it is suited for human consumption (edible). For these traits, the cyanobacterium Arthrospira sp. PCC 8005 was selected by the European Space Agency (ESA) as part of the life support system MELiSSA for recycling oxygen, water, and food during future long-haul space missions. However, during such extended missions, Arthrospira sp. PCC 8005 will be exposed to continuous artificial illumination and harmful cosmic radiation. The aim of this study was to investigate how Arthrospira will react and behave when exposed to such stress environment. The cyanobacterium Arthrospira sp. PCC 8005 was exposed to high gamma rays doses in order to unravel in details the response of this bacterium following such stress. Test results showed that after acute exposure to high doses of 60Co gamma radiation upto 3200 Gy, Arthrospira filaments were still able to restart photosynthesis and proliferate normally. Doses above 3200 Gy, did have a detrimental effect on the cells, and delayed post-irradiation proliferation. The photosystem activity, measured as the PSII quantum yield immediately after irradiation, decreased significantly at radiation doses above 3200 Gy. Likewise through pigment content analysis a significant decrease in phycocyanin was observed following exposure to 3200 Gy. The high tolerance of this bacterium to 60Co gamma rays (i.e. ca. 1000x more resistant than human cells for example) raised our interest to investigate in details the cellular and molecular mechanisms behind this amazing resistance. Optimised DNA, RNA and protein extraction methods and a new microarray chip specific for Arthrospira sp. PCC 8005 were developed to identify the global cellular and molecular response following exposure to 3200 Gy and 5000 Gy A total of 15,29 % and 30,18 % genes were found differentially expressed in RNA

  20. Lack of Methylated Hopanoids Renders the Cyanobacterium Nostoc punctiforme Sensitive to Osmotic and pH Stress.

    Science.gov (United States)

    Garby, Tamsyn J; Matys, Emily D; Ongley, Sarah E; Salih, Anya; Larkum, Anthony W D; Walter, Malcolm R; Summons, Roger E; Neilan, Brett A

    2017-07-01

    To investigate the function of 2-methylhopanoids in modern cyanobacteria, the hpnP gene coding for the radical S -adenosyl methionine (SAM) methylase protein that acts on the C-2 position of hopanoids was deleted from the filamentous cyanobacterium Nostoc punctiforme ATCC 29133S. The resulting Δ hpnP mutant lacked all 2-methylhopanoids but was found to produce much higher levels of two bacteriohopanepentol isomers than the wild type. Growth rates of the Δ hpnP mutant cultures were not significantly different from those of the wild type under standard growth conditions. Akinete formation was also not impeded by the absence of 2-methylhopanoids. The relative abundances of the different hopanoid structures in akinete-dominated cultures of the wild-type and Δ hpnP mutant strains were similar to those of vegetative cell-dominated cultures. However, the Δ hpnP mutant was found to have decreased growth rates under both pH and osmotic stress, confirming a role for 2-methylhopanoids in stress tolerance. Evidence of elevated photosystem II yield and NAD(P)H-dependent oxidoreductase activity in the Δ hpnP mutant under stress conditions, compared to the wild type, suggested that the absence of 2-methylhopanoids increases cellular metabolic rates under stress conditions. IMPORTANCE As the first group of organisms to develop oxygenic photosynthesis, Cyanobacteria are central to the evolutionary history of life on Earth and the subsequent oxygenation of the atmosphere. To investigate the origin of cyanobacteria and the emergence of oxygenic photosynthesis, geobiologists use biomarkers, the remnants of lipids produced by different organisms that are found in geologic sediments. 2-Methylhopanes have been considered indicative of cyanobacteria in some environmental settings, with the parent lipids 2-methylhopanoids being present in many contemporary cyanobacteria. We have created a Nostoc punctiforme Δ hpnP mutant strain that does not produce 2-methylhopanoids to assess the

  1. [Growth and metabolite production of the marine cyanobacterium Synechococcus sp. (Chroococcales) in function to irradiance].

    Science.gov (United States)

    Rosales-Loaiza, Néstor; Guevara, Miguel; Lodeiros, César; Morales, Ever

    2008-06-01

    Changes in salinity, temperature and irradiance during wet and dry seasons have induced metabolic versatility in cyanobacteria from saline environments. Cyanobacteria from these environments have biotechnological potential for the production of metabolites with pharmaceutical and industrial interest. We studied the growth, dry mass and metabolite production of the cyanobacterium Synechococcus sp. MOF-03 in function of irradiance (78, 156 and 234 micromol q m(-2) s(-1)). All batch cultures were maintained by triplicate in constant aeration, 12:12 h photoperiod, 30 +/- 2 degrees C and 35% per hundred. Maximum values of protein, carbohydrates and lipids, of 530.19 +/- 11.16, 408.94 +/- 4.27 and 56.20 +/- 1.17 microg ml(-1), respectively, were achieved at 78 micromol q m(-2) s(-1). Pigments, analyzed by HPLC, showed maximum values at 78 micromol q m(-2) s(-1) for chlorophyll a with 7.72 +/- 0.16 microg ml(-1), and at 234 micromol q m(-2) s(-1) for beta-carotene and zeaxanthin with 0.70 +/- 0.01 and 0.67 +/- 0.05 microg ml(-1). Chlorophyll a:beta-carotene ratio decreased from 17.15 to 6.91 at 78 and 234 micromol q m(-2) s(-'1); whereas beta-carotene:zeaxanthin ratio showed no changes between 78 and 156 micromol q m(-2) s(-1), around 1.21, and decreased at 234 micromol q m(-2) s(-1), to 1.04. Also, this cyanobacterium produced the greatest cell density and dry mass at 156 micromol q m(-2) s(-1), with 406.13 +/- 21.74 x l0(6) cell ml(-1) and 1.49 +/- 0.11 mg ml(-1), respectively. Exopolysaccharide production was stable between 156 y 234 micromol q m(-2) s(-1), around 110 microg ml(-1). This Synechococcus strain shows a great potential for the production of enriched biomass with high commercial value metabolites.

  2. Evidence for paralytic shellfish poisons in the freshwater cyanobacterium Lyngbya wollei (Farlow ex Gomont) comb. nov.

    Science.gov (United States)

    Carmichael, W W; Evans, W R; Yin, Q Q; Bell, P; Moczydlowski, E

    1997-08-01

    Lyngbya wollei (Farlow ex Gomont) comb. nov., a perennial mat-forming filamentous cyanobacterium prevalent in lakes and reservoirs of the southeastern United States, was found to produce a potent, acutely lethal neurotoxin when tested in the mouse bioassay. Signs of poisoning were similar to those of paralytic shellfish poisoning. As part of the Tennessee Valley Authority master plan for Guntersville Reservoir, the mat-forming filamentous cyanobacterium L. wollei, a species that had recently invaded from other areas of the southern United States, was studied to determine if it could produce any of the known cyanotoxins. Of the 91 field samples collected at 10 locations at Guntersville Reservoir, Ala., on the Tennessee River, over a 3-year period, 72.5% were toxic. The minimum 100% lethal doses of the toxic samples ranged from 150 to 1,500 mg kg of lyophilized L. wollei cells-1, with the majority of samples being toxic at 500 mg kg-1. Samples bioassayed for paralytic shellfish toxins by the Association of Official Analytical Chemists method exhibited saxitoxin equivalents ranging from 0 to 58 micrograms g (dry weight)-1. Characteristics of the neurotoxic compound(s), such as the lack of adsorption by C18 solid-phase extraction columns, the short retention times on C18 high-performance liquid chromatography (HPLC) columns, the interaction of the neurotoxins with saxiphilin (a soluble saxitoxin-binding protein), and external blockage of voltage-sensitive sodium channels, led to our discovery that this neurotoxin(s) is related to the saxitoxins, the compounds responsible for paralytic shellfish poisonings. The major saxitoxin compounds thus far identified by comparison of HPLC fluorescence retention times are decarbamoyl gonyautoxins 2 and 3. There was no evidence of paralytic shellfish poison C toxins being produced by L. wollei. Fifty field samples were placed in unialgal culture and grown under defined culture conditions. Toxicity and signs of poisoning for these

  3. Molecular Cloning and Biochemical Characterization of the Iron Superoxide Dismutase from the Cyanobacterium Nostoc punctiforme ATCC 29133 and Its Response to Methyl Viologen-Induced Oxidative Stress.

    Science.gov (United States)

    Moirangthem, Lakshmipyari Devi; Ibrahim, Kalibulla Syed; Vanlalsangi, Rebecca; Stensjö, Karin; Lindblad, Peter; Bhattacharya, Jyotirmoy

    2015-12-01

    Superoxide dismutase (SOD) detoxifies cell-toxic superoxide radicals and constitutes an important component of antioxidant machinery in aerobic organisms, including cyanobacteria. The iron-containing SOD (SodB) is one of the most abundant soluble proteins in the cytosol of the nitrogen-fixing cyanobacterium Nostoc punctiforme ATCC 29133, and therefore, we investigated its biochemical properties and response to oxidative stress. The putative SodB-encoding open reading frame Npun_R6491 was cloned and overexpressed in Escherichia coli as a C-terminally hexahistidine-tagged protein. The purified recombinant protein had a SodB specific activity of 2560 ± 48 U/mg protein at pH 7.8 and was highly thermostable. The presence of a characteristic iron absorption peak at 350 nm, and its sensitivity to H2O2 and azide, confirmed that the SodB is an iron-containing SOD. Transcript level of SodB in nitrogen-fixing cultures of N. punctiforme decreased considerably (threefold) after exposure to an oxidative stress-generating herbicide methyl viologen for 4 h. Furthermore, in-gel SOD activity analysis of such cultures grown at increasing concentrations of methyl viologen also showed a loss of SodB activity. These results suggest that SodB is not the primary scavenger of superoxide radicals induced by methyl viologen in N. punctiforme.

  4. Chromatic regulation in cyanobacterium as studies by HPLC quantitation of photosynthetic pigments. Kogosei shikiso no HPLC teiryo ni motozuku ranso no hikari tekio process tsuiseki

    Energy Technology Data Exchange (ETDEWEB)

    Maeda, H.; Watanabe, T. (The Univ. Tokyo, Tokyo (Japan). Inst. of Industrial Science)

    1991-08-01

    Plants higher than Cyanobacterium have two kinds of resction centers(RC) which convert photon energy to a flow of electrons and whose photosensitive spectral regions are slightly deviated from each other. In the photosynthetic process, the ratio of numbers between these two kinds of reaction centers is adaptively varied so as to allow the overall flow of electrons to proceed in a well-balanced manner. It is important to rapidly and exactly determine the ratio of RC numbers between the two photochemical systems in order to investigate such photoadaptive process. The report describes the quantitative determination using high performance liquid chromatography(HPLC) for this purpose. Pigments were extracted from Cyanobacteria which are in different adaptive processes brought by being cultured in the environments differing in the quantity of light or in the environment of varying quantity of light, and subjected to quantitave determination in consideration of the fact that the reaction centers, I and II, have the respective special kinds of chlorophyl derivatives Chl-a, Chl-a{prime}. As the results, it was confirmed that validity can be given to the estimation of the numbers of reaction centers in terms of the quantities of Chl-a and Chl-a prime and the proposed method is drastically faster and simpler than the conventional methods. 14 refs., 5 figs..

  5. Growth Kinetics of Diazotrophic Bacillus sphaericus UPMB10 Cultured Using Different Types and Concentrations of Carbon and Nitrogen Sources

    Directory of Open Access Journals (Sweden)

    Ooi, T. C.

    2008-01-01

    Full Text Available Growth kinetics of newly isolated diazotrophic Bacillus sphaericus UPMB10 grown in various carbon (lactate, acetate, glycerol, malate, fructose, xylose and sucrose and nitrogen (glutamate, yeast extract, arginine, hystadine, glycine, polypeptone, tryptophan, lysine, NH4Cl and urea sources was investigated using 2 L stirred tank fermenter. The highest growth was obtained in a medium containing lactate as a carbon source, which gave the highest maximum cell concentration of 2.30 g/L, which is corresponding to maximum viable cell count of 4.60 x 10^9 cfu/mL. However, the highest cell yield (1.06 g cell/g carbon consumed was obtained in cultivation using glycerol though slightly lower maximum viable cell count was obtained (3.22 x 10^9 cfu/mL. In addition, cost for the production of live cell using glycerol was about 15 times lower than the cost using lactate. Growth performance of this bacterium when yeast extract was used as a nitrogen source was comparable to the use of pure amino acid. The medium containing 1.8 g/L glycerol and 2 g/L yeast extract was suggested as optimal for growth of this bacterium, which gave carbon to nitrogen ratio (C/N of 10:1. The maximum viable cell count obtained in cultivation using optimised medium in 2 L stirred tank fermenter was 3.34 x 10^9 cfu/mL and the cells maintained its capacity for N2 fixation at 18 nmol C2H2/h.mL.

  6. Effect of carbon and nitrogen assimilation on chlorophyll fluorescence emission by the cyanobacterium Anacystis nidulans

    Energy Technology Data Exchange (ETDEWEB)

    Romero, J.M.; Lara, C. (Instituto de Bioquimica Vegetal y Fotosintesis, Univ. de Sevilla y CSIC, Sevilla (ES)); Sivak, M.N. (Dept. of Biochemistry, Michigan State Univ., East Lansing (US))

    1992-01-01

    O{sub 2} evolution and chlorophyll A fluorescence emission have been monitored in intact cells of the cyanobacterium Anacystis nidulans 1402-1 to study the influence of carbon and nitrogen assimilation on the operation of the photosynthetic apparatus. The pattern of fluorescence induction in dark-adapted cyanobacterial cells was different from that of higher plants. Cyanobacteria undergo large, rapid state transitions upon illumination, which lead to marked changes in the fluorescence yield, complicating the estimation of quenching coefficients. The Kautsky effect was not evident, although it could be masked by a state II-state I transition, upon illumination with actinic light. The use of inhibitors of carbon assimilation such as D,L-glyceraldehyde or iodoacetamide allowed us to relate changes in variable fluorescence to active CO{sub 2} fixation. Ammonium, but not nitrate, induced non-photochemical fluorescence quenching, in agreement with a previous report on green algae, indicative of an ammonium-induced state i transition. (au).

  7. Anoxygenic Photosynthesis Controls Oxygenic Photosynthesis in a Cyanobacterium from a Sulfidic Spring

    KAUST Repository

    Klatt, Judith M.

    2015-03-15

    Before the Earth\\'s complete oxygenation (0.58 to 0.55 billion years [Ga] ago), the photic zone of the Proterozoic oceans was probably redox stratified, with a slightly aerobic, nutrient-limited upper layer above a light-limited layer that tended toward euxinia. In such oceans, cyanobacteria capable of both oxygenic and sulfide-driven anoxygenic photosynthesis played a fundamental role in the global carbon, oxygen, and sulfur cycle. We have isolated a cyanobacterium, Pseudanabaena strain FS39, in which this versatility is still conserved, and we show that the transition between the two photosynthetic modes follows a surprisingly simple kinetic regulation controlled by this organism\\'s affinity for H2S. Specifically, oxygenic photosynthesis is performed in addition to anoxygenic photosynthesis only when H2S becomes limiting and its concentration decreases below a threshold that increases predictably with the available ambient light. The carbon-based growth rates during oxygenic and anoxygenic photosynthesis were similar. However, Pseudanabaena FS39 additionally assimilated NO3 - during anoxygenic photosynthesis. Thus, the transition between anoxygenic and oxygenic photosynthesis was accompanied by a shift of the C/N ratio of the total bulk biomass. These mechanisms offer new insights into the way in which, despite nutrient limitation in the oxic photic zone in the mid-Proterozoic oceans, versatile cyanobacteria might have promoted oxygenic photosynthesis and total primary productivity, a key step that enabled the complete oxygenation of our planet and the subsequent diversification of life.

  8. Genetic analysis of amino acid transport in the facultatively heterotrophic cyanobacterium Synechocystis sp. Strain 6803

    International Nuclear Information System (INIS)

    Labarre, J.; Thuriaux, P.; Chauvat, F.

    1987-01-01

    The existence of active transport systems (permeases) operating on amino acids in the photoautotrophic cyanobacterium Synechocystis sp. strain 6803 was demonstrated by following the initial rates of uptake with 14 C-labeled amino acids, measuring the intracellular pools of amino acids, and isolating mutants resistant to toxic amino acids. One class of mutants (Pfa1) corresponds to a regulatory defect in the biosynthesis of the aromatic amino acids, but two other classes (Can1 and Aza1) are defective in amino acid transport. The Can1 mutants are defective in the active transport of three basic amino acids (arginine, histidine, and lysine) and in one of two transport systems operating on glutamine. The Aza1 mutants are not affected in the transport of the basic amino acids but have lost the capacity to transport all other amino acids except glutamate. The latter amino acid is probably transported by a third permease which could be identical to the Can1-independent transport operating on glutamine. Thus, genetic evidence suggests that strain 6803 has only a small number of amino acid transport systems with fairly broad specificity and that, with the exception of glutamine, each amino acid is accumulated by only one major transport system. Compared with heterotrophic bacteria such as Escherichia coli, these permeases are rather inefficient in terms of affinity (apparent K/sub m/ ranging from 6 to 60 μM) and of V/sub max/

  9. Effect of solar radiation on photosynthesis and pigmentation in the cyanobacterium microcoleus chtihonoplastes

    International Nuclear Information System (INIS)

    Annan, J.N.; Galyuon, I. K. A.; Donkor, V.A.

    2007-01-01

    The effects of solar radiation on the photosynthetic oxygen production and pigmentation were investigated in the marine filamentous cyanobacterium. Microcoleus chthonoplastes harvested from the intertidal zone of the Biriwa coast in Ghana. The organism was exposed to unfiltered solar radiation (UV-B. UV-A and PAR) and solar radiation filtered through optical filters. WG320 (UV-A and PAR), GG400 (PAR only), and UG5 (only UV-B and UV-A), Photosynthetic oxygen production was impaired. The reduction in the rate of photosynthetic oxygen production took over 2 hours to occur. The photoinhibition due to unfiltered solar radiation and combined UV-A and PAR were most severe. Absorption spectra of the crude extracts of M. chthonoplastes, indicated the presence of chlorophyll a, carotenoids, phycoerythrin and phycocyanin as the photosynthetic pigments, which were significantly bleached under the various solar radiation wavelengths. Generally, the phycobilins were affected most. Fluorescence measurements showed peaks that decreased significantly in amplitude and also underwent a shift towards shorter wavelengths, with prolonged exposure time, indicating that energy transfer from the accessory pigments was adversely affected. The implication is that increased solar radiation may have severe consequences on the marine ecosystem. (au)

  10. Anoxygenic photosynthesis controls oxygenic photosynthesis in a cyanobacterium from a sulfidic spring.

    Science.gov (United States)

    Klatt, Judith M; Al-Najjar, Mohammad A A; Yilmaz, Pelin; Lavik, Gaute; de Beer, Dirk; Polerecky, Lubos

    2015-03-01

    Before the Earth's complete oxygenation (0.58 to 0.55 billion years [Ga] ago), the photic zone of the Proterozoic oceans was probably redox stratified, with a slightly aerobic, nutrient-limited upper layer above a light-limited layer that tended toward euxinia. In such oceans, cyanobacteria capable of both oxygenic and sulfide-driven anoxygenic photosynthesis played a fundamental role in the global carbon, oxygen, and sulfur cycle. We have isolated a cyanobacterium, Pseudanabaena strain FS39, in which this versatility is still conserved, and we show that the transition between the two photosynthetic modes follows a surprisingly simple kinetic regulation controlled by this organism's affinity for H2S. Specifically, oxygenic photosynthesis is performed in addition to anoxygenic photosynthesis only when H2S becomes limiting and its concentration decreases below a threshold that increases predictably with the available ambient light. The carbon-based growth rates during oxygenic and anoxygenic photosynthesis were similar. However, Pseudanabaena FS39 additionally assimilated NO3 (-) during anoxygenic photosynthesis. Thus, the transition between anoxygenic and oxygenic photosynthesis was accompanied by a shift of the C/N ratio of the total bulk biomass. These mechanisms offer new insights into the way in which, despite nutrient limitation in the oxic photic zone in the mid-Proterozoic oceans, versatile cyanobacteria might have promoted oxygenic photosynthesis and total primary productivity, a key step that enabled the complete oxygenation of our planet and the subsequent diversification of life. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  11. A Novel Epiphytic Chlorophyll d-containing Cyanobacterium Isolated from a Mangrove-associated Red Alga.

    Science.gov (United States)

    Larkum, Anthony W D; Chen, Min; Li, Yaqiong; Schliep, Martin; Trampe, Erik; West, John; Salih, Anya; Kühl, Michael

    2012-12-01

    A new habitat and a new chlorophyll (Chl) d-containing cyanobacterium belonging to the genus Acaryochloris are reported in this study. Hyperspectral microscopy showed the presence of Chl d-containing microorganisms in epiphytic biofilms on a red alga (Gelidium caulacantheum) colonizing the pneumato-phores of a temperate mangrove (Avicennia marina). The presence of Chl d was further proven by high performance liquid chromatography (HPLC)-based pigment analysis and by confocal imaging of cultured cells. Enrichment of mangrove biofilm samples under near-infrared radiation (NIR) yielded the new Acaryochloris sp. MPGRS1, which was closely related in terms of 16S rRNA gene sequence to an isolate from the hypertrophic Salton Sea, USA. The new isolate used Chl d as its major photopigment; Chl d and Chl a contents were ~98% and 1%-2% of total cellular chlorophyll, respectively. These findings expand the variety of ecological niches known to harbor Chl d-containing cyanobacteria and support our working hypothesis that such oxyphototrophs may be ubiquitous in habitats depleted of visible light, but with sufficient NIR exposure. © 2012 Phycological Society of America.

  12. Anti-MRSA-acting carbamidocyclophanes H-L from the Vietnamese cyanobacterium Nostoc sp. CAVN2.

    Science.gov (United States)

    Preisitsch, Michael; Harmrolfs, Kirsten; Pham, Hang T L; Heiden, Stefan E; Füssel, Anna; Wiesner, Christoph; Pretsch, Alexander; Swiatecka-Hagenbruch, Monika; Niedermeyer, Timo H J; Müller, Rolf; Mundt, Sabine

    2015-03-01

    The methanol extract of the Vietnamese freshwater cyanobacterium Nostoc sp. CAVN2 exhibited cytotoxic effects against MCF-7 and 5637 cancer cell lines as well as against nontumorigenic FL and HaCaT cells and was active against methicillin-resistant Staphylococcus aureus (MRSA) and Streptococcus pneumoniae. High-resolution mass spectrometric analysis indicated the presence of over 60 putative cyclophane-like compounds in an antimicrobially active methanol extract fraction. A paracyclophanes-focusing extraction and separation methodology led to the isolation of 5 new carbamidocyclophanes (1-5) and 11 known paracyclophanes (6-16). The structures and their stereochemical configurations were elucidated by a combination of spectrometric and spectroscopic methods including HRMS, 1D and 2D NMR analyses and detailed comparative CD analysis. The newly described monocarbamoylated [7.7]paracyclophanes (1, 2, 4 and 5) differ by a varying degree of chlorination in the side chains. Carbamidocyclophane J (3) is the very first reported carbamidocyclophane bearing a single halogenation in both butyl residues. Based on previous studies a detailed phylogenetic examination of cyclophane-producing cyanobacteria was carried out. The biological evaluation of 1-16 against various clinical pathogens highlighted a remarkable antimicrobial activity against MRSA with MICs of 0.1-1.0 μM, and indicated that the level of antibacterial activity is related to the presence of carbamoyl moieties.

  13. Metabolomic approach to optimizing and evaluating antibiotic treatment in the axenic culture of cyanobacterium Nostoc flagelliforme.

    Science.gov (United States)

    Han, Pei-pei; Jia, Shi-ru; Sun, Ying; Tan, Zhi-lei; Zhong, Cheng; Dai, Yu-jie; Tan, Ning; Shen, Shi-gang

    2014-09-01

    The application of antibiotic treatment with assistance of metabolomic approach in axenic isolation of cyanobacterium Nostoc flagelliforme was investigated. Seven antibiotics were tested at 1-100 mg L(-1), and order of tolerance of N. flagelliforme cells was obtained as kanamycin > ampicillin, tetracycline > chloromycetin, gentamicin > spectinomycin > streptomycin. Four antibiotics were selected based on differences in antibiotic sensitivity of N. flagelliforme and associated bacteria, and their effects on N. flagelliforme cells including the changes of metabolic activity with antibiotics and the metabolic recovery after removal were assessed by a metabolomic approach based on gas chromatography-mass spectrometry combined with multivariate analysis. The results showed that antibiotic treatment had affected cell metabolism as antibiotics treated cells were metabolically distinct from control cells, but the metabolic activity would be recovered via eliminating antibiotics and the sequence of metabolic recovery time needed was spectinomycin, gentamicin > ampicillin > kanamycin. The procedures of antibiotic treatment have been accordingly optimized as a consecutive treatment starting with spectinomycin, then gentamicin, ampicillin and lastly kanamycin, and proved to be highly effective in eliminating the bacteria as examined by agar plating method and light microscope examination. Our work presented a strategy to obtain axenic culture of N. flagelliforme and provided a method for evaluating and optimizing cyanobacteria purification process through diagnosing target species cellular state.

  14. Simultaneous Production of Anabaenopeptins and Namalides by the Cyanobacterium Nostoc sp. CENA543.

    Science.gov (United States)

    Shishido, Tânia K; Jokela, Jouni; Fewer, David P; Wahlsten, Matti; Fiore, Marli F; Sivonen, Kaarina

    2017-11-17

    Anabaenopeptins are a diverse group of cyclic peptides, which contain an unusual ureido linkage. Namalides are shorter structural homologues of anabaenopeptins, which also contain an ureido linkage. The biosynthetic origins of namalides are unknown despite a strong resemblance to anabaenopeptins. Here, we show the cyanobacterium Nostoc sp. CENA543 strain producing new (nostamide B-E (2, 4, 5, and 6)) and known variants of anabaenopeptins (schizopeptin 791 (1) and anabaenopeptin 807 (3)). Surprisingly, Nostoc sp. CENA543 also produced namalide B (8) and the new namalides D (7), E (9), and F (10) in similar amounts to anabaenopeptins. Analysis of the complete Nostoc sp. CENA543 genome sequence indicates that both anabaenopeptins and namalides are produced by the same biosynthetic pathway through module skipping during biosynthesis. This unique process involves the skipping of two modules present in different nonribosomal peptide synthetases during the namalide biosynthesis. This skipping is an efficient mechanism since both anabaenopeptins and namalides are synthesized in similar amounts by Nostoc sp. CENA543. Consequently, gene skipping may be used to increase and possibly broaden the chemical diversity of related peptides produced by a single biosynthetic gene cluster. Genome mining demonstrated that the anabaenopeptin gene clusters are widespread in cyanobacteria and can also be found in tectomicrobia bacteria.

  15. Merocyclophanes C and D from the Cultured Freshwater Cyanobacterium Nostoc sp. (UIC 10110).

    Science.gov (United States)

    May, Daniel S; Chen, Wei-Lun; Lantvit, Daniel D; Zhang, Xiaoli; Krunic, Aleksej; Burdette, Joanna E; Eustaquio, Alessandra; Orjala, Jimmy

    2017-04-28

    Merocyclophanes C and D (1 and 2) were isolated from the cell extract of the cultured cyanobacterium UIC 10110. The structures were determined by one-dimensional nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectrometry and confirmed by 2D NMR techniques. The absolute configurations were determined using electronic circular dichroism spectroscopy. Merocyclophanes C and D represent the first known analogues of the merocyclophane core structure, a recently discovered scaffold of [7,7] paracyclophanes characterized by an α-branched methyl at C-1/C-14; 1 and 2 showed antiproliferative activity against the MDA-MB-435 cell line with IC 50 values of 1.6 and 0.9 μM, respectively. Partial 16S analysis determined UIC 10110 to be a Nostoc sp., and it was found to clade with UIC 10062 Nostoc sp., the only other strain known to produce merocyclophanes. The genome of UIC 10110 was sequenced, and a biosynthetic gene cluster was identified that is proposed to encode type I and type III polyketide synthases that are potentially responsible for production of the merocyclophanes; however, further experiments will be required to verify the true function of the gene cluster. The gene cluster provides a genetic basis for the observed structural differences of the [7,7] paracyclophane core structures.

  16. Nostopeptolide plays a governing role during cellular differentiation of the symbiotic cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Liaimer, Anton; Helfrich, Eric J N; Hinrichs, Katrin; Guljamow, Arthur; Ishida, Keishi; Hertweck, Christian; Dittmann, Elke

    2015-02-10

    Nostoc punctiforme is a versatile cyanobacterium that can live either independently or in symbiosis with plants from distinct taxa. Chemical cues from plants and N. punctiforme were shown to stimulate or repress, respectively, the differentiation of infectious motile filaments known as hormogonia. We have used a polyketide synthase mutant that accumulates an elevated amount of hormogonia as a tool to understand the effect of secondary metabolites on cellular differentiation of N. punctiforme. Applying MALDI imaging to illustrate the reprogramming of the secondary metabolome, nostopeptolides were identified as the predominant difference in the pks2(-) mutant secretome. Subsequent differentiation assays and visualization of cell-type-specific expression of nostopeptolides via a transcriptional reporter strain provided evidence for a multifaceted role of nostopeptolides, either as an autogenic hormogonium-repressing factor or as a chemoattractant, depending on its extracellular concentration. Although nostopeptolide is constitutively expressed in the free-living state, secreted levels dynamically change before, during, and after the hormogonium differentiation phase. The metabolite was found to be strictly down-regulated in symbiosis with Gunnera manicata and Blasia pusilla, whereas other metabolites are up-regulated, as demonstrated via MALDI imaging, suggesting plants modulate the fine-balanced cross-talk network of secondary metabolites within N. punctiforme.

  17. Structural elucidation and molecular docking of a novel antibiotic compound from cyanobacterium Nostoc sp. MGL001

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    Niveshika No Name

    2016-11-01

    Full Text Available Cyanobacteria are rich source of array of bioactive compounds. The present study reports a novel antibacterial bioactive compound purified from cyanobacterium Nostoc sp. MGL001 using various chromatographic techniques viz. thin layer chromatography (TLC and high performance liquid chromatography (HPLC. Further characterization was done using electrospray ionisation mass spectroscopy (ESIMS and nuclear magnetic resonance (NMR and predicted structure of bioactive compound was 9-Ethyliminomethyl-12-(morpholin - 4 - ylmethoxy -5, 8, 13, 16 – tetraaza – hexacene - 2, 3 dicarboxylic acid (EMTAHDCA. Structure of EMTAHDCA clearly indicated that it is a novel compound that was not reported in literature or natural product database. The compound exhibited growth inhibiting effects mainly against the gram negative bacterial strains and produced maximum zone of inhibition at 150 μg/mL concentration. The compound was evaluated through in silico studies for its ability to bind 30S ribosomal fragment (PDB ID: 1YRJ, 1MWL, 1J7T and 1LC4 and OmpF porin protein (4GCP, 4GCQ and 4GCS which are the common targets of various antibiotic drugs. Comparative molecular docking study revealed that EMTAHDCA has strong binding affinity for these selected targets in comparison to a number of most commonly used antibiotics. The ability of EMTAHDCA to bind the active sites on the proteins and 30S ribosomal fragments where the antibiotic drugs generally bind indicated that it is functionally similar to the commercially available drugs.

  18. Semicontinuous cultivation of the Cyanobacterium Spirulina platensis in a closed photobioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Reichert, C.C.; Costa, J.A.V. [Fundacao Universidade Federal do Rio Grande (FURG), Rio Grande, RS (Brazil). Dept. de Quimica], Email: dqmjorge@furg.br; Reinehr, C.O. [Universidade de Passo Fundo, RS (Brazil). Centro de Pesquisa em Alimentacao], Email: reinehr@upf.br

    2006-01-15

    The cultivation of photosynthetic microorganisms such as the cyanobacterium Spirulina platensis has been studied by researchers in many countries because these organisms can produce products with industrial potential. We studied the specific growth rate ({mu}{sub x}, day{sup -1}) and productivity (P{sub x}, in mg/L/day of Spirulina platensis biomass, dry weight basis) of two S. platensis strains (LEB-52 and Paracas) growing in aerated semicontinuous culture in two-liter Erlenmeyer flasks for 90 days (2160 h) at 30 deg C under 2500 lux of illumination in a 12 h photoperiod. Independent of the S. platensis strain used we found that low biomass concentrations (0.50 g/L) and high renewal rates (50% v/v) resulted in a high specific growth rate ({mu}{sub x} = 0.111 day{sup -1}) and high productivity (P{sub x} = 42.3 mg/L/day). These values are two to four times higher than those obtained in simple batch cultivation and indicate that the semicontinuous cultivation of S. platensis is viable. (author)

  19. Semicontinuous cultivation of the cyanobacterium Spirulina platensis in a closed photobioreactor

    Directory of Open Access Journals (Sweden)

    C. C. Reichert

    2006-03-01

    Full Text Available The cultivation of photosynthetic microorganisms such as the cyanobacterium Spirulina platensis has been studied by researchers in many countries because these organisms can produce products with industrial potential. We studied the specific growth rate (µx, day-1 and productivity (Px, in mg/L/day of Spirulina platensis biomass, dry weight basis of two S. platensis strains (LEB-52 and Paracas growing in aerated semicontinuous culture in two-liter Erlenmeyer flasks for 90 days (2160 h at 30°C under 2500 lux of illumination in a 12 h photoperiod. Independent of the S. platensis strain used we found that low biomass concentrations (0.50 g/L and high renewal rates (50% v/v resulted in a high specific growth rate (µx = 0.111 day-1 and high productivity (Px = 42.3 mg/L/day. These values are two to four times higher than those obtained in simple batch cultivation and indicate that the semicontinuous cultivation of S. platensis is viable.

  20. Anoxygenic Photosynthesis Controls Oxygenic Photosynthesis in a Cyanobacterium from a Sulfidic Spring

    KAUST Repository

    Klatt, Judith M.; Alnajjar, Mohammad Ahmad; Yilmaz, Pelin; Lavik, Gaute; de Beer, Dirk; Polerecky, Lubos

    2015-01-01

    Before the Earth's complete oxygenation (0.58 to 0.55 billion years [Ga] ago), the photic zone of the Proterozoic oceans was probably redox stratified, with a slightly aerobic, nutrient-limited upper layer above a light-limited layer that tended toward euxinia. In such oceans, cyanobacteria capable of both oxygenic and sulfide-driven anoxygenic photosynthesis played a fundamental role in the global carbon, oxygen, and sulfur cycle. We have isolated a cyanobacterium, Pseudanabaena strain FS39, in which this versatility is still conserved, and we show that the transition between the two photosynthetic modes follows a surprisingly simple kinetic regulation controlled by this organism's affinity for H2S. Specifically, oxygenic photosynthesis is performed in addition to anoxygenic photosynthesis only when H2S becomes limiting and its concentration decreases below a threshold that increases predictably with the available ambient light. The carbon-based growth rates during oxygenic and anoxygenic photosynthesis were similar. However, Pseudanabaena FS39 additionally assimilated NO3 - during anoxygenic photosynthesis. Thus, the transition between anoxygenic and oxygenic photosynthesis was accompanied by a shift of the C/N ratio of the total bulk biomass. These mechanisms offer new insights into the way in which, despite nutrient limitation in the oxic photic zone in the mid-Proterozoic oceans, versatile cyanobacteria might have promoted oxygenic photosynthesis and total primary productivity, a key step that enabled the complete oxygenation of our planet and the subsequent diversification of life.

  1. Proteomic analysis of the cyanobacterium of the Azolla symbiosis: identity, adaptation, and NifH modification.

    Science.gov (United States)

    Ekman, Martin; Tollbäck, Petter; Bergman, Birgitta

    2008-01-01

    Cyanobacteria are able to form stable nitrogen-fixing symbioses with diverse eukaryotes. To extend our understanding of adaptations imposed by plant hosts, two-dimensional gel electrophoresis and mass spectrometry (MS) were used for comparative protein expression profiling of a cyanobacterium (cyanobiont) dwelling in leaf cavities of the water-fern Azolla filiculoides. Homology-based protein identification using peptide mass fingerprinting [matrix-assisted laser desorption ionization-time of flight (MALDI-TOF-MS)], tandem MS analyses, and sequence homology searches resulted in an identification success rate of 79% of proteins analysed in the unsequenced cyanobiont. Compared with a free-living strain, processes related to energy production, nitrogen and carbon metabolism, and stress-related functions were up-regulated in the cyanobiont while photosynthesis and metabolic turnover rates were down-regulated, stressing a slow heterotrophic mode of growth, as well as high heterocyst frequencies and nitrogen-fixing capacities. The first molecular data set on the nature of the NifH post-translational modification in cyanobacteria was also obtained: peptide mass spectra of the protein demonstrated the presence of a 300-400 Da protein modification localized to a specific 13 amino acid sequence, within the part of the protein that is ADP-ribosylated in other bacteria and close to the active site of nitrogenase. Furthermore, the distribution of the highest scoring database hits for the identified proteins points to the possibility of using proteomic data in taxonomy.

  2. Crystal Structure of Allophycocyanin from Marine Cyanobacterium Phormidium sp. A09DM.

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    Ravi Raghav Sonani

    Full Text Available Isolated phycobilisome (PBS sub-assemblies have been widely subjected to X-ray crystallography analysis to obtain greater insights into the structure-function relationship of this light harvesting complex. Allophycocyanin (APC is the phycobiliprotein always found in the PBS core complex. Phycocyanobilin (PCB chromophores, covalently bound to conserved Cys residues of α- and β- subunits of APC, are responsible for solar energy absorption from phycocyanin and for transfer to photosynthetic apparatus. In the known APC structures, heterodimers of α- and β- subunits (known as αβ monomers assemble as trimer or hexamer. We here for the first time report the crystal structure of APC isolated from a marine cyanobacterium (Phormidium sp. A09DM. The crystal structure has been refined against all the observed data to the resolution of 2.51 Å to Rwork (Rfree of 0.158 (0.229 with good stereochemistry of the atomic model. The Phormidium protein exists as a trimer of αβ monomers in solution and in crystal lattice. The overall tertiary structures of α- and β- subunits, and trimeric quaternary fold of the Phormidium protein resemble the other known APC structures. Also, configuration and conformation of the two covalently bound PCB chromophores in the marine APC are same as those observed in fresh water cyanobacteria and marine red algae. More hydrophobic residues, however, constitute the environment of the chromophore bound to α-subunit of the Phormidium protein, owing mainly to amino acid substitutions in the marine protein.

  3. Detection of weed algae in open pond cultures of Cyanobacterium aponinum using PAM

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    Dominik Winckelmann

    2016-02-01

    Full Text Available Abstract The potential use of non-arable land in the al-Wusta region of the Sultanate of Oman for the production of algae biomass was examined. Brackish cleaned production water from oil production supplemented with commercial fertilizer was used as growth medium. The indigenous isolate Cyanobacterium aponinum WP7(1 was grown in open ponds using batch or semi-continuous cultivation. Biomass production rates of 15–24 g/m2/day were achieved. The change of salinity due to evaporation, which was thought to be a major challenge, did not exceed 35 ppt. All cultures showed contaminations with weed algae. Contaminations with green algae or diatoms were detectable using fluorescence pattern excited by four different wavelengths using a pulse-amplitude-modulation chlorophyll fluorometer (PAM. It is possible to estimate the health level and the mayor groups of which a culture is composed using the PAM method. Therefore, the fluorescence of the photosynthetically inactive sample is compared with the fluorescence after all copies of photosystem II were closed by exposing the sample to a high-intensity light beam. A detection limit of one weed algae cell in a hundred cells was achieved.

  4. Anilofos tolerance and its mineralization by the cyanobacterium Synechocystis sp. strain PUPCCC 64.

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    D P Singh

    Full Text Available This study deals with anilofos tolerance and its mineralization by the common rice field cyanobacterium Synechocystis sp. strain PUPCCC 64. The organism tolerated anilofos up to 25 mg L(-1. The herbicide caused inhibitory effects on photosynthetic pigments of the test organism in a dose-dependent manner. The organism exhibited 60, 89, 96, 85 and 79% decrease in chlorophyll a, carotenoids, phycocyanin, allophycocyanin and phycoerythrin, respectively, in 20 mg L(-1 anilofos on day six. Activities of superoxide dismutase, catalase and peroxidase increased by 1.04 to 1.80 times over control cultures in presence of 20 mg L(-1 anilofos. Glutathione content decreased by 26% while proline content was unaffected by 20 mg L(-1 anilofos. The test organism showed intracellular uptake and metabolized the herbicide. Uptake of herbicide by test organism was fast during initial six hours followed by slow uptake until 120 hours. The organism exhibited maximum anilofos removal at 100 mg protein L(-1, pH 8.0 and 30°C. Its growth in phosphate deficient basal medium in the presence of anilofos (2.5 mg L(-1 indicated that herbicide was used by the strain PUPCCC 64 as a source of phosphate.

  5. Novel toxic effects associated with a tropical Limnothrix/Geitlerinema-like cyanobacterium.

    Science.gov (United States)

    Bernard, Catherine; Froscio, Suzanne; Campbell, Rebecca; Monis, Paul; Humpage, Andrew; Fabbro, Larelle

    2011-06-01

    The presence of a toxic strain of a fine filamentous cyanobacterium belonging to the Oscillatorialean family Pseudanabaenacea was detected during a survey of cyanobacterial taxa associated with the presence of cylindrospermopsin in dams in Central Queensland (Australia). The strain, AC0243, was isolated and cultured, its genomic DNA extracted and 16S RNA gene sequenced. Phylogenetic analysis placed AC0243 with Limnothrix species, although this genus appears polyphyletic. Moreover, not all morphological characters are consistent with this genus but more closely fit the description of Geitlerinema unigranulatum (R.N. Singh) Komárek and Azevedo. The potential toxic effects of AC0243 extract were assessed chemically and biologically. Cell free protein synthesis was inhibited by the extract. Exposure of Vero cells to the extract resulted in a significant reduction in cellular ATP levels following 24-72 h incubation. The presence of cylindrospermopsin was excluded based on the nature of responses obtained in cell and cell-free assays; in addition, (i) it could not be detected by HPLC, LC-MS, or immunological assay, and (ii) no genes currently associated with the production of cylindrospermopsin were found in the genome. Other known cyanobacterial toxins were not detected. The apparent novelty of this toxin is discussed. Copyright © 2009 Wiley Periodicals, Inc.

  6. ORGANIZATION OF THE nif GENES OF THE NONHETEROCYSTOUS CYANOBACTERIUM TRICHODESMIUM SP. IMS101.

    Science.gov (United States)

    Dominic, Benny; Zani, Sabino; Chen, Yi-Bu; Mellon, Mark T; Zehr, Jonathan P

    2000-08-26

    An approximately 16-kb fragment of the Trichodesmium sp. IMS101 (a nonheterocystous filamentous cyanobacterium) "conventional"nif gene cluster was cloned and sequenced. The gene organization of the Trichodesmium and Anabaena variabilis vegetative (nif 2) nitrogenase gene clusters spanning the region from nif B to nif W are similar except for the absence of two open reading frames (ORF3 and ORF1) in Trichodesmium. The Trichodesmium nif EN genes encode a fused Nif EN polypeptide that does not appear to be processed into individual Nif E and Nif N polypeptides. Fused nif EN genes were previously found in the A. variabilis nif 2 genes, but we have found that fused nif EN genes are widespread in the nonheterocystous cyanobacteria. Although the gene organization of the nonheterocystous filamentous Trichodesmium nif gene cluster is very similar to that of the A. variabilis vegetative nif 2 gene cluster, phylogenetic analysis of nif sequences do not support close relatedness of Trichodesmium and A. variabilis vegetative (nif 2) nitrogenase genes.

  7. Proteome-wide analysis and diel proteomic profiling of the cyanobacterium Arthrospira platensis PCC 8005.

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    Sabine Matallana-Surget

    Full Text Available The filamentous cyanobacterium Arthrospira platensis has a long history of use as a food supply and it has been used by the European Space Agency in the MELiSSA project, an artificial microecosystem which supports life during long-term manned space missions. This study assesses progress in the field of cyanobacterial shotgun proteomics and light/dark diurnal cycles by focusing on Arthrospira platensis. Several fractionation workflows including gel-free and gel-based protein/peptide fractionation procedures were used and combined with LC-MS/MS analysis, enabling the overall identification of 1306 proteins, which represents 21% coverage of the theoretical proteome. A total of 30 proteins were found to be significantly differentially regulated under light/dark growth transition. Interestingly, most of the proteins showing differential abundance were related to photosynthesis, the Calvin cycle and translation processes. A novel aspect and major achievement of this work is the successful improvement of the cyanobacterial proteome coverage using a 3D LC-MS/MS approach, based on an immobilized metal affinity chromatography, a suitable tool that enabled us to eliminate the most abundant protein, the allophycocyanin. We also demonstrated that cell growth follows a light/dark cycle in A. platensis. This preliminary proteomic study has highlighted new characteristics of the Arthrospira platensis proteome in terms of diurnal regulation.

  8. Effect of Selected Plant Extracts and D- and L-Lysine on the Cyanobacterium Microcystis aeruginosa

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    Miquel Lürling

    2014-06-01

    Full Text Available We tested extracts from Fructus mume, Salvia miltiorrhiza and Moringa oleifera as well as L-lysine and D-Lysine as curative measures to rapidly suppress the cyanobacterium Microcystis aeruginosa NIVA-CYA 43. We tested these compounds under similar conditions to facilitate comparisons. We hypothesized that for each compound, relatively low concentrations—i.e., 5–50 mg L−1, would reduce M. aeruginosa biomass. At these low concentrations, only L-lysine caused a decline in M. aeruginosa biomass at ≥4.3 mg L−1. F. mume extract was effective to do so at high concentrations, i.e., at ≥240 mg L−1, but the others were virtually non-effective. Low pH caused by organic acids is a probable explanation for the effect of F. mume extract. No complete wipe-outs of the experimental population were achieved as Photosystem II efficiency showed a recovery after six days. L-lysine may be effective at low concentrations—meaning low material costs. However, the effect of L-lysine seems relatively short-lived. Overall, the results of our study did not support the use of the tested plant extracts and amino-acid as promising candidates for curative application in M. aeruginosa bloom control.

  9. Theoretical investigation of biomass productivities achievable in solar rectangular photobioreactors for the cyanobacterium Arthrospira platensis.

    Science.gov (United States)

    Pruvost, Jeremy; Cornet, J F; Goetz, Vincent; Legrand, Jack

    2012-01-01

    Modeling was done to simulate whole-year running of solar rectangular photobioreactors (PBRs). Introducing the concept of ideal reactor, the maximal biomass productivity that could be achieved on Earth on nitrate as N-source was calculated. Two additional factors were also analyzed with respect to dynamic calculations over the whole year: the effect of PBR location and the effects of given operating conditions on the resulting decrease in productivity compared with the ideal one. Simulations were conducted for the cyanobacterium Arthospira platensis, giving an ideal productivity (upper limit) in the range 55-60 tX ha(-1) year(-1) for a sun tracking system (and around 35-40 tX ha(-1) year(-1) for a fixed horizontal PBR). For an implantation in France (Nantes, west coast), the modification in irradiation conditions resulted in a decrease in biomass productivity of 40%. Various parameters were investigated, with special emphasis on the influence of the incident angle of solar illumination on resulting productivities, affecting both light capture and light transfer inside the bulk culture. It was also found that with appropriate optimization of the residence time as permitted by the model, productivities close to maximal could be achieved for a given location. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

  10. Acclimation to and recovery from cadmium and zinc exposure by a freshwater cyanobacterium, Microcystis aeruginosa

    International Nuclear Information System (INIS)

    Zeng Jin; Yang Liuyan; Wang Wenxiong

    2009-01-01

    To understand the metal tolerance of a bloom-forming cyanobacterium, Microcystis aeruginosa, we investigated its acclimation to and recovery from cadmium (Cd) and zinc (Zn) exposure. The intracellular Cd and Zn (intra-Cd and intra-Zn) quotas increased upon acclimation to increased metal concentrations and were reduced following 1-day or 5-day recovery. Different acclimation to varying metal concentrations or durations (5 days or 15 days) did not have significant effects on the short-term uptake of Cd or Zn, whereas a 1-day recovery period promoted Cd or Zn uptake significantly. The values of median growth-inhibition concentrations (free ion concentration or intracellular quota) increased when the cyanobacterial cells were acclimated to higher Cd or Zn concentrations, indicating that M. aeruginosa became more tolerant to these metals. Consistent with the significant increase in metal uptake, the cyanobacteria become very sensitive to metals following 1-day recovery. A longer recovery (5 days) led to comparable uptake and toxicity responses to the controls. The efflux rate constants were not significantly different following metal acclimation. In the subcellular metal measurements, Cd was mostly distributed in the soluble fraction, whereas Zn was distributed evenly in the adsorbed, insoluble and soluble fractions of the cells. This study suggested the strong ability of these cyanobacteria to acclimate to different environments.

  11. A new antibiotic produced by the cyanobacterium-symbiotic fungus Simplicillium lanosoniveum.

    Science.gov (United States)

    Dong, Qinglin; Dong, Rongzhen; Xing, Xiangying; Li, Yukuan

    2018-06-01

    The culture broth of the cyanobacterium-symbiotic fungus Simplicillium lanosoniveum var. Tianjinienss Q. L. Dong exhibited unanticipated antibacterial activities against the Gram-positive bacteria, particularly the pathogenic bacterium Staphylococcus aureus, indicating the secretion of antibiotic-like metabolite, for which the modified Sabouraud medium was the suitable medium. The antibiotic-like metabolite was separated with macroporous resins CT-12 (absorption) and 95% ethanol (desorption), purified by ion-exchange resins D301T and displayed a characteristic absorption peak at 228 nm, suggesting the presence of nitrogen. The negative biuret and ninhydrin tests confirmed the absence of -NH 2 and -COOH groups. Further, HPLC and mass spectrometry analyses showed that the retention time and molecular weight of the antibiotic-like metabolite were 4.1031 min and 163.0182 (Δ ± 2.3 ppm), respectively. Taking together, we speculated that the antibiotic-like metabolite was a new antibiotic structurally similar to alkaloid, which was the first one isolated from the species of Simplicillium genus.

  12. Bioprocess Engineering Aspects of Biopolymer Production by the Cyanobacterium Spirulina Strain LEB 18

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    Roberta Guimarães Martins

    2014-01-01

    Full Text Available Microbial biopolymers can replace environmentally damaging plastics derived from petrochemicals. We investigated biopolymer synthesis by the cyanobacterium Spirulina strain LEB 18. Autotrophic culture used unmodified Zarrouk medium or modified Zarrouk medium in which the NaNO3 content was reduced to 0.25 g L−1 and the NaHCO3 content reduced to 8.4 g L−1 or increased to 25.2 g L−1. Heterotrophic culture used modified Zarrouk medium containing 0.25 g L−1 NaNO3 with the NaHCO3 replaced by 0.2 g L−1, 0.4 g L−1, or 0.6 g L−1 of glucose (C6H12O6 or sodium acetate (CH3COONa. Mixotrophic culture used modified Zarrouk medium containing 0.25 g L−1 NaNO3 plus 16.8 g L−1 NaHCO3 with the addition of 0.2 g L−1, 0.4 g L−1, or 0.6 g L−1 of glucose or sodium acetate. The highest biopolymer yield was 44% when LEB 18 was growing autotrophically in media containing 0.25 g L−1 NaNO3 and 8.4 g L−1 NaHCO3.

  13. Inhibitory effects of sanguinarine against the cyanobacterium Microcystis aeruginosa NIES-843 and possible mechanisms of action

    Energy Technology Data Exchange (ETDEWEB)

    Shao, Jihai [College of Resources and Environment, Hunan Agricultural University, Changsha 410128 (China); Hunan Provincial Key Laboratory of Farmland Pollution Control and Agricultural Resources Use, Hunan Agricultural University, Changsha 410128 (China); Liu, Deming [State Key Laboratory Breeding Base of Crop Germplasm Innovation and Resource Utilization, Hunan Agricultural University, Changsha 410128 (China); Gong, Daoxin; Zeng, Qingru; Yan, Zhiyong [College of Resources and Environment, Hunan Agricultural University, Changsha 410128 (China); Gu, Ji-Dong, E-mail: jdgu@hku.hk [Hunan Provincial Key Laboratory of Farmland Pollution Control and Agricultural Resources Use, Hunan Agricultural University, Changsha 410128 (China); Laboratory of Environmental Microbiology and Toxicology, School of Biological Sciences, The University of Hong Kong, Hong Kong SAR (China)

    2013-10-15

    Highlights: •Sanguinarine was found as a strong algicidal biologically derived substance. •Sanguinarine can induce oxidative stress in the cells of Microcystis aeruginosa. •Photosystem is a target of toxicity of sanguinarine on M. aeruginosa. •Sanguinarine can induce DNA damage and inhibit cell division. -- Abstract: Sanguinarine showed strong inhibitory effect against Microcystis aeruginosa, a typical water bloom-forming and microcystins-producing cyanobacterium. The EC50 of sanguinarine against the growth of M. aeruginosa NIES-843 was 34.54 ± 1.17 μg/L. Results of chlorophyll fluorescence transient analysis indicated that all the electron donating side, accepting side, and the reaction center of the Photosystem II (PS II) were the targets of sanguinarine against M. aeruginosa NIES-843. The elevation of reactive oxygen species (ROS) level in the cells of M. aeruginosa NIES-843 upon exposure indicated that sanguinarine induced oxidative stress in the active growing cells of M. aeruginosa NIES-843. Further results of gene expression analysis indicated that DNA damage and cell division inhibition were also involved in the inhibitory action mechanism of sanguinarine against M. aeruginosa NIES-843. The inhibitory characteristics of sanguinarine against M. aeruginosa suggest that the ecological- and public health-risks need to be evaluated before its application in cyanobacterial bloom control to avoid devastating events irreversibly.

  14. Nice to meet you: genetic, epigenetic and metabolic controls of plant perception of beneficial associative and endophytic diazotrophic bacteria in non-leguminous plants.

    Science.gov (United States)

    Carvalho, T L G; Ballesteros, H G F; Thiebaut, F; Ferreira, P C G; Hemerly, A S

    2016-04-01

    A wide range of rhizosphere diazotrophic bacteria are able to establish beneficial associations with plants, being able to associate to root surfaces or even endophytically colonize plant tissues. In common, both associative and endophytic types of colonization can result in beneficial outcomes to the plant leading to plant growth promotion, as well as increase in tolerance against biotic and abiotic stresses. An intriguing question in such associations is how plant cell surface perceives signals from other living organisms, thus sorting pathogens from beneficial ones, to transduce this information and activate proper responses that will finally culminate in plant adaptations to optimize their growth rates. This review focuses on the recent advances in the understanding of genetic and epigenetic controls of plant-bacteria signaling and recognition during beneficial associations with associative and endophytic diazotrophic bacteria. Finally, we propose that "soil-rhizosphere-rhizoplane-endophytes-plant" could be considered as a single coordinated unit with dynamic components that integrate the plant with the environment to generate adaptive responses in plants to improve growth. The homeostasis of the whole system should recruit different levels of regulation, and recognition between the parties in a given environment might be one of the crucial factors coordinating these adaptive plant responses.

  15. Effects of tungsten and titanium oxide nanoparticles on the diazotrophic growth and metals acquisition by Azotobacter vinelandii under molybdenum limiting condition.

    Science.gov (United States)

    Allard, Patrick; Darnajoux, Romain; Phalyvong, Karine; Bellenger, Jean-Philippe

    2013-02-19

    The acquisition of essential metals, such as the metal cofactors (molybdenum (Mo) and iron (Fe)) of the nitrogenase, the enzyme responsible for the reduction of dinitrogen (N(2)) to ammonium, is critical to N(2) fixing bacteria in soil. The release of metal nanoparticles (MNPs) to the environment could be detrimental to N(2) fixing bacteria by introducing a new source of toxic metals and by interfering with the acquisition of essential metals such as Mo. Since Mo has been reported to limit nonsymbiotic N(2) fixation in many ecosystems from tropical to cold temperate, this question is particularly acute in the context of Mo limitation. Using a combination of microbiology and analytical chemistry techniques, we have evaluated the effect of titanium (Ti) and tungsten (W) oxide nanoparticles on the diazotrophic growth and metals acquisition in pure culture of the ubiquitous N(2) fixing bacterium Azotobacter vinelandii under Mo replete and Mo limiting conditions. We report that under our conditions (≤10 mg·L(-1)) TiO(2) NPs have no effects on the diazotrophic growth of A. vinelandii while WO(3) NPs are highly detrimental to the growth especially under Mo limiting conditions. Our results show that the toxicity of WO(3) NPs to A. vinelandii is due to an interference with the catechol-metalophores assisted uptake of Mo.

  16. The success of the cyanobacterium Cylindrospermopsis raciborskii in freshwaters is enhanced by the combined effects of light intensity and temperature

    Directory of Open Access Journals (Sweden)

    Sylvia Bonilla

    2016-06-01

    Full Text Available Toxic cyanobacterial blooms in freshwaters are thought to be a consequence of the combined effects of anthropogenic eutrophication and climate change. It is expected that climate change will affect water mixing regimes that alter the water transparency and ultimately the light environment for phytoplankton. Blooms of the potentially toxic cyanobacterium Cylindrospermopsis raciborskii are expanding from tropical towards temperate regions. Several hypotheses have been proposed to explain this expansion, including an increase in water temperature due to climate change and the high phenotypic plasticity of the species that allows it to exploit different light environments. We performed an analysis based on eight lakes in tropical, subtropical and temperate regions to examine the distribution and abundance of C. raciborskii in relation to water temperature and transparency. We then conducted a series of short-term factorial experiments that combined three temperatures and two light intensity levels using C. raciborskii cultures alone and in interaction with another cyanobacterium to identify its growth capacity. Our results from the field, in contrast to predictions, showed no differences in dominance (>40% to the total biovolume of C. raciborskii between climate regions. C. raciborskii was able to dominate the phytoplankton in a wide range of light environments (euphotic zone = 1.5 to 5 m, euphotic zone/mixing zone ratio <0.5 to >1.5. Moreover, C. raciborskii was capable of dominating the phytoplankton at low temperatures (<15°C. Our experimental results showed that C. raciborskii growing in interaction was enhanced by the increase of the temperature and light intensity. C. raciborskii growth in high light intensities and at a wide range of temperatures, suggests that any advantage that this species may derive from climate change that favors its dominance in the phytoplankton is likely due to changes in the light environment rather than changes in

  17. Diversity of the Marine Cyanobacterium Trichodesmium: Characterization of the Woods Hole Culture Collection and Quantification of Field Populations

    Science.gov (United States)

    2009-09-01

    stations on all four cruises; Fig. 5-4 C), and Plec - tonema sp. (North Pacific MP09-19, North Atlantic EN361-1, 2, 3; Fig. 5-4 D). Calothrix sp. was found in...under a chlorophyll filter; (C) ELF-labeled heterotrophic bacteria (Sta. MP09-16, 11 Aug 03, puff); (D) ELF-labeled cyanobacterium Plec - tonema sp...epiphyte of the pelagic diatom Chaetoceros (Foster and Zehr, 2006). While Plec - tonema is known to fix N2 only in lowered oxygen tension (Rippka et al

  18. Strong Regionality and Dominance of Anaerobic Bacterial Taxa Characterize Diazotrophic Bacterial Communities of the Arcto-Alpine Plant Species Oxyria digyna and Saxifraga oppositifolia.

    Science.gov (United States)

    Kumar, Manoj; van Elsas, Jan Dirk; Nissinen, Riitta

    2017-01-01

    Arctic and alpine biomes are most often strongly nitrogen-limited, and hence biological nitrogen fixation is a strong driver of these ecosystems. Both biomes are characterized by low temperatures and short growing seasons, but they differ in seasonality of solar radiation and in soil water balance due to underlying permafrost in the Arctic. Arcto-alpine plant species are well-adapted to the low temperatures that prevail in their habitats, and plant growth is mainly limited by the availability of nutrients, in particular nitrogen, due to slow mineralization. Nitrogen fixing bacteria are likely important for plant growth in these habitats, but very little is known of these bacteria or forces shaping their communities. In this study, we characterized the potential nitrogen fixing bacterial (PNFB) communities associated with two arcto-alpine pioneer plant species, Oxyria digyna (mountain sorrel) and Saxifraga oppositifolia (blue saxifrage), in three climate regions. Both of these plants readily colonize low nutrient mineral soils. Our goal was to investigate how climate (region) and, on the other hand, host plant and plant species shape these communities. To our knowledge, this is the first comprehensive study describing PNFB communities associated with pioneer plants in different arcto-alpine biomes. Replicate samples were taken from two arctic regions, Kilpisjärvi and Ny-Ålesund, and one alpine region, Mayrhofen. In these, the PNFB communities in the bulk and rhizosphere soils and the plant endospheres were characterized by nifH -targeted PCR and massive parallel sequencing. The data revealed strong effects of climatic region on the dominating nitrogen fixers. Specifically, nifH sequences related to Geobacter (δ- Proteobacteria ) were present in high relative abundances in the nitrogen-fixing communities in the Mayrhofen and Kilpisjärvi regions, while members of the Clostridiales prevailed in the Kilpisjärvi and Ny-Ålesund regions. The bulk and rhizosphere soil

  19. Strong Regionality and Dominance of Anaerobic Bacterial Taxa Characterize Diazotrophic Bacterial Communities of the Arcto-Alpine Plant Species Oxyria digyna and Saxifraga oppositifolia

    Directory of Open Access Journals (Sweden)

    Manoj Kumar

    2017-10-01

    Full Text Available Arctic and alpine biomes are most often strongly nitrogen-limited, and hence biological nitrogen fixation is a strong driver of these ecosystems. Both biomes are characterized by low temperatures and short growing seasons, but they differ in seasonality of solar radiation and in soil water balance due to underlying permafrost in the Arctic. Arcto-alpine plant species are well-adapted to the low temperatures that prevail in their habitats, and plant growth is mainly limited by the availability of nutrients, in particular nitrogen, due to slow mineralization. Nitrogen fixing bacteria are likely important for plant growth in these habitats, but very little is known of these bacteria or forces shaping their communities. In this study, we characterized the potential nitrogen fixing bacterial (PNFB communities associated with two arcto-alpine pioneer plant species, Oxyria digyna (mountain sorrel and Saxifraga oppositifolia (blue saxifrage, in three climate regions. Both of these plants readily colonize low nutrient mineral soils. Our goal was to investigate how climate (region and, on the other hand, host plant and plant species shape these communities. To our knowledge, this is the first comprehensive study describing PNFB communities associated with pioneer plants in different arcto-alpine biomes. Replicate samples were taken from two arctic regions, Kilpisjärvi and Ny-Ålesund, and one alpine region, Mayrhofen. In these, the PNFB communities in the bulk and rhizosphere soils and the plant endospheres were characterized by nifH-targeted PCR and massive parallel sequencing. The data revealed strong effects of climatic region on the dominating nitrogen fixers. Specifically, nifH sequences related to Geobacter (δ-Proteobacteria were present in high relative abundances in the nitrogen-fixing communities in the Mayrhofen and Kilpisjärvi regions, while members of the Clostridiales prevailed in the Kilpisjärvi and Ny-Ålesund regions. The bulk and

  20. Highly plastic genome of Microcystis aeruginosa PCC 7806, a ubiquitous toxic freshwater cyanobacterium

    Directory of Open Access Journals (Sweden)

    Latifi Amel

    2008-06-01

    Full Text Available Abstract Background The colonial cyanobacterium Microcystis proliferates in a wide range of freshwater ecosystems and is exposed to changing environmental factors during its life cycle. Microcystis blooms are often toxic, potentially fatal to animals and humans, and may cause environmental problems. There has been little investigation of the genomics of these cyanobacteria. Results Deciphering the 5,172,804 bp sequence of Microcystis aeruginosa PCC 7806 has revealed the high plasticity of its genome: 11.7% DNA repeats containing more than 1,000 bases, 6.8% putative transposases and 21 putative restriction enzymes. Compared to the genomes of other cyanobacterial lineages, strain PCC 7806 contains a large number of atypical genes that may have been acquired by lateral transfers. Metabolic pathways, such as fermentation and a methionine salvage pathway, have been identified, as have genes for programmed cell death that may be related to the rapid disappearance of Microcystis blooms in nature. Analysis of the PCC 7806 genome also reveals striking novel biosynthetic features that might help to elucidate the ecological impact of secondary metabolites and lead to the discovery of novel metabolites for new biotechnological applications. M. aeruginosa and other large cyanobacterial genomes exhibit a rapid loss of synteny in contrast to other microbial genomes. Conclusion Microcystis aeruginosa PCC 7806 appears to have adopted an evolutionary strategy relying on unusual genome plasticity to adapt to eutrophic freshwater ecosystems, a property shared by another strain of M. aeruginosa (NIES-843. Comparisons of the genomes of PCC 7806 and other cyanobacterial strains indicate that a similar strategy may have also been used by the marine strain Crocosphaera watsonii WH8501 to adapt to other ecological niches, such as oligotrophic open oceans.

  1. Using oxidized liquid and solid human waste as nutrients for Chlorella vulgaris and cyanobacterium Oscillatoria deflexa

    Science.gov (United States)

    Trifonov, Sergey V.; Kalacheva, Galina; Tirranen, Lyalya; Gribovskaya, Iliada

    At stationary terrestrial and space stations with closed and partially closed substance exchange not only plants, but also algae can regenerate atmosphere. Their biomass can be used for feeding Daphnia and Moina species, which, in their turn, serve as food for fish. In addition, it is possible to use algae for production of biological fuel. We suggested two methods of human waste mineralization: dry (evaporation with subsequent incineration in a muffle furnace) and wet (oxidation in a reactor using hydrogen peroxide). The research task was to prepare nutrient media for green alga Chlorella vulgaris and cyanobacterium Oscillatoria deflexa using liquid human waste mineralized by dry method, and to prepare media for chlorella on the basis of 1) liquid and 2) liquid and solid human waste mineralized by wet method. The algae were grown in batch culture in a climate chamber with the following parameters: illumination 7 klx, temperature 27-30 (°) C, culture density 1-2 g/l of dry weight. The control for chlorella was Tamiya medium, pH-5, and for oscillstoria — Zarrouk medium, pH-10. Maximum permissible concentrations of NaCl, Cl, urea (NH _{2}) _{2}CO, and native urine were established for algae. Missing ingredients (such as salts and acids) for experimental nutrient media were determined: their addition made it possible to obtain the biomass production not less than that in the control. The estimation was given of the mineral and biochemical composition of algae grown on experimental media. Microbiological test revealed absence of foreign microbial flora in experimental cultures.

  2. Ethylene Regulates the Physiology of the Cyanobacterium Synechocystis sp. PCC 6803 via an Ethylene Receptor.

    Science.gov (United States)

    Lacey, Randy F; Binder, Brad M

    2016-08-01

    Ethylene is a plant hormone that plays a crucial role in the growth and development of plants. The ethylene receptors in plants are well studied, and it is generally assumed that they are found only in plants. In a search of sequenced genomes, we found that many bacterial species contain putative ethylene receptors. Plants acquired many proteins from cyanobacteria as a result of the endosymbiotic event that led to chloroplasts. We provide data that the cyanobacterium Synechocystis (Synechocystis sp. PCC 6803) has a functional receptor for ethylene, Synechocystis Ethylene Response1 (SynEtr1). We first show that SynEtr1 directly binds ethylene. Second, we demonstrate that application of ethylene to Synechocystis cells or disruption of the SynEtr1 gene affects several processes, including phototaxis, type IV pilus biosynthesis, photosystem II levels, biofilm formation, and spontaneous cell sedimentation. Our data suggest a model where SynEtr1 inhibits downstream signaling and ethylene inhibits SynEtr1. This is similar to the inverse-agonist model of ethylene receptor signaling proposed for plants and suggests a conservation of structure and function that possibly originated over 1 billion years ago. Prior research showed that SynEtr1 also contains a light-responsive phytochrome-like domain. Thus, SynEtr1 is a bifunctional receptor that mediates responses to both light and ethylene. To our knowledge, this is the first demonstration of a functional ethylene receptor in a nonplant species and suggests that that the perception of ethylene is more widespread than previously thought. © 2016 American Society of Plant Biologists. All Rights Reserved.

  3. Hopanoids play a role in stress tolerance and nutrient storage in the cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Ricci, J N; Morton, R; Kulkarni, G; Summers, M L; Newman, D K

    2017-01-01

    Hopanes are abundant in ancient sedimentary rocks at discrete intervals in Earth history, yet interpreting their significance in the geologic record is complicated by our incomplete knowledge of what their progenitors, hopanoids, do in modern cells. To date, few studies have addressed the breadth of diversity of physiological functions of these lipids and whether those functions are conserved across the hopanoid-producing bacterial phyla. Here, we generated mutants in the filamentous cyanobacterium, Nostoc punctiforme, that are unable to make all hopanoids (shc) or 2-methylhopanoids (hpnP). While the absence of hopanoids impedes growth of vegetative cells at high temperature, the shc mutant grows faster at low temperature. This finding is consistent with hopanoids acting as membrane rigidifiers, a function shared by other hopanoid-producing phyla. Apart from impacting fitness under temperature stress, hopanoids are dispensable for vegetative cells under other stress conditions. However, hopanoids are required for stress tolerance in akinetes, a resting survival cell type. While 2-methylated hopanoids do not appear to contribute to any stress phenotype, total hopanoids and to a lesser extent 2-methylhopanoids were found to promote the formation of cyanophycin granules in akinetes. Finally, although hopanoids support symbiotic interactions between Alphaproteobacteria and plants, they do not appear to facilitate symbiosis between N. punctiforme and the hornwort Anthoceros punctatus. Collectively, these findings support interpreting hopanes as general environmental stress biomarkers. If hopanoid-mediated enhancement of nitrogen-rich storage products turns out to be a conserved phenomenon in other organisms, a better understanding of this relationship may help us parse the enrichment of 2-methylhopanes in the rock record during episodes of disrupted nutrient cycling. © 2016 John Wiley & Sons Ltd.

  4. Isolation and characterization of the small subunit of the uptake hydrogenase from the cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Raleiras, Patrícia; Kellers, Petra; Lindblad, Peter; Styring, Stenbjörn; Magnuson, Ann

    2013-06-21

    In nitrogen-fixing cyanobacteria, hydrogen evolution is associated with hydrogenases and nitrogenase, making these enzymes interesting targets for genetic engineering aimed at increased hydrogen production. Nostoc punctiforme ATCC 29133 is a filamentous cyanobacterium that expresses the uptake hydrogenase HupSL in heterocysts under nitrogen-fixing conditions. Little is known about the structural and biophysical properties of HupSL. The small subunit, HupS, has been postulated to contain three iron-sulfur clusters, but the details regarding their nature have been unclear due to unusual cluster binding motifs in the amino acid sequence. We now report the cloning and heterologous expression of Nostoc punctiforme HupS as a fusion protein, f-HupS. We have characterized the anaerobically purified protein by UV-visible and EPR spectroscopies. Our results show that f-HupS contains three iron-sulfur clusters. UV-visible absorption of f-HupS has bands ∼340 and 420 nm, typical for iron-sulfur clusters. The EPR spectrum of the oxidized f-HupS shows a narrow g = 2.023 resonance, characteristic of a low-spin (S = ½) [3Fe-4S] cluster. The reduced f-HupS presents complex EPR spectra with overlapping resonances centered on g = 1.94, g = 1.91, and g = 1.88, typical of low-spin (S = ½) [4Fe-4S] clusters. Analysis of the spectroscopic data allowed us to distinguish between two species attributable to two distinct [4Fe-4S] clusters, in addition to the [3Fe-4S] cluster. This indicates that f-HupS binds [4Fe-4S] clusters despite the presence of unusual coordinating amino acids. Furthermore, our expression and purification of what seems to be an intact HupS protein allows future studies on the significance of ligand nature on redox properties of the iron-sulfur clusters of HupS.

  5. Isolation and in silico analysis of Fe-superoxide dismutase in the cyanobacterium Nostoc commune.

    Science.gov (United States)

    Kesheri, Minu; Kanchan, Swarna; Richa; Sinha, Rajeshwar P

    2014-12-15

    Cyanobacteria are known to endure various stress conditions due to the inbuilt potential for oxidative stress alleviation owing to the presence of an array of antioxidants. The present study shows that Antarctic cyanobacterium Nostoc commune possesses two antioxidative enzymes viz., superoxide dismutase (SOD) and catalase that jointly cope with environmental stresses prevailing at its natural habitat. Native-PAGE analysis illustrates the presence of a single prominent isoform recognized as Fe-SOD and three distinct isoforms of catalase. The protein sequence of Fe-SOD in N. commune retrieved from NCBI protein sequence database was used for in silico analysis. 3D structure of N. commune was predicted by comparative modeling using MODELLER 9v11. Further, this model was validated for its quality by Ramachandran plot, ERRAT, Verify 3D and ProSA-web which revealed good structure quality of the model. Multiple sequence alignment showed high conservation in N and C-terminal domain regions along with all metal binding positions in Fe-SOD which were also found to be highly conserved in all 28 cyanobacterial species under study, including N. commune. In silico prediction of isoelectric point and molecular weight of Fe-SOD was found to be 5.48 and 22,342.98Da respectively. The phylogenetic tree revealed that among 28 cyanobacterial species, Fe-SOD in N. commune was the closest evolutionary homolog of Fe-SOD in Nostoc punctiforme as evident by strong bootstrap value. Thus, N. commune may serve as a good biological model for studies related to survival of life under extreme conditions prevailing at the Antarctic region. Moreover cyanobacteria may be exploited for biochemical and biotechnological applications of enzymatic antioxidants. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. DL-7-azatryptophan and citrulline metabolism in the cyanobacterium Anabaena sp. strain 1F

    International Nuclear Information System (INIS)

    Chen, C.H.; Van Baalen, C.; Tabita, F.R.

    1987-01-01

    An alternative route for the primary assimilation of ammonia proceeds via glutamine synthetase-carbamyl phosphate synthetase and its inherent glutaminase activity in Anabaena sp. strain 1F, a marine filamentous, heterocystous cyanobacterium. Evidence for the presence of this possible alternative route to glutamate was provided by the use of amino acid analogs as specific enzyme inhibitors, enzymological studies, and radioistopic labeling experiments. The amino acid pool patterns of continuous cultures of Anabaena sp. strain 1F were markedly influenced by the nitrogen source. A relatively high concentration of glutamate was maintained in the amino acid pools of all cultures irrespective of the nitrogen source, reflecting the central role of glutamate in nitrogen metabolism. The addition of 1.0 microM azaserine increased the intracellular pools of glutamate and glutamine. All attempts to detect any enzymatic activity for glutamate synthase by measuring the formation of L-[ 14 C]glutamate from 2-keto-[1- 14 C]glutarate and glutamine failed. The addition of 10 microM DL-7-azatryptophan caused a transient accumulation of intracellular citrulline and alanine which was not affected by the presence of chloramphenicol. The in vitro activity of carbamyl phosphate synthetase and glutaminase increased severalfold in the presence of azatryptophan. Results from radioisotopic labeling experiments with [ 14 C]bicarbonate and L-[1- 14 C]ornithine also indicated that citrulline was formed via carbamyl phosphate synthetase and ornithine transcarbamylase. In addition to its effects on nitrogen metabolism, azatryptophan also affected carbon metabolism by inhibiting photosynthetic carbon assimilation and photosynthetic oxygen evolution

  7. A Salt-Inducible Mn-Catalase (KatB) Protects Cyanobacterium from Oxidative Stress.

    Science.gov (United States)

    Chakravarty, Dhiman; Banerjee, Manisha; Bihani, Subhash C; Ballal, Anand

    2016-02-01

    Catalases, enzymes that detoxify H2O2, are widely distributed in all phyla, including cyanobacteria. Unlike the heme-containing catalases, the physiological roles of Mn-catalases remain inadequately characterized. In the cyanobacterium Anabaena, pretreatment of cells with NaCl resulted in unusually enhanced tolerance to oxidative stress. On exposure to H2O2, the NaCl-treated Anabaena showed reduced formation of reactive oxygen species, peroxides, and oxidized proteins than the control cells (i.e. not treated with NaCl) exposed to H2O2. This protective effect correlated well with the substantial increase in production of KatB, a Mn-catalase. Addition of NaCl did not safeguard the katB mutant from H2O2, suggesting that KatB was indeed responsible for detoxifying the externally added H2O2. Moreover, Anabaena deficient in KatB was susceptible to oxidative effects of salinity stress. The katB gene was strongly induced in response to osmotic stress or desiccation. Promoter-gfp analysis showed katB to be expressed only in the vegetative cells but not in heterocysts. Biochemically, KatB was an efficient, robust catalase that remained active in the presence of high concentrations of NaCl. Our findings unravel the role of Mn-catalase in acclimatization to salt/oxidative stress and demonstrate that the oxidative stress resistance of an organism can be enhanced by a simple compound such as NaCl. © 2016 American Society of Plant Biologists. All Rights Reserved.

  8. Compost biofortification with diazotrophic and P-solubilizing bacteria improves maturation process and P availability.

    Science.gov (United States)

    Busato, Jader G; Zandonadi, Daniel B; Mól, Alan R; Souza, Rafaela S; Aguiar, Kamilla P; Júnior, Fábio B Reis; Olivares, Fábio L

    2017-02-01

    Phosphorus-containing fertilizers play an important role in tropical agriculture owing to the well documented shortage of plant-available P in soils. Traditional P fertilizer production is based on chemical processing of insoluble rock phosphate (RP), which includes an acid treatment at high temperature. Processing the RP increases fertilizer costs, making it unavailable for undercapitalized and typically family-based farmers. Biotechnological methods have been proposed as an alternative to increase phosphate availability in RP. In this study, Burkholderia silvatlantica and Herbaspirillum seropedicae were co-inoculated into an RP-enriched compost with the aim of determining the effects of this technology on the levels of phosphatase activities and release of plant-available P. Inoculation of both microorganisms resulted in higher organic matter decomposition and higher humic acid formation in composting. Herbaspirillum seropedicae was the most promising microorganism for the production of acid and alkaline phosphatase enzymes. Both microorganisms presented potential to increase the supply of P from poorly soluble sources owing to increased levels of water-soluble P and citric acid P. Burkholderia silvatlantica and H. seropedicae in RP-enriched compost may represent an important biotechnological tool to reduce the overall time required for composting and increase the supply of P from poorly soluble sources. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  9. Evidence regarding the possible role of c-phycoerythrin in ultraviolet-B tolerance in a thermophilic cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Wingard, C.E.; Castenholz, R.W. [Oregon Univ., Eugene, OR (United States). Dept. of Biology; Schiller, J.R. [Bowdoin College, Brunswick, ME (United States)

    1997-05-01

    It was recently reported that a strain of Nostoc spongiaeforme (cyanobacteria) with the photopigment c-phycoerythrin (c-PE) may be more tolerant of the adverse effects of UVB radiation than the same strain lacking c-PE due to chromatic adaptation (CA) (Tyagi et al., Photochem. Photobiol. 55, 401-407, 1992). It was proposed that this increased UVB tolerance may be due to the presence of c-PE, perhaps as a function of the ability of strains with c-PE to chromatically adapt. We tested the role of c-PE in UVB tolerance by comparing the short- and long-term effects of UVB exposure on photosynthesis pigmentation and the protein contents of four experimental cultures of the thermophilic cyanobacterium Oscillatoria cf. amphigranulata. These cultures consisted of a wild-type strain that produces c-PE, a green pigment variant (subcloned from the parent wild-type strain) incapable of producing c-PE and two chromatically adapted color forms of the wild-type strain that varied with regard to their total c-PE content. There were no significant results suggesting a role for c-PE in UVB tolerance. It is concluded that the photopigment c-PE does not confer enhanced resistance to the deleterious effects of UVB radiation on photosynthesis in this cyanobacterium. (author).

  10. The effects of the exopolysaccharide and growth rate on the morphogenesis of the terrestrial filamentous cyanobacterium Nostoc flagelliforme

    Directory of Open Access Journals (Sweden)

    Lijuan Cui

    2017-09-01

    Full Text Available The terrestrial cyanobacterium Nostoc flagelliforme, which contributes to carbon and nitrogen supplies in arid and semi-arid regions, adopts a filamentous colony form. Owing to its herbal and dietary values, this species has been overexploited. Largely due to the lack of understanding on its morphogenesis, artificial cultivation has not been achieved. Additionally, it may serve as a useful model for recognizing the morphological adaptation of colonial cyanobacteria in terrestrial niches. However, it shows very slow growth in native habitats and is easily disintegrated under laboratory conditions. Thus, a novel experimental system is necessary to explore its morphogenetic mechanism. Liquid-cultured N. flagelliforme has been well developed for exopolysaccharide (EPS production, in which microscopic colonies (micro-colonies are generally formed. In this study, we sought to gain some insight into the morphogenesis of N. flagelliforme by examining the effects of two external factors, the EPS and environmental stress-related growth rate, on the morphological shaping of micro-colonies. Our findings indicate that the EPS matrix could act as a basal barrier, leading to the bending of trichomes during their elongation, while very slow growth is conducive to their straight elongation. These findings will guide future cultivation and application of this cyanobacterium for ecological improvement.

  11. Arabinogalactan proteins occur in the free-living cyanobacterium genus Nostoc and in plant-Nostoc symbioses.

    Science.gov (United States)

    Jackson, Owen; Taylor, Oliver; Adams, David G; Knox, J Paul

    2012-10-01

    Arabinogalactan proteins (AGP) are a diverse family of proteoglycans associated with the cell surfaces of plants. AGP have been implicated in a wide variety of plant cell processes, including signaling in symbioses. This study investigates the existence of putative AGP in free-living cyanobacterial cultures of the nitrogen-fixing, filamentous cyanobacteria Nostoc punctiforme and Nostoc sp. strain LBG1 and at the symbiotic interface in the symbioses between Nostoc spp. and two host plants, the angiosperm Gunnera manicata (in which the cyanobacterium is intracellular) and the liverwort Blasia pusilla (in which the cyanobacterium is extracellular). Enzyme-linked immunosorbent assay, immunoblotting, and immunofluorescence analyses demonstrated that three AGP glycan epitopes (recognized by monoclonal antibodies LM14, MAC207, and LM2) are present in free-living Nostoc cyanobacterial species. The same three AGP glycan epitopes are present at the Gunnera-Nostoc symbiotic interface and the LM2 epitope is detected during the establishment of the Blasia-Nostoc symbiosis. Bioinformatic analysis of the N. punctiforme genome identified five putative AGP core proteins that are representative of AGP classes found in plants. These results suggest a possible involvement of AGP in cyanobacterial-plant symbioses and are also suggestive of a cyanobacterial origin of AGP.

  12. Evidence regarding the possible role of c-phycoerythrin in ultraviolet-B tolerance in a thermophilic cyanobacterium

    International Nuclear Information System (INIS)

    Wingard, C.E.; Castenholz, R.W.

    1997-01-01

    It was recently reported that a strain of Nostoc spongiaeforme (cyanobacteria) with the photopigment c-phycoerythrin (c-PE) may be more tolerant of the adverse effects of UVB radiation than the same strain lacking c-PE due to chromatic adaptation (CA) (Tyagi et al., Photochem. Photobiol. 55, 401-407, 1992). It was proposed that this increased UVB tolerance may be due to the presence of c-PE, perhaps as a function of the ability of strains with c-PE to chromatically adapt. We tested the role of c-PE in UVB tolerance by comparing the short- and long-term effects of UVB exposure on photosynthesis pigmentation and the protein contents of four experimental cultures of the thermophilic cyanobacterium Oscillatoria cf. amphigranulata. These cultures consisted of a wild-type strain that produces c-PE, a green pigment variant (subcloned from the parent wild-type strain) incapable of producing c-PE and two chromatically adapted color forms of the wild-type strain that varied with regard to their total c-PE content. There were no significant results suggesting a role for c-PE in UVB tolerance. It is concluded that the photopigment c-PE does not confer enhanced resistance to the deleterious effects of UVB radiation on photosynthesis in this cyanobacterium. (author)

  13. A Nostoc punctiforme Sugar Transporter Necessary to Establish a Cyanobacterium-Plant Symbiosis1[C][W

    Science.gov (United States)

    Ekman, Martin; Picossi, Silvia; Campbell, Elsie L.; Meeks, John C.; Flores, Enrique

    2013-01-01

    In cyanobacteria-plant symbioses, the symbiotic nitrogen-fixing cyanobacterium has low photosynthetic activity and is supplemented by sugars provided by the plant partner. Which sugars and cyanobacterial sugar uptake mechanism(s) are involved in the symbiosis, however, is unknown. Mutants of the symbiotically competent, facultatively heterotrophic cyanobacterium Nostoc punctiforme were constructed bearing a neomycin resistance gene cassette replacing genes in a putative sugar transport gene cluster. Results of transport activity assays using 14C-labeled fructose and glucose and tests of heterotrophic growth with these sugars enabled the identification of an ATP-binding cassette-type transporter for fructose (Frt), a major facilitator permease for glucose (GlcP), and a porin needed for the optimal uptake of both fructose and glucose. Analysis of green fluorescent protein fluorescence in strains of N. punctiforme bearing frt::gfp fusions showed high expression in vegetative cells and akinetes, variable expression in hormogonia, and no expression in heterocysts. The symbiotic efficiency of N. punctiforme sugar transport mutants was investigated by testing their ability to infect a nonvascular plant partner, the hornwort Anthoceros punctatus. Strains that were specifically unable to transport glucose did not infect the plant. These results imply a role for GlcP in establishing symbiosis under the conditions used in this work. PMID:23463784

  14. Characterization of the coccoid cyanobacterium Myxosarcina sp. KIOST-1 isolated from mangrove forest in Chuuk State, Federated States of Micronesia

    Science.gov (United States)

    Kim, Ji Hyung; Lee, JunMo; Affan, Md-Abu; Lee, Dae-Won; Kang, Do-Hyung

    2017-09-01

    Mangrove forests are known to be inhabited by diverse symbiotic cyanobacterial communities that are capable of N2 fixation. To investigate its biodiversity, root sediments were collected from a mangrove forest in Chuuk State, Federated States of Micronesia (FSM), and an entangled yellow-brown coccoid cyanobacterium was isolated. The isolated cyanobacterium was reproduced by multiple fission and eventually produced baeocytes. Phylogenetic analysis revealed that the isolate was most similar to the genera Myxosarcina and Chroococcidiopsis in the order Pleurocapsales. Compositions of protein, lipid and carbohydrate in the cyanobacterial cells were estimated to be 19.4 ± 0.1%, 18.8 ± 0.4% and 31.5 ± 0.1%, respectively. Interestingly, total fatty acids in the isolate were mainly composed of saturated fatty acids and monounsaturated fatty acids, whereas polyunsaturated fatty acids were not detected. Based on the molecular and biochemical characteristics, the isolate was finally classified in the genus Myxosarcina, and designated as Myxosarcina sp. KIOST-1. These results will contribute to better understanding of cyanobacterial biodiversity in the mangrove forest in FSM as well as the genus Myxosarcina, and also will allow further exploitation of its biotechnological potential on the basis of its cellular characteristics.

  15. Characterization of the cytochrome c oxidase in isolated and purified plasma membranes from the cyanobacterium Anacystis nidulans

    International Nuclear Information System (INIS)

    Peschek, G.A.; Wastyn, M.; Trnka, M.; Molitor, V.; Fry, I.V.; Packer, L.

    1989-01-01

    Functionally intact plasma membranes were isolated from the cyanobacterium (blue-green alga) Anacystis nidulans through French pressure cell extrusion of lysozyme/EDTA-treated cells, separated from thylakoid membranes by discontinuous sucrose density gradient centrifugation, and purified by repeated recentrifugation. Origin and identity of the chlorophyll-free plasma membrane fraction were confirmed by labeling of intact cells with impermeant protein markers, [ 35 S]diazobenzenesulfonate and fluorescamine, prior to membrane isolation. Rates of oxidation of reduced horse heart cytochrome c by purified plasma and thylakoid membranes were 90 and 2 nmol min -1 (mg of protein) -1 , respectively. The cytochrome oxidase in isolated plasma membranes was identified as a copper-containing aa 3 -type enzyme from the properties of its redox-active and EDTA-resistant Cu 2+ ESR signal, the characteristic inhibition profile, reduced minus oxidized difference spectra, carbon monoxide difference spectra, photoaction and photodissociation spectra of the CO-inhibited enzyme, and immunological cross-reaction of two subunits of the enzyme with antibodies against subunits I and II, and the holoenzyme, of Paracoccus denitrificans aa 3 -type cytochrome oxidase. The data presented are the first comprehensive evidence for the occurrence of aa 3 -type cytochrome oxidase in the plasma membrane of a cyanobacterium similar to the corresponding mitochondrial enzyme

  16. Importance measures

    International Nuclear Information System (INIS)

    Gomez Cobo, A.

    1997-01-01

    The presentation discusses the following: general concepts of importance measures; example fault tree, used to illustrate importance measures; Birnbaum's structural importance; criticality importance; Fussel-Vesely importance; upgrading function; risk achievement worth; risk reduction worth

  17. HupW Protease Specifically Required for Processing of the Catalytic Subunit of the Uptake Hydrogenase in the Cyanobacterium Nostoc sp. Strain PCC 7120

    Science.gov (United States)

    Lindberg, Pia; Devine, Ellenor; Stensjö, Karin

    2012-01-01

    The maturation process of [NiFe] hydrogenases includes a proteolytic cleavage of the large subunit. We constructed a mutant of Nostoc strain PCC 7120 in which hupW, encoding a putative hydrogenase-specific protease, is inactivated. Our results indicate that the protein product of hupW selectively cleaves the uptake hydrogenase in this cyanobacterium. PMID:22020512

  18. Concerted changes in gene expression and cell physiology of the cyanobacterium Synechocystis sp. strain PCC 6803 during transitions between nitrogen and light-limited growth

    NARCIS (Netherlands)

    Aquirre von Wobeser, E.; Ibelings, B.W.; Bok, J.M.; Krasikov, V.; Huisman, J.; Matthijs, H.C.P.

    2011-01-01

    Physiological adaptation and genome-wide expression profiles of the cyanobacterium Synechocystis sp. strain PCC 6803 in response to gradual transitions between nitrogen-limited and light-limited growth conditions were measured in continuous cultures. Transitions induced changes in pigment

  19. The Genome Sequence of the Cyanobacterium Oscillatoria sp. PCC 6506 Reveals Several Gene Clusters Responsible for the Biosynthesis of Toxins and Secondary Metabolites▿

    Science.gov (United States)

    Méjean, Annick; Mazmouz, Rabia; Mann, Stéphane; Calteau, Alexandra; Médigue, Claudine; Ploux, Olivier

    2010-01-01

    We report a draft sequence of the genome of Oscillatoria sp. PCC 6506, a cyanobacterium that produces anatoxin-a and homoanatoxin-a, two neurotoxins, and cylindrospermopsin, a cytotoxin. Beside the clusters of genes responsible for the biosynthesis of these toxins, we have found other clusters of genes likely involved in the biosynthesis of not-yet-identified secondary metabolites. PMID:20675499

  20. THE STRUCTURE OF PHOTOSYSTEM-I FROM THE THERMOPHILIC CYANOBACTERIUM SYNECHOCOCCUS SP DETERMINED BY ELECTRON-MICROSCOPY OF 2-DIMENSIONAL CRYSTALS

    NARCIS (Netherlands)

    BOTTCHER, B; GRABER, P; BOEKEMA, EJ

    1992-01-01

    The structure of the Photosystem I (PS I) complex from the thermophilic cyanobacterium Synechococcus sp. has been investigated by electron microscopy and image analysis of two-dimensional crystals. Crystals were obtained from isolated PS I by removal of detergents with Bio-Beads. After negative

  1. Enhancement of human adaptive immune responses by administration of a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensis

    DEFF Research Database (Denmark)

    Pedersen, Morten Løbner; Walsted, Anette; Larsen, Rune

    2008-01-01

    The effect of consumption of Immulina, a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensis, on adaptive immune responses was investigated by evaluation of changes in leukocyte responsiveness to two foreign recall antigens, Candida albicans (CA) and tetanus...

  2. Optical characterization of the oceanic unicellular cyanobacterium Synechococcus grown under a day-night cycle in natural irradiance

    Science.gov (United States)

    Stramski, Dariusz; Shalapyonok, Alexi; Reynolds, Rick A.

    1995-01-01

    The optical properties of the ocenanic cyanobacterium Synechococcus (clone WH8103) were examined in a nutrient-replete laboratory culture grown under a day-night cycle in natural irradiance. Measurements of the spectral absorption and beam attenuation coefficients, the size distribution of cells in suspension, and microscopic analysis of samples were made at intervals of 2-4 hours for 2 days. These measurements were used to calculate the optical properties at the level of a single 'mean' cell representative of the acutal population, specifically, the optical cross sections for spectral absorption bar-(sigma(sub a)), scattering bar-sigma(sub b))(lambda), and attentuation bar-(sigma(sub c))(lambda). In addition, concurrent determinations of chlorophyll a and particulate organic carbon allowed calculation of the Chl a- and C-specific optical coefficients. The refractive index of cells was derived from the observed data using a theory of light absorption and scattering by homogeneous spheres. Low irradiance because of cloudy skies resulted in slow division rates of cells in the culture. The percentage of dividing cells was unusually high (greater than 30%) throughout the experiment. The optical cross sections varied greatly over a day-night cycle, with a minimum near dawn or midmorning and maximum near dusk. During daylight hours, bar-(sigma(sub b)) and bar-(sigma(sub c)) can increase more than twofold and bar-(sigma(sub a) by as much as 45%. The real part of the refractive index n increaed during the day; changes in n had equal or greater effect than the varying size distribution on changes in bar-(sigma(sub c)) and bar-(sigma(sub b)). The contribution of changes in n to the increase of bar-(sigma(sub c))(660) during daylight hours was 65.7% and 45.1% on day 1 and 2, respectively. During the dark period, when bar-(sigma(sub c))(660) decreased by a factor of 2.9, the effect of decreasing n was dominant (86.3%). With the exception of a few hours during the second light

  3. Transcription and Regulation of the Bidirectional Hydrogenase in the Cyanobacterium Nostoc sp. Strain PCC 7120▿

    Science.gov (United States)

    Sjöholm, Johannes; Oliveira, Paulo; Lindblad, Peter

    2007-01-01

    The filamentous, heterocystous cyanobacterium Nostoc sp. strain PCC 7120 (Anabaena sp. strain PCC 7120) possesses an uptake hydrogenase and a bidirectional enzyme, the latter being capable of catalyzing both H2 production and evolution. The completely sequenced genome of Nostoc sp. strain PCC 7120 reveals that the five structural genes encoding the bidirectional hydrogenase (hoxEFUYH) are separated in two clusters at a distance of approximately 8.8 kb. The transcription of the hox genes was examined under nitrogen-fixing conditions, and the results demonstrate that the cluster containing hoxE and hoxF can be transcribed as one polycistronic unit together with the open reading frame alr0750. The second cluster, containing hoxU, hoxY, and hoxH, is transcribed together with alr0763 and alr0765, located between the hox genes. Moreover, alr0760 and alr0761 form an additional larger operon. Nevertheless, Northern blot hybridizations revealed a rather complex transcription pattern in which the different hox genes are expressed differently. Transcriptional start points (TSPs) were identified 66 and 57 bp upstream from the start codon of alr0750 and hoxU, respectively. The transcriptions of the two clusters containing the hox genes are both induced under anaerobic conditions concomitantly with the induction of a higher level of hydrogenase activity. An additional TSP, within the annotated alr0760, 244 bp downstream from the suggested translation start codon, was identified. Electrophoretic mobility shift assays with purified LexA from Nostoc sp. strain PCC 7120 demonstrated specific interactions between the transcriptional regulator and both hox promoter regions. However, when LexA from Synechocystis sp. strain PCC 6803 was used, the purified protein interacted only with the promoter region of the alr0750-hoxE-hoxF operon. A search of the whole Nostoc sp. strain PCC 7120 genome demonstrated the presence of 216 putative LexA binding sites in total, including recA and rec

  4. The persistence and ecological impacts of a cyanobacterium genetically engineered to express mosquitocidal Bacillus thuringiensis toxins.

    Science.gov (United States)

    Ketseoglou, Irene; Bouwer, Gustav

    2016-05-10

    The cyanobacterium Anabaena PCC 7120#11 has been genetically engineered to act as a delivery vehicle for Bacillus thuringiensis subspecies israelensis mosquitocidal toxins. To address ecological concerns about releasing this genetically engineered microorganism into the environment for mosquito larva control, the persistence and ecological impacts of PCC 7120#11 was evaluated using multi-species, standardized aquatic microcosms. The microcosms were set up as described in ASTM E1366-02 (Standard Practice for Standardized Aquatic Microcosms: Fresh Water), with a few modifications. The treatment group microcosms were inoculated with PCC 7120#11 and key water quality parameters and non-target effects were compared between the treatment and control groups over a period of 35 days. PCC 7120#11 decreased from a concentration of 4.50 × 10(6) cells/ml (at inoculation) to 1.32 × 10(3) cells/ml after 4 weeks and larvicidal activity against third instar larvae of Anopheles arabiensis was only evident for two weeks after treatment. Both treatment and the interaction of treatment and time had a significant effect on nitrate, phosphate and photosynthetic microorganism concentrations. Treatment with PCC 7120#11 caused a temporary spike in ammonia in the microcosms a week after treatment, but the concentrations were well below acute and chronic criteria values for ammonia in freshwater ecosystems. Cyprinotus vidua concentrations were not significantly different between PCC 7120#11 and control microcosms. In PCC 7120#11 microcosms, Daphnia pulex concentrations were significantly lower than control concentrations between days 18 and 25. By the end of the experiment, none of the measured variables were significantly different between the treatment groups. The standard aquatic microcosm experiments provided more data on the ecological impacts of PCC 7120#11 than single-organism assessments would have. On the basis of the relatively minor, short-term effects that PCC 7120

  5. Diazotrophic microbial mats

    NARCIS (Netherlands)

    Severin, I.; Stal, L.J.; Seckbach, J.; Oren, A.

    2010-01-01

    Microbial mats have been the focus of scientific research for a few decades. These small-scale ecosystems are examples of versatile benthic communities of microorganisms, usually dominated by phototrophic bacteria (e.g., Krumbein et al., 1977; Jørgensen et al., 1983). They develop as vertically

  6. Photosynthetic and Behavioral Versatility of the Cyanobacterium Oscillatoria-Boryana in a Sulfide-Rich Microbial Mat

    DEFF Research Database (Denmark)

    CASTENHOLZ, RW; JØRGENSEN, BB; DAMELIO, E.

    1991-01-01

    resulted in a downward retreat. The result was a lowered irradiance level for the Oscillatoria but, nevertheless, a high rate of oxygenic photosynthesis. O. boryana is a versatile cyanobacterium that appears to avoid photoinhibitory conditions and to optimize its light intensity for photosynthesis...... with dense O. boryana populations were used to make vertical profiles at intervals of 0.1-0.2 mm and also to estimate rates of oxygenic and anoxygenic photosynthesis during rapid light-dark transitions. In addition, attenuation of irradiance was measured in mats with O. boryana by a spectroradiometer...... with mini-fiber optic probe. Light-dependent incorporation of [C-14]-bicarbonate and [C-14]-acetate was measured in collected field populations of O. boryana. The combined results led to the conclusion that populations of O. boryana typically employed sulfide-dependent anoxygenic photosynthesis in early...

  7. Photoreactivation and excision repair of UV induced pyrimidine dimers in the unicellular cyanobacterium Gloeocapsa alpicola (Synechocystis PCC 6308)

    International Nuclear Information System (INIS)

    O'Brien, P.A.; Houghton, J.A.

    1982-01-01

    The survival curve obtained after UV irradiation of the unicellular cyanobacterium Synechocystis is typical of a DNA repair competent organism. Inhibition of DNA replication, by incubating cells in the dark, increased resistance to the lethal effects of UV at higher fluences. Exposure of irradiated cells to near ultraviolet light (350-500 nm) restored viability to pre-irradiation levels. In order to measure DNA repair activity, techniques have been developed for the chromatographic analysis of pyrimidine dimers in synechocystis. The specificity of this method was established using a haploid strain of Saccharomyces cerevisiae. In accordance with the physiological responses of irradiated cells to photoreactivating light, pyrimidine dimers were not detected after photoreactivation treatment. Incubation of irradiated cells under non-photoreactivating growth conditions for 15h resulted in complete removal of pyrimidine dimers. It is concluded that Synechocystis contains photoreactivation and excision repair systems for the removal of pyrimidine dimers. (author)

  8. Phosphorus addition reverses the positive effect of zebra mussels (Dreissena polymorpha) on the toxic cyanobacterium, Microcystis aeruginosa.

    Science.gov (United States)

    Sarnelle, Orlando; White, Jeffrey D; Horst, Geoffrey P; Hamilton, Stephen K

    2012-07-01

    We tested the hypothesis that zebra mussels (Dreissena polymorpha) have positive effects on the toxin-producing cyanobacterium, Microcystis aeruginosa, at low phosphorus (P) concentrations, but negative effects on M. aeruginosa at high P, with a large-scale enclosure experiment in an oligotrophic lake. After three weeks, mussels had a significantly positive effect on M. aeruginosa at ambient P (total phosphorus, TP ∼10 μg L⁻¹), and a significantly negative effect at high P (simulating a TP of ∼40 μg L⁻¹ in lakes). Positive and negative effects were strong and very similar in magnitude. Thus, we were able to ameliorate a negative effect of Dreissena invasion on water quality (i.e., promotion of Microcystis) by adding P to water from an oligotrophic lake. Our results are congruent with many field observations of Microcystis response to Dreissena invasion across ecosystems of varying P availability. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. Optimization and effects of different culture conditions on growth of Halomicronema hongdechloris – a filamentous cyanobacterium containing chlorophyll f

    Directory of Open Access Journals (Sweden)

    Yaqiong eLi

    2014-02-01

    Full Text Available A chlorophyll f containing cyanobacterium, Halomicronema hongdechloris (H. hongdechloris was isolated from a stromatolite cyanobacterial community. However, the extremely slower growth rate of H. hongdechloris culture became a critical factor, hindering the research on this newly isolated cyanobacterium and the investigation of chlorophyll f-photosynthesis. Therefore, optimizing H. hongdechloris culture conditions has become an essential requirement for future research. This work investigated the effects of various culture conditions, essential nutrients and light environments to determine the optimal growth conditions for H. hongdechloris and the biosynthetic rate of chlorophyll f. Based on the total chlorophyll concentration, an optimal growth rate of 0.22 ± 0.02 day-1 (doubling time: 3.1 ± 0.3 days was observed when cells were grown under continuous illumination with far-red light with an intensity of 20 µE at 32°C in modified K+ES seawater (pH 8.0 with additional supplements of 11.75 mM NaNO3 and 0.15 mM K2HPO4. High performance liquid chromatography on H. hongdechloris pigments confirmed that chlorophyll a is the major chlorophyll and chlorophyll f constitutes approximately 10% of the total chlorophyll from cells grown under far-red light. Fluorescence confocal image analysis demonstrated changes of photosynthetic membranes and the distribution of photopigments in response to different light conditions. The total photosynthetic oxygen evolution yield per cell showed no changes under different light conditions, which confirms the involvement of chlorophyll f in oxygenic photosynthesis. The implications of the presence of chlorophyll f in H. hongdechloris and its relationship to light environment are discussed.

  10. Photosystem Trap Energies and Spectrally-Dependent Energy-Storage Efficiencies in the Chl d-Utilizing Cyanobacterium, Acaryochloris Marina

    Science.gov (United States)

    Mielke, Steven P.; Kiang, Nancy Y.; Blankenship, Robert E.; Mauzerall, David

    2012-01-01

    Acaryochloris marina is the only species known to utilize chlorophyll (Chl) d as a principal photopigment. The peak absorption wavelength of Chl d is redshifted approx. 40 nm in vivo relative to Chl a, enabling this cyanobacterium to perform oxygenic phototrophy in niche environments enhanced in far-red light. We present measurements of the in vivo energy-storage (E-S) efficiency of photosynthesis in A. marina, obtained using pulsed photoacoustics (PA) over a 90-nm range of excitation wavelengths in the red and far-red. Together with modeling results, these measurements provide the first direct observation of the trap energies of PSI and PSII, and also the photosystem-specific contributions to the total E-S efficiency. We find the maximum observed efficiency in A. marina (40+/-1% at 735 nm) is higher than in the Chl a cyanobacterium Synechococcus leopoliensis (35+/-1% at 690 nm). The efficiency at peak absorption wavelength is also higher in A. marina (36+/-1% at 710 nm vs. 31+/-1% at 670 nm). In both species, the trap efficiencies are approx. 40% (PSI) and approx. 30% (PSII). The PSI trap in A. marina is found to lie at 740+/-5 nm, in agreement with the value inferred from spectroscopic methods. The best fit of the model to the PA data identifies the PSII trap at 723+/-3 nm, supporting the view that the primary electron-donor is Chl d, probably at the accessory (ChlD1) site. A decrease in efficiency beyond the trap wavelength, consistent with uphill energy transfer, is clearly observed and fit by the model. These results demonstrate that the E-S efficiency in A. marina is not thermodynamically limited, suggesting that oxygenic photosynthesis is viable in even redder light environments.

  11. Proteomic strategy for the analysis of the polychlorobiphenyl-degrading cyanobacterium Anabaena PD-1 exposed to Aroclor 1254.

    Directory of Open Access Journals (Sweden)

    Hangjun Zhang

    Full Text Available The cyanobacterium Anabaena PD-1, which was originally isolated from polychlorobiphenyl (PCB-contaminated paddy soils, has capabilities for dechlorinatin and for degrading the commercial PCB mixture Aroclor 1254. In this study, 25 upregulated proteins were identified using 2D electrophoresis (2-DE coupled with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS. These proteins were involved in (i PCB degradation (i.e., 3-chlorobenzoate-3,4-dioxygenase; (ii transport processes [e.g., ATP-binding cassette (ABC transporter substrate-binding protein, amino acid ABC transporter substrate-binding protein, peptide ABC transporter substrate-binding protein, putrescine-binding protein, periplasmic solute-binding protein, branched-chain amino acid uptake periplasmic solute-binding protein, periplasmic phosphate-binding protein, phosphonate ABC transporter substrate-binding protein, and xylose ABC transporter substrate-binding protein]; (iii energetic metabolism (e.g., methanol/ethanol family pyrroloquinoline quinone (PQQ-dependent dehydrogenase, malate-CoA ligase subunit beta, enolase, ATP synthase β subunit, FOF1 ATP synthase subunit beta, ATP synthase α subunit, and IMP cyclohydrolase; (iv electron transport (cytochrome b6f complex Fe-S protein; (v general stress response (e.g., molecular chaperone DnaK, elongation factor G, and translation elongation factor thermostable; (vi carbon metabolism (methanol dehydrogenase and malate-CoA ligase subunit beta; and (vii nitrogen reductase (nitrous oxide reductase. The results of real-time polymerase chain reaction showed that the genes encoding for dioxygenase, ABC transporters, transmembrane proteins, electron transporter, and energetic metabolism proteins were significantly upregulated during PCB degradation. These genes upregulated by 1.26- to 8.98-fold. These findings reveal the resistance and adaptation of cyanobacterium to the presence of PCBs, shedding light on the

  12. Microenvironmental Ecology of the Chlorophyll b-containing Symbiotic Cyanobacterium Prochloron in the Didemnid Ascidian Lissoclinum patella

    Directory of Open Access Journals (Sweden)

    Michael eKühl

    2012-11-01

    Full Text Available The discovery of the cyanobacterium Prochloron was the first finding of a bacterial oxyphototroph with chlorophyll (Chl b, in addition to Chl a. It was first described as Prochloron didemni but a number of clades have since been described. Prochloron is a conspicuously large (7-25 µm unicellular cyanobacterium living in a symbiotic relationship, primarily with (sub- tropical didemnid ascidians; it has resisted numerous cultivation attempts and appears truly obligatory symbiotic. Recently, a Prochloron draft genome was published, revealing no lack of metabolic genes that could explain the apparent inability to reproduce and sustain photosynthesis in a free-living stage. Possibly, the unsuccessful cultivation is partly due to a lack of knowledge about the microenvironmental conditions and ecophysiology of Prochloron in its natural habitat. We used microsensors, variable chlorophyll fluorescence imaging and imaging of O2 and pH to obtain a detailed insight to the microenvironmental ecology and photobiology of Prochloron in hospite in the didemnid ascidian Lissoclinum patella. The microenvironment within ascidians is characterized by steep gradients of light and chemical parameters that change rapidly with varying irradiances. The interior zone of the ascidians harboring Prochloron thus became anoxic and acidic within a few min of darkness, while the same zone exhibited O2 super-saturation and strongly alkaline pH after a few min of illumination. Photosynthesis showed lack of photoinhibition even at high irradiances equivalent to full sunlight, and photosynthesis recovered rapidly after periods of anoxia. We discuss these new insights on the ecological niche of Prochloron and possible interactions with its host and other microbes in light of its recently published genome and a recent study of the overall microbial diversity and metagenome of L. patella.

  13. Antagonism at combined effects of chemical fertilizers and carbamate insecticides on the rice-field N2-fixing cyanobacterium Cylindrospermum sp. in vitro

    OpenAIRE

    Padhy Rabindra N.; Nayak Nabakishore; Rath Shakti

    2014-01-01

    Effects of chemical fertilizers (urea, super phosphate and potash) on toxicities of two carbamate insecticides, carbaryl and carbofuran, individually to the N2-fixing cyanobacterium, Cylindrospermum sp. were studied in vitro at partially lethal levels (below highest permissive concentrations) of each insecticide. The average number of vegetative cells between two polar heterocysts was 16.3 in control cultures, while the mean value of filament length increased in the presence of chemical ferti...

  14. Draft Genome Sequence of Cyanobacterium sp. Strain HL-69, Isolated from a Benthic Microbial Mat from a Magnesium Sulfate-Dominated Hypersaline Lake

    Energy Technology Data Exchange (ETDEWEB)

    Mobberley, J. M.; Romine, M. F.; Cole, J. K.; Maezato, Y.; Lindemann, S. R.; Nelson, W. C.

    2018-02-08

    ABSTRACT

    The complete genome sequence ofCyanobacteriumsp. strain HL-69 consists of 3,155,247 bp and contains 2,897 predicted genes comprising a chromosome and two plasmids. The genome is consistent with a halophilic nondiazotrophic phototrophic lifestyle, and this organism is able to synthesize most B vitamins and produces several secondary metabolites.

  15. IMPORTANT NOTIFICATION

    CERN Multimedia

    HR Department

    2009-01-01

    Green plates, removals and importation of personal effects Please note that, as from 1 April 2009, formalities relating to K and CD special series French vehicle plates (green plates), removals and importation of personal effects into France and Switzerland will be dealt with by GS Department (Building 73/3-014, tel. 73683/74407). Importation and purchase of tax-free vehicles in Switzerland, as well as diplomatic privileges, will continue to be dealt with by the Installation Service of HR Department (Building 33/1-011, tel. 73962). HR and GS Departments

  16. Transcriptional Activities of the Microbial Consortium Living with the Marine Nitrogen-Fixing Cyanobacterium Trichodesmium Reveal Potential Roles in Community-Level Nitrogen Cycling.

    Science.gov (United States)

    Lee, Michael D; Webb, Eric A; Walworth, Nathan G; Fu, Fei-Xue; Held, Noelle A; Saito, Mak A; Hutchins, David A

    2018-01-01

    Trichodesmium is a globally distributed cyanobacterium whose nitrogen-fixing capability fuels primary production in warm oligotrophic oceans. Like many photoautotrophs, Trichodesmium serves as a host to various other microorganisms, yet little is known about how this associated community modulates fluxes of environmentally relevant chemical species into and out of the supraorganismal structure. Here, we utilized metatranscriptomics to examine gene expression activities of microbial communities associated with Trichodesmium erythraeum (strain IMS101) using laboratory-maintained enrichment cultures that have previously been shown to harbor microbial communities similar to those of natural populations. In enrichments maintained under two distinct CO 2 concentrations for ∼8 years, the community transcriptional profiles were found to be specific to the treatment, demonstrating a restructuring of overall gene expression had occurred. Some of this restructuring involved significant increases in community respiration-related transcripts under elevated CO 2 , potentially facilitating the corresponding measured increases in host nitrogen fixation rates. Particularly of note, in both treatments, community transcripts involved in the reduction of nitrate, nitrite, and nitrous oxide were detected, suggesting the associated organisms may play a role in colony-level nitrogen cycling. Lastly, a taxon-specific analysis revealed distinct ecological niches of consistently cooccurring major taxa that may enable, or even encourage, the stable cohabitation of a diverse community within Trichodesmium consortia. IMPORTANCE Trichodesmium is a genus of globally distributed, nitrogen-fixing marine cyanobacteria. As a source of new nitrogen in otherwise nitrogen-deficient systems, these organisms help fuel carbon fixation carried out by other more abundant photoautotrophs and thereby have significant roles in global nitrogen and carbon cycling. Members of the Trichodesmium genus tend to

  17. HYDROGEN PRODUCTION BY THE CYANOBACTERIUM PLECTONEMA BORYANUM: EFFECTS OF INITIAL NITRATE CONCENTRATION, LIGHT INTENSITY, AND INHIBITION OF PHOTOSYSTEM II BY DCMU

    Energy Technology Data Exchange (ETDEWEB)

    Carter, B.; Huesemann, M.

    2008-01-01

    The alarming rate at which atmospheric carbon dioxide levels are increasing due to the burning of fossil fuels will have incalculable consequences if disregarded. Fuel cells, a source of energy that does not add to carbon dioxide emissions, have become an important topic of study. Although signifi cant advances have been made related to fuel cells, the problem of cheap and renewable hydrogen production still remains. The cyanobacterium Plectonema boryanum has demonstrated potential as a resolution to this problem by producing hydrogen under nitrogen defi cient growing conditions. Plectonema boryanum cultures were tested in a series of experiments to determine the effects of light intensity, initial nitrate concentration, and photosystem II inhibitor DCMU (3-(3,4- dichlorophenyl)-1,1-dimethylurea) upon hydrogen production. Cultures were grown in sterile Chu. No. 10 medium within photobioreactors constantly illuminated by halogen lights. Because the enzyme responsible for hydrogen production is sensitive to oxygen, the medium was continuously sparged with argon/CO2 (99.7%/0.3% vol/vol) by gas dispersion tubes immersed in the culture. Hydrogen production was monitored by using a gas chromatograph equipped with a thermal conductivity detector. In the initial experiment, the effects of initial nitrate concentration were tested and results revealed cumulative hydrogen production was maximum at an initial nitrate concentration of 1 mM. A second experiment was then conducted at an initial nitrate concentration of 1 mM to determine the effects of light intensity at 50, 100, and 200 μmole m-2 s-1. Cumulative hydrogen production increased with increasing light intensity. A fi nal experiment, conducted at an initial nitrate concentration of 2 mM, tested the effects of high light intensity at 200 and 400 μmole m-2 s-1. Excessive light at 400 μmole m-2 s-1 decreased cumulative hydrogen production. Based upon all experiments, cumulative hydrogen production rates were optimal

  18. Synthesis of chlorophyll-binding proteins in a fully-segregated ∆ycf54 strain of the cyanobacterium Synechocystis PCC 6803

    Directory of Open Access Journals (Sweden)

    Sarah eHollingshead

    2016-03-01

    Full Text Available In the chlorophyll (Chl biosynthesis pathway the formation of protochlorophyllide is catalyzed by Mg-protoporphyrin IX methyl ester (MgPME cyclase. The Ycf54 protein was recently shown to form a complex with another component of the oxidative cyclase, Sll1214 (CycI, and partial inactivation of the ycf54 gene leads to Chl deficiency in cyanobacteria and plants. The exact function of the Ycf54 is not known, however, and further progress depends on construction and characterisation of a mutant cyanobacterial strain with a fully inactivated ycf54 gene. Here, we report the complete deletion of the ycf54 gene in the cyanobacterium Synechocystis 6803; the resulting ycf54 strain accumulates huge concentrations of the cyclase substrate MgPME together with another pigment, which we identified using nuclear magnetic resonance as 3-formyl MgPME. The detection of a small amount (~13% of Chl in the ycf54 mutant provides clear evidence that the Ycf54 protein is important, but not essential, for activity of the oxidative cyclase. The greatly reduced formation of protochlorophyllide in the ycf54 strain provided an opportunity to use 35S protein labelling combined with 2D electrophoresis to examine the synthesis of all known Chl-binding protein complexes under drastically restricted de novo Chl biosynthesis. We show that although the ycf54 strain synthesizes very limited amounts of photosystem I and the CP47 and CP43 subunits of photosystem II (PSII, the synthesis of PSII D1 and D2 subunits and their assembly into the reaction centre (RCII assembly intermediate were not affected. Furthermore, the levels of other Chl complexes such as cytochrome b6f and the HliD– Chl synthase remained comparable to wild-type. These data demonstrate that the requirement for de novo Chl molecules differs completely for each Chl-binding protein. Chl traffic and recycling in the cyanobacterial cell as well as the function of Ycf54 are discussed.

  19. Cell surface acid-base properties of the cyanobacterium Synechococcus: Influences of nitrogen source, growth phase and N:P ratios

    Science.gov (United States)

    Liu, Yuxia; Alessi, D. S.; Owttrim, G. W.; Kenney, J. P. L.; Zhou, Qixing; Lalonde, S. V.; Konhauser, K. O.

    2016-08-01

    The distribution of many trace metals in the oceans is controlled by biological uptake. Recently, Liu et al. (2015) demonstrated the propensity for a marine cyanobacterium to adsorb cadmium from seawater, suggesting that cell surface reactivity might also play an important role in the cycling of metals in the oceans. However, it remains unclear how variations in cyanobacterial growth rates and nutrient supply might affect the chemical properties of their cellular surfaces. In this study we used potentiometric titrations and Fourier Transform Infrared (FT-IR) spectrometry to profile the key metabolic changes and surface chemical responses of a Synechococcus strain, PCC 7002, during different growth regimes. This included testing various nitrogen (N) to phosphorous (P) ratios (both nitrogen and phosphorous dependent), nitrogen sources (nitrate, ammonium and urea) and growth stages (exponential, stationary, and death phase). FT-IR spectroscopy showed that varying the growth substrates on which Synechococcus cells were cultured resulted in differences in either the type or abundance of cellular exudates produced or a change in the cell wall components. Potentiometric titration data were modeled using three distinct proton binding sites, with resulting pKa values for cells of the various growth conditions in the ranges of 4.96-5.51 (pKa1), 6.67-7.42 (pKa2) and 8.13-9.95 (pKa3). According to previous spectroscopic studies, these pKa ranges are consistent with carboxyl, phosphoryl, and amine groups, respectively. Comparisons between the titration data (for the cell surface) and FT-IR spectra (for the average cellular changes) generally indicate (1) that the nitrogen source is a greater determinant of ligand concentration than growth phase, and (2) that phosphorus limitation has a greater impact on Synechococcus cellular and extracellular properties than does nitrogen limitation. Taken together, these techniques indicate that nutritional quality during cell growth can

  20. Higher production of C-phycocyanin by nitrogen-free (diazotrophic) cultivation of Nostoc sp. NK and simplified extraction by dark-cold shock.

    Science.gov (United States)

    Lee, Na Kyeong; Oh, Hee-Mock; Kim, Hee-Sik; Ahn, Chi-Yong

    2017-03-01

    Nostoc sp. NK (KCTC 12772BP) was isolated and cultivated in a BG11 medium and a nitrate-free BG11 medium (BG11 0 ). To enhance C-phycocyanin (C-PC) content in the cells, different fluorescent lamps (white, plant, and red) were used as light sources for complementary chromatic adaptation (CCA). The maximum biomass productivity was 0.42g/L/d and 0.32g/L/d under BG11 and BG11 0 conditions, respectively. The maximum C-PC contents were 8.4% (w/w) under white lamps, 13.6% (w/w) under plant lamps, and 18% (w/w) under BG11 0 and the red light condition. The maximum C-PC productivity was 57.4mg/L/d in BG11 0 under the red lamp condition. These results indicate that a higher C-PC content could be obtained under a diazotrophic condition and a CCA reaction. The C-PC could be released naturally from cells without any extraction processes, when Nostoc sp. NK was cultivated in the BG11 0 medium with CO 2 aeration and put in dark conditions at 5°C. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Organic matter inoculated with diazotrophic bacterium Beijerinckia indica and Cunninghamella elegans fungus containing chitosan on banana “Williams” in field

    Directory of Open Access Journals (Sweden)

    Newton Pereira Stamford

    2017-01-01

    Full Text Available The production of biofertilizers from rocks increases nutrients for plant nutrition without environmental pollution. The aim of this study was to evaluate the effectiveness of biofertilizers from phosphate and potassium rocks mixed with organic matter (earthworm compound inoculated with free living diazotrophic bacteria (NFB 10001 and Cunninghamella elegans (fungus with chitosan on yield, characteristics, and nutrient uptake of banana (cv. Williams, and attributes of a Red Yellow Argisoil of the rainforest Zone of Pernambuco, Brazil. The experimental design included two biofertilizers: (a PK rock biofertilizers plus organic matter (NPKB and (b bioprotector (NPKP applied at 50, 100 and 150% of the recommended rate for banana, which were compared with soluble mineral fertilizers (NPKF applied at the recommended rate, and earthworm compound (20 ton ha-1. The best results of the plant parameters were obtained with NPKB and NPKP applied at the highest rates (150% RR. A normal yield was produced when NPKB and NPKP were applied at the highest rates and NPKF at the recommended rate. The available P and K in the soil showed a significant fertilization effect, especially when NPKB and NPKP were applied at the highest rates. The biofertilizer and bioprotector may be alternatives to mineral soluble fertilizers.

  2. Identities of epilithic hydrocarbon-utilizing diazotrophic bacteria from the Arabian Gulf Coasts, and their potential for oil bioremediation without nitrogen supplementation.

    Science.gov (United States)

    Radwan, Samir; Mahmoud, Huda; Khanafer, Majida; Al-Habib, Aamar; Al-Hasan, Redha

    2010-08-01

    Gravel particles from four sites along the Arabian Gulf coast in autumn, winter, and spring were naturally colonized with microbial consortia containing between 7 and 400 × 10(2) cm(-2) of cultivable oil-utilizing bacteria. The 16S rRNA gene sequences of 70 representatives of oil-utilizing bacteria revealed that they were predominantly affiliated with the Gammaproteobacteria and the Actinobacteria. The Gammaproteobacteria comprised among others, the genera Pseudomonas, Pseudoalteromonas, Shewanella, Marinobacter, Psychrobacter, Idiomarina, Alcanivorax, Cobetia, and others. Actinobacteria comprised the genera Dietzia, Kocuria, Isoptericola, Rhodococcus, Microbacterium, and others. In autumn, Firmicutes members were isolated from bay and nonbay stations while Alphaproteobacteria were detected only during winter from Anjefa bay station. Fingerprinting by denaturing gradient gel electrophoresis of amplified 16S rRNA genes of whole microbial consortia confirmed the culture-based bacterial diversities in the various epilithons in various sites and seasons. Most of the representative oil-utilizing bacteria isolated from the epilithons were diazotrophic and could attenuate oil also in nitrogen-rich (7.9-62%) and nitrogen-free (4-54%) cultures, which, makes the microbial consortia suitable for oil bioremediation in situ, without need for nitrogen supplementation. This was confirmed in bench-scale experiments in which unfertilized oily seawater was bioremediated by epilithon-coated gravel particles.

  3. Structural Interaction Between GFP-Labeled Diazotrophic Endophytic Bacterium Herbaspirillum seropedicae RAM10 and Pineapple Plantlets ‘VitóRia’

    Science.gov (United States)

    Estrela Borges Baldotto, Lílian; Lopes Olivares, Fábio; Bressan-Smith, Ricardo

    2011-01-01

    The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets ‘Vitória’ were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae. PMID:24031612

  4. An assessment of the usefulness of the cyanobacterium Synechococcus subsalsus as a source of biomass for biofuel production

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    Bruno R.S. Setta

    2014-05-01

    Full Text Available Nowadays algal biofuels are considered one of the most promising solutions of global energy crisis and climate change for the years to come. By manipulation of the culture conditions, many algal species can be induced to accumulate high concentrations of particular biomolecules and can be directed to the desired output for each fuel. In this context, the present study involved the assessment of the effects of CO2 availability and nitrogen starvation on growth and chemical composition of the cyanobacterium Synechococcus subsalsus, testing a fast-growing native strain. The control experiments were performed with Conway culture medium in 12-day batch cultures, in 6-liter flasks and 12 h photoperiod, with addition of 2 L min-1 filtered air to each flask. Other two experimental conditions were also tested: (i the placement into the cultures of additional dissolved nutrients except nitrogen, one week after the start of growth (N-, and (ii the input of pure CO2 into the flasks from the 5th day of growth (C+. In all cultures, daily cell counts were done throughout the cultivation, as well as measurements of pH and cell biovolumes. Maximum cell yield were found in N-experiments, while cell yields of C+ and control were similar. Dissolved nitrogen was exhausted before the end of the experiments, but dissolved phosphorus was not totally consumed. Protein and chlorophyll-a concentrations decreased from the exponential to the stationary growth phase of all experiments, except for protein in the control. In all experiments, carbohydrate, lipid and total carotenoid increased from the exponential to the stationary growth phase, as an effect of nitrogen limitation. Increments in carbohydrate concentrations were remarkable, achieving more than 42% of the dry weight (dw, but concentrations of lipid were always lower than 13% dw. The addition of pure CO2 did not cause a significant increase in biomass of S. subsalsus nor generated more lipid and carbohydrate than

  5. Transcriptional analysis of the jamaicamide gene cluster from the marine cyanobacterium Lyngbya majuscula and identification of possible regulatory proteins

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    Dorrestein Pieter C

    2009-12-01

    Full Text Available Abstract Background The marine cyanobacterium Lyngbya majuscula is a prolific producer of bioactive secondary metabolites. Although biosynthetic gene clusters encoding several of these compounds have been identified, little is known about how these clusters of genes are transcribed or regulated, and techniques targeting genetic manipulation in Lyngbya strains have not yet been developed. We conducted transcriptional analyses of the jamaicamide gene cluster from a Jamaican strain of Lyngbya majuscula, and isolated proteins that could be involved in jamaicamide regulation. Results An unusually long untranslated leader region of approximately 840 bp is located between the jamaicamide transcription start site (TSS and gene cluster start codon. All of the intergenic regions between the pathway ORFs were transcribed into RNA in RT-PCR experiments; however, a promoter prediction program indicated the possible presence of promoters in multiple intergenic regions. Because the functionality of these promoters could not be verified in vivo, we used a reporter gene assay in E. coli to show that several of these intergenic regions, as well as the primary promoter preceding the TSS, are capable of driving β-galactosidase production. A protein pulldown assay was also used to isolate proteins that may regulate the jamaicamide pathway. Pulldown experiments using the intergenic region upstream of jamA as a DNA probe isolated two proteins that were identified by LC-MS/MS. By BLAST analysis, one of these had close sequence identity to a regulatory protein in another cyanobacterial species. Protein comparisons suggest a possible correlation between secondary metabolism regulation and light dependent complementary chromatic adaptation. Electromobility shift assays were used to evaluate binding of the recombinant proteins to the jamaicamide promoter region. Conclusion Insights into natural product regulation in cyanobacteria are of significant value to drug discovery

  6. Engineering a cyanobacterium as the catalyst for the photosynthetic conversion of CO2 to 1,2-propanediol

    Directory of Open Access Journals (Sweden)

    Li Han

    2013-01-01

    Full Text Available Abstract Background The modern society primarily relies on petroleum and natural gas for the production of fuels and chemicals. One of the major commodity chemicals 1,2-propanediol (1,2-PDO, which has an annual production of more than 0.5 million tons in the United States, is currently produced by chemical processes from petroleum derived propylene oxide, which is energy intensive and not sustainable. In this study, we sought to achieve photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Compared to the previously reported biological 1,2-PDO production processes which used sugar or glycerol as the substrates, direct chemical production from CO2 in photosynthetic organisms recycles the atmospheric CO2 and will not compete with food crops for arable land. Results In this study, we reported photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Introduction of the genes encoding methylglyoxal synthase (mgsA, glycerol dehydrogenase (gldA, and aldehyde reductase (yqhD resulted in the production of ~22mg/L 1,2-PDO from CO2. However, a comparable amount of the pathway intermediate acetol was also produced, especially during the stationary phase. The production of 1,2-PDO requires a robust input of reducing equivalents from cellular metabolism. To take advantage of cyanobacteria’s NADPH pool, the synthetic pathway of 1,2-PDO was engineered to be NADPH-dependent by exploiting the NADPH-specific secondary alcohol dehydrogenases which have not been reported for 1,2-PDO production previously. This optimization strategy resulted in the production of ~150mg/L 1,2-PDO and minimized the accumulation of the incomplete reduction product, acetol. Conclusion This work demonstrated that cyanobacteria can be engineered as a catalyst for the photosynthetic conversion of CO2 to 1,2-PDO. This work also characterized two NADPH

  7. Unravelling the cross-talk between iron starvation and oxidative stress responses highlights the key role of PerR (alr0957) in peroxide signalling in the cyanobacterium Nostoc PCC 7120.

    Science.gov (United States)

    Yingping, Fan; Lemeille, Sylvain; Talla, Emmanuel; Janicki, Annick; Denis, Yann; Zhang, Cheng-Cai; Latifi, Amel

    2014-10-01

    The cyanobacterial phylum includes oxygenic photosynthetic prokaryotes of a wide variety of morphologies, metabolisms and ecologies. Their adaptation to their various ecological niches is mainly achieved by sophisticated regulatory mechanisms and depends on a fine cross-talk between them. We assessed the global transcriptomic response of the filamentous cyanobacterium Nostoc PCC 7120 to iron starvation and oxidative stress. More than 20% of the differentially expressed genes in response to iron stress were also responsive to oxidative stress. These transcripts include antioxidant proteins-encoding genes that confirms that iron depletion leads to reactive oxygen accumulation. The activity of the Fe-superoxide dismutase was not significantly decreased under iron starvation, indicating that the oxidative stress generated under iron deficiency is not a consequence of (SOD) deficiency. The transcriptional data indicate that the adaptation of Nostoc to iron-depleted conditions displays important differences with what has been shown in unicellular cyanobacteria. While the FurA protein that regulates the response to iron deprivation has been well characterized in Nostoc, the regulators in charge of the oxidative stress response are unknown. Our study indicates that the alr0957 (perR) gene encodes the master regulator of the peroxide stress. PerR is a peroxide-sensor repressor that senses peroxide by metal-catalysed oxidation.

  8. Importance classification

    International Nuclear Information System (INIS)

    Mizumachi, Wataru; Kobayashi, Masahide

    2008-01-01

    Conventionally, the design of a nuclear reactor has been performed from a viewpoint of a safety function and the importance on earthquake-proof on the basis of not giving off the mainly included radioactivity outside. In this Niigataken-Chuetsuoki earthquake, there is almost no damage to the system, components and structure on safe also in the earthquake beyond assumption, and the validity of the design was checked. But, the situation peculiar to a big earthquake was also generated. The emergency plan room which should serve as a connection center with the exterior was not able to open a door and use at the beginning. Fire-extinguishing system piping fractured and self-defense fire fighting was not made. And so on. Discussion from the following three viewpoints was performed. 1st: The importance from a viewpoint which should maintain a function also with the disaster in case of an earthquake like an emergency plan room etc. 2nd: In the earthquake, since the safe system and un-safe system was influenced, the importance from a viewpoint which may have influence safely inquired when the un-safe system broke down. 3rd: Although it was not directly related safely, discussion from a viewpoint which influences fear of insecurity, such as taking out smoke, for example, was performed (author)

  9. Santacruzamate A, a potent and selective histone deacetylase inhibitor from the Panamanian marine cyanobacterium cf. Symploca sp.

    Science.gov (United States)

    Pavlik, Christopher M; Wong, Christina Y B; Ononye, Sophia; Lopez, Dioxelis D; Engene, Niclas; McPhail, Kerry L; Gerwick, William H; Balunas, Marcy J

    2013-11-22

    A dark brown tuft-forming cyanobacterium, morphologically resembling the genus Symploca, was collected during an expedition to the Coiba National Park, a UNESCO World Heritage Site on the Pacific coast of Panama. Phylogenetic analysis of its 16S rRNA gene sequence indicated that it is 4.5% divergent from the type strain for Symploca and thus is likely a new genus. Fractionation of the crude extract led to the isolation of a new cytotoxin, designated santacruzamate A (1), which has several structural features in common with suberoylanilide hydroxamic acid [(2), SAHA, trade name Vorinostat], a clinically approved histone deacetylase (HDAC) inhibitor used to treat refractory cutaneous T-cell lymphoma. Recognition of the structural similarly of 1 and SAHA led to the characterization of santacruzamate A as a picomolar level selective inhibitor of HDAC2, a Class I HDAC, with relatively little inhibition of HDAC4 or HDAC6, both Class II HDACs. As a result, chemical syntheses of santacruzamate A as well as a structurally intriguing hybrid molecule, which blends aspects of both agents (1 and 2), were achieved and evaluated for their HDAC activity and specificity.

  10. Santacruzamate A, a Potent and Selective Histone Deacetylase (HDAC) Inhibitor from the Panamanian Marine Cyanobacterium cf. Symploca sp.

    Science.gov (United States)

    Pavlik, Christopher M.; Wong, Christina Y.B.; Ononye, Sophia; Lopez, Dioxelis D.; Engene, Niclas; McPhail, Kerry L.; Gerwick, William H.; Balunas, Marcy J.

    2013-01-01

    A dark-brown tuft-forming cyanobacterium, morphologically resembling the genus Symploca, was collected during an expedition to the Coiba National Park, a UNESCO World Heritage Site on the Pacific coast of Panama. Phylogenetic analysis of its 16S rRNA gene sequence indicated that it is 4.5% divergent from the type strain for Symploca, and thus is likely a new genus. Fractionation of the crude extract led to the isolation of a new cytotoxin, designated santacruzamate A (1), which has several structural features in common with suberoylanilide hydroxamic acid [(2), SAHA, trade name Vorinostat®], a clinically approved histone deacetylase (HDAC) inhibitor used to treat refractory cutaneous T-cell lymphoma. Recognition of the structural similarly of 1 and SAHA led to the characterization of santacruzamate A as a picomolar level selective inhibitor of HDAC2, a Class I HDAC, with relatively little inhibition of HDAC4 or HDAC6, both Class II HDACs. As a result, chemical syntheses of santacruzamate A as well as a structurally intriguing hybrid molecule, which blends aspects of both agents (1 and 2), were achieved and evaluated for their HDAC activity and specificity. PMID:24164245

  11. Effects of Hydrogen Peroxide and Ultrasound on Biomass Reduction and Toxin Release in the Cyanobacterium, Microcystis aeruginosa

    Directory of Open Access Journals (Sweden)

    Miquel Lürling

    2014-12-01

    Full Text Available Cyanobacterial blooms are expected to increase, and the toxins they produce threaten human health and impair ecosystem services. The reduction of the nutrient load of surface waters is the preferred way to prevent these blooms; however, this is not always feasible. Quick curative measures are therefore preferred in some cases. Two of these proposed measures, peroxide and ultrasound, were tested for their efficiency in reducing cyanobacterial biomass and potential release of cyanotoxins. Hereto, laboratory assays with a microcystin (MC-producing cyanobacterium (Microcystis aeruginosa were conducted. Peroxide effectively reduced M. aeruginosa biomass when dosed at 4 or 8 mg L−1, but not at 1 and 2 mg L−1. Peroxide dosed at 4 or 8 mg L−1 lowered total MC concentrations by 23%, yet led to a significant release of MCs into the water. Dissolved MC concentrations were nine-times (4 mg L−1 and 12-times (8 mg L−1 H2O2 higher than in the control. Cell lysis moreover increased the proportion of the dissolved hydrophobic variants, MC-LW and MC-LF (where L = Leucine, W = tryptophan, F = phenylalanine. Ultrasound treatment with commercial transducers sold for clearing ponds and lakes only caused minimal growth inhibition and some release of MCs into the water. Commercial ultrasound transducers are therefore ineffective at controlling cyanobacteria.

  12. Soft x-ray imaging of intracellular granules of filamentous cyanobacterium generating musty smell in Lake Biwa

    International Nuclear Information System (INIS)

    Takemoto, K; Mizuta, G; Namba, H; Yamamoto, A; Kihara, H; Yoshimura, M; Ichise, S

    2013-01-01

    A planktonic blue-green algae, which are currently identified as Phormidium tenue, was observed by a soft x-ray microscopy (XM) for comparing a musty smell generating green strain (PTG) and a non-smell brown strain (PTB). By XM, cells were clearly imaged, and several intracellular granules which could not be observed under a light microscope were visualized. The diameter of granules was about 0.5–1 μm, and one or a few granules were seen in a cell. XM analyses showed that width of cells and sizes of intracellular granules were quite different between PTG and PTB strains. To study the granules observed by XM, transmission in more detail, transmission electron microscopy (TEM) and indirect fluorescent-antibody technique (IFA) were applied. By TEM, carboxysomes, thylakoids and polyphosphate granules were observed. IFA showed the presence of carboxysomes. Results lead to the conclusion that intracellular granules observed under XM are carboxysomes or polyphosphate granules. These results demonstrate that soft XM is effective for analyzing fine structures of small organisms such as cyanobacterium, and for discriminating the strains which generates musty smells from others

  13. Creation of glyphosate-resistant Brassica napus L. plants expressing DesC desaturase of cyanobacterium Synechococcus vulcanus

    Directory of Open Access Journals (Sweden)

    Goldenkova-Pavlova I. V.

    2012-12-01

    Full Text Available Aim. Creation of glyphosate-resistant canola plants expressing bifunctional hybrid desC::licBM3 gene. In the hybrid gene the sequence of DesC desaturase of cyanobacterium S. vulcanus without plastid targeting was fused with the sequence of thermostable lichenase reporter LicBM3 gene. Methods. Agrobacterium tumefaciens-mediated transformation, PCR, quantitative and qualitative determination of lichenase activity, genetic analysis. Results. Transgenic canola plants, carring the enolpyruvat shikimat phosphate syntase gene (epsps, conferring on plants resistance to phosphonomethyl glycine herbicides (Roundup, as well as the desC::licBM3 gene, were selected. The presence of transgenes was confimed by multiplex PCR. The epsps gene expression in canola was shown at the transcription level, during in vitro growth and after greenhouse herbicide treatment. Activity of the licBM3 gene product as a part of hybrid protein allowed quantitative and qualitative estimation of the desaturase gene expression. Inheritance of heterologous genes and their expression in the first generation were investigated. Conclusions. Transgenic canola plants were obtained, the presence of trangenes in plant genome was proved and expression of the target genes was detected.

  14. Degradative crystal–chemical transformations of clay minerals under the influence of cyanobacterium-actinomycetal symbiotic associations

    Directory of Open Access Journals (Sweden)

    Ekaterina Ivanova

    2014-04-01

    Full Text Available Cyanobacteria and actinomycetes are essential components of soil microbial community and play an active role in ash elements leaching from minerals of the parent rock. Content and composition of clay minerals in soil determine the sorption properties of the soil horizons, water-holding capacity of the soil, stickiness, plasticity, etc. The transformative effect of cyanobacterial–actinomycetes associations on the structure of clay minerals – kaolinite, vermiculite, montmorillonite, biotite and muscovite – was observed, with the greatest structural lattice transformation revealed under the influence of association in comparison with monocultures of cyanobacterium and actinomycete. The range of the transformative effect depended both on the type of biota (component composition of association and on the crystal–chemical parameters of the mineral itself (trioctahedral mica – biotite, was more prone to microbial degradation than the dioctahedral – muscovite. The formation of the swelling phase – the product of biotite transformation into the mica–vermicullite mixed-layered formation was revealed as a result of association cultivation. Crystal chemical transformation of vermiculite was accompanied by the removal of potassium (К, magnesium (Mg and aluminum (Al from the crystal lattice. The study of such prokaryotic communities existed even in the early stages of the Earth's history helps to understand the causes and nature of the transformations undergone by the atmosphere, hydrosphere and lithosphere of the planet.contribution of treatments on structure induces and model parameters are discussed in the paper.

  15. Sulfate-driven elemental sparing is regulated at the transcriptional and posttranscriptional levels in a filamentous cyanobacterium.

    Science.gov (United States)

    Gutu, Andrian; Alvey, Richard M; Bashour, Sami; Zingg, Daniel; Kehoe, David M

    2011-03-01

    Sulfur is an essential nutrient that can exist at growth-limiting concentrations in freshwater environments. The freshwater cyanobacterium Fremyella diplosiphon (also known as Tolypothrix sp. PCC 7601) is capable of remodeling the composition of its light-harvesting antennae, or phycobilisomes, in response to changes in the sulfur levels in its environment. Depletion of sulfur causes these cells to cease the accumulation of two forms of a major phycobilisome protein called phycocyanin and initiate the production of a third form of phycocyanin, which possesses a minimal number of sulfur-containing amino acids. Since phycobilisomes make up approximately 50% of the total protein in these cells, this elemental sparing response has the potential to significantly influence the fitness of this species under low-sulfur conditions. This response is specific for sulfate and occurs over the physiological range of sulfate concentrations likely to be encountered by this organism in its natural environment. F. diplosiphon has two separate sulfur deprivation responses, with low sulfate levels activating the phycobilisome remodeling response and low sulfur levels activating the chlorosis or bleaching response. The phycobilisome remodeling response results from changes in RNA abundance that are regulated at both the transcriptional and posttranscriptional levels. The potential of this response, and the more general bleaching response of cyanobacteria, to provide sulfur-containing amino acids during periods of sulfur deprivation is examined.

  16. Effect of nitrogen on cellular production and release of the neurotoxin anatoxin-a in a nitrogen-fixing cyanobacterium

    Directory of Open Access Journals (Sweden)

    Alexis eGagnon

    2012-06-01

    Full Text Available Anatoxin-a (ANTX is a neurotoxin produced by several freshwater cyanobacteria and implicated in lethal poisonings of domesticated animals and wildlife. The factors leading to its production in nature and in culture are not well understood. Resource availability may influence its cellular production as suggested by the carbon-nutrient hypothesis, which links the amount of secondary metabolites produced by plants or microbes to the relative abundance of nutrients. We tested the effects of nitrogen supply on ANTX production and release in a toxic strain of the cyanobacterium Aphanizomenon issatschenkoi (Nostocales. We hypothesized that nitrogen deficiency might constrain the production of ANTX. However, the total concentration and more significantly the cellular content of anatoxin-a peaked (max. 146 µg/L and 1683 µg•g-1 dry weight at intermediate levels of nitrogen supply when N-deficiency was evident based on phycocyanin to chlorophyll a and carbon to nitrogen ratios. The results suggest that the cellular production of anatoxin-a may be stimulated by moderate nutrient stress as described recently for another cyanotoxin (microcystin.

  17. Evidence regarding the UV sunscreen role of a mycosporine-like compound in the cyanobacterium Gloeocapsa sp

    International Nuclear Information System (INIS)

    Garcia-Pichel, F.; Wingard, C.E.; Castenholz, R.W.

    1993-01-01

    The mycosporine-like amino acids (MAAs) have been thought to serve a UV sunscreen role in organisms that produce or contain them because MAAs present strong absorbance in the UV region and because there is no other apparent biological function. The researchers used the cyanobacterium Gloeocapsa sp. to assess the possible sunscreen role of MAAs. Five conditions are evaluated: (1) absorption of radiation high enough to provide benefit to the organisms; (2) correlation of presence of the compound with enhansed fitness under UV; (3) concentration of the compound and resistance to UV still present under physiological inactivity; (4) effect maximal at wavelengths of maximal absorption; (5) loss of protection after artificial removal of compound. The results indicate that only a small sunscreen effect can be ascribed to the MAA in the Gloecapsa sp. under these experimental conditions. It is possible however, that in the typical undisturbed colonial growth form, MAAs and their screening action may become major factors in resistance to UV radiation. 25 refs., 7 figs., 1 tab

  18. Acute toxicity of excess mercury on the photosynthetic performance of cyanobacterium, S. platensis--assessment by chlorophyll fluorescence analysis.

    Science.gov (United States)

    Lu, C M; Chau, C W; Zhang, J H

    2000-07-01

    Measurement of chlorophyll fluorescence has been shown to be a rapid, non-invasive, and reliable method to assess photosynthetic performance in a changing environment. In this study, acute toxicity of excess Hg on the photosynthetic performance of the cyanobacterium S. platensis, was investigated by use of chlorophyll fluorescence analysis after cells were exposed to excess Hg (up to 20 microM) for 2 h. The results determined from the fast fluorescence kinetics showed that Hg induced a significant increase in the proportion of the Q(B)-non-reducing PSII reaction centers. The fluorescence parameters measured under the steady state of photosynthesis demonstrated that the increase of Hg concentration led to a decrease in the maximal efficiency of PSII photochemistry, the efficiency of excitation energy capture by the open PSII reaction centers, and the quantum yield of PSII electron transport. Mercury also resulted in a decrease in the coefficients of photochemical and non-photochemical quenching. Mercury may have an acute toxicity on cyanobacteria by inhibiting the quantum yield of photosynthesis sensitively and rapidly. Such changes occurred before any other visible damages that may be evaluated by other conventional measurements. Our results also demonstrated that chlorophyll fluorescence analysis can be used as a useful physiological tool to assess early stages of change in photosynthetic performance of algae in response to heavy metal pollution.

  19. Dynamic localization of HmpF regulates type IV pilus activity and directional motility in the filamentous cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Cho, Ye Won; Gonzales, Alfonso; Harwood, Thomas V; Huynh, Jessica; Hwang, Yeji; Park, Jun Sang; Trieu, Anthony Q; Italia, Parth; Pallipuram, Vivek K; Risser, Douglas D

    2017-10-01

    Many cyanobacteria exhibit surface motility powered by type 4 pili (T4P). In the model filamentous cyanobacterium Nostoc punctiforme, the T4P systems are arrayed in static, bipolar rings in each cell. The chemotaxis-like Hmp system is essential for motility and the coordinated polar accumulation of PilA on cells in motile filaments, while the Ptx system controls positive phototaxis. Using transposon mutagenesis, a gene, designated hmpF, was identified as involved in motility. Synteny among filamentous cyanobacteria and the similar expression patterns for hmpF and hmpD imply that HmpF is part of the Hmp system. Deletion of hmpF produced a phenotype distinct from other hmp genes, but indistinguishable from pilB or pilQ. Both an HmpF-GFPuv fusion protein, and PilA, as assessed by in situ immunofluorescence, displayed coordinated, unipolar localization at the leading pole of each cell. Reversals were modulated by changes in light intensity and preceded by the migration of HmpF-GFPuv to the lagging cell poles. These results are consistent with a model where direct interaction between HmpF and the T4P system activates pilus extension, the Hmp system facilitates coordinated polarity of HmpF to establish motility, and the Ptx system modulates HmpF localization to initiate reversals in response to changes in light intensity. © 2017 John Wiley & Sons Ltd.

  20. Gene expression of a two-component regulatory system associated with sunscreen biosynthesis in the cyanobacterium Nostoc punctiforme ATCC 29133.

    Science.gov (United States)

    Janssen, Jacob; Soule, Tanya

    2016-01-01

    Long-wavelength ultraviolet radiation (UVA) can damage cells through photooxidative stress, leading to harmful photosensitized proteins and pigments in cyanobacteria. To mitigate damage, some cyanobacteria secrete the UVA-absorbing pigment scytonemin into their extracellular sheath. Comparative genomic analyses suggest that scytonemin biosynthesis is regulated by the two-component regulatory system (TCRS) proteins encoded by Npun_F1277 and Npun_F1278 in the cyanobacterium Nostoc punctiforme ATCC 29133. To understand the dynamics of these genes, their expression was measured following exposure to UVA, UVB, high visible (VIS) irradiance and oxidative stress for 20, 40 and 60 min. Overall, both genes had statistically similar patterns of expression for all four conditions and were generally upregulated, except for those exposed to UVB by 60 min and for the cells under oxidative stress. The greatest UVA response was an upregulation by 20 min, while the response to UVB was the most dramatic and persisted through 40 min. High VIS irradiance resulted in a modest upregulation, while oxidative stress caused a slight downregulation. Both genes were also found to occur on the same transcript. These results demonstrate that these genes are positively responding to several light-associated conditions, which suggests that this TCRS may regulate more than just scytonemin biosynthesis under UVA stress. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Dark hydrogen production in nitrogen atmosphere - An approach for sustainability by marine cyanobacterium Leptolyngbya valderiana BDU 20041

    Energy Technology Data Exchange (ETDEWEB)

    Prabaharan, D.; Arun Kumar, D.; Uma, L.; Subramanian, G. [National Facility for Marine Cyanobacteria (Sponsored by DBT, Govt. of India), Department of Marine Biotechnology, Bharathidasan University, Tiruchirapalli 620 024 (India)

    2010-10-15

    Biological hydrogen production is an ideal system for three main reasons i) forms a renewable energy source, ii) gives clean fuel and iii) serves as a good supplement to oil reserves. The major challenges faced in biological hydrogen production are the presence of uptake hydrogenase and lack of sustainability in the cyanobacterial hydrogen production system. Three different marine cyanobacterial species viz. Leptolyngbya valderiana BDU 20041, Dichothrix baueriana BDU 40481 and Nostoc calcicola BDU 40302 were studied for their potential use in hydrogen production. Among these, L. valderiana BDU 20041, was found to produce hydrogen even in 100% nitrogen atmosphere which was 85% of the hydrogen produced in argon atmosphere. This is the first report of such a high rate of production of hydrogen in a nitrogen atmosphere by a cyanobacterium, which makes it possible to develop sustained hydrogen production systems. L. valderiana BDU 20041, a dark hydrogen producer uses the reductant essentially supplied by the respiratory pathway for hydrogen production. Using inhibitors, this organism was found to produce hydrogen due to the activities of both nitrogenase and bidirectional hydrogenase, while it had no 'uptake' hydrogenase activity. The other two organisms though had low levels of bidirectional hydrogenase, possessed considerable 'uptake' hydrogenase activity and hence could not release much hydrogen either in argon or nitrogen atmosphere. (author)

  2. The non-metabolizable sucrose analog sucralose is a potent inhibitor of hormogonium differentiation in the filamentous cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Splitt, Samantha D; Risser, Douglas D

    2016-03-01

    Nostoc punctiforme is a filamentous cyanobacterium which forms nitrogen-fixing symbioses with several different plants and fungi. Establishment of these symbioses requires the formation of motile hormogonium filaments. Once infected, the plant partner is thought to supply a hormogonium-repressing factor (HRF) to maintain the cyanobacteria in a vegetative, nitrogen-fixing state. Evidence implies that sucrose may serve as a HRF. Here, we tested the effects of sucralose, a non-metabolizable sucrose analog, on hormogonium differentiation. Sucralose inhibited hormogonium differentiation at a concentration approximately one-tenth that of sucrose. This result implies that: (1) sucrose, not a sucrose catabolite, is perceived by the cell and (2) inhibition is not due to a more general osmolarity-dependent effect. Additionally, both sucrose and sucralose induced the accrual of a polysaccharide sheath which bound specifically to the lectin ConA, indicating the presence of α-D-mannose and/or α-D-glucose. A ConA-specific polysaccharide was also found to be expressed in N. punctiforme colonies from tissue sections of the symbiotically grown hornwort Anthoceros punctatus. These findings imply that plant-derived sucrose or sucrose analogs may have multiple effects on N. punctiforme, including both repression of hormogonia and the induction of a polysaccharide sheath that may be essential to establish and maintain the symbiotic state.

  3. Elevated Atmospheric CO2 and Warming Stimulates Growth and Nitrogen Fixation in a Common Forest Floor Cyanobacterium under Axenic Conditions

    Directory of Open Access Journals (Sweden)

    Zoë Lindo

    2017-03-01

    Full Text Available The predominant input of available nitrogen (N in boreal forest ecosystems originates from moss-associated cyanobacteria, which fix unavailable atmospheric N2, contribute to the soil N pool, and thereby support forest productivity. Alongside climate warming, increases in atmospheric CO2 concentrations are expected in Canada’s boreal region over the next century, yet little is known about the combined effects of these factors on N fixation by forest floor cyanobacteria. Here we assess changes in N fixation in a common forest floor, moss-associated cyanobacterium, Nostoc punctiforme Hariot, under elevated CO2 conditions over 30 days and warming combined with elevated CO2 over 90 days. We measured rates of growth and changes in the number of specialized N2 fixing heterocyst cells, as well as the overall N fixing activity of the cultures. Elevated CO2 stimulated growth and N fixation overall, but this result was influenced by the growth stage of the cyanobacteria, which in turn was influenced by our temperature treatments. Taken together, climate change factors of warming and elevated CO2 are expected to stimulate N2 fixation by moss-associated cyanobacteria in boreal forest systems.

  4. Discovery of Rare and Highly Toxic Microcystins from Lichen-Associated Cyanobacterium Nostoc sp. Strain IO-102-I

    Science.gov (United States)

    Oksanen, Ilona; Jokela, Jouni; Fewer, David P.; Wahlsten, Matti; Rikkinen, Jouko; Sivonen, Kaarina

    2004-01-01

    The production of hepatotoxic cyclic heptapeptides, microcystins, is almost exclusively reported from planktonic cyanobacteria. Here we show that a terrestrial cyanobacterium Nostoc sp. strain IO-102-I isolated from a lichen association produces six different microcystins. Microcystins were identified with liquid chromatography-UV mass spectrometry by their retention times, UV spectra, mass fragmentation, and comparison to microcystins from the aquatic Nostoc sp. strain 152. The dominant microcystin produced by Nostoc sp. strain IO-102-I was the highly toxic [ADMAdda5]microcystin-LR, which accounted for ca. 80% of the total microcystins. We assigned a structure of [DMAdda5]microcystin-LR and [d-Asp3,ADMAdda5]microcystin-LR and a partial structure of three new [ADMAdda5]-XR type of microcystin variants. Interestingly, Nostoc spp. strains IO-102-I and 152 synthesized only the rare ADMAdda and DMAdda subfamilies of microcystin variants. Phylogenetic analyses demonstrated congruence between genes involved directly in microcystin biosynthesis and the 16S rRNA and rpoC1 genes of Nostoc sp. strain IO-102-I. Nostoc sp. strain 152 and the Nostoc sp. strain IO-102-I are distantly related, revealing a sporadic distribution of toxin production in the genus Nostoc. Nostoc sp. strain IO-102-I is closely related to Nostoc punctiforme PCC 73102 and other symbiotic Nostoc strains and most likely belongs to this species. Together, this suggests that other terrestrial and aquatic strains of the genus Nostoc may have retained the genes necessary for microcystin biosynthesis. PMID:15466511

  5. Culture temperature affects gene expression and metabolic pathways in the 2-methylisoborneol-producing cyanobacterium Pseudanabaena galeata.

    Science.gov (United States)

    Kakimoto, Masayuki; Ishikawa, Toshiki; Miyagi, Atsuko; Saito, Kazuaki; Miyazaki, Motonobu; Asaeda, Takashi; Yamaguchi, Masatoshi; Uchimiya, Hirofumi; Kawai-Yamada, Maki

    2014-02-15

    A volatile metabolite, 2-methylisoborneol (2-MIB), causes an unpleasant taste and odor in tap water. Some filamentous cyanobacteria produce 2-MIB via a two-step biosynthetic pathway: methylation of geranyl diphosphate (GPP) by methyl transferase (GPPMT), followed by the cyclization of methyl-GPP by monoterpene cyclase (MIBS). We isolated the genes encoding GPPMT and MIBS from Pseudanabaena galeata, a filamentous cyanobacterium known to be a major causal organism of 2-MIB production in Japanese lakes. The predicted amino acid sequence showed high similarity with that of Pseudanabaena limnetica (96% identity in GPPMT and 97% identity in MIBS). P. galeata was cultured at different temperatures to examine the effect of growth conditions on the production of 2-MIB and major metabolites. Gas chromatograph-mass spectrometry (GC-MS) measurements showed higher accumulation of 2-MIB at 30 °C than at 4 °C or 20 °C after 24 h of culture. Real-time-RT PCR analysis showed that the expression levels of the genes encoding GPPMT and MIBS decreased at 4 °C and increased at 30 °C, compared with at 20 °C. Furthermore, metabolite analysis showed dramatic changes in primary metabolite concentrations in cyanobacteria grown at different temperatures. The data indicate that changes in carbon flow in the TCA cycle affect 2-MIB biosynthesis at higher temperatures. Copyright © 2013 Elsevier GmbH. All rights reserved.

  6. In vivo toxicity of the culturable marine cyanobacterium Geitlerinema pseudacutissimum CNP 1019 extract on male Swiss albino mice (Mus musculus).

    Science.gov (United States)

    Maruthanayagam, Veerabadhran; Nagarajan, Manivel; Sundararaman, Muthuraman

    2014-01-01

    In this study, we investigated the in vivo toxicity of Geitlerinema pseudacutissimum CNP 1019 organic extract in a murine host. A single intraperitoneal injection of 1 g extract kg⁻¹ body weight (BW) did not exhibit mortality, whereas 3 g extract kg⁻¹ BW (approximate lethal dose) resulted in mortality within 5 days. To perform subchronic exposure toxicity analyses (i.e., daily exposure for a total of 14 days), a maximum concentration of ≤1 g extract kg⁻¹ BW was used. Subchronic toxicity studies in the treated mice, showed fluctuations of feed intake, loss of body weight, increase in specific activity of serum lactate dehydrogenase, alanine aminotransferase and decrease in whole serum protein concentration. LDH isoenzyme expression was found, and levels of the various isoforms were decreased as a result of the treatment. Histopathology studies in liver, kidney, and spleen isolated from the treated mice showed the presence of necrotic debris, hemorrhage, and micronuclei revealing the toxicity of the extract. The dose-dependent alterations in biochemical parameters in conjunction with the histological lesions noted in the animals treated with the prepared extract illustrate the likely potential toxicity to mammals from any encounters with the studied cyanobacterium.

  7. Amino Acid Transporters and Release of Hydrophobic Amino Acids in the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120

    Directory of Open Access Journals (Sweden)

    Rafael Pernil

    2015-04-01

    Full Text Available Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium that can use inorganic compounds such as nitrate or ammonium as nitrogen sources. In the absence of combined nitrogen, it can fix N2 in differentiated cells called heterocysts. Anabaena also shows substantial activities of amino acid uptake, and three ABC-type transporters for amino acids have been previously characterized. Seven new loci encoding predicted amino acid transporters were identified in the Anabaena genomic sequence and inactivated. Two of them were involved in amino acid uptake. Locus alr2535-alr2541 encodes the elements of a hydrophobic amino acid ABC-type transporter that is mainly involved in the uptake of glycine. ORF all0342 encodes a putative transporter from the dicarboxylate/amino acid:cation symporter (DAACS family whose inactivation resulted in an increased uptake of a broad range of amino acids. An assay to study amino acid release from Anabaena filaments to the external medium was set up. Net release of the alanine analogue α-aminoisobutyric acid (AIB was observed when transport system N-I (a hydrophobic amino acid ABC-type transporter was engaged in the uptake of a specific substrate. The rate of AIB release was directly proportional to the intracellular AIB concentration, suggesting leakage from the cells by diffusion.

  8. Carbon-free production of 2-deoxy-scyllo-inosose (DOI) in cyanobacterium Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Watanabe, Satoru; Ozawa, Hiroaki; Kato, Hiroaki; Nimura-Matsune, Kaori; Hirayama, Toshifumi; Kudo, Fumitaka; Eguchi, Tadashi; Kakinuma, Katsumi; Yoshikawa, Hirofumi

    2018-01-01

    Owing to their photosynthetic capabilities, there is increasing interest in utilizing cyanobacteria to convert solar energy into biomass. 2-Deoxy-scyllo-inosose (DOI) is a valuable starting material for the benzene-free synthesis of catechol and other benzenoids. DOI synthase (DOIS) is responsible for the formation of DOI from d-glucose-6-phosphate (G6P) in the biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics such as neomycin and butirosin. DOI fermentation using a recombinant Escherichia coli strain has been reported, although a carbon source is necessary for high-yield DOI production. We constructed DOI-producing cyanobacteria toward carbon-free and sustainable DOI production. A DOIS gene derived from the butirosin producer strain Bacillus circulans (btrC) was introduced and expressed in the cyanobacterium Synechococcus elongatus PCC 7942. We ultimately succeeded in producing 400 mg/L of DOI in S. elongatus without using a carbon source. DOI production by cyanobacteria represents a novel and efficient approach for producing benzenoids from G6P synthesized by photosynthesis.

  9. Metabolic engineering of the pentose phosphate pathway for enhanced limonene production in the cyanobacterium Synechocysti s sp. PCC 6803.

    Science.gov (United States)

    Lin, Po-Cheng; Saha, Rajib; Zhang, Fuzhong; Pakrasi, Himadri B

    2017-12-13

    Isoprenoids are diverse natural compounds, which have various applications as pharmaceuticals, fragrances, and solvents. The low yield of isoprenoids in plants makes them difficult for cost-effective production, and chemical synthesis of complex isoprenoids is impractical. Microbial production of isoprenoids has been considered as a promising approach to increase the yield. In this study, we engineered the model cyanobacterium Synechocystis sp. PCC 6803 for sustainable production of a commercially valuable isoprenoid, limonene. Limonene synthases from the plants Mentha spicata and Citrus limon were expressed in cyanobacteria for limonene production. Production of limonene was two-fold higher with limonene synthase from M. spicata than that from C. limon. To enhance isoprenoid production, computational strain design was conducted by applying the OptForce strain design algorithm on Synechocystis 6803. Based on the metabolic interventions suggested by this algorithm, genes (ribose 5-phosphate isomerase and ribulose 5-phosphate 3-epimerase) in the pentose phosphate pathway were overexpressed, and a geranyl diphosphate synthase from the plant Abies grandis was expressed to optimize the limonene biosynthetic pathway. The optimized strain produced 6.7 mg/L of limonene, a 2.3-fold improvement in productivity. Thus, this study presents a feasible strategy to engineer cyanobacteria for photosynthetic production of isoprenoids.

  10. In-situ optical and acoustical measurements of the buoyant cyanobacterium p. Rubescens: spatial and temporal distribution patterns.

    Directory of Open Access Journals (Sweden)

    Hilmar Hofmann

    Full Text Available Optical (fluorescence and acoustic in-situ techniques were tested in their ability to measure the spatial and temporal distribution of plankton in freshwater ecosystems with special emphasis on the harmful and buoyant cyanobacterium P. rubescens. Fluorescence was measured with the multi-spectral FluoroProbe (Moldaenke FluoroProbe, MFP and a Seapoint Chlorophyll Fluorometer (SCF. In-situ measurements of the acoustic backscatter strength (ABS were conducted with three different acoustic devices covering multiple acoustic frequencies (614 kHz ADCP, 2 MHz ADP, and 6 MHz ADV. The MFP provides a fast and reliable technique to measure fluorescence at different wavelengths in situ, which allows discriminating between P. rubescens and other phytoplankton species. All three acoustic devices are sensitive to P. rubescens even if other scatterers, e.g., zooplankton or suspended sediment, are present in the water column, because P. rubescens containing gas vesicles has a strong density difference and hence acoustic contrast to the ambient water and other scatterers. After calibration, the combination of optical and acoustical measurements not only allows qualitative and quantitative observation of P. rubescens, but also distinction between P. rubescens, other phytoplankton, and zooplankton. As the measuring devices can sample in situ at high rates they enable assessment of plankton distributions at high temporal (minutes and spatial (decimeters resolution or covering large temporal (seasonal and spatial (basin scale scales.

  11. Roles of xanthophyll carotenoids in protection against photoinhibition and oxidative stress in the cyanobacterium Synechococcus sp. strain PCC 7002.

    Science.gov (United States)

    Zhu, Yuehui; Graham, Joel E; Ludwig, Marcus; Xiong, Wei; Alvey, Richard M; Shen, Gaozhong; Bryant, Donald A

    2010-12-01

    Synechococcus sp. strain PCC 7002 is a robust, genetically tractable cyanobacterium that produces six different xanthophyll carotenoids (zeaxanthin, cryptoxanthin, myxoxanthophyll (myxol-2'-fucoside), echinenone, 3'-hydroxyechinenone, and synechoxanthin) and tolerates many environmental stresses, including high light intensities. Targeted mutations were introduced to block the branches of the carotenoid biosynthetic pathway leading to specific xanthophylls, and a mutant lacking all xanthophylls was constructed. Some of the mutants showed severe growth defects at high light intensities, and multi-locus mutants had somewhat lower chlorophyll contents and lower photosystem I levels. The results suggested that xanthophylls, particularly zeaxanthin and echinenone, might play regulatory roles in thylakoid biogenesis. Measurements of reactive oxygen (ROS) and nitrogen (RNS) species in the mutants showed that all xanthophylls participate in preventing ROS/RNS accumulation and that a mutant lacking all xanthophylls accumulated very high levels of ROS/RNS. Results from transcription profiling showed that mRNA levels for most genes encoding the enzymes of carotenogenesis are significantly more abundant after exposure to high light. These studies indicated that all xanthophylls contribute to protection against photo-oxidative stress. Copyright © 2010 Elsevier Inc. All rights reserved.

  12. Acute Exposure to Microcystin-Producing Cyanobacterium Microcystis aeruginosa Alters Adult Zebrafish (Danio rerio Swimming Performance Parameters

    Directory of Open Access Journals (Sweden)

    Luiza Wilges Kist

    2011-01-01

    Full Text Available Microcystins (MCs are toxins produced by cyanobacteria (blue-green algae, primarily Microcystis aeruginosa, forming water blooms worldwide. When an organism is exposed to environmental perturbations, alterations in normal behavioral patterns occur. Behavioral repertoire represents the consequence of a diversity of physiological and biochemical alterations. In this study, we assessed behavioral patterns and whole-body cortisol levels of adult zebrafish (Danio rerio exposed to cell culture of the microcystin-producing cyanobacterium M. aeruginosa (MC-LR, strain RST9501. MC-LR exposure (100 μg/L decreased by 63% the distance traveled and increased threefold the immobility time when compared to the control group. Interestingly, no significant alterations in the number of line crossings were found at the same MC-LR concentration and time of exposure. When animals were exposed to 50 and 100 μg/L, MC-LR promoted a significant increase (around 93% in the time spent in the bottom portion of the tank, suggesting an anxiogenic effect. The results also showed that none of the MC-LR concentrations tested promoted significant alterations in absolute turn angle, path efficiency, social behavior, or whole-body cortisol level. These findings indicate that behavior is susceptible to MC-LR exposure and provide evidence for a better understanding of the ecological consequences of toxic algal blooms.

  13. Effects of the Atrazine herbicide over the diazotrophic microorganisms associated with corn in a Saldana soil (Tolima); Efecto del herbicida Atrazina sobre los microorganismos diazotrofos asociados a un cultivo de maiz en un suelo de Saldana (Tolima)

    Energy Technology Data Exchange (ETDEWEB)

    Ordonez, Alba R; Lozano de Yunda, A; Fuentes, C

    1999-12-01

    In this study, it was evaluated, at greenhouse in soil pots, the effect of the herbicide atrazine in different dose (corresponding to o, 0.75, 1.5 and 3.0 kg of soil hectare) over time (15, 30, 45, 60 and 90 days after the application) on the diazotrophic micro-organisms associated with rhizosphere soil and root of a corn cultivation in a clay loam soil of Saldana - Tolima. It was determined the population of diazotrophic microorganisms in soil by the most probable number technique; also it was observed the characteristic growth in the specific cultivation semisolid Nfb-malic acid medium and was evaluated the nitrogenase activity by means of the acetylene reduction technique. Of the same form, it was observed the characteristic growth and it was evaluated the nitrogenase activity of direct fragment sowings of root in the Nfb-malic acid medium. The 60.8% of soil isolates presented characteristic cultural growth in the form of whitish pellicle under the surface of the medium. in the root 83.3% showed this type of growth. The population of diazotrophic microorganisms of soil did not present significant differences ({alpha} = 0.05) between treatments neither over time the nitrogenase activity of soil dilutions demonstrated that there is no a significant effect ({alpha} = 0.05) of the treatments (dose) but if there is significant effect of the time. The highest values were presented to 45 days of application of the herbicide, with the application of the highest dose. the nitrogenase activity in the root to what is long of the time was different from rhizosphere soil. The treatment without application of atrazine presented greater values to what is long of the time, however, also in this case were not presented significant differences ({alpha} = 0.05) between the treatments (dose) neither to what is long of the time. For so much, as a conclusion, they were not presented effects on the population and the nitrogenase activity of the diazotrophic microorganisms until 90

  14. Unconventional imports

    International Nuclear Information System (INIS)

    Yamaguchi, N.D.

    2001-01-01

    This article focuses on bitumens and bitumen products from Canadian oil sands and explores how they will affect the Canadian oil industry and the US refining industry. The falling production of crude, the growing demand for it, and the stagnating refining capacity in the US are reported, and Canadian and Mexican exports to the US, the definition of bitumens and bitumen quality, and the position of Canada as world leader in bitumen resources are considered. Bitumen production techniques, sales of bitumens and synthetic crude, the production outlook, and the quality and refining of bitumen and synthetic crude are examined. Plots illustrating North American crude refining capacity, production and demand for 1980-2000; US crude imports from Canada and Mexico (1981-2000), world proven oil reserves (2001), world bitumen resources, and Canadian oil production (1998-2000) are provided. Details of the composition of crudes and bitumens, and recent synthetic crude production are tabulated

  15. Tolerância de bactérias diazotróficas simbióticas à salinidade in vitro Tolerance of diazotrophic symbiotic bacteria to salinity

    Directory of Open Access Journals (Sweden)

    Rafaela Simão Abrahão Nóbrega

    2004-08-01

    Full Text Available A busca de estirpes de bactérias diazotróficas simbióticas tolerantes à salinidade, em conjunto com a seleção de hospedeiros, tem por objetivo aumentar o desempenho da simbiose e manter a produtividade vegetal de forma sustentada em condições de estresse salino. Doze estirpes de bactérias diazotróficas de diferentes procedências foram estudadas quanto à tolerância a diferentes concentrações de NaCl em meios de cultura 79 e LB. As estirpes que apresentaram maior tolerância à salinidade (30 g L-1 em meio 79 foram: BR 6806 e BR 4007, ambas de crescimento rápido isoladas de solos do nordeste brasileiro, UFLA 03-51 e UFLA 03-65, de crescimento rápido e UFLA 03-84, de crescimento lento, sendo as três últimas oriundas da região Amazônica. Essas também estiveram entre as mais tolerantes em meio de cultura LB. A estirpe UFLA 03-84, selecionada em trabalhos prévios por sua alta eficiência para caupi (Vigna unguiculata, pode ser indicada para estudos de inoculação, visando a aumentar o rendimento dessa cultura em solos salinos. Os meios de cultura 79 e LB foram eficientes para avaliar a tolerância relativa de rizóbio a NaCl, mas o meio 79 é mais indicado, pois permite o crescimento de todas as estirpes. Pelos resultados, infere-se haver relação entre tolerância in vitro à salinidade e origem dos microssimbiontes, pois as estirpes BR 4007 e BR 6806, oriundas do Ceará, estiveram entre as mais tolerantes.Tolerance to salinity of diazotrophic bacteriaand their host species could increase plant growth and sustainability in salt stressed soils. Twelve diazotrophic bacteria strains were studied regarding their tolerance to different NaCl concentrations in 79 and LB media. More tolerant (30 g L-1 strains in 79 medium were: the fast growers BR 6806 and BR 4007, isolated from northeast Brazil, UFLA 03-51 and UFLA 03-65, and slow growing strain UFLA03-84, being these last three isolated from Amazon region. These strains were among

  16. Dynamics of the diazotroph Bacillus polymyxa in the rhizosphere of wheat (Triticum aestivum L.) after inoculation and its effect on uptake of 15N-labelled fertilizer

    International Nuclear Information System (INIS)

    Gouzou, L.; Cheneby, D.; Nicolardot, B.; Heulin, T.

    1995-01-01

    An experiment under glasshouse conditions with a sandy soil was conducted to evaluate the effect of the inoculation of wheat (Triticum aestivum L.) roots with Bacillus polymyxa on N uptake by plants and to investigate the effect of N fertilization on the dynamics of this strain. Different treatments were considered: (i) amended (100 kg N ha −1 ) or not with 15 N labelled ammonium nitrate; (ii) inoculated (2.5 10 7 spores per seed and 0.5 10 9 spores kg −1 soil) or not with B. polymyxa CF43. No promoting effect on wheat growth (dry yield) or N uptake was observed after inoculation under these experimental conditions; 62 per cent of the N fertilizer was assimilated by plants during the first 33 days. Inoculation had no effect on the quantity of soil adhering to the roots. Soluble organic C and inorganic N concentrations were higher in the soil adhering to the roots than in bulk soil. However, the nitrate concentration in soil adhering to the roots was influenced by inoculation at the beginning of plant growth, nitrate supply to the root system being delayed by its probable retention in the rhizosphere. Furthermore, total bacteria in roots, soil adhering to the roots and bulk soil was not affected by N fertilization. The number of B. polymyxa cells measured by an AB-ELISA method increased first in the bulk soil and later in the soil adhering to the roots after inoculation. Fertilization with N decreased the B. polymyxa population in adhering soil, competitive suppression of diazotrophs by non-fixing bacteria probably being induced by the presence of large amounts of nutrients in the rhizosphere when N was added. (author)

  17. Characterization of the extracellular polysaccharide produced by a marine cyanobacterium, Cyanothece sp. ATCC 51142, and its exploitation toward metal removal from solutions.

    Science.gov (United States)

    Shah, V; Ray, A; Garg, N; Madamwar, D

    2000-04-01

    Cyanobacterium, Cyanothece sp. ATCC 51142 produces an exopolysaccharide at a high level. Physical analysis of the exopolysaccharide (EPS), such as nuclear magnetic resonance, infrared spectrum, were done to determine its possible structure. Thermal gravimetric analysis, differential scanning calorimeter, and differential thermal analysis of the polymer were done to find out the thermal behavior. Calcium content within the sample was found out. Some of the physicochemical properties, such as relative viscosity, specific viscosity, and intrinsic viscosity of the EPS were studied under different conditions. The phenomenon of gel formation by the EPS was investigated for its potential application in metal removal from solutions.

  18. A Putative O-Linked β-N-Acetylglucosamine Transferase Is Essential for Hormogonium Development and Motility in the Filamentous Cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Khayatan, Behzad; Bains, Divleen K; Cheng, Monica H; Cho, Ye Won; Huynh, Jessica; Kim, Rachelle; Omoruyi, Osagie H; Pantoja, Adriana P; Park, Jun Sang; Peng, Julia K; Splitt, Samantha D; Tian, Mason Y; Risser, Douglas D

    2017-05-01

    Most species of filamentous cyanobacteria are capable of gliding motility, likely via a conserved type IV pilus-like system that may also secrete a motility-associated polysaccharide. In a subset of these organisms, motility is achieved only after the transient differentiation of hormogonia, which are specialized filaments that enter a nongrowth state dedicated to motility. Despite the fundamental importance of hormogonia to the life cycles of many filamentous cyanobacteria, the molecular regulation of hormogonium development is largely undefined. To systematically identify genes essential for hormogonium development and motility in the model heterocyst-forming filamentous cyanobacterium Nostoc punctiforme , a forward genetic screen was employed. The first gene identified using this screen, designated ogtA , encodes a putative O-linked β- N -acetylglucosamine transferase (OGT). The deletion of ogtA abolished motility, while ectopic expression of ogtA induced hormogonium development even under hormogonium-repressing conditions. Transcription of ogtA is rapidly upregulated (1 h) following hormogonium induction, and an OgtA-GFPuv fusion protein localized to the cytoplasm. In developing hormogonia, accumulation of PilA but not HmpD is dependent on ogtA Reverse transcription-quantitative PCR (RT-qPCR) analysis indicated equivalent levels of pilA transcript in the wild-type and Δ ogtA mutant strains, while a reporter construct consisting of the intergenic region in the 5' direction of pilA fused to gfp produced lower levels of fluorescence in the Δ ogtA mutant strain than in the wild type. The production of hormogonium polysaccharide in the Δ ogtA mutant strain is reduced compared to that in the wild type but comparable to that in a pilA deletion strain. Collectively, these results imply that O -GlcNAc protein modification regulates the accumulation of PilA via a posttranscriptional mechanism in developing hormogonia. IMPORTANCE Filamentous cyanobacteria are among

  19. Construction of new synthetic biology tools for the control of gene expression in the cyanobacterium Synechococcus sp. strain PCC 7002.

    Science.gov (United States)

    Zess, Erin K; Begemann, Matthew B; Pfleger, Brian F

    2016-02-01

    Predictive control of gene expression is an essential tool for developing synthetic biological systems. The current toolbox for controlling gene expression in cyanobacteria is a barrier to more in-depth genetic analysis and manipulation. Towards relieving this bottleneck, this work describes the use of synthetic biology to construct an anhydrotetracycline-based induction system and adapt a trans-acting small RNA (sRNA) system for use in the cyanobacterium Synechococcus sp. strain PCC 7002. An anhydrotetracycline-inducible promoter was developed to maximize intrinsic strength and dynamic range. The resulting construct, PEZtet , exhibited tight repression and a maximum 32-fold induction upon addition of anhydrotetracycline. Additionally, a sRNA system based on the Escherichia coli IS10 RNA-IN/OUT regulator was adapted for use in Synechococcus sp. strain PCC 7002. This system exhibited 70% attenuation of target gene expression, providing a demonstration of the use of sRNAs for differential gene expression in cyanobacteria. These systems were combined to produce an inducible sRNA system, which demonstrated 59% attenuation of target gene expression. Lastly, the role of Hfq, a critical component of sRNA systems in E. coli, was investigated. Genetic studies showed that the Hfq homolog in Synechococcus sp. strain PCC 7002 did not impact repression by the engineered sRNA system. In summary, this work describes new synthetic biology tools that can be applied to physiological studies, metabolic engineering, or sRNA platforms in Synechococcus sp. strain PCC 7002. © 2015 Wiley Periodicals, Inc.

  20. Ethylene Regulates the Physiology of the Cyanobacterium Synechocystis sp. PCC 6803 via an Ethylene Receptor1[OPEN

    Science.gov (United States)

    2016-01-01

    Ethylene is a plant hormone that plays a crucial role in the growth and development of plants. The ethylene receptors in plants are well studied, and it is generally assumed that they are found only in plants. In a search of sequenced genomes, we found that many bacterial species contain putative ethylene receptors. Plants acquired many proteins from cyanobacteria as a result of the endosymbiotic event that led to chloroplasts. We provide data that the cyanobacterium Synechocystis (Synechocystis sp. PCC 6803) has a functional receptor for ethylene, Synechocystis Ethylene Response1 (SynEtr1). We first show that SynEtr1 directly binds ethylene. Second, we demonstrate that application of ethylene to Synechocystis cells or disruption of the SynEtr1 gene affects several processes, including phototaxis, type IV pilus biosynthesis, photosystem II levels, biofilm formation, and spontaneous cell sedimentation. Our data suggest a model where SynEtr1 inhibits downstream signaling and ethylene inhibits SynEtr1. This is similar to the inverse-agonist model of ethylene receptor signaling proposed for plants and suggests a conservation of structure and function that possibly originated over 1 billion years ago. Prior research showed that SynEtr1 also contains a light-responsive phytochrome-like domain. Thus, SynEtr1 is a bifunctional receptor that mediates responses to both light and ethylene. To our knowledge, this is the first demonstration of a functional ethylene receptor in a nonplant species and suggests that that the perception of ethylene is more widespread than previously thought. PMID:27246094

  1. The response regulator Npun_F1278 is essential for scytonemin biosynthesis in the cyanobacterium Nostoc punctiforme ATCC 29133.

    Science.gov (United States)

    Naurin, Sejuti; Bennett, Janine; Videau, Patrick; Philmus, Benjamin; Soule, Tanya

    2016-08-01

    Following exposure to long-wavelength ultraviolet radiation (UVA), some cyanobacteria produce the indole-alkaloid sunscreen scytonemin. The genomic region associated with scytonemin biosynthesis in the cyanobacterium Nostoc punctiforme includes 18 cotranscribed genes. A two-component regulatory system (Npun_F1277/Npun_F1278) directly upstream from the biosynthetic genes was identified through comparative genomics and is likely involved in scytonemin regulation. In this study, the response regulator (RR), Npun_F1278, was evaluated for its ability to regulate scytonemin biosynthesis using a mutant strain of N. punctiforme deficient in this gene, hereafter strain Δ1278. Following UVA radiation, the typical stimulus to initiate scytonemin biosynthesis, Δ1278 was incapable of producing scytonemin. A phenotypic characterization of Δ1278 suggests that aside from the ability to produce scytonemin, the deletion of the Npun_F1278 gene does not affect the cellular morphology, cellular differentiation capability, or lipid-soluble pigment complement of Δ1278 compared to the wildtype. The mutant, however, had a slower specific growth rate under white light and produced ~2.5-fold more phycocyanin per cell under UVA than the wildtype. Since Δ1278 does not produce scytonemin, this study demonstrates that the RR gene, Npun_F1278, is essential for scytonemin biosynthesis in N. punctiforme. While most of the evaluated effects of this gene appear to be specific for scytonemin, this regulator may also influence the overall health of the cell and phycobiliprotein synthesis, directly or indirectly. This is the first study to identify a regulatory gene involved in the biosynthesis of the sunscreen scytonemin and posits a link between cell growth, pigment synthesis, and sunscreen production. © 2016 Phycological Society of America.

  2. Effects of UV-B Radiation and Periodic Desiccation on the Morphogenesis of the Edible Terrestrial Cyanobacterium Nostoc flagelliforme

    Science.gov (United States)

    Feng, Yan-Na; Zhang, Zhong-Chun; Feng, Jun-Li

    2012-01-01

    The terrestrial cyanobacterium Nostoc flagelliforme Berk. et M. A. Curtis has been a popular food and herbal ingredient for hundreds of years. To meet great market demand and protect the local ecosystem, for decades researchers have tried to cultivate N. flagelliforme but have failed to get macroscopic filamentous thalli. In this study, single trichomes with 50 to 200 vegetative cells were induced from free-living cells by low light and used to investigate the morphogenesis of N. flagelliforme under low UV-B radiation and periodic desiccation. Low-fluence-rate UV-B (0.1 W m−2) did not inhibit trichome growth; however, it significantly increased the synthesis of extracellular polysaccharides and mycosporine-like amino acids and promoted sheath formation outside the trichomes. Under low UV-B radiation, single trichomes developed into filamentous thalli more than 1 cm long after 28 days of cultivation, most of which grew separately in liquid BG11 medium. With periodic desiccation treatment, the single trichomes formed flat or banded thalli that grew up to 2 cm long after 3 months on solid BG11 medium. When trichomes were cultivated on solid BG11 medium with alternate treatments of low UV-B and periodic desiccation, dark and scraggly filamentous thalli that grew up to about 3 cm in length after 40 days were obtained. In addition, the cultivation of trichomes on nitrogen-deficient solid BG11 medium (BG110) suggested that nitrogen availability could affect the color and lubricity of newly developed thalli. This study provides promising techniques for artificial cultivation of N. flagelliforme in the future. PMID:22865081

  3. Identification of OmpR-family response regulators interacting with thioredoxin in the Cyanobacterium Synechocystis sp. PCC 6803.

    Directory of Open Access Journals (Sweden)

    Taro Kadowaki

    Full Text Available The redox state of the photosynthetic electron transport chain is known to act as a signal to regulate the transcription of key genes involved in the acclimation responses to environmental changes. We hypothesized that the protein thioredoxin (Trx acts as a mediator connecting the redox state of the photosynthetic electron transport chain and transcriptional regulation, and established a screening system to identify transcription factors (TFs that interact with Trx. His-tagged TFs and S-tagged mutated form of Trx, TrxMC35S, whose active site cysteine 35 was substituted with serine to trap the target interacting protein, were co-expressed in E. coli cells and Trx-TF complexes were detected by immuno-blotting analysis. We examined the interaction between Trx and ten OmpR family TFs encoded in the chromosome of the cyanobacterium Synechocystis sp. PCC 6803 (S.6803. Although there is a highly conserved cysteine residue in the receiver domain of all OmpR family TFs, only three, RpaA (Slr0115, RpaB (Slr0947 and ManR (Slr1837, were identified as putative Trx targets [corrected].The recombinant forms of wild-type TrxM, RpaA, RpaB and ManR proteins from S.6803 were purified following over-expression in E. coli and their interaction was further assessed by monitoring changes in the number of cysteine residues with free thiol groups. An increase in the number of free thiols was observed after incubation of the oxidized TFs with Trx, indicating the reduction of cysteine residues as a consequence of interaction with Trx. Our results suggest, for the first time, the possible regulation of OmpR family TFs through the supply of reducing equivalents from Trx, as well as through the phospho-transfer from its cognate sensor histidine kinase.

  4. Novel derivatives of 9,10-anthraquinone are selective algicides against the musty-odor cyanobacterium Oscillatoria perornata.

    Science.gov (United States)

    Schrader, Kevin K; Nanayakkara, N P Dhammika; Tucker, Craig S; Rimando, Agnes M; Ganzera, Markus; Schaneberg, Brian T

    2003-09-01

    Musty "off-flavor" in pond-cultured channel catfish (Ictalurus punctatus) costs the catfish production industry in the United States at least 30 million US dollars annually. The cyanobacterium Oscillatoria perornata (Skuja) is credited with being the major cause of musty off-flavor in farm-raised catfish in Mississippi. The herbicides diuron and copper sulfate, currently used by catfish producers as algicides to help mitigate musty off-flavor problems, have several drawbacks, including broad-spectrum toxicity towards the entire phytoplankton community that can lead to water quality deterioration and subsequent fish death. By use of microtiter plate bioassays, a novel group of compounds derived from the natural compound 9,10-anthraquinone have been found to be much more selectively toxic towards O. perornata than diuron and copper sulfate. In efficacy studies using limnocorrals placed in catfish production ponds, application rates of 0.3 micro M (125 micro g/liter) of the most promising anthraquinone derivative, 2-[methylamino-N-(1'-methylethyl)]-9,10-anthraquinone monophosphate (anthraquinone-59), dramatically reduced the abundance of O. perornata and levels of 2-methylisoborneol, the musty compound produced by O. perornata. The abundance of green algae and diatoms increased dramatically 2 days after application of a 0.3 micro M concentration of anthraquinone-59 to pond water within the limnocorrals. The half-life of anthraquinone-59 in pond water was determined to be 19 h, making it much less persistent than diuron. Anthraquinone-59 appears to be promising for use as a selective algicide in catfish aquaculture.

  5. Physiology, Fe(II oxidation, and Fe mineral formation by a marine planktonic cyanobacterium grown under ferruginous conditions

    Directory of Open Access Journals (Sweden)

    Elizabeth D. Swanner

    2015-10-01

    Full Text Available Evidence for Fe(II oxidation and deposition of Fe(III-bearing minerals from anoxic or redox-stratified Precambrian oceans has received support from decades of sedimentological and geochemical investigation of Banded Iron Formations (BIF. While the exact mechanisms of Fe(II oxidation remains equivocal, reaction with O2 in the marine water column, produced by cyanobacteria or early oxygenic phototrophs, was likely. In order to understand the role of cyanobacteria in the deposition of Fe(III minerals to BIF, we must first know how planktonic marine cyanobacteria respond to ferruginous (anoxic and Fe(II-rich waters in terms of growth, Fe uptake and homeostasis, and Fe mineral formation. We therefore grew the common marine cyanobacterium Synechococcus PCC 7002 in closed bottles that began anoxic, and contained Fe(II concentrations that span the range of possible concentrations in Precambrian seawater. These results, along with cell suspension experiments, indicate that Fe(II is likely oxidized by this strain via chemical oxidation with oxygen produced during photosynthesis, and not via any direct enzymatic or photosynthetic pathway. Imaging of the cell-mineral aggregates with scanning electron microscopy (SEM and confocal laser scanning microscopy (CLSM are consistent with extracellular precipitation of Fe(III (oxyhydroxide minerals, but that >10% of Fe(III sorbs to cell surfaces rather than precipitating. Proteomic experiments support the role of reactive oxygen species (ROS in Fe(II toxicity to Synechococcus PCC 7002. The proteome expressed under low Fe conditions included multiple siderophore biosynthesis and siderophore and Fe transporter proteins, but most siderophores are not expressed during growth with Fe(II. These results provide a mechanistic and quantitative framework for evaluating the geochemical consequences of perhaps life’s greatest metabolic innovation, i.e. the evolution and activity of oxygenic photosynthesis, in ferruginous

  6. Novel Derivatives of 9,10-Anthraquinone Are Selective Algicides against the Musty-Odor Cyanobacterium Oscillatoria perornata

    Science.gov (United States)

    Schrader, Kevin K.; Dhammika Nanayakkara, N. P.; Tucker, Craig S.; Rimando, Agnes M.; Ganzera, Markus; Schaneberg, Brian T.

    2003-01-01

    Musty “off-flavor” in pond-cultured channel catfish (Ictalurus punctatus) costs the catfish production industry in the United States at least $30 million annually. The cyanobacterium Oscillatoria perornata (Skuja) is credited with being the major cause of musty off-flavor in farm-raised catfish in Mississippi. The herbicides diuron and copper sulfate, currently used by catfish producers as algicides to help mitigate musty off-flavor problems, have several drawbacks, including broad-spectrum toxicity towards the entire phytoplankton community that can lead to water quality deterioration and subsequent fish death. By use of microtiter plate bioassays, a novel group of compounds derived from the natural compound 9,10-anthraquinone have been found to be much more selectively toxic towards O. perornata than diuron and copper sulfate. In efficacy studies using limnocorrals placed in catfish production ponds, application rates of 0.3 μM (125 μg/liter) of the most promising anthraquinone derivative, 2-[methylamino-N-(1′-methylethyl)]-9,10-anthraquinone monophosphate (anthraquinone-59), dramatically reduced the abundance of O. perornata and levels of 2-methylisoborneol, the musty compound produced by O. perornata. The abundance of green algae and diatoms increased dramatically 2 days after application of a 0.3 μM concentration of anthraquinone-59 to pond water within the limnocorrals. The half-life of anthraquinone-59 in pond water was determined to be 19 h, making it much less persistent than diuron. Anthraquinone-59 appears to be promising for use as a selective algicide in catfish aquaculture. PMID:12957919

  7. First description of a cyanophage infecting the cyanobacterium Arthrospira platensis (Spirulina)

    Digital Repository Service at National Institute of Oceanography (India)

    Jacquet, S.; Zhong, X.; Parvathi, A.; Ram, A.S.P.

    Cyanobacteria constitute a versatile group of photosynthetic bacteria of immense commercial and ecological importance. Some species of this group are cultivated and sold as food because of their high nutritional value. This is typically the case...

  8. Antagonism at combined effects of chemical fertilizers and carbamate insecticides on the rice-field N2-fixing cyanobacterium Cylindrospermum sp. in vitro

    Directory of Open Access Journals (Sweden)

    Padhy Rabindra N.

    2014-03-01

    Full Text Available Effects of chemical fertilizers (urea, super phosphate and potash on toxicities of two carbamate insecticides, carbaryl and carbofuran, individually to the N2-fixing cyanobacterium, Cylindrospermum sp. were studied in vitro at partially lethal levels (below highest permissive concentrations of each insecticide. The average number of vegetative cells between two polar heterocysts was 16.3 in control cultures, while the mean value of filament length increased in the presence of chemical fertilizers, individually. Urea at the 10 ppm level was growth stimulatory and at the 50 ppm level it was growth inhibitory in control cultures, while at 100 ppm it was antagonistic, i.e. toxicity-enhancing along with carbaryl, individually to the cyanobacterium, antagonism was recorded. Urea at 50 ppm had toxicity reducing effect with carbaryl or carbofuran. At 100 and 250 ppm carbofuran levels, 50 ppm urea only had a progressive growth enhancing effect, which was marked well at 250 ppm carbofuran level, a situation of synergism. Super phosphate at the 10 ppm level only was growth promoting in control cultures, but it was antagonistic at its higher levels (50 and 100 ppm along with both insecticides, individually. Potash (100, 200, 300 and 400 ppm reduced toxicity due to carbaryl 20 and carbofuran 250 ppm levels, but potash was antagonistic at the other insecticide levels. The data clearly showed that the chemical fertilizers used were antagonistic with both the insecticides during toxicity to Cylindrospermum sp.

  9. Antagonism at combined effects of chemical fertilizers and carbamate insecticides on the rice-field N2-fixing cyanobacterium Cylindrospermum sp. in vitro

    Science.gov (United States)

    Nayak, Nabakishore; Rath, Shakti

    2014-01-01

    Effects of chemical fertilizers (urea, super phosphate and potash) on toxicities of two carbamate insecticides, carbaryl and carbofuran, individually to the N2-fixing cyanobacterium, Cylindrospermum sp. were studied in vitro at partially lethal levels (below highest permissive concentrations) of each insecticide. The average number of vegetative cells between two polar heterocysts was 16.3 in control cultures, while the mean value of filament length increased in the presence of chemical fertilizers, individually. Urea at the 10 ppm level was growth stimulatory and at the 50 ppm level it was growth inhibitory in control cultures, while at 100 ppm it was antagonistic, i.e. toxicity-enhancing along with carbaryl, individually to the cyanobacterium, antagonism was recorded. Urea at 50 ppm had toxicity reducing effect with carbaryl or carbofuran. At 100 and 250 ppm carbofuran levels, 50 ppm urea only had a progressive growth enhancing effect, which was marked well at 250 ppm carbofuran level, a situation of synergism. Super phosphate at the 10 ppm level only was growth promoting in control cultures, but it was antagonistic at its higher levels (50 and 100 ppm) along with both insecticides, individually. Potash (100, 200, 300 and 400 ppm) reduced toxicity due to carbaryl 20 and carbofuran 250 ppm levels, but potash was antagonistic at the other insecticide levels. The data clearly showed that the chemical fertilizers used were antagonistic with both the insecticides during toxicity to Cylindrospermum sp. PMID:26038669

  10. CO2 Removal from Biogas by Cyanobacterium Leptolyngbya sp. CChF1 Isolated from the Lake Chapala, Mexico: Optimization of the Temperature and Light Intensity.

    Science.gov (United States)

    Choix, Francisco J; Snell-Castro, Raúl; Arreola-Vargas, Jorge; Carbajal-López, Alberto; Méndez-Acosta, Hugo O

    2017-12-01

    In the present study, the capacity of the cyanobacterium Leptolyngbya sp. CChF1 to remove CO 2 from real and synthetic biogas was evaluated. The identification of the cyanobacterium, isolated from the lake Chapala, was carried out by means of morphological and molecular analyses, while its potential for CO 2 removal from biogas streams was evaluated by kinetic experiments and optimized by a central composite design coupled to a response surface methodology. Results demonstrated that Leptolyngbya sp. CChF1 is able to remove CO 2 and grow indistinctly in real or synthetic biogas streams, showing tolerance to high concentrations of CO 2 and CH 4 , 25 and 75%, respectively. The characterization of the biomass composition at the end of the kinetic assays revealed that the main accumulated by-products under both biogas streams were lipids, followed by proteins and carbohydrates. Regarding the optimization experiments, light intensity and temperature were the studied variables, while synthetic biogas was the carbon source. Results showed that light intensity was significant for CO 2 capture efficiency (p = 0.0290), while temperature was significant for biomass production (p = 0.0024). The predicted CO 2 capture efficiency under optimal conditions (27.1 °C and 920 lx) was 93.48%. Overall, the results of the present study suggest that Leptolyngbya sp. CChF1 is a suitable candidate for biogas upgrading.

  11. The NADPH thioredoxin reductase C functions as an electron donor to 2-Cys peroxiredoxin in a thermophilic cyanobacterium Thermosynechococcus elongatus BP-1

    International Nuclear Information System (INIS)

    Sueoka, Keigo; Yamazaki, Teruaki; Hiyama, Tetsuo; Nakamoto, Hitoshi

    2009-01-01

    An NADPH thioredoxin reductase C was co-purified with a 2-Cys peroxiredoxin by the combination of anion exchange chromatography and electroelution from gel slices after native PAGE from a thermophilic cyanobacterium Thermosynechococcus elongatus as an NAD(P)H oxidase complex induced by oxidative stress. The result provided a strong evidence that the NADPH thioredoxin reductase C interacts with the 2-Cys peroxiredoxin in vivo. An in vitro reconstitution assay with purified recombinant proteins revealed that both proteins were essential for an NADPH-dependent reduction of H 2 O 2 . These results suggest that the reductase transfers the reducing power from NADPH to the peroxiredoxin, which reduces peroxides in the cyanobacterium under oxidative stress. In contrast with other NADPH thioredoxin reductases, the NADPH thioredoxin reductase C contains a thioredoxin-like domain in addition to an NADPH thioredoxin reductase domain in the same polypeptide. Each domain contains a conserved CXYC motif. A point mutation at the CXYC motif in the NADPH thioredoxin reductase domain resulted in loss of the NADPH oxidation activity, while a mutation at the CXYC motif in the thioredoxin-like domain did not affect the electron transfer, indicating that this motif is not essential in the electron transport from NADPH to the 2-Cys peroxiredoxin.

  12. Novel glycosylated mycosporine-like amino acid, 13-O-(β-galactosyl)-porphyra-334, from the edible cyanobacterium Nostoc sphaericum-protective activity on human keratinocytes from UV light.

    Science.gov (United States)

    Ishihara, Kenji; Watanabe, Ryuichi; Uchida, Hajime; Suzuki, Toshiyuki; Yamashita, Michiaki; Takenaka, Hiroyuki; Nazifi, Ehsan; Matsugo, Seiichi; Yamaba, Minami; Sakamoto, Toshio

    2017-07-01

    A UV-absorbing compound was purified and identified as a novel glycosylated mycosporine-like amino acid (MAA), 13-O-β-galactosyl-porphyra-334 (β-Gal-P334) from the edible cyanobacterium Nostoc sphaericum, known as "ge xian mi" in China and "cushuro" in Peru. Occurrence of the hexosylated derivative of shinorine (hexosyl-shinorine) was also supported by LC-MS/MS analysis. β-Gal-P334 accounted for about 86.5% of total MAA in N. sphaericum, followed by hexosyl-shinorine (13.2%) and porphyra-334 (0.2%). β-Gal-P334 had an absorption maximum at 334nm and molecular absorption coefficient was 46,700 at 334nm. Protection activity of β-Gal-P334 from UVB and UVA+8-methoxypsoralen induced cell damage on human keratinocytes (HaCaT) was assayed in comparison with other MAA (porphyra-334, shinorine, palythine and mycosporine-glycine). The UVB protection activity was highest in mycosporine-glycine, followed by palythine, β-Gal-P334, porphyra-334 and shinorine in order. β-Gal-P334 had highest protection activity from UVA+8-methoxypsoralen induced cell damage followed by porphyra-334, shinorine, mycosporine-glycine and palythine. We also found an antioxidant (radical-scavenging) activity of β-Gal-P334 by colorimetric and ESR methods. From these findings, β-Gal-P334 was suggested to play important roles in stress tolerant mechanisms such as UV and oxidative stress in N. sphaericum as a major MAA. We also consider that the newly identified MAA, β-Gal-P334 has a potential for use as an ingredient of cosmetics and toiletries. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. RNA-seq analyses reveal insights into the function of respiratory nitrate reductase of the diazotroph Herbaspirillum seropedicae.

    Science.gov (United States)

    Bonato, Paloma; Batista, Marcelo B; Camilios-Neto, Doumit; Pankievicz, Vânia C S; Tadra-Sfeir, Michelle Z; Monteiro, Rose Adele; Pedrosa, Fabio O; Souza, Emanuel M; Chubatsu, Leda S; Wassem, Roseli; Rigo, Liu Un

    2016-09-01

    Herbaspirillum seropedicae is a nitrogen-fixing β-proteobacterium that associates with roots of gramineous plants. In silico analyses revealed that H. seropedicae genome has genes encoding a putative respiratory (NAR) and an assimilatory nitrate reductase (NAS). To date, little is known about nitrate metabolism in H. seropedicae, and, as this bacterium cannot respire nitrate, the function of NAR remains unknown. This study aimed to investigate the function of NAR in H. seropedicae and how it metabolizes nitrate in a low aerated-condition. RNA-seq transcriptional profiling in the presence of nitrate allowed us to pinpoint genes important for nitrate metabolism in H. seropedicae, including nitrate transporters and regulatory proteins. Additionally, both RNA-seq data and physiological characterization of a mutant in the catalytic subunit of NAR (narG mutant) showed that NAR is not required for nitrate assimilation but is required for: (i) production of high levels of nitrite, (ii) production of NO and (iii) dissipation of redox power, which in turn lead to an increase in carbon consumption. In addition, wheat plants showed an increase in shoot dry weight only when inoculated with H. seropedicae wild type, but not with the narG mutant, suggesting that NAR is important to H. seropedicae-wheat interaction. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  14. Identification and characterization of a carboxysomal γ-carbonic anhydrase from the cyanobacterium Nostoc sp. PCC 7120.

    Science.gov (United States)

    de Araujo, Charlotte; Arefeen, Dewan; Tadesse, Yohannes; Long, Benedict M; Price, G Dean; Rowlett, Roger S; Kimber, Matthew S; Espie, George S

    2014-09-01

    Carboxysomes are proteinaceous microcompartments that encapsulate carbonic anhydrase (CA) and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco); carboxysomes, therefore, catalyze reversible HCO3 (-) dehydration and the subsequent fixation of CO2. The N- and C-terminal domains of the β-carboxysome scaffold protein CcmM participate in a network of protein-protein interactions that are essential for carboxysome biogenesis, organization, and function. The N-terminal domain of CcmM in the thermophile Thermosynechococcus elongatus BP-1 is also a catalytically active, redox regulated γ-CA. To experimentally determine if CcmM from a mesophilic cyanobacterium is active, we cloned, expressed and purified recombinant, full-length CcmM from Nostoc sp. PCC 7120 as well as the N-terminal 209 amino acid γ-CA-like domain. Both recombinant proteins displayed ethoxyzolamide-sensitive CA activity in mass spectrometric assays, as did the carboxysome-enriched TP fraction. NstCcmM209 was characterized as a moderately active and efficient γ-CA with a k cat of 2.0 × 10(4) s(-1) and k cat/K m of 4.1 × 10(6) M(-1) s(-1) at 25 °C and pH 8, a pH optimum between 8 and 9.5 and a temperature optimum spanning 25-35 °C. NstCcmM209 also catalyzed the hydrolysis of the CO2 analog carbonyl sulfide. Circular dichroism and intrinsic tryptophan fluorescence analysis demonstrated that NstCcmM209 was progressively and irreversibly denatured above 50 °C. NstCcmM209 activity was inhibited by the reducing agent tris(hydroxymethyl)phosphine, an effect that was fully reversed by a molar excess of diamide, a thiol oxidizing agent, consistent with oxidative activation being a universal regulatory mechanism of CcmM orthologs. Immunogold electron microscopy and Western blot analysis of TP pellets indicated that Rubisco and CcmM co-localize and are concentrated in Nostoc sp. PCC 7120 carboxysomes.

  15. Diversidade de bactérias diazotróficas endofíticas associadas a plantas de milho Diversity of diazotrophic endophytic bacteria associated with maize plants

    Directory of Open Access Journals (Sweden)

    Luiz Fernando Wurdig Roesch

    2007-12-01

    capazes de colonizar o interior de plantas de milho e que as diferentes condições edafoclimáticas estão correlacionadas com a diversidade dos genes nifH.Endophytic diazotrophic bacteria are capable of promoting maize growth through biological nitrogen fixation (BNF or by the production of plant hormones. The aim of this study was to characterize diversity of endophytic bacteria in maize at sites with different climate and soil conditions in Rio Grande do Sul, Brazil. A PCR-RFLP approach and sequence analysis of nifH Cluster I clone libraries were used to assess diversity in maize plants. The Shannon-Weaver and Equitability indices were calculated to estimate the diazotroph diversity as well as the nucleotide diversity and the average sequence divergence to estimate genetic diversity. To evaluate the variability in populations we performed the F ST test. A greater variation in bacterial communities was detected between rather than within regions, particularly among communities of different soil types and varying water regimes and geographical regions. The Shannon-Weaver index indicated a high difference in terms of diversity of taxonomic units among the communities. The diversity of the communities in the northern region, with higher precipitation and clay content, tended to be higher than that in the south. The Equitability index indicated that there was a dominant taxonomic unit within each community. All clones grouped into nifH gene cluster I. The nifH sequence types from Alpha, Beta and Gammaproteobacteria were recovered. These results demonstrate that there is a large diversity of endophytic nitrogen-fixing bacteria able to colonize maize tissue and that nifH diversity is correlated with the different environmental conditions.

  16. Identification of a transporter Slr0982 involved in ethanol tolerance in cyanobacterium Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Yanan eZhang

    2015-05-01

    Full Text Available Cyanobacteria have been engineered to produce ethanol through recent synthetic biology efforts. However, one major challenge to the cyanobacterial systems for high-efficiency ethanol production is their low tolerance to the ethanol toxicity. With a major goal to identify novel transporters involved in ethanol tolerance, we constructed gene knockout mutants for 58 transporter-encoding genes of Synechocystis sp. PCC 6803 and screened their tolerance change under ethanol stress. The efforts allowed discovery of a mutant of slr0982 gene encoding an ATP-binding cassette transporter which grew poorly in BG11 medium supplemented with 1.5% (v/v ethanol when compared with the wild type, and the growth loss could be recovered by complementing slr0982 in the ∆slr0982 mutant, suggesting that slr0982 is involved in ethanol tolerance in Synechocystis. To decipher the tolerance mechanism involved, a comparative metabolomic and network-based analysis of the wild type and the ethanol-sensitive ∆slr0982 mutant was performed. The analysis allowed the identification of four metabolic modules related to slr0982 deletion in the ∆slr0982 mutant, among which metabolites like sucrose and L-pyroglutamic acid which might be involved in ethanol tolerance, were found important for slr0982 deletion in the ∆slr0982 mutant. This study reports on the first transporter related to ethanol tolerance in Synechocystis, which could be a useful target for further tolerance engineering. In addition, metabolomic and network analysis provides important findings for better understanding of the tolerance mechanism to ethanol stress in Synechocystis.

  17. Biochemical Validation of the Glyoxylate Cycle in the Cyanobacterium Chlorogloeopsis fritschii Strain PCC 9212.

    Science.gov (United States)

    Zhang, Shuyi; Bryant, Donald A

    2015-05-29

    Cyanobacteria are important photoautotrophic bacteria with extensive but variable metabolic capacities. The existence of the glyoxylate cycle, a variant of the TCA cycle, is still poorly documented in cyanobacteria. Previous studies reported the activities of isocitrate lyase and malate synthase, the key enzymes of the glyoxylate cycle in some cyanobacteria, but other studies concluded that these enzymes are missing. In this study the genes encoding isocitrate lyase and malate synthase from Chlorogloeopsis fritschii PCC 9212 were identified, and the recombinant enzymes were biochemically characterized. Consistent with the presence of the enzymes of the glyoxylate cycle, C. fritschii could assimilate acetate under both light and dark growth conditions. Transcript abundances for isocitrate lyase and malate synthase increased, and C. fritschii grew faster, when the growth medium was supplemented with acetate. Adding acetate to the growth medium also increased the yield of poly-3-hydroxybutyrate. When the genes encoding isocitrate lyase and malate synthase were expressed in Synechococcus sp. PCC 7002, the acetate assimilation capacity of the resulting strain was greater than that of wild type. Database searches showed that the genes for the glyoxylate cycle exist in only a few other cyanobacteria, all of which are able to fix nitrogen. This study demonstrates that the glyoxylate cycle exists in a few cyanobacteria, and that this pathway plays an important role in the assimilation of acetate for growth in one of those organisms. The glyoxylate cycle might play a role in coordinating carbon and nitrogen metabolism under conditions of nitrogen fixation. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. Looking at the stability of life-support microorganisms in space : the MELGEN activity highlights the cyanobacterium Arthrospira sp. PCC8005

    Science.gov (United States)

    Morin, Nicolas

    The MELGEN activity (MELiSSA Genetic Stability Study) mainly covers the molecular aspects of the regenerative life-support system MELiSSA (Micro-Ecological Life Support System Alternative) of the European Space Agency (ESA). The general objective of MELGEN is to establish and validate methods and the related hardware in order to detect genetic instability and microbial contaminants in the MELISSA compartments. This includes (1) a genetic description of the MELISSA strains, (2) studies of microbial behavior and genetic stability in bioreactors and (3) the detection of chemical, genetical and biological contamination and their effect on microbial metabolism. Selected as oxygen producer and complementary food source, the cyanobacterium Arthrospira sp. PCC8005 plays a major role within the MELiSSA loop. As the genomic information on this organism was insufficient, sequencing of its genome was proposed at the French National Sequencing Center, Genoscope, as a joint effort between ESA and different laboratories. So far, a preliminary assembly of 16 contigs representing circa 6.3 million basepairs was obtained. Even though the finishing of the genome is on its way, automatic annotation of the contigs has already been performed on the MaGe annotation platform, and curation of the sequence is currently being carried out, with a special focus on biosynthesis pathways, photosynthesis, and maintenance processes of the cell. According to the index of repetitiveness described by Haubold and Wiehe (2006), we discovered that the genome of Arthrospira sp. is among the 50 most repeated bacterial genomes sequenced to date. Thanks to the sequencing project, we have identified and catalogued mobile genetics elements (MGEs) dispersed throughout the unique chromosome of this cyanobacterium. They represent a quite large proportion of the genome, as genes identified as putative transposases are indeed found in circa 5 Results : We currently have a first draft of the complete genome of

  19. Kinetic Modeling of Arsenic Cycling by a Freshwater Cyanobacterium as Influenced by N:P Ratios: A Potential Biologic Control in an Iron-Limited Drainage Basin

    Science.gov (United States)

    Markley, C. T.; Herbert, B. E.

    2004-12-01

    Elevated As levels are common in South Texas surface waters, where As is derived from the natural weathering of geogenic sources and a byproduct of historical uranium mining. The impacted surface waters of the Nueces River drainage basin supply Lake Corpus Christi (LCC), a major drinking water reservoir for the Corpus Christi area. The soils and sediments of the Nueces River drainage basin generally have low levels of reactive iron (average concentration of 2780 mg/kg), limiting the control of iron oxyhydroxides on As geochemistry and bioavailability. Given these conditions, biologic cycling of As may have a large influence on As fate and transport in LCC. Sediment cores from LCC show evidence for cyanobacterial blooms after reservoir formation based upon stable isotopes, total organic matter and specific elemental correlations. While algae have been shown to accumulate and reduce inorganic As(V), few studies have reported biologic cycling of As by cyanobacteria. Therefore, As(V) uptake, accumulation, reduction, and excretion in a 1.0 μ M As(V) solution by the freshwater cyanobacterium, Anabaena sp. Strain PCC 7120, was measured over time as a function of low, middle and high N:P ratios (1.2, 12, 120) to determine nutrient effects on As cycling by the cyanobacterium. Total As(V) reduction was observed in all three conditions upon completion of the ten-day experiment. Maximum As(V) reduction rates ranged from (0.013 mmol g C-1 day-1) in the low N:P solution to (0.398 mmol g C-1 day-1) in the high N:P solution. Increased cell biomass in the low N:P ratio solution compensated for the low maximum reduction rate to allow total As(V) reduction. Kinetic equations commonly used to model algal-nutrient interactions were utilized in modeling the current data. The Michaelis-Menten enzyme saturation equation modified with a competitive inhibition term adequately modeled As(III) excretion in the high and middle N:P ratio test conditions. The low N:P test condition further

  20. Dynamics and controls of heterotrophic prokaryotic production in the western tropical South Pacific Ocean: links with diazotrophic and photosynthetic activity

    Science.gov (United States)

    Van Wambeke, France; Gimenez, Audrey; Duhamel, Solange; Dupouy, Cécile; Lefevre, Dominique; Pujo-Pay, Mireille; Moutin, Thierry

    2018-05-01

    Heterotrophic prokaryotic production (BP) was studied in the western tropical South Pacific (WTSP) using the leucine technique, revealing spatial and temporal variability within the region. Integrated over the euphotic zone, BP ranged from 58 to 120 mg C m-2 d-1 within the Melanesian Archipelago, and from 31 to 50 mg C m-2 d-1 within the western subtropical gyre. The collapse of a bloom was followed during 6 days in the south of Vanuatu using a Lagrangian sampling strategy. During this period, rapid evolution was observed in the three main parameters influencing the metabolic state: BP, primary production (PP) and bacterial growth efficiency. With N2 fixation being one of the most important fluxes fueling new production, we explored relationships between BP, PP and N2 fixation rates over the WTSP. The contribution of N2 fixation rates to bacterial nitrogen demand ranged from 3 to 81 %. BP variability was better explained by the variability of N2 fixation rates than by that of PP in surface waters of the Melanesian Archipelago, which were characterized by N-depleted layers and low DIP turnover times (TDIP 100 h), deeper in the Melanesian Archipelago, or within the entire euphotic zone in the subtropical gyre. The bacterial carbon demand to gross primary production ratio ranged from 0.75 to 3.1. These values are discussed in the framework of various assumptions and conversion factors used to estimate this ratio, including the methodological errors, the daily variability of BP, the bacterial growth efficiency and one bias so far not considered: the ability for Prochlorococcus to assimilate leucine in the dark.

  1. Protein Network Signatures Associated with Exogenous Biofuels Treatments in Cyanobacterium Synechocystis sp. PCC 6803

    International Nuclear Information System (INIS)

    Pei, Guangsheng; Chen, Lei; Wang, Jiangxin; Qiao, Jianjun; Zhang, Weiwen

    2014-01-01

    Although recognized as a promising microbial cell factory for producing biofuels, current productivity in cyanobacterial systems is low. To make the processes economically feasible, one of the hurdles, which need to be overcome is the low tolerance of hosts to toxic biofuels. Meanwhile, little information is available regarding the cellular responses to biofuels stress in cyanobacteria, which makes it challenging for tolerance engineering. Using large proteomic datasets of Synechocystis under various biofuels stress and environmental perturbation, a protein co-expression network was first constructed and then combined with the experimentally determined protein–protein interaction network. Proteins with statistically higher topological overlap in the integrated network were identified as common responsive proteins to both biofuels stress and environmental perturbations. In addition, a weighted gene co-expression network analysis was performed to distinguish unique responses to biofuels from those to environmental perturbations and to uncover metabolic modules and proteins uniquely associated with biofuels stress. The results showed that biofuel-specific proteins and modules were enriched in several functional categories, including photosynthesis, carbon fixation, and amino acid metabolism, which may represent potential key signatures for biofuels stress responses in Synechocystis. Network-based analysis allowed determination of the responses specifically related to biofuels stress, and the results constituted an important knowledge foundation for tolerance engineering against biofuels in Synechocystis.

  2. Elevated CO2 causes changes in the photosynthetic apparatus of a toxic cyanobacterium, Cylindrospermopsis raciborskii.

    Science.gov (United States)

    Pierangelini, Mattia; Stojkovic, Slobodanka; Orr, Philip T; Beardall, John

    2014-07-15

    We studied the physiological acclimation of growth, photosynthesis and CO2-concentrating mechanism (CCM) in Cylindrospermopsis raciborskii exposed to low (present day; L-CO2) and high (1300ppm; H-CO2) pCO2. Results showed that under H-CO2 the cell specific division rate (μc) was higher and the CO2- and light-saturated photosynthetic rates (Vmax and Pmax) doubled. The cells' photosynthetic affinity for CO2 (K0.5CO2) was halved compared to L-CO2 cultures. However, no significant differences were found in dark respiration rates (Rd), pigment composition and light harvesting efficiency (α). In H-CO2 cells, non-photochemical quenching (NPQ), associated with state transitions of the electron transport chain (ETC), was negligible. Simultaneously, a reorganisation of PSII features including antenna connectivity (JconPSIIα), heterogeneity (PSIIα/β) and effective absorption cross sectional area (σPSIIα/β) was observed. In relation to different activities of the CCM, our findings suggest that for cells grown under H-CO2: (1) there is down-regulation of CCM activity; (2) the ability of cells to use the harvested light energy is altered; (3) the occurrence of state transitions is likely to be associated with changes of electron flow (cyclic vs linear) through the ETC; (4) changes in PSII characteristics are important in regulating state transitions. Copyright © 2014 Elsevier GmbH. All rights reserved.

  3. Function and Regulation of Ferredoxins in the Cyanobacterium, Synechocystis PCC6803: Recent Advances.

    Science.gov (United States)

    Cassier-Chauvat, Corinne; Chauvat, Franck

    2014-11-07

    Ferredoxins (Fed), occurring in most organisms, are small proteins that use their iron-sulfur cluster to distribute electrons to various metabolic pathways, likely including hydrogen production. Here, we summarize the current knowledge on ferredoxins in cyanobacteria, the prokaryotes regarded as important producers of the oxygenic atmosphere and biomass for the food chain, as well as promising cell factories for biofuel production. Most studies of ferredoxins were performed in the model strain, Synechocystis PCC6803, which possesses nine highly-conserved ferredoxins encoded by monocistronic or operonic genes, some of which are localized in conserved genome regions. Fed1, encoded by a light-inducible gene, is a highly abundant protein essential to photosynthesis. Fed2-Fed9, encoded by genes differently regulated by trophic conditions, are low-abundant proteins that play prominent roles in the tolerance to environmental stresses. Concerning the selectivity/redundancy of ferredoxin, we report that Fed1, Fed7 and Fed9 belong to ferredoxin-glutaredoxin-thioredoxin crosstalk pathways operating in the protection against oxidative and metal stresses. Furthermore, Fed7 specifically interacts with a DnaJ-like protein, an interaction that has been conserved in photosynthetic eukaryotes in the form of a composite protein comprising DnaJ- and Fed7-like domains. Fed9 specifically interacts with the Flv3 flavodiiron protein acting in the photoreduction of O2 to H2O.

  4. Function and Regulation of Ferredoxins in the Cyanobacterium, Synechocystis PCC6803: Recent Advances

    Directory of Open Access Journals (Sweden)

    Corinne Cassier-Chauvat

    2014-11-01

    Full Text Available Ferredoxins (Fed, occurring in most organisms, are small proteins that use their iron-sulfur cluster to distribute electrons to various metabolic pathways, likely including hydrogen production. Here, we summarize the current knowledge on ferredoxins in cyanobacteria, the prokaryotes regarded as important producers of the oxygenic atmosphere and biomass for the food chain, as well as promising cell factories for biofuel production. Most studies of ferredoxins were performed in the model strain, Synechocystis PCC6803, which possesses nine highly-conserved ferredoxins encoded by monocistronic or operonic genes, some of which are localized in conserved genome regions. Fed1, encoded by a light-inducible gene, is a highly abundant protein essential to photosynthesis. Fed2-Fed9, encoded by genes differently regulated by trophic conditions, are low-abundant proteins that play prominent roles in the tolerance to environmental stresses. Concerning the selectivity/redundancy of ferredoxin, we report that Fed1, Fed7 and Fed9 belong to ferredoxin-glutaredoxin-thioredoxin crosstalk pathways operating in the protection against oxidative and metal stresses. Furthermore, Fed7 specifically interacts with a DnaJ-like protein, an interaction that has been conserved in photosynthetic eukaryotes in the form of a composite protein comprising DnaJ- and Fed7-like domains. Fed9 specifically interacts with the Flv3 flavodiiron protein acting in the photoreduction of O2 to H2O.

  5. Protein Network Signatures Associated with Exogenous Biofuels Treatments in Cyanobacterium Synechocystis sp. PCC 6803

    Energy Technology Data Exchange (ETDEWEB)

    Pei, Guangsheng; Chen, Lei; Wang, Jiangxin; Qiao, Jianjun, E-mail: jianjunq@tju.edu.cn; Zhang, Weiwen, E-mail: jianjunq@tju.edu.cn [Laboratory of Synthetic Microbiology, School of Chemical Engineering and Technology, Tianjin University, Tianjin (China); Key Laboratory of Systems Bioengineering, Ministry of Education of China, Tianjin (China); SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin (China)

    2014-11-03

    Although recognized as a promising microbial cell factory for producing biofuels, current productivity in cyanobacterial systems is low. To make the processes economically feasible, one of the hurdles, which need to be overcome is the low tolerance of hosts to toxic biofuels. Meanwhile, little information is available regarding the cellular responses to biofuels stress in cyanobacteria, which makes it challenging for tolerance engineering. Using large proteomic datasets of Synechocystis under various biofuels stress and environmental perturbation, a protein co-expression network was first constructed and then combined with the experimentally determined protein–protein interaction network. Proteins with statistically higher topological overlap in the integrated network were identified as common responsive proteins to both biofuels stress and environmental perturbations. In addition, a weighted gene co-expression network analysis was performed to distinguish unique responses to biofuels from those to environmental perturbations and to uncover metabolic modules and proteins uniquely associated with biofuels stress. The results showed that biofuel-specific proteins and modules were enriched in several functional categories, including photosynthesis, carbon fixation, and amino acid metabolism, which may represent potential key signatures for biofuels stress responses in Synechocystis. Network-based analysis allowed determination of the responses specifically related to biofuels stress, and the results constituted an important knowledge foundation for tolerance engineering against biofuels in Synechocystis.

  6. An integrative approach to energy, carbon, and redox metabolism in the cyanobacterium Synechocystis sp. PCC 6803

    Energy Technology Data Exchange (ETDEWEB)

    Overbeek, Ross; Fonstein, Veronika; Osterman, Andrei; Gerdes, Svetlana; Vassieva, Olga; Zagnitko, Olga; Rodionov, Dmitry

    2005-02-15

    The team of the Fellowship for Interpretation of Genomes (FIG) under the leadership of Ross Overbeek, began working on this Project in November 2003. During the previous year, the Project was performed at Integrated Genomics Inc. A transition from the industrial environment to the public domain prompted us to adjust some aspects of the Project. Notwithstanding the challenges, we believe that these adjustments had a strong positive impact on our deliverables. Most importantly, the work of the research team led by R. Overbeek resulted in the deployment of a new open source genomic platform, the SEED (Specific Aim 1). This platform provided a foundation for the development of CyanoSEED a specialized portal to comparative analysis and metabolic reconstruction of all available cyanobacterial genomes (Specific Aim 3). The SEED represents a new generation of software for genome analysis. Briefly, it is a portable and extendable system, containing one of the largest and permanently growing collections of complete and partial genomes. The complete system with annotations and tools is freely available via browsing or via installation on a user's Mac or Linux computer. One of the important unique features of the SEED is the support of metabolic reconstruction and comparative genome analysis via encoding and projection of functional subsystems. During the project period, the FIG research team has validated the new software by developing a significant number of core subsystems, covering many aspects of central metabolism (Specific Aim 2), as well as metabolic areas specific for cyanobacteria and other photoautotrophic organisms (Specific Aim 3). In addition to providing a proof of technology and a starting point for further community-based efforts, these subsystems represent a valuable asset. An extensive coverage of central metabolism provides the bulk of information required for metabolic modeling in Synechocystis sp.PCC 6803. Detailed analysis of several subsystems

  7. Distinguishing the Roles of Thylakoid Respiratory Terminal Oxidases in the Cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Ermakova, Maria; Huokko, Tuomas; Richaud, Pierre; Bersanini, Luca; Howe, Christopher J; Lea-Smith, David J; Peltier, Gilles; Allahverdiyeva, Yagut

    2016-06-01

    Various oxygen-utilizing electron sinks, including the soluble flavodiiron proteins (Flv1/3), and the membrane-localized respiratory terminal oxidases (RTOs), cytochrome c oxidase (Cox) and cytochrome bd quinol oxidase (Cyd), are present in the photosynthetic electron transfer chain of Synechocystis sp. PCC 6803. However, the role of individual RTOs and their relative importance compared with other electron sinks are poorly understood, particularly under light. Via membrane inlet mass spectrometry gas exchange, chlorophyll a fluorescence, P700 analysis, and inhibitor treatment of the wild type and various mutants deficient in RTOs, Flv1/3, and photosystem I, we investigated the contribution of these complexes to the alleviation of excess electrons in the photosynthetic chain. To our knowledge, for the first time, we demonstrated the activity of Cyd in oxygen uptake under light, although it was detected only upon inhibition of electron transfer at the cytochrome b6f site and in ∆flv1/3 under fluctuating light conditions, where linear electron transfer was drastically inhibited due to impaired photosystem I activity. Cox is mostly responsible for dark respiration and competes with P700 for electrons under high light. Only the ∆cox/cyd double mutant, but not single mutants, demonstrated a highly reduced plastoquinone pool in darkness and impaired gross oxygen evolution under light, indicating that thylakoid-based RTOs are able to compensate partially for each other. Thus, both electron sinks contribute to the alleviation of excess electrons under illumination: RTOs continue to function under light, operating on slower time ranges and on a limited scale, whereas Flv1/3 responds rapidly as a light-induced component and has greater capacity. © 2016 American Society of Plant Biologists. All Rights Reserved.

  8. Approach to improve the productivity of bioactive compounds of the cyanobacterium Anabaena oryzae using factorial design

    Directory of Open Access Journals (Sweden)

    Ragaa A. Hamouda

    2017-09-01

    Full Text Available Cyanobacteria are one of the richest sources of biomedical relevant compounds with extensive therapeutic pharmaceutical applications and are also known as producer of intracellular and extracellular metabolites with diverse biological activities. The genus Anabaena sp. is known to produce antimicrobial compounds, like phycocyanin and others. The goal of this study was to optimize the production of these bioactive compounds. The Plackett–Burman experimental design was used to screen and evaluate the important medium components that influence the production of bioactive compounds. In this present study, eight independent factors including NaNO3, K2HPO4, MgSO4·7H2O, CaCl2, citric acid, ammonium ferric citrate, ethylene diamine tetraacetic acid disodium magnesium salt (EDTA-Na2Mg and Na2CO3 were surveyed and the effective variables for algal components production of Anabaena oryzae were determined using two-levels Plackett–Burman design. Results analysis showed that the best medium components were NaNO3 (2.25 g l−1; K2HPO4 (0.02 g l−1; MgSO4 (0.0375 g l−1; CaCl2 (0.018 g l−1; citric acid (0.009 g l−1; ammonium ferric citrate (0.009 g l−1 and EDTA-Na2 (0.0015 g l−1 respectively. The total chlorophyll-a, carotenoids, phenol, tannic acid and flavonoid contents in crude extract of Anabaena oryzae were determined. They were 47.7, 4.11, 0.256, 1.046 and 1.83 μg/ml, respectively. The antioxidant capacity was 62.81%.

  9. Global Proteomic Analysis Reveals an Exclusive Role of Thylakoid Membranes in Bioenergetics of a Model Cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Liberton, Michelle; Saha, Rajib; Jacobs, Jon M.; Nguyen, Amelia Y.; Gritsenko, Marina A.; Smith, Richard D.; Koppenaal, David W.; Pakrasi, Himadri B.

    2016-04-07

    Cyanobacteria are photosynthetic microbes with highly differentiated membrane systems. These organisms contain an outer membrane, plasma membrane, and an internal system of thylakoid membranes where the photosynthetic and respiratory machinery are found. This existence of compartmentalization and differentiation of membrane systems poses a number of challenges for cyanobacterial cells in terms of organization and distribution of proteins to the correct membrane system. Proteomics studies have long sought to identify the components of the different membrane systems, and to date about 450 different proteins have been attributed to either the plasma membrane or thylakoid membrane. Given the complexity of these membranes, many more proteins remain to be identified in these membrane systems, and a comprehensive catalog of plasma membrane and thylakoid membrane proteins is needed. Here we describe the identification of 635 proteins in Synechocystis sp. PCC 6803 by quantitative iTRAQ isobaric labeling; of these, 459 proteins were localized to the plasma membrane and 176 were localized to the thylakoid membrane. Surprisingly, we found over 2.5 times the number of unique proteins identified in the plasma membrane compared to the thylakoid membrane. This suggests that the protein composition of the thylakoid membrane is more homogeneous than the plasma membrane, consistent with the role of the plasma membrane in diverse cellular processes including protein trafficking and nutrient import, compared to a more specialized role for the thylakoid membrane in cellular energetics. Overall, the protein composition of the Synechocystis 6803 plasma membrane and thylakoid membrane is quite similar to the E.coli plasma membrane and Arabidopsis thylakoid membrane, respectively. Synechocystis 6803 can therefore be described as a gram-negative bacterium that has an additional internal membrane system that fulfils the energetic requirements of the cell.

  10. Global Proteomic Analysis Reveals an Exclusive Role of Thylakoid Membranes in Bioenergetics of a Model Cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Liberton, Michelle; Saha, Rajib; Jacobs, Jon M.; Nguyen, Amelia Y.; Gritsenko, Marina A.; Smith, Richard D.; Koppenaal, David W.; Pakrasi, Himadri B.

    2016-04-07

    Cyanobacteria are photosynthetic microbes with highlydifferentiated membrane systems. These organisms contain an outer membrane, plasma membrane, and an internal system of thylakoid membranes where the photosynthetic and respiratory machinery are found. This existence of compartmentalization and differentiation of membrane systems poses a number of challenges for cyanobacterial cells in terms of organization and distribution of proteins to the correct membrane system. Proteomics studies have long sought to identify the components of the different membrane systems in cyanobacteria, and to date about 450 different proteins have been attributed to either the plasma membrane or thylakoid membrane. Given the complexity of these membranes, many more proteins remain to be identified, and a comprehensive catalogue of plasma membrane and thylakoid membrane proteins is needed. Here we describe the identification of 635 differentially localized proteins in Synechocystis sp. PCC 6803 by quantitative iTRAQ isobaric labeling; of these, 459 proteins were localized to the plasma membrane and 176 were localized to the thylakoid membrane. Surprisingly, we found over 2.5 times the number of unique proteins identified in the plasma membrane compared with the thylakoid membrane. This suggests that the protein composition of the thylakoid membrane is more homogeneous than the plasma membrane, consistent with the role of the plasma membrane in diverse cellular processes including protein trafficking and nutrient import, compared with a more specialized role for the thylakoid membrane in cellular energetics. Thus, our data clearly define the two membrane systems with distinct functions. Overall, the protein compositions of the Synechocystis 6803 plasma membrane and thylakoid membrane are quite similar to that of the plasma membrane of Escherichia coli and thylakoid membrane of Arabidopsis chloroplasts, respectively. Synechocystis 6803 can therefore be described as a Gram

  11. Ocorrência e efeito de bactérias diazotróficas em genótipos de trigo Occurrence and effect of diazotrophic bacteria in wheat genotypes

    Directory of Open Access Journals (Sweden)

    Valéria Marino Rodrigues Sala

    2005-06-01

    Full Text Available A pesquisa sobre bactérias diazotróficas na cultura do trigo tem demonstrado a necessidade de associar bactérias eficientes a genótipos promissores, os quais se beneficiariam dessa associação. Em um experimento com parcelas subdivididas, instalado em condições de campo, em Mococa (SP, empregando os tratamentos: três doses de N (0, 60 e 120 kg ha-1 e três genótipos de trigo (IAC-24, ITD-19 e IAC-355, foi avaliada a ocorrência de microrganismos diazotróficos endofíticos em raízes desinfestadas superficialmente, utilizando-se três meios de cultivo distintos, NFb, JNFb e LGI-P. Somente para o genótipo IAC-355, houve um ajuste linear ascendente da quantidade de bactérias diazotróficas com o aumento na quantidade de N adicionada, apesar de o mesmo genótipo apresentar o menor número de bactérias diazotróficas endofíticas nos três meios de cultivo utilizados para quantificação. Foram obtidos oito isolados bacterianos do meio NFb com as características de Azospirillum e doze do meio JNFb com as características de Herbaspirillum. Esses isolados foram testados "in vitro", nos genótipos dos quais foram originalmente isolados, ou seja, ITD-19 e IAC-24. Todos os isolados testados no genótipo ITD-19 causaram maior crescimento radicular que a testemunha e apenas um isolado do meio JNFb propiciou aumento significativo do N acumulado na parte aérea. A interação planta-bactéria diazotrófica associativa indicou que é possível obter benefícios desta associação.Research on wheat root diazotrophic bacteria has demonstrated the need to associate effective bacteria to promising genotypes, which would benefit from this association. A field experiment was carried out in Mococa County, State of São Paulo, Brazil, using a split-plot design. Treatments consisted of 3 wheat genotypes (IAC-24, ITD-19 and IAC-355 under 3 nitrogen doses (0, 60 and 120 kg ha-1. The occurrence of diazotrophic bacteria was evaluated in three growth media

  12. Resposta de plantas micropropagadas de abacaxizeiro à inoculação de bactérias diazotróficas em casa de vegetação Response of pineapple plantlets to inoculation with diazotrophic bacteria in greenhouse

    Directory of Open Access Journals (Sweden)

    Olmar Baller Weber

    2003-12-01

    Full Text Available O objetivo deste trabalho foi selecionar e avaliar bactérias diazotróficas isoladas de abacaxizeiro (Ananas comosus (L. Merril no desenvolvimento de cultivares micropropagadas da mesma espécie em casa de vegetação. Plantas da cultivar Perolera foram submetidas à inoculação com Asaia bogorensis (AB219 e cultivadas em tubetes, durante 145 dias, com as misturas: casca de arroz carbonizada, folha de carnaubeira triturada e vermicomposto; casca de arroz carbonizada, pó da casca do coco maduro e vermicomposto; casca de arroz carbonizada, vermiculita e vermicomposto. Plantas da cultivar Primavera receberam inóculos com o AB219 e bactérias relacionadas a Burkholderia cepacia (AB202 e AB213, enquanto plantas das cultivares Pérola e Smooth Cayenne receberam AB219 e AB213, sendo cultivadas, por 140 dias, em tubetes com a mistura de vermicomposto e vermiculita. A colonização dos abacaxizeiros pelas bactérias diazotróficas foi confirmada. As plantas da cultivar Perolera cresceram melhor em casca de arroz carbonizada, vermiculita e vermicomposto e responderam positivamente ao AB219. Já as plantas da cultivar Primavera não apresentaram resposta significativa à inoculação com AB219, AB202 e AB213. Houve incremento de 23,1% a 38,5% na matéria seca de raízes das plantas da cultivar Pérola na presença de AB213 e AB219, respectivamente. A presença de AB213 incrementou em 15,2% a matéria seca da parte aérea das plantas da cultivar Smooth Cayenne. Os resultados revelam a eficiência de bactérias diazotróficas na promoção do crescimento de abacaxizeiros.The objective of this work was to select and to evaluate isolates of diazotrophic bacteria associated to pineapple (Ananas comosus (L. Merril plants in the development of micropropagated plantlets of cultivars pineapple in greenhouse. Plantlets from the Perolera cultivar have been submitted to inoculation with Asaia bogorensis (AB219 and were cultivated during 145 days on the following

  13. Efficiency of Photosynthesis in a Chl d-Utilizing Cyanobacterium is Comparable to or Higher than that in Chl a-Utilizing Oxygenic Species

    Science.gov (United States)

    Mielke, S. P.; Kiang, N. Y.; Blankenship, R. E.; Gunner, M. R.; Mauzerall, D.

    2011-01-01

    The cyanobacterium Acaryochloris marina uses chlorophyll d to carry out oxygenic photosynthesis in environments depleted in visible and enhanced in lower-energy, far-red light. However, the extent to which low photon energies limit the efficiency of oxygenic photochemistry in A. marina is not known. Here, we report the first direct measurements of the energy-storage efficiency of the photosynthetic light reactions in A. marina whole cells,and find it is comparable to or higher than that in typical, chlorophyll a-utilizing oxygenic species. This finding indicates that oxygenic photosynthesis is not fundamentally limited at the photon energies employed by A. marina, and therefore is potentially viable in even longer-wavelength light environments.

  14. Discovery and Synthesis of Caracolamide A, an Ion Channel Modulating Dichlorovinylidene Containing Phenethylamide from a Panamanian Marine Cyanobacterium cf. Symploca Species.

    Science.gov (United States)

    Naman, C Benjamin; Almaliti, Jehad; Armstrong, Lorene; Caro-Díaz, Eduardo J; Pierce, Marsha L; Glukhov, Evgenia; Fenner, Amanda; Spadafora, Carmenza; Debonsi, Hosana M; Dorrestein, Pieter C; Murray, Thomas F; Gerwick, William H

    2017-08-25

    A recent untargeted metabolomics investigation into the chemical profile of 10 organic extracts from cf. Symploca spp. revealed several interesting chemical leads for further natural product drug discovery. Subsequent target-directed isolation efforts with one of these, a Panamanian marine cyanobacterium cf. Symploca sp., yielded a phenethylamide metabolite that terminates in a relatively rare gem-dichlorovinylidene moiety, caracolamide A (1), along with a known isotactic polymethoxy-1-alkene (2). Detailed NMR and HRESIMS analyses were used to determine the structures of these molecules, and compound 1 was confirmed by a three-step synthesis. Pure compound 1 was shown to have in vitro calcium influx and calcium channel oscillation modulatory activity when tested as low as 10 pM using cultured murine cortical neurons, but was not cytotoxic to NCI-H460 human non-small-cell lung cancer cells in vitro (IC 50 > 10 μM).

  15. Recobrimento de sementes de milho com ácidos húmicos e bactérias diazotróficas endofíticas Corn seed coating with humic acids and endophytic diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Patrícia Marluci da Conceição

    2008-04-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito do recobrimento de sementes de milho com ácidos húmicos (AH, bactérias diazotróficas endofíticas e o uso em conjunto de AH e bactérias diazotróficas endofíticas, na estimulação do crescimento vegetal e na população de bactérias estabelecidas na planta hospedeira. A adição de AH, bactérias e o uso em conjunto estimularam o crescimento vegetal. Os AH utilizados no recobrimento de sementes de milho têm menor capacidade de estimular o crescimento radicular, em comparação ao uso em solução. O recobrimento de sementes é uma opção de inoculação de bactérias diazotróficas endofíticas da espécie Herbaspirillum seropedicae (Z67.The objective of this work was to evaluate the effect of seed coating of maize with humic acid (HA, endophytic diazotrophic bacteria, and the combination of both, on plant growth stimulation and bacteria population establishment in roots of inoculated plant host. The addition of HA, bacteria, and the combined use of bacteria and HA stimulated plant growth. Humic acids used in the coated seed formulation show diminished capacity for stimulation of root growth compared with its use in solution. Seed coat is an option for inoculation of endophytic diazotrophic bacteria like Herbaspirillum seropedicae (Z67.

  16. Influence of Extractive Solvents on Lipid and Fatty Acids Content of Edible Freshwater Algal and Seaweed Products, the Green Microalga Chlorella kessleri and the Cyanobacterium Spirulina platensis

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    Jarmila Vavra Ambrozova

    2014-02-01

    Full Text Available Total lipid contents of green (Chlorella pyrenoidosa, C, red (Porphyra tenera, N; Palmaria palmata, D, and brown (Laminaria japonica, K; Eisenia bicyclis, A; Undaria pinnatifida, W, WI; Hizikia fusiformis, H commercial edible algal and cyanobacterial (Spirulina platensis, S products, and autotrophically cultivated samples of the green microalga Chlorella kessleri (CK and the cyanobacterium Spirulina platensis (SP were determined using a solvent mixture of methanol/chloroform/water (1:2:1, v/v/v, solvent I and n-hexane (solvent II. Total lipid contents ranged from 0.64% (II to 18.02% (I by dry weight and the highest total lipid content was observed in the autotrophically cultivated cyanobacterium Spirulina platensis. Solvent mixture I was found to be more effective than solvent II. Fatty acids were determined by gas chromatography of their methyl esters (% of total FAMEs. Generally, the predominant fatty acids (all results for extractions with solvent mixture I were saturated palmitic acid (C16:0; 24.64%–65.49%, monounsaturated oleic acid (C18:1(n-9; 2.79%–26.45%, polyunsaturated linoleic acid (C18:2(n-6; 0.71%–36.38%, α-linolenic acid (C18:3(n-3; 0.00%–21.29%, γ-linolenic acid (C18:3(n-6; 1.94%–17.36%, and arachidonic acid (C20:4(n-6; 0.00%–15.37%. The highest content of ω-3 fatty acids (21.29% was determined in Chlorella pyrenoidosa using solvent I, while conversely, the highest content of ω-6 fatty acids (41.42% was observed in Chlorella kessleri using the same solvent.

  17. The marine cyanobacterium

    NARCIS (Netherlands)

    Pade, N.; Compaoré, J.; Klähn, S.; Stal, L.J.; Hagemann, M.

    2012-01-01

    Compatible solutes are small organic molecules that are involved in the acclimation to various stresses such as temperature and salinity. Marine or moderate halotolerant cyanobacteria accumulate glucosylglycerol, while cyanobacteria with low salt tolerance (freshwater strains) usually accumulate

  18. Carotenóides da cianobactéria Synechocystis pevalekii produzida em condições normais e sob limitação de nutrientes Carotenoids of the cyanobacterium Synechocystis pevalekii produced under normal conditions and under nutrient limitation

    Directory of Open Access Journals (Sweden)

    Marcos Coelho Müller

    2003-12-01

    -²-carotene, echinenone, ²-cryptoxanthin, 3-hydroxy-4'-ketocarotenoid, zeaxanthin and 3,3-dihydroxy-4'-ketocarotenoid were identified in the cyanobacterium Synechocystis pevalekii. The cianobacterium was green because of the presence of chlorophylls. When cultivated under stress (80% reduction of nutrient content of the original Conway medium the chlorophylls disappeared and the cyanobacterium assumed an orange color. ²-Carotene decreased from 307 to 248 µg/g and ²-cryptoxanthin from 94 to 13 µg/g. On the other hand, zeaxanthin increased from 29 to 220 µg/g. Thus, S. pevalekii appears to have commercial potential as source of zeaxanthin, which is implicated in the reduction of the risk of macular degeneration and cataract, together with lutein. The results also showed that conditions for the production of the cyanobacterium can be established so that the biosynthesis of carotenoids important to human health, but difficult to obtain, can be favored. There are already several commercial sources of ²-carotene, but sources of zeaxanthin are rare.

  19. Effects of lindane on the photosynthetic apparatus of the cyanobacterium Anabaena: fluorescence induction studies and immunolocalization of ferredoxin-NADP+ reductase.

    Science.gov (United States)

    Bueno, Marta; Fillat, Maria F; Strasser, Reto J; Maldonado-Rodriguez, Ronald; Marina, Nerea; Smienk, Henry; Gómez-Moreno, Carlos; Barja, Francisco

    2004-01-01

    Cyanobacteria have the natural ability to degrade moderate amounts of organic pollutants. However, when pollutant concentration exceeds the level of tolerance, bleaching of the cells and death occur within 24 hours. Under stress conditions, cyanobacterial response includes the short-term adaptation of the photosynthetic apparatus to light quality, named state transitions. Moreover, prolonged stresses produce changes in the functional organization of phycobilisomes and in the core-complexes of both photosystems, which can result in large changes in the PS II fluorescence yield. The localization of ferredoxin-NADP+ reductase (FNR) at the ends of some peripheral rods of the cyanobacterial phycobilisomes, makes this protein a useful marker to check phycobilisome integrity. The goal of this work is to improve the knowledge of the mechanism of action of a very potent pesticide, lindane (gamma-hexaclorociclohexane), in the cyanobacterium Anabaena sp., which can be considered a potential candidate for bioremediation of pesticides. We have studied the effect of lindane on the photosynthetic apparatus of Anabaena using fluorescence induction studies. As ferredoxin-NADP+ reductase plays a key role in the response to oxidative stress in several systems, changes in synthesis, degradation and activity of FNR were analyzed. Immunolocalization of this enzyme was used as a marker of phycobilisome integrity. The knowledge of the changes caused by lindane in the photosynthetic apparatus is essential for rational further design of genetically-modified cyanobacteria with improved biorremediation abilities. Polyphasic chlorophyll a fluorescence rise measurements (OJIP) have been used to evaluate the vitality and stress adaptation of the nitrogen-fixing cyanobacterium Anabaena PCC 7119 in the presence of increasing concentrations of lindane. Effects of the pesticide on the ultrastructure have been investigated by electron microscopy, and FNR has been used as a marker of phycobilisome

  20. Ocorrência de bactérias diazotróficas e fungos micorrízicos arbusculares na cultura da mandioca Occurrence of diazotrophic bacteria and arbuscular mycorrhizal fungi on the cassava crop

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    Elcio Liborio Balota

    1999-07-01

    Full Text Available Este trabalho teve como objetivo avaliar a ocorrência, isolar e identificar fungos micorrízicos arbusculares associados à cultura da mandioca (Manihot esculenta. Amostras de solo rizosférico e de várias partes da planta (raízes, tubérculos, manivas e folhas de locais nos Estados do Rio de Janeiro, São Paulo e Paraná, foram inoculadas nos meios LGI-P, NFb-malato e NFb-GOC, avaliando-se o número mais provável de células e a atividade de redução de acetileno. Bactérias diazotróficas foram isoladas de todas as partes da planta, com exceção das folhas, sendo identificadas como Klebsiella sp., Azospirillum lipoferum e uma bactéria denominada "E", provavelmente pertencente ao gênero Burkholderia. A Bactéria E acumulou de 7,63 mg a 14,84 mg de N/g de C em meio semi-sólido, isento de N, e conseguiu manter a capacidade de fixação biológica de N, mesmo após uma dezena de repicagens consecutivas. A colonização micorrízica variou de 31% a 69%, e a densidade de esporos de 10 a 384 esporos/100 mL de solo, predominando as espécies Entrophospora colombiana e Acaulospora scrobiculata no Rio de Janeiro, A. scrobiculata e Scutellospora heterogama no Paraná e em Piracicaba (São Paulo e A. appendicula e S. pellucida em Campinas (São Paulo.This study was performed to evaluate the occurrence and to isolate and identify diazotrophic bacteria and arbuscular mycorrhizal fungi associated with the cassava (Manihot esculenta crop. Samples from rhizospherical soil, roots, tubers, stems and leaves from several localities of the States of Rio de Janeiro, São Paulo and Paraná, in Brazil, were inoculated in three media specific for diazotrophic associative bacteria, LGI-P, NFb-malate and NFb-GOC, evaluating the most probable number of cells and the acetylene-reducing activity. Diazotrophic bacteria were detected in all plant parts except for the leaves, and were identified as Klebsiella sp., Azospirillum lipoferum and a bacterium called "E

  1. Bactérias diazotróficas associadas a coqueiros na região de baixada litorânea em Sergipe Diazotroph bacteria associated to coconut palms in a coastal lowland region in Sergipe State, Brazil

    Directory of Open Access Journals (Sweden)

    Marcelo Ferreira Fernandes

    2001-12-01

    Full Text Available Objetivou-se neste trabalho, isolar, identificar e quantificar bactérias diazotróficas existentes nas raízes e folhas de coqueiro (híbrido PB121 cultivados na baixada litorânea de Sergipe. As populações de bactérias diazotróficas nesses órgãos foram quantificadas pela técnica do número mais provável (NMP em meios semi-sólidos JNFb e NFb, e identificadas por meio de avaliações microscópicas, culturais e de testes bioquímicos dos sistemas API 20 - E e API 20 - NE. O conteúdo de N em meios semi-sólidos NFb e JNFb foi determinado colorimetricamente, após oito dias de incubação das bactérias, a 32ºC, para estimar a capacidade de fixação biológica dos isolados. Enterobactérias pertencentes ao gênero Enterobacter e bactérias com características semelhantes às do gênero Pseudomonas (pseudomonadas foram predominantes entre os isolados obtidos. As enterobactérias e pseudomonadas isoladas de folhas foram predominantemente endofíticas. Quanto às diazotróficas isoladas de raízes, observou-se predominância de enterobactérias na sua superfície, ao passo que as pseudomonadas ocorreram em proporções semelhantes na superfície e no interior desse órgão. Após oito dias de incubação, os conteúdos de N nos meios com inoculação das bactérias pseudomonadas foram maiores do que nos meios com inoculação das enterobactérias.The aim of this work was to isolate, identify and quantify diazotrophic bacteria existing in roots and leaves of coconut palms (hybrid PB121 grown in a coastal lowland of Sergipe, in Brazil. Diazotrophic populations in these organs were quantified by the most probable number (MNP technique on NFb and JNFb semi-solid media and identified by the evaluation of microscopic, cultural and biochemical characteristics of the isolates. Biochemical characteristics were evaluated using the API 20 - E and API 20 - NE galleries. Nitrogen content in NFb and JNFb semi-solid media inoculated with each of the

  2. CARACTERIZAÇÃO FENOTÍPICA DE BACTÉRIAS DIAZOTRÓFICAS ENDOFÍTICAS ISOLADAS DE CANA DE AÇÚCAR PHENOTYPIC CHARACTERIZATION OF ENDOPHYTIC DIAZOTROPHIC BACTERIAS ISOLATED OF SUGARCANE

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    Robson Cavalcante de Lima

    2011-06-01

    Full Text Available A utilização de inoculantes contendo microrganismos diazotróficos na cana-de-açúcar pode contribuir para a promoção do crescimento destas plantas e promover ganhos significativos na produtividade. Objetivou-se no presente trabalho caracterizar bactérias diazotróficas isoladas de colmos cana-de-açúcar. Foram utilizados os meios de cultivo semi-sólidos NFb e JNFb sem adição de nitrogênio. Foram isoladas cinco estirpes nativas de bactérias diazotróficas as quais foram avaliadas microscopicamente pela coloração de Gram e em relação à resistência intrínseca aos antibióticos; fungicida furadan (i.a. carbofuran e inseticida regente (i.a. fipronil, e à capacidade de fixar o nitrogênio em condições de casa-de-vegetação e campo. Os resultados obtidos permitiram observar que todos os isolados apresentaram características de bacilos Gram-negativos, e ampla resistência aos antibióticos. A estirpe nativa UCCBc5 apresentou resistência ao fungicida furadan e ao inseticida regente. As bactérias isoladas UCCBc1 e UCCBc5 apresentam capacidade e eficiência de fixar o nitrogênio atmosférico em condições de casa-de-vegetação. Verificou-se que a estirpe UCCBc5, resistente ao furadan e ao regente apresentou capacidade fixadora do nitrogênio atmosférico, em condições de campo. Na avaliação de doses de inoculante observou-se que existe relação dose de inoculante/ eficiência de fixação de nitrogênio. Estes resultados permitiram afirmar que as bactérias diazotróficas endofíticas podem ser utilizadas, em alguns casos, como substituto da adubação nitrogenada, na cultura de cana-de- açúcar.The use of inoculants containing diazotrophic microorganism in sugarcane can contribute to promoting the growth of these plants and tp promote significant gains in productivity. The objective of this study was to characterize diazotrophic bacteria isolated from stalks of sugarcane. We used the means of a semi-solid NFB and JNFb

  3. Diversidade de bactérias diazotróficas endofíticas dos gêneros Herbaspirillum e Burkholderia na cultura do arroz inundado Diversity of endophytic diazotrophic bacteria of the genus Herbaspirillum and Burkholderia in wetland rice

    Directory of Open Access Journals (Sweden)

    Luciana da Silva Rodrigues

    2006-02-01

    Full Text Available O objetivo deste trabalho foi avaliar a diversidade de bactérias diazotróficas endofíticas, dos gêneros Herbaspirillum e Burkholderia, em duas variedades de arroz, consideradas de alta (IR 42 e baixa (IAC 4440 eficiência de fixação biológica de nitrogênio. Foram realizados dois experimentos em casa de vegetação, em vasos com dois tipos de solos, provenientes dos Estados de Goiás e do Rio de Janeiro. Foi feita a contagem do número de bactérias e o isolamento em diferentes partes e estágios de desenvolvimento das plantas, mediante o uso de meios de cultivo JNFb e JMV. Os isolados bacterianos foram caracterizados a partir de aspectos morfológicos das colônias, com o crescimento em meios de cultivo, e de testes fisiológicos (uso de fontes de carbono e atividade de redução de acetileno. A contagem revelou grande número de bactérias diazotróficas (10(6 células g-1 matéria fresca, presentes em ambas as variedades de arroz, principalmente nas amostras radiculares. Os dados, obtidos na matriz de similaridade, mostram a presença de representantes da espécie Herbaspirillum seropedicae, bem como a diversidade entre isolados pertencentes ao gênero Burkholderia.The objective of this work was to evaluate the diversity of endophytic diazotrophic bacteria of the genera Herbaspirillum and Burkholderia, in two rice varieties, considered of high (IR 42 and low (IAC 4440 contribution on BNF. Two experiments were conducted in greenhouse conditions, in order to study the association of endophytic diazotrophic bacteria with wetland rice varieties, which were planted in two types of soil: one from Rio de Janeiro State and another from Goiás State, Brazil. Bacterial population (in different parts and physiological stages of the plants were evaluated, followed by the both genera strains isolation using culture media. The isolated bacteria were characterized based on morphological and physiological aspects. High bacterial counts were detected

  4. The microbiome of medicinal plants: diversity and importance for plant growth, quality and health

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    Martina eKöberl

    2013-12-01

    Full Text Available Past medicinal plant research primarily focused on bioactive phytochemicals, however the focus is currently shifting due to the recognition that a significant number of phytotherapeutic compounds are actually produced by associated microbes or through interaction with their host. Medicinal plants provide an enormous bioresource of potential use in modern medicine and agriculture, yet their microbiome is largely unknown. The objective of this review is i to introduce novel insights into the plant microbiome with a focus on medicinal plants, ii to provide details about plant- and microbe-derived ingredients of medicinal plants, and iii to discuss possibilities for plant growth promotion and plant protection for commercial cultivation of medicinal plants. In addition, we also present a case study performed both to analyse the microbiome of three medicinal plants (Matricaria chamomilla L., Calendula officinalis L. and Solanum distichum Schumach. and Thonn. cultivated on organically managed Egyptian desert farm and to develop biological control strategies. The soil microbiome of the desert ecosystem was comprised of a high abundance of Gram-positive bacteria of prime importance for pathogen suppression under arid soil conditions. For all three plants, we observed a clearly plant-specific selection of the microbes as well as highly specific diazotrophic communities that overall identify plant species as important drivers in structural and functional diversity. Lastly, native Bacillus spec. div. strains were able to promote plant growth and elevate the plants’ flavonoid production. These results underline the numerous links between the plant-associated microbiome and the plant metabolome.

  5. The microbiome of medicinal plants: diversity and importance for plant growth, quality and health.

    Science.gov (United States)

    Köberl, Martina; Schmidt, Ruth; Ramadan, Elshahat M; Bauer, Rudolf; Berg, Gabriele

    2013-12-20

    Past medicinal plant research primarily focused on bioactive phytochemicals, however, the focus is currently shifting due to the recognition that a significant number of phytotherapeutic compounds are actually produced by associated microbes or through interaction with their host. Medicinal plants provide an enormous bioresource of potential use in modern medicine and agriculture, yet their microbiome is largely unknown. The objective of this review is (i) to introduce novel insights into the plant microbiome with a focus on medicinal plants, (ii) to provide details about plant- and microbe-derived ingredients of medicinal plants, and (iii) to discuss possibilities for plant growth promotion and plant protection for commercial cultivation of medicinal plants. In addition, we also present a case study performed both to analyse the microbiome of three medicinal plants (Matricaria chamomilla L., Calendula officinalis L., and Solanum distichum Schumach. and Thonn.) cultivated on organically managed Egyptian desert farm and to develop biological control strategies. The soil microbiome of the desert ecosystem was comprised of a high abundance of Gram-positive bacteria of prime importance for pathogen suppression under arid soil conditions. For all three plants, we observed a clearly plant-specific selection of the microbes as well as highly specific diazotrophic communities that overall identify plant species as important drivers in structural and functional diversity. Lastly, native Bacillus spec. div. strains were able to promote plant growth and elevate the plants' flavonoid production. These results underline the numerous links between the plant-associated microbiome and the plant metabolome.

  6. Effects of the filamentous cyanobacterium Nodularia on fitness and feeding behavior of young-of-the-year (YOY) Eurasian perch (Perca fluviatilis).

    Science.gov (United States)

    Persson, Karl-Johan; Stenroth, Patrik; Legrand, Catherine

    2011-06-01

    This study reveals that both cyanobacterial toxicity and turbidity have the potential to reduce the growth and energy storage of young-of-the-year (YOY) perch and thereby influence survival rates. During the 1990's a reduction in recruitment of YOY perch (Perca fluviatilis) occurred along the Swedish East coast. Concurrently, large blooms of filamentous cyanobacteria have increased in the Baltic Proper and in coastal waters. This study examined whether extended exposure to toxic and non-toxic filamentous cyanobacterium Nodularia affect YOY perch growth and feeding behavior under simulated bloom conditions (30 days at 50 μg Chl a L(-1)). Specific growth rate (SGR), the somatic condition index (SCI) and the lipid content of YOY perch (10-12 weeks old) were significantly lower in perch exposed to Nodularia compared to fed controls (no Nodularia). YOY perch exposed to non-toxic Nodularia displayed a higher attack rate than perch living in Nodularia free controls in 2 out of 3 trials. Reductions in growth and energy storage, mediated by cyanobacteria, increase the risk of starvation and predation and could locally influence recruitment of YOY perch. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. Biochemical and Molecular Phylogenetic Study of Agriculturally Useful Association of a Nitrogen-Fixing Cyanobacterium and Nodule Sinorhizobium with Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    E. V. Karaushu

    2015-01-01

    Full Text Available Seed inoculation with bacterial consortium was found to increase legume yield, providing a higher growth than the standard nitrogen treatment methods. Alfalfa plants were inoculated by mono- and binary compositions of nitrogen-fixing microorganisms. Their physiological and biochemical properties were estimated. Inoculation by microbial consortium of Sinorhizobium meliloti T17 together with a new cyanobacterial isolate Nostoc PTV was more efficient than the single-rhizobium strain inoculation. This treatment provides an intensification of the processes of biological nitrogen fixation by rhizobia bacteria in the root nodules and an intensification of plant photosynthesis. Inoculation by bacterial consortium stimulates growth of plant mass and rhizogenesis and leads to increased productivity of alfalfa and to improving the amino acid composition of plant leaves. The full nucleotide sequence of the rRNA gene cluster and partial sequence of the dinitrogenase reductase (nifH gene of Nostoc PTV were deposited to GenBank (JQ259185.1, JQ259186.1. Comparison of these gene sequences of Nostoc PTV with all sequences present at the GenBank shows that this cyanobacterial strain does not have 100% identity with any organisms investigated previously. Phylogenetic analysis showed that this cyanobacterium clustered with high credibility values with Nostoc muscorum.

  8. Plasmid stability in dried cells of the desert cyanobacterium Chroococcidiopsis and its potential for GFP imaging of survivors on Earth and in space.

    Science.gov (United States)

    Billi, Daniela

    2012-06-01

    Two GFP-based plasmids, namely pTTQ18-GFP-pDU1(mini) and pDUCA7-GFP, of about 7 kbp and 15 kbp respectively, able to replicate in Chroococcidiopsis sp. CCMEE 029 and CCMEE 123, were developed. Both plasmids were maintained in Chroococcidiopsis cells after 18 months of dry storage as demonstrated by colony PCR, plasmid restriction analysis, GFP imaging and colony-forming ability under selection of dried transformants; thus suggesting that strategies employed by this cyanobacterium to stabilize dried chromosomal DNA, must have protected plasmid DNA. The suitability of pDU1(mini)-plasmid for GFP tagging in Chroococcidiopsis was investigated by using the RecA homolog of Synechocystis sp. PCC 6803. After 2 months of dry storage, the presence of dried cells with a GFP-RecA(Syn) distribution resembling that of hydrated cells, supported its capability of preventing desiccation-induced genome damage, whereas the rewetted cells with filamentous GFP-RecA(Syn) structures revealed sub-lethal DNA damage. The long-term stability of plasmid DNA in dried Chroococcidiopsis has implication for space research, for example when investigating the recovery of dried cells after Martian and space simulations or when developing life support systems based on phototrophs with genetically enhanced stress tolerance and stored in the dry state for prolonged periods.

  9. The interplay between siderophore secretion and coupled iron and copper transport in the heterocyst-forming cyanobacterium Anabaena sp. PCC 7120.

    Science.gov (United States)

    Nicolaisen, Kerstin; Hahn, Alexander; Valdebenito, Marianne; Moslavac, Suncana; Samborski, Anastazia; Maldener, Iris; Wilken, Corinna; Valladares, Ana; Flores, Enrique; Hantke, Klaus; Schleiff, Enrico

    2010-11-01

    Iron uptake is essential for Gram-negative bacteria including cyanobacteria. In cyanobacteria, however, the iron demand is higher than in proteobacteria due to the function of iron as a cofactor in photosynthesis and nitrogen fixation, but our understanding of iron uptake by cyanobacteria stands behind the knowledge in proteobacteria. Here, two genes involved in this process in the heterocyst-forming cyanobacterium Anabaena sp. PCC 7120 were identified. ORF all4025 encodes SchE, a putative cytoplasmic membrane-localized transporter involved in TolC-dependent siderophore secretion. Inactivation of schE resulted in an enhanced sensitivity to high metal concentrations and decreased secretion of hydroxamate-type siderophores. ORF all4026 encodes a predicted outer membrane-localized TonB-dependent iron transporter, IacT. Inactivation of iacT resulted in decreased sensitivity to elevated iron and copper levels. Expression of iacT from the artificial trc promoter (P(trc)) resulted in sensitization against tested metals. Further analysis showed that iron and copper effects are synergistic because a decreased supply of iron induced a significant decrease of copper levels in the iacT insertion mutant but an increase of those levels in the strain carrying P(trc)-iacT. Our results unravel a link between iron and copper homeostasis in Anabaena sp. PCC 7120. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Intricate interactions between the bloom-forming cyanobacterium Microcystis aeruginosa and foreign genetic elements, revealed by diversified clustered regularly interspaced short palindromic repeat (CRISPR) signatures.

    Science.gov (United States)

    Kuno, Sotaro; Yoshida, Takashi; Kaneko, Takakazu; Sako, Yoshihiko

    2012-08-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) confer sequence-dependent, adaptive resistance in prokaryotes against viruses and plasmids via incorporation of short sequences, called spacers, derived from foreign genetic elements. CRISPR loci are thus considered to provide records of past infections. To describe the host-parasite (i.e., cyanophages and plasmids) interactions involving the bloom-forming freshwater cyanobacterium Microcystis aeruginosa, we investigated CRISPR in four M. aeruginosa strains and in two previously sequenced genomes. The number of spacers in each locus was larger than the average among prokaryotes. All spacers were strain specific, except for a string of 11 spacers shared in two closely related strains, suggesting diversification of the loci. Using CRISPR repeat-based PCR, 24 CRISPR genotypes were identified in a natural cyanobacterial community. Among 995 unique spacers obtained, only 10 sequences showed similarity to M. aeruginosa phage Ma-LMM01. Of these, six spacers showed only silent or conservative nucleotide mutations compared to Ma-LMM01 sequences, suggesting a strategy by the cyanophage to avert CRISPR immunity dependent on nucleotide identity. These results imply that host-phage interactions can be divided into M. aeruginosa-cyanophage combinations rather than pandemics of population-wide infectious cyanophages. Spacer similarity also showed frequent exposure of M. aeruginosa to small cryptic plasmids that were observed only in a few strains. Thus, the diversification of CRISPR implies that M. aeruginosa has been challenged by diverse communities (almost entirely uncharacterized) of cyanophages and plasmids.

  11. An alternative methionine aminopeptidase, MAP-A, is required for nitrogen starvation and high-light acclimation in the cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Drath, Miriam; Baier, Kerstin; Forchhammer, Karl

    2009-05-01

    Methionine aminopeptidases (MetAPs or MAPs, encoded by map genes) are ubiquitous and pivotal enzymes for protein maturation in all living organisms. Whereas most bacteria harbour only one map gene, many cyanobacterial genomes contain two map paralogues, the genome of Synechocystis sp. PCC 6803 even three. The physiological function of multiple map paralogues remains elusive so far. This communication reports for the first time differential MetAP function in a cyanobacterium. In Synechocystis sp. PCC 6803, the universally conserved mapC gene (sll0555) is predominantly expressed in exponentially growing cells and appears to be a housekeeping gene. By contrast, expression of mapA (slr0918) and mapB (slr0786) genes increases during stress conditions. The mapB paralogue is only transiently expressed, whereas the widely distributed mapA gene appears to be the major MetAP during stress conditions. A mapA-deficient Synechocystis mutant shows a subtle impairment of photosystem II properties even under non-stressed conditions. In particular, the binding site for the quinone Q(B) is affected, indicating specific N-terminal methionine processing requirements of photosystem II components. MAP-A-specific processing becomes essential under certain stress conditions, since the mapA-deficient mutant is severely impaired in surviving conditions of prolonged nitrogen starvation and high light exposure.

  12. NanoSIMS Analyses of Mo Indicate Nitrogenase Activity and Help Solve a N and C Fixation Puzzle in a Marine Cyanobacterium

    Science.gov (United States)

    Pett-Ridge, J.; Weber, P. K.; Finzi, J.; Hutcheon, I. D.; Capone, D. G.

    2006-12-01

    Diazotrophic cyanobacteria are capable of both CO2 and N2 fixation, yet must separate these two functions because the nitrogenase enzymes used in N2 fixation are strongly inhibited by O2 produced during photosynthesis. Some lineages, such as Anabaena, use specialized cells (heterocysts) to maintain functional segregation. However the mechanism of this segregation is poorly understood in Trichodesmium, a critical component of marine primary production in the tropical and subtropical North Atlantic. While some Trichodesmium studies suggest a temporal segregation of the nitrogen and carbon fixing processes, others indicate nitrogen fixation is spatially isolated in differentiated cells called diazocytes. In order to isolate the intracellular location of N fixation in both species, we used a combination of TEM, SEM and NanoSIMS analysis to map the distribution of C, N and Mo (a critical nitrogenase co-factor) isotopes in intact cells. NanoSIMS is a powerful surface analysis tool which combines nanometer-scale imaging resolution with the high sensitivity of mass spectrometry. Using cells grown in a 13CO^2 and 15N2 enriched atmosphere, our analyses indicate that in Anabaena, heterocysts are consistently enriched in Mo, and Mo accumulation suggests active N fixation (as opposed to N storage). In the non- heterocystous Trichodesmium, Mo is concentrated in sub-regions of individual cells, and is not associated with regions of N storage (cyanophycin granules). We suggest that NanoSIMS mapping of metal enzyme co- factors is a unique method of identifying physiological and morphological characteristics within individual bacterial cells. This combination of NanoSIMS analysis and high resolution microscopy allows isotopic analysis to be linked to morphological features and holds great promise for fine-scale studies of bacteria metabolism.

  13. Cell wall amidase AmiC1 is required for cellular communication and heterocyst development in the cyanobacterium Anabaena PCC 7120 but not for filament integrity.

    Science.gov (United States)

    Berendt, Susanne; Lehner, Josef; Zhang, Yao Vincent; Rasse, Tobias M; Forchhammer, Karl; Maldener, Iris

    2012-10-01

    Filamentous cyanobacteria of the order Nostocales display typical properties of multicellular organisms. In response to nitrogen starvation, some vegetative cells differentiate into heterocysts, where fixation of N(2) takes place. Heterocysts provide a micro-oxic compartment to protect nitrogenase from the oxygen produced by the vegetative cells. Differentiation involves fundamental remodeling of the gram-negative cell wall by deposition of a thick envelope and by formation of a neck-like structure at the contact site to the vegetative cells. Cell wall-hydrolyzing enzymes, like cell wall amidases, are involved in peptidoglycan maturation and turnover in unicellular bacteria. Recently, we showed that mutation of the amidase homologue amiC2 gene in Nostoc punctiforme ATCC 29133 distorts filament morphology and function. Here, we present the functional characterization of two amiC paralogues from Anabaena sp. strain PCC 7120. The amiC1 (alr0092) mutant was not able to differentiate heterocysts or to grow diazotrophically, whereas the amiC2 (alr0093) mutant did not show an altered phenotype under standard growth conditions. In agreement, fluorescence recovery after photobleaching (FRAP) studies showed a lack of cell-cell communication only in the AmiC1 mutant. Green fluorescent protein (GFP)-tagged AmiC1 was able to complement the mutant phenotype to wild-type properties. The protein localized in the septal regions of newly dividing cells and at the neck region of differentiating heterocysts. Upon nitrogen step-down, no mature heterocysts were developed in spite of ongoing heterocyst-specific gene expression. These results show the dependence of heterocyst development on amidase function and highlight a pivotal but so far underestimated cellular process, the remodeling of peptidoglycan, for the biology of filamentous cyanobacteria.

  14. Permutation importance: a corrected feature importance measure.

    Science.gov (United States)

    Altmann, André; Toloşi, Laura; Sander, Oliver; Lengauer, Thomas

    2010-05-15

    In life sciences, interpretability of machine learning models is as important as their prediction accuracy. Linear models are probably the most frequently used methods for assessing feature relevance, despite their relative inflexibility. However, in the past years effective estimators of feature relevance have been derived for highly complex or non-parametric models such as support vector machines and RandomForest (RF) models. Recently, it has been observed that RF models are biased in such a way that categorical variables with a large number of categories are preferred. In this work, we introduce a heuristic for normalizing feature importance measures that can correct the feature importance bias. The method is based on repeated permutations of the outcome vector for estimating the distribution of measured importance for each variable in a non-informative setting. The P-value of the observed importance provides a corrected measure of feature importance. We apply our method to simulated data and demonstrate that (i) non-informative predictors do not receive significant P-values, (ii) informative variables can successfully be recovered among non-informative variables and (iii) P-values computed with permutation importance (PIMP) are very helpful for deciding the significance of variables, and therefore improve model interpretability. Furthermore, PIMP was used to correct RF-based importance measures for two real-world case studies. We propose an improved RF model that uses the significant variables with respect to the PIMP measure and show that its prediction accuracy is superior to that of other existing models. R code for the method presented in this article is available at http://www.mpi-inf.mpg.de/ approximately altmann/download/PIMP.R CONTACT: altmann@mpi-inf.mpg.de, laura.tolosi@mpi-inf.mpg.de Supplementary data are available at Bioinformatics online.

  15. Promoção de enraizamento de microtoletes de cana-de-açúcar pelo uso conjunto de substâncias húmicas e bactérias diazotróficas endofíticas Rooting of micro seed pieces by combined use of humic substances and endophytic diazotrophic bacteria in sugar cane

    Directory of Open Access Journals (Sweden)

    Roberto Batista Marques Júnior

    2008-06-01

    éficos da inoculação de estirpes selecionadas de H. seropedicae, já para ácidos húmicos, as respostas positivas independem da termoterapia. A promoção do crescimento radicular por AH e a fixação biológica de N2 podem representar uma estratégia inovadora para produção sustentada em sistemas agrícolas.Besides the direct nutritional effect of mineralization of organic matter or by biological nitrogen fixation activity, the humic substances and endophytic diazotrophic bacteria can directly influence plant metabolism, modifying plant growth and development patterns. The purpose of this study was to evaluate the effect of the combined use of humic acid (HA and Herbaspirillum seropedicae, an endophytic nitrogen-fixing bacteria, on the root growth of seed pieces - heat-treated or not - of the sugarcane variety RB 72 454. After hot water treatment, the cane cuttings were immersed for 12 hours as follows: in water (control plant, in HA solution (20 mg L-1 of C from vermicompost, in bacterial inoculant of H. seropedicae, strain HRC54 (10(8 cells mL-1, and in a combination of bacteria and HA. Root growth was improved by 60 to 118 % in length and from 33 % to 233 % in surface area on sugarcane plant treatments compared to control, with more pronounced effect in plants under heat treatment. Likewise, the positive effect of the combinded treatment (bacteria inoculation and humic acid on shoot and root biomass was significant compared to the control with heat-treated cuts. For non-heated seed pieces, bacteria inoculation did not result in a positive plant growth effect, but only in the presence of humic acid. The combined or isolated use of both HA and bacteria did not significantly modify the bacteria population in the root tissue of heat-untreated sugarcane. For heat-treated cuts, bacteria inoculation, combined or not with HA, increased the size of diazotrophic bacteria population on roots. The results highlight the importance of thermotherapy to increase the positive

  16. Characterization of nifB, nifS, and nifU genes in the cyanobacterium Anabaena variabilis: NifB is required for the vanadium-dependent nitrogenase.

    OpenAIRE

    Lyons, E M; Thiel, T

    1995-01-01

    Anabaena variabilis ATCC 29413 is a heterotrophic, nitrogen-fixing cyanobacterium containing both a Mo-dependent nitrogenase encoded by the nif genes and V-dependent nitrogenase encoded by the vnf genes. The nifB, nifS, and nifU genes of A. variabilis were cloned, mapped, and partially sequenced. The fdxN gene was between nifB and nifS. Growth and acetylene reduction assays using wild-type and mutant strains indicated that the nifB product (NifB) was required for nitrogen fixation not only by...

  17. Metabolic engineering of the Chl d-dominated cyanobacterium Acaryochloris marina: production of a novel Chl species by the introduction of the chlorophyllide a oxygenase gene.

    Science.gov (United States)

    Tsuchiya, Tohru; Mizoguchi, Tadashi; Akimoto, Seiji; Tomo, Tatsuya; Tamiaki, Hitoshi; Mimuro, Mamoru

    2012-03-01

    In oxygenic photosynthetic organisms, the properties of photosynthetic reaction systems primarily depend on the Chl species used. Acquisition of new Chl species with unique optical properties may have enabled photosynthetic organisms to adapt to various light environments. The artificial production of a new Chl species in an existing photosynthetic organism by metabolic engineering provides a model system to investigate how an organism responds to a newly acquired pigment. In the current study, we established a transformation system for a Chl d-dominated cyanobacterium, Acaryochloris marina, for the first time. The expression vector (constructed from a broad-host-range plasmid) was introduced into A. marina by conjugal gene transfer. The introduction of a gene for chlorophyllide a oxygenase, which is responsible for Chl b biosynthesis, into A. marina resulted in a transformant that synthesized a novel Chl species instead of Chl b. The content of the novel Chl in the transformant was approximately 10% of the total Chl, but the level of Chl a, another Chl in A. marina, did not change. The chemical structure of the novel Chl was determined to be [7-formyl]-Chl d(P) by mass spectrometry and nuclear magnetic resonance spectroscopy. [7-Formyl]-Chl d(P) is hypothesized to be produced by the combined action of chlorophyllide a oxygenase and enzyme(s) involved in Chl d biosynthesis. These results demonstrate the flexibility of the Chl biosynthetic pathway for the production of novel Chl species, indicating that a new organism with a novel Chl might be discovered in the future.

  18. Transcription profiling of the model cyanobacterium Synechococcus sp. strain PCC 7002 by NextGen (SOLiD™ Sequencing of cDNA

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    Marcus eLudwig

    2011-03-01

    Full Text Available The genome of the unicellular, euryhaline cyanobacterium Synechococcus sp. PCC 7002 encodes about 3200 proteins. Transcripts were detected for nearly all annotated open reading frames by a global transcriptomic analysis by Next-Generation (SOLiDTM sequencing of cDNA. In the cDNA samples sequenced, ~90% of the mapped sequences were derived from the 16S and 23S ribosomal RNAs and ~10% of the sequences were derived from mRNAs. In cells grown photoautotrophically under standard conditions (38 °C, 1% (v/v CO2 in air, 250 µmol photons m-2 s-1, the highest transcript levels (up to 2% of the total mRNA for the most abundantly transcribed genes (e. g., cpcAB, psbA, psaA were generally derived from genes encoding structural components of the photosynthetic apparatus. High light exposure for one hour caused changes in transcript levels for genes encoding proteins of the photosynthetic apparatus, Type-1 NADH dehydrogenase complex and ATP synthase, whereas dark incubation for one hour resulted in a global decrease in transcript levels for photosynthesis-related genes and an increase in transcript levels for genes involved in carbohydrate degradation. Transcript levels for pyruvate kinase and the pyruvate dehydrogenase complex decreased sharply in cells incubated in the dark. Under dark anoxic (fermentative conditions, transcript changes indicated a global decrease in transcripts for respiratory proteins and suggested that cells employ an alternative phosphoenolpyruvate degradation pathway via phosphoenolpyruvate synthase (ppsA and the pyruvate:ferredoxin oxidoreductase (nifJ. Finally, the data suggested that an apparent operon involved in tetrapyrrole biosynthesis and fatty acid desaturation, acsF2-ho2-hemN2-desF, may be regulated by oxygen concentration.

  19. Unraveling the Physiological Roles of the Cyanobacterium Geitlerinema sp. BBD and Other Black Band Disease Community Members through Genomic Analysis of a Mixed Culture

    Science.gov (United States)

    Den Uyl, Paul A.; Richardson, Laurie L.; Jain, Sunit

    2016-01-01

    Black band disease (BBD) is a cyanobacterial-dominated polymicrobial mat that propagates on and migrates across coral surfaces, necrotizing coral tissue. Culture-based laboratory studies have investigated cyanobacteria and heterotrophic bacteria isolated from BBD, but the metabolic potential of various BBD microbial community members and interactions between them remain poorly understood. Here we report genomic insights into the physiological and metabolic potential of the BBD-associated cyanobacterium Geitlerinema sp. BBD 1991 and six associated bacteria that were also present in the non-axenic culture. The essentially complete genome of Geitlerinema sp. BBD 1991 contains a sulfide quinone oxidoreductase gene for oxidation of sulfide, suggesting a mechanism for tolerating the sulfidic conditions of BBD mats. Although the operon for biosynthesis of the cyanotoxin microcystin was surprisingly absent, potential relics were identified. Genomic evidence for mixed-acid fermentation indicates a strategy for energy metabolism under the anaerobic conditions present in BBD during darkness. Fermentation products may supply carbon to BBD heterotrophic bacteria. Among the six associated bacteria in the culture, two are closely related to organisms found in culture-independent studies of diseased corals. Their metabolic pathways for carbon and sulfur cycling, energy metabolism, and mechanisms for resisting coral defenses suggest adaptations to the coral surface environment and biogeochemical roles within the BBD mat. Polysulfide reductases were identified in a Flammeovirgaceae genome (Bacteroidetes) and the sox pathway for sulfur oxidation was found in the genome of a Rhodospirillales bacterium (Alphaproteobacteria), revealing mechanisms for sulfur cycling, which influences virulence of BBD. Each genomic bin possessed a pathway for conserving energy from glycerol degradation, reflecting adaptations to the glycerol-rich coral environment. The presence of genes for detoxification

  20. Inactivation of uptake hydrogenase leads to enhanced and sustained hydrogen production with high nitrogenase activity under high light exposure in the cyanobacterium Anabaena siamensis TISTR 8012

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    Khetkorn Wanthanee

    2012-10-01

    Full Text Available Abstract Background Biohydrogen from cyanobacteria has attracted public interest due to its potential as a renewable energy carrier produced from solar energy and water. Anabaena siamensis TISTR 8012, a novel strain isolated from rice paddy field in Thailand, has been identified as a promising cyanobacterial strain for use as a high-yield hydrogen producer attributed to the activities of two enzymes, nitrogenase and bidirectional hydrogenase. One main obstacle for high hydrogen production by A. siamensis is a light-driven hydrogen consumption catalyzed by the uptake hydrogenase. To overcome this and in order to enhance the potential for nitrogenase based hydrogen production, we engineered a hydrogen uptake deficient strain by interrupting hupS encoding the small subunit of the uptake hydrogenase. Results An engineered strain lacking a functional uptake hydrogenase (∆hupS produced about 4-folds more hydrogen than the wild type strain. Moreover, the ∆hupS strain showed long term, sustained hydrogen production under light exposure with 2–3 folds higher nitrogenase activity compared to the wild type. In addition, HupS inactivation had no major effects on cell growth and heterocyst differentiation. Gene expression analysis using RT-PCR indicates that electrons and ATP molecules required for hydrogen production in the ∆hupS strain may be obtained from the electron transport chain associated with the photosynthetic oxidation of water in the vegetative cells. The ∆hupS strain was found to compete well with the wild type up to 50 h in a mixed culture, thereafter the wild type started to grow on the relative expense of the ∆hupS strain. Conclusions Inactivation of hupS is an effective strategy for improving biohydrogen production, in rates and specifically in total yield, in nitrogen-fixing cultures of the cyanobacterium Anabaena siamensis TISTR 8012.

  1. The biomechanical role of overall-shape transformation in a primitive multicellular organism: A case study of dimorphism in the filamentous cyanobacterium Arthrospira platensis.

    Science.gov (United States)

    Chaiyasitdhi, Atitheb; Miphonpanyatawichok, Wirat; Riehle, Mathis Oliver; Phatthanakun, Rungrueang; Surareungchai, Werasak; Kundhikanjana, Worasom; Kuntanawat, Panwong

    2018-01-01

    Morphological transformations in primitive organisms have long been observed; however, its biomechanical roles are largely unexplored. In this study, we investigate the structural advantages of dimorphism in Arthrospira platensis, a filamentous multicellular cyanobacterium. We report that helical trichomes, the default shape, have a higher persistence length (Lp), indicating a higher resistance to bending or a large value of flexural rigidity (kf), the product of the local cell stiffness (E) and the moment of inertia of the trichomes' cross-section (I). Through Atomic Force Microscopy (AFM), we determined that the E of straight and helical trichomes were the same. In contrast, our computational model shows that I is greatly dependent on helical radii, implying that trichome morphology is the major contributor to kf variation. According to our estimation, increasing the helical radii alone can increase kf by 2 orders of magnitude. We also observe that straight trichomes have improved gliding ability, due to its structure and lower kf. Our study shows that dimorphism provides mechanical adjustability to the organism and may allow it to thrive in different environmental conditions. The higher kf provides helical trichomes a better nutrient uptake through advection in aquatic environments. On the other hand, the lower kf improves the gliding ability of straight trichomes in aquatic environments, enabling it to chemotactically relocate to more favorable territories when it encounters certain environmental stresses. When more optimal conditions are encountered, straight trichomes can revert to their original helical form. Our study is one of the first to highlight the biomechanical role of an overall-shape transformation in cyanobacteria.

  2. Differential sensitivity of five cyanobacterial strains to ammonium toxicity and its inhibitory mechanism on the photosynthesis of rice-field cyanobacterium Ge-Xian-Mi (Nostoc)

    International Nuclear Information System (INIS)

    Dai Guozheng; Deblois, Charles P.; Liu Shuwen; Juneau, Philippe; Qiu Baosheng

    2008-01-01

    Effects of two fertilizers, NH 4 Cl and KCl, on the growth of the edible cyanobacterium Ge-Xian-Mi (Nostoc) and four other cyanobacterial strains were compared at pH 8.3 ± 0.2 and 25 deg. C. Their growth was decreased by at least 65% at 10 mmol L -1 NH 4 Cl but no inhibitory effect was observed at the same level of KCl. Meanwhile, the strains exhibited a great variation of sensitivity to NH 4 + toxicity in the order: Ge-Xian-Mi > Anabaena azotica FACHB 118 > Microcystis aeruginosa FACHB 905 > M. aeruginosa FACHB 315 > Synechococcus FACHB 805. The 96-h EC 50 value for relative growth rate with regard to NH 4 + for Ge-Xian-Mi was 1.105 mmol L -1 , which was much less than the NH 4 + concentration in many agricultural soils (2-20 mmol L -1 ). This indicated that the use of ammonium as nitrogen fertilizer was responsible for the reduced resource of Ge-Xian-Mi in the paddy field. After 96 h exposure to 1 mmol L -1 NH 4 Cl, the photosynthetic rate, F v /F m value, saturating irradiance for photosynthesis and PSII activity of Ge-Xian-Mi colonies were remarkably decreased. The chlorophyll synthesis of Ge-Xian-Mi was more sensitive to NH 4 + toxicity than phycobiliproteins. Thus, the functional absorption cross section of Ge-Xian-Mi PSII was increased markedly at NH 4 Cl levels ≥1 mmol L -1 and the electron transport on the acceptor side of PSII was significantly accelerated by NH 4 Cl addition ≥3 mmol L -1 . Dark respiration of Ge-Xian-Mi was significantly increased by 246% and 384% at 5 and 10 mmol L -1 NH 4 Cl, respectively. The rapid fluorescence rise kinetics indicated that the oxygen-evolving complex of PSII was the inhibitory site of NH 4 +

  3. Single-cell confocal spectrometry of a filamentous cyanobacterium Nostoc at room and cryogenic temperature. Diversity and differentiation of pigment systems in 311 cells.

    Science.gov (United States)

    Sugiura, Kana; Itoh, Shigeru

    2012-08-01

    The fluorescence spectrum at 298 and 40 K and the absorption spectrum at 298 K of each cell of the filamentous cyanobacterium Nostoc sp. was measured by single-cell confocal laser spectroscopy to study the differentiation of cell pigments. The fluorescence spectra of vegetative (veg) and heterocyst (het) cells of Nostoc formed separate groups with low and high PSII to PSI ratios, respectively. The fluorescence spectra of het cells at 40 K still contained typical PSII bands. The PSII/PSI ratio estimated for the veg cells varied between 0.4 and 1.2, while that of het cells varied between 0 and 0.22 even in the same culture. The PSII/PSI ratios of veg cells resembled each other more closely in the same filament. 'pro-het' cells, which started to differentiate into het cells, were identified from the small but specific difference in the PSII/PSI ratio. The allophycocyanin (APC)/PSII ratio was almost constant in both veg and het cells, indicating their tight couplings. Phycocyanin (PC) showed higher fluorescence in most het cells, suggesting the uncoupling from PSII. Veg cells seem to vary their PSI contents to give different PSII/PSI ratios even in the same culture, and to suppress the synthesis of PSII, APC and PC to differentiate into het cells. APC and PC are gradually liberated from membranes in het cells with the uncoupling from PSII. Single-cell spectrometry will be useful to study the differentiation of intrinsic pigments of cells and chloroplasts, and to select microbes from natural environments.

  4. Unraveling the Physiological Roles of the Cyanobacterium Geitlerinema sp. BBD and Other Black Band Disease Community Members through Genomic Analysis of a Mixed Culture.

    Science.gov (United States)

    Den Uyl, Paul A; Richardson, Laurie L; Jain, Sunit; Dick, Gregory J

    2016-01-01

    Black band disease (BBD) is a cyanobacterial-dominated polymicrobial mat that propagates on and migrates across coral surfaces, necrotizing coral tissue. Culture-based laboratory studies have investigated cyanobacteria and heterotrophic bacteria isolated from BBD, but the metabolic potential of various BBD microbial community members and interactions between them remain poorly understood. Here we report genomic insights into the physiological and metabolic potential of the BBD-associated cyanobacterium Geitlerinema sp. BBD 1991 and six associated bacteria that were also present in the non-axenic culture. The essentially complete genome of Geitlerinema sp. BBD 1991 contains a sulfide quinone oxidoreductase gene for oxidation of sulfide, suggesting a mechanism for tolerating the sulfidic conditions of BBD mats. Although the operon for biosynthesis of the cyanotoxin microcystin was surprisingly absent, potential relics were identified. Genomic evidence for mixed-acid fermentation indicates a strategy for energy metabolism under the anaerobic conditions present in BBD during darkness. Fermentation products may supply carbon to BBD heterotrophic bacteria. Among the six associated bacteria in the culture, two are closely related to organisms found in culture-independent studies of diseased corals. Their metabolic pathways for carbon and sulfur cycling, energy metabolism, and mechanisms for resisting coral defenses suggest adaptations to the coral surface environment and biogeochemical roles within the BBD mat. Polysulfide reductases were identified in a Flammeovirgaceae genome (Bacteroidetes) and the sox pathway for sulfur oxidation was found in the genome of a Rhodospirillales bacterium (Alphaproteobacteria), revealing mechanisms for sulfur cycling, which influences virulence of BBD. Each genomic bin possessed a pathway for conserving energy from glycerol degradation, reflecting adaptations to the glycerol-rich coral environment. The presence of genes for detoxification

  5. Unraveling the Physiological Roles of the Cyanobacterium Geitlerinema sp. BBD and Other Black Band Disease Community Members through Genomic Analysis of a Mixed Culture.

    Directory of Open Access Journals (Sweden)

    Paul A Den Uyl

    Full Text Available Black band disease (BBD is a cyanobacterial-dominated polymicrobial mat that propagates on and migrates across coral surfaces, necrotizing coral tissue. Culture-based laboratory studies have investigated cyanobacteria and heterotrophic bacteria isolated from BBD, but the metabolic potential of various BBD microbial community members and interactions between them remain poorly understood. Here we report genomic insights into the physiological and metabolic potential of the BBD-associated cyanobacterium Geitlerinema sp. BBD 1991 and six associated bacteria that were also present in the non-axenic culture. The essentially complete genome of Geitlerinema sp. BBD 1991 contains a sulfide quinone oxidoreductase gene for oxidation of sulfide, suggesting a mechanism for tolerating the sulfidic conditions of BBD mats. Although the operon for biosynthesis of the cyanotoxin microcystin was surprisingly absent, potential relics were identified. Genomic evidence for mixed-acid fermentation indicates a strategy for energy metabolism under the anaerobic conditions present in BBD during darkness. Fermentation products may supply carbon to BBD heterotrophic bacteria. Among the six associated bacteria in the culture, two are closely related to organisms found in culture-independent studies of diseased corals. Their metabolic pathways for carbon and sulfur cycling, energy metabolism, and mechanisms for resisting coral defenses suggest adaptations to the coral surface environment and biogeochemical roles within the BBD mat. Polysulfide reductases were identified in a Flammeovirgaceae genome (Bacteroidetes and the sox pathway for sulfur oxidation was found in the genome of a Rhodospirillales bacterium (Alphaproteobacteria, revealing mechanisms for sulfur cycling, which influences virulence of BBD. Each genomic bin possessed a pathway for conserving energy from glycerol degradation, reflecting adaptations to the glycerol-rich coral environment. The presence of genes

  6. The tryptophan-rich sensory protein (TSPO is involved in stress-related and light-dependent processes in the cyanobacterium Fremyella diplosiphon

    Directory of Open Access Journals (Sweden)

    Andrea eBusch

    2015-12-01

    Full Text Available The tryptophan-rich sensory protein (TSPO is a membrane protein, which is a member of the 18 kilodalton translocator protein/peripheral-type benzodiazepine receptor (MBR family of proteins that is present in most organisms and is also referred to as Translocator protein 18 kDa. Although TSPO is associated with stress- and disease-related processes in organisms from bacteria to mammals, full elucidation of the functional role of the TSPO protein is lacking for most organisms in which it is found. In this study, we describe the regulation and function of a TSPO homolog in the cyanobacterium Fremyella diplosiphon, designated FdTSPO. Accumulation of the FdTSPO transcript is upregulated by green light and in response to nutrient deficiency and stress. A F. diplosiphon TSPO deletion mutant (i.e., ΔFdTSPO showed altered responses compared to the wild type strain under stress conditions, including salt treatment, osmotic stress and induced oxidative stress. Under salt stress, the FdTSPO transcript is upregulated and a ΔFdTSPO mutant accumulates lower levels of reactive oxygen species (ROS and displays increased growth compared to WT. In response to osmotic stress, FdTSPO transcript levels are upregulated and ΔFdTSPO mutant cells exhibit impaired growth compared to the wild type. By comparison, methyl viologen-induced oxidative stress results in higher ROS levels in the ΔFdTSPO mutant compared to the wild type strain. Taken together, our results provide support for the involvement of membrane-localized FdTSPO in mediating cellular responses to stress in F. diplosiphon and represent detailed functional analysis of a cyanobacterial TSPO. This study advances our understanding of the functional roles of TSPO homologs in vivo.

  7. Enzyme kinetics, inhibitors, mutagenesis and electron paramagnetic resonance analysis of dual-affinity nitrate reductase in unicellular N(2)-fixing cyanobacterium Cyanothece sp. PCC 8801.

    Science.gov (United States)

    Wang, Tung-Hei; Chen, Yung-Han; Huang, Jine-Yung; Liu, Kang-Cheng; Ke, Shyue-Chu; Chu, Hsiu-An

    2011-11-01

    The assimilatory nitrate reductase (NarB) of N(2)-fixing cyanobacterium Cyanothece sp. PCC 8801 is a monomeric enzyme with dual affinity for substrate nitrate. We purified the recombinant NarB of Cyanothece sp. PCC 8801 and further investigated it by enzyme kinetics analysis, site-directed mutagenesis, inhibitor kinetics analysis, and electron paramagnetic resonance (EPR) spectroscopy. The NarB showed 2 kinetic regimes at pH 10.5 or 8 and electron-donor conditions methyl viologen or ferredoxin (Fd). Fd-dependent NR assay revealed NarB with very high affinity for nitrate (K(m)1, ∼1μM; K(m)2, ∼270μM). Metal analysis and EPR results showed that NarB contains a Mo cofactor and a [4Fe-4S] cluster. In addition, the R352A mutation on the proposed nitrate-binding site of NarB greatly altered both high- and low-affinity kinetic components. Furthermore, the effect of azide on the NarB of Cyanothece sp. PCC 8801 was more complex than that on the NarB of Synechococcus sp. PCC 7942 with its single kinetic regime. With 1mM azide, the kinetics of the wild-type NarB was transformed from 2 kinetic regimes to hyperbolic kinetics, and its activity was enhanced significantly under medium nitrate concentrations. Moreover, EPR results also suggested a structural difference between the two NarBs. Taken together, our results show that the NarB of Cyanothece sp. PCC 8801 contains only a single Mo-catalytic center, and we rule out that the enzyme has 2 independent, distinct catalytic sites. In addition, the NarB of Cyanothece sp. PCC 8801 may have a regulatory nitrate-binding site. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  8. Induction of the Nitrate Assimilation nirA Operon and Protein-Protein Interactions in the Maturation of Nitrate and Nitrite Reductases in the Cyanobacterium Anabaena sp. Strain PCC 7120.

    Science.gov (United States)

    Frías, José E; Flores, Enrique

    2015-07-01

    Nitrate is widely used as a nitrogen source by cyanobacteria, in which the nitrate assimilation structural genes frequently constitute the so-called nirA operon. This operon contains the genes encoding nitrite reductase (nirA), a nitrate/nitrite transporter (frequently an ABC-type transporter; nrtABCD), and nitrate reductase (narB). In the model filamentous cyanobacterium Anabaena sp. strain PCC 7120, which can fix N2 in specialized cells termed heterocysts, the nirA operon is expressed at high levels only in media containing nitrate or nitrite and lacking ammonium, a preferred nitrogen source. Here we examined the genes downstream of the nirA operon in Anabaena and found that a small open reading frame of unknown function, alr0613, can be cotranscribed with the operon. The next gene in the genome, alr0614 (narM), showed an expression pattern similar to that of the nirA operon, implying correlated expression of narM and the operon. A mutant of narM with an insertion mutation failed to produce nitrate reductase activity, consistent with the idea that NarM is required for the maturation of NarB. Both narM and narB mutants were impaired in the nitrate-dependent induction of the nirA operon, suggesting that nitrite is an inducer of the operon in Anabaena. It has previously been shown that the nitrite reductase protein NirA requires NirB, a protein likely involved in protein-protein interactions, to attain maximum activity. Bacterial two-hybrid analysis confirmed possible NirA-NirB and NarB-NarM interactions, suggesting that the development of both nitrite reductase and nitrate reductase activities in cyanobacteria involves physical interaction of the corresponding enzymes with their cognate partners, NirB and NarM, respectively. Nitrate is an important source of nitrogen for many microorganisms that is utilized through the nitrate assimilation system, which includes nitrate/nitrite membrane transporters and the nitrate and nitrite reductases. Many cyanobacteria

  9. The Combined Use of in Silico, in Vitro, and in Vivo Analyses to Assess Anti-cancerous Potential of a Bioactive Compound from Cyanobacterium Nostoc sp. MGL001

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    Niveshika

    2017-11-01

    Full Text Available Escalating incidences of cancer, especially in developed and developing countries, demand evaluation of potential unexplored natural drug resources. Here, anticancer potential of 9-Ethyliminomethyl-12-(morpholin-4-ylmethoxy-5,8,13,16-tetraaza -hexacene-2,3-dicarboxylic acid (EMTAHDCA isolated from fresh water cyanobacterium Nostoc sp. MGL001 was screened through in silico, in vitro, and in vivo studies. For in silico analysis, EMTAHDCA was selected as ligand and 11 cancer related proteins (Protein Data Bank ID: 1BIX, 1NOW, 1TE6, 2RCW, 2UVL, 2VCJ, 3CRY, 3HQU, 3NMQ, 5P21, and 4B7P which are common targets of various anticancer drugs were selected as receptors. The results obtained from in silico analysis showed that EMTAHDCA has strong binding affinity for all the 11 target protein receptors. The ability of EMTAHDCA to bind active sites of cancer protein targets indicated that it is functionally similar to commercially available anticancer drugs. For assessing cellular metabolic activities, in vitro studies were performed by using calorimetric assay viz. 3-(4,5-dimethylthiazol-2-yl-2,5 diphenyltetrazolium bromide (MTT. Results showed that EMTAHDCA induced significant cytotoxic response against Dalton's lymphoma ascites (DLA cells in a dose and time dependent manner with an inhibitory concentration (IC50 value of 372.4 ng/mL after 24 h of incubation. However, in case of normal bone marrow cells, the EMTAHDCA did not induce cytotoxicity as the IC50 value was not obtained even with higher dose of 1,000 ng/mL EMTAHDCA. Further, in vivo studies revealed that the median life span/survival days of tumor bearing mice treated with EMTAHDCA increased significantly with a fold change of ~1.9 and 1.81 corresponding to doses of 5 and 10 mg/kg body weight (B.W. of EMTAHDCA respectively, as compared to the DL group. Our results suggest that 5 mg/kg B.W. is effective since the dose of 10 mg/kg B.W. did not show any significant difference as compared to 5 mg/kg B