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Sample records for alpha-proteobacterium sinorhizobium meliloti

  1. Sulfite oxidation in Sinorhizobium meliloti.

    Science.gov (United States)

    Wilson, Jeremy J; Kappler, Ulrike

    2009-12-01

    Sulfite-oxidizing enzymes (SOEs) are crucial for the metabolism of many cells and are particularly important in bacteria oxidizing inorganic or organic sulfur compounds. However, little is known about SOE diversity and metabolic roles. Sinorhizobium meliloti contains four candidate genes encoding SOEs of three different types, and in this work we have investigated the role of SOEs in S. meliloti and their possible link to the metabolism of the organosulfonate taurine. Low level SOE activity (approximately 1.4 U/mg) was present under all conditions tested while growth on taurine and thiosulfate induced high activities (5.5-8.8 U/mg) although S. meliloti cannot metabolize thiosulfate. Protein purification showed that although expression of two candidate genes matched SOE activity patterns, only a single group 2 SOE, SorT (SMc04049), is responsible for this activity. SorT is a heme-free, periplasmic homodimer (78 kDa) that has low homology to other bacterial SOEs. SorT has an apparent k(cat) of 343 s(-1) and high affinities for both sulfite (K(Mapp_pH8) 15.5 microM) and ferricyanide (K(Mapp_pH8) 3.44 microM), but not cytochrome c, suggesting a need for a high redox potential natural electron acceptor. K(Mapp_sulfite) was nearly invariant with pH which is in contrast to all other well characterized SOEs. SorT is part of an operon (SMc04049-04047) also containing a gene for a cytochrome c and an azurin, and these might be the natural electron acceptors for the enzyme. Phylogenetic analysis of SorT-related SOEs and enzymes of taurine degradation indicate that there is no link between the two processes.

  2. PCR Analysis of "expR" Gene Regulating Biosynthesis of Exopolysaccharides in "Sinorhizobium Meliloti"

    Science.gov (United States)

    Sorroche, Fernando G.; Giordano, Walter

    2012-01-01

    Exopolysaccharide (EPS) production by the rhizobacterium "Sinorhizobium meliloti" is essential for root nodule formation on its legume host (alfalfa), and for establishment of a nitrogen-fixing symbiosis between the two partners. Production of EPS II (galactoglucan) by certain "S. meliloti" strains results in a mucoid colony phenotype. Other…

  3. Three way interactions between Thymus vulgaris, Medicago truncatula and Sinorhizobium meliloti

    OpenAIRE

    Grøndahl, Eva; Ehlers, Bodil Kirstine

    2012-01-01

    Thymus vulgaris is a dominating component of the Mediterranean garrigue vegetation. It produces aromatic oil, containing monoterpenes, which affects the performance (growth, survival) of other plants, and microorganisms. Annual plant species of the genus Medicago are commonly found in Mediterranean thyme communities; in fact they often grow very close to thyme plants (within 1 square meter). Medicago has a symbiosis with the nitrogen fixing bacteria Sinorhizobium meliloti – which is essential...

  4. Sinorhizobium meliloti Cells Require Biotin and either Cobalt or Methionine for Growth

    OpenAIRE

    Watson, Robert J.; Heys, Roselyn; Martin, Teresa; Savard, Marc

    2001-01-01

    Sinorhizobium meliloti is usually cultured in rich media containing yeast extract. It has been suggested that some components of yeast extract are also required for growth in minimal medium. We tested 27 strains of this bacterium and found that none were able to grow in minimal medium when methods to limit carryover of yeast extract were used during inoculation. By fractionation of yeast extract, two required growth factors were identified. Biotin was found to be absolutely required for growt...

  5. Biodegradable plastics from Sinorhizobium meliloti as plastics compatible with the environment and human health

    Directory of Open Access Journals (Sweden)

    Mehrdad Hashemi Beidokhti

    2016-03-01

    Full Text Available Introduction: Polyhydroxyalkanoates (PHAs are natural polyesters and biodegradable plastics that are stored as intracellular inclusion bodies by a great variety of bacteria. The aim of this study was to extract polyhydroxyalkanoate from native Sinorhizobium meliloti in Iran. Materials and methods: Sinorhizobium meliloti isolates were collected from roots of alfalfa plants and were identified by Gram staining, biochemical experiments and amplification of 1500 bp fragment of 16Sr DNA gene. PHA granules were detected by microscopic examination. PHA production was evaluated in nutrient deficient medium and its amount was determined by conversion of PHA into crotonic acid by sulphuric acid treatment. The effect of various temperatures, agitation rate and carbon source (sucrose, mannitol, and maltose were evaluated on dry cell weight and polyhydroxybutyrate (PHB production. Results: The maximum amount of polymer production (43.10% was seen in basal mineral medium at 29°C, pH~7 and 215 revolutions per minute (rpm. The results of this research showed that the S5 isolate was capable to produce maximum poly3- hydroxybutyrate. The produced polymer was analyzed for its purity by GC- mass (gas chromatography- mass spectroscopy and confirmed to be PHB compared with the standard polymer. Discussion and conclusion: Native strains of Sinorhizobium can be used in the production of biodegradable plastics and the results of present study showed that S. meliloti S5 was capable to produce maximum PHB at 29°C, agitation rate of 215 rpm, and pH~7. 

  6. Exogenous Camp upregulates the expression of glnII and glnK-amtB genes in Sinorhizobium meliloti 1021

    Institute of Scientific and Technical Information of China (English)

    TIAN Zhexian; MAO Xianjun; SU Wei; LI Jian; BECKER Anke; WANG Yiping

    2006-01-01

    The existence of multiple adenylate cyclase encoding genes implies the importance of Camp in Sinorhizobium meliloti 1021. In this study, as a pioneer step of understanding Camp roles, microarray analysis on S. Meliloti was carried out for the function of exogenous Camp. To our surprise, the result showed that the transcriptions of glnII and glnK genes were significantly upshifted in the presence of exogenous Camp in S. Meliloti. This phenomenon is further confirmed in S. Meliloti that the expression of either glnII or glnK promoter-lacZ translational fusion is higher in the presence of exogenous Camp.Therefore, for the first time, we have identified genes from S. Meliloti whose expression is activated by Camp. The potential physiological role of upregulation of glnII and glnK by Camp is discussed.

  7. Expression of the Sinorhizobium meliloti C4-dicarboxylate transport gene during symbiosis with the Medicago host plant

    NARCIS (Netherlands)

    Boesten, B.

    1999-01-01

    During symbiosis between Sinorhizobium meliloti and the Medicago host plant, the energy required to fix atmospheric nitrogen, is derived from the plant photosynthate. Current evidence indicates that C 4 -dicarboxylates (dCA) are the major and probably only source of carbon provided to the

  8. Cell Cycle Control by the Master Regulator CtrA in Sinorhizobium meliloti.

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    Francesco Pini

    2015-05-01

    Full Text Available In all domains of life, proper regulation of the cell cycle is critical to coordinate genome replication, segregation and cell division. In some groups of bacteria, e.g. Alphaproteobacteria, tight regulation of the cell cycle is also necessary for the morphological and functional differentiation of cells. Sinorhizobium meliloti is an alphaproteobacterium that forms an economically and ecologically important nitrogen-fixing symbiosis with specific legume hosts. During this symbiosis S. meliloti undergoes an elaborate cellular differentiation within host root cells. The differentiation of S. meliloti results in massive amplification of the genome, cell branching and/or elongation, and loss of reproductive capacity. In Caulobacter crescentus, cellular differentiation is tightly linked to the cell cycle via the activity of the master regulator CtrA, and recent research in S. meliloti suggests that CtrA might also be key to cellular differentiation during symbiosis. However, the regulatory circuit driving cell cycle progression in S. meliloti is not well characterized in both the free-living and symbiotic state. Here, we investigated the regulation and function of CtrA in S. meliloti. We demonstrated that depletion of CtrA cause cell elongation, branching and genome amplification, similar to that observed in nitrogen-fixing bacteroids. We also showed that the cell cycle regulated proteolytic degradation of CtrA is essential in S. meliloti, suggesting a possible mechanism of CtrA depletion in differentiated bacteroids. Using a combination of ChIP-Seq and gene expression microarray analysis we found that although S. meliloti CtrA regulates similar processes as C. crescentus CtrA, it does so through different target genes. For example, our data suggest that CtrA does not control the expression of the Fts complex to control the timing of cell division during the cell cycle, but instead it negatively regulates the septum-inhibiting Min system. Our

  9. Cloning-free genome engineering in Sinorhizobium meliloti advances applications of Cre/loxP site-specific recombination.

    Science.gov (United States)

    Döhlemann, Johannes; Brennecke, Meike; Becker, Anke

    2016-09-10

    The soil-dwelling α-proteobacterium Sinorhizobium meliloti serves as model for studies of symbiotic nitrogen fixation, a highly important process in sustainable agriculture. Here, we report advancements of the genetic toolbox accelerating genome editing in S. meliloti. The hsdMSR operon encodes a type-I restriction-modification (R-M) system. Transformation of S. meliloti is counteracted by the restriction endonuclease HsdR degrading DNA which lacks the appropriate methylation pattern. We provide a stable S. meliloti hsdR deletion mutant showing enhanced transformation with Escherichia coli-derived plasmid DNA and demonstrate that using an E. coli plasmid donor, expressing S. meliloti methyl transferase genes, is an alternative strategy of increasing the transformation efficiency of S. meliloti. Furthermore, we devise a novel cloning-free genome editing (CFGE) method for S. meliloti, Agrobacterium tumefaciens and Xanthomonas campestris, and demonstrate the applicability of this method for intricate applications of the Cre/lox recombination system in S. meliloti. An enhanced Cre/lox system, allowing for serial deletions of large genomic regions, was established. An assay of lox spacer mutants identified a set of lox sites mediating specific recombination. The availability of several non-promiscuous Cre recognition sites enables simultaneous specific Cre/lox recombination events. CFGE combined with Cre/lox recombination is put forward as powerful approach for targeted genome editing, involving serial steps of manipulation to expedite the genetic accessibility of S. meliloti as chassis. PMID:27393468

  10. Quantitative Proteomic Analysis of the Hfq-Regulon in Sinorhizobium meliloti 2011

    Science.gov (United States)

    Sobrero, Patricio; Schlüter, Jan-Philip; Lanner, Ulrike; Schlosser, Andreas; Becker, Anke; Valverde, Claudio

    2012-01-01

    Riboregulation stands for RNA-based control of gene expression. In bacteria, small non-coding RNAs (sRNAs) are a major class of riboregulatory elements, most of which act at the post-transcriptional level by base-pairing target mRNA genes. The RNA chaperone Hfq facilitates antisense interactions between target mRNAs and regulatory sRNAs, thus influencing mRNA stability and/or translation rate. In the α-proteobacterium Sinorhizobium meliloti strain 2011, the identification and detection of multiple sRNAs genes and the broadly pleitropic phenotype associated to the absence of a functional Hfq protein both support the existence of riboregulatory circuits controlling gene expression to ensure the fitness of this bacterium in both free living and symbiotic conditions. In order to identify target mRNAs subject to Hfq-dependent riboregulation, we have compared the proteome of an hfq mutant and the wild type S. meliloti by quantitative proteomics following protein labelling with 15N. Among 2139 univocally identified proteins, a total of 195 proteins showed a differential abundance between the Hfq mutant and the wild type strain; 65 proteins accumulated ≥2-fold whereas 130 were downregulated (≤0.5-fold) in the absence of Hfq. This profound proteomic impact implies a major role for Hfq on regulation of diverse physiological processes in S. meliloti, from transport of small molecules to homeostasis of iron and nitrogen. Changes in the cellular levels of proteins involved in transport of nucleotides, peptides and amino acids, and in iron homeostasis, were confirmed with phenotypic assays. These results represent the first quantitative proteomic analysis in S. meliloti. The comparative analysis of the hfq mutant proteome allowed identification of novel strongly Hfq-regulated genes in S. meliloti. PMID:23119037

  11. Complementation analyses of Sinorhizobium meliloti nifA mutant with different originated nifA genes

    Institute of Scientific and Technical Information of China (English)

    YAO Zhenhua; R(U)VERG Silvia; WANG Yiping; ZOU Huasong; TIAN Zhexian; DAI Xiaomi; BECKER Anke; LI Jian; YAN Haiqin; XIAO Yan; ZHU Jiabi; YU Guanqiao

    2006-01-01

    A previous work inferred that the nifA gene of Enterobacter cloacae did not restore the symbiotic phenotype of Sinorhizobium meliloti nifA mutant. In the present study, two nifA genes of Bradyrhizobium japonicum and Mesorhizobium huakuii also did not restore the symbiotic phenotype of S.meliloti nifA mutant. In whole genomic microarray experiments, 238 genes were found to be differentially expressed after S. meliloti nifA had been constitutively expressed in its nifA mutant. In contrast,only 20, 7 and 9 genes changed their transcriptional levels when expressing B. japonium, M. huakuii and Enterobacter cloacae nifA genes in Sm nifA mutant,separately. These genes were classified into several functional groups including house keeping, energy and central intermediary metabolism, transport systems and symbiosis. Interestingly, the genes that of nifH operons showed high expression levels in the presence of either B. japonium or M. huakuii NifA,which was confirmed by subsequent lacZ fusion experiments.

  12. nifH Promoter Activity Is Regulated by DNA Supercoiling in Sinorhizobium meliloti

    Institute of Scientific and Technical Information of China (English)

    Yan-Jie LIU; Biao HU; Jia-Bi ZHU; Shan-Jiong SHEN; Guan-Qiao YU

    2005-01-01

    In prokaryotes, DNA supercoiling regulates the expression of many genes; for example, the expression of Klebsiella pneumoniae nifLA operon depends on DNA negative supercoiling in anaerobically grown cells, which indicates that DNA supercoiling might play a role in gene regulation of the anaerobic response. Since the expression of the nifH promoter in Sinorhizobium meliloti is not repressed by oxygen, it is proposed that the status of DNA supercoiling may not affect the expression of the nifH promoter. We tested this hypothesis by analyzing nifH promoter activity in wild-type and gyr- Escherichia coli in the presence and absence of DNA gyrase inhibitors. Our results show that gene expression driven by the S.meliloti nifH promoter requires the presence of active DNA gyrase. Because DNA gyrase increases the number of negative superhelical turns in DNA in the presence of ATP, our data indicate that negative supercoiling is also important for nifH promoter activity. Our study also shows that the DNA supercoilingdependent S. meliloti nifH promoter activity is related to the trans-acting factors NtrC and NifA that activate it. DNA supercoiling appeared to have a stronger effect on NtrC-activated nifH promoter activity than on NifA-activated promoter activity. Collectively, these results from the S. meliloti nifH promoter model system seem to indicate that, in addition to regulating gene expression during anaerobic signaling, DNA supercoiling may also provide a favorable topology for trans-acting factor binding and promoter activation regardless of oxygen status.

  13. The Sinorhizobium meliloti RNA chaperone Hfq influences central carbon metabolism and the symbiotic interaction with alfalfa

    Directory of Open Access Journals (Sweden)

    Jiménez-Zurdo José I

    2010-03-01

    Full Text Available Abstract Background The bacterial Hfq protein is able to interact with diverse RNA molecules, including regulatory small non-coding RNAs (sRNAs, and thus it is recognized as a global post-transcriptional regulator of gene expression. Loss of Hfq has an extensive impact in bacterial physiology which in several animal pathogens influences virulence. Sinorhizobium meliloti is a model soil bacterium known for its ability to establish a beneficial nitrogen-fixing intracellular symbiosis with alfalfa. Despite the predicted general involvement of Hfq in the establishment of successful bacteria-eukaryote interactions, its function in S. meliloti has remained unexplored. Results Two independent S. meliloti mutants, 2011-3.4 and 1021Δhfq, were obtained by disruption and deletion of the hfq gene in the wild-type strains 2011 and 1021, respectively, both exhibiting similar growth defects as free-living bacteria. Transcriptomic profiling of 1021Δhfq revealed a general down-regulation of genes of sugar transporters and some enzymes of the central carbon metabolism, whereas transcripts specifying the uptake and metabolism of nitrogen sources (mainly amino acids were more abundant than in the wild-type strain. Proteomic analysis of the 2011-3.4 mutant independently confirmed these observations. Symbiotic tests showed that lack of Hfq led to a delayed nodulation, severely compromised bacterial competitiveness on alfalfa roots and impaired normal plant growth. Furthermore, a large proportion of nodules (55%-64% elicited by the 1021Δhfq mutant were non-fixing, with scarce content in bacteroids and signs of premature senescence of endosymbiotic bacteria. RT-PCR experiments on RNA from bacteria grown under aerobic and microoxic conditions revealed that Hfq contributes to regulation of nifA and fixK1/K2, the genes controlling nitrogen fixation, although the Hfq-mediated regulation of fixK is only aerobiosis dependent. Finally, we found that some of the recently

  14. Transcriptome analysis of Sinorhizobium meliloti nodule bacteria in nifA mutant background

    Institute of Scientific and Technical Information of China (English)

    TIAN Zhexian; WANG Yiping; ZOU Huasong; LI Jian; ZHANG Yuantao; LIU Ying; YU Guanqiao; ZHU Jiabi; R(U)BERG Silvia; BECKER Anke

    2006-01-01

    Gene expression profiles of a Sinorhizobium meliloti 1021 nifA mutant and wild type nodule bacteria were compared using whole genome microarrays. The results revealed a large scale alteration of gene expression (601 genes) in the nifA minus background. The loss of NifA altered the expression of many functional groups of genes (macromolecular metabolism, TCA cycle and respiration,nodulation and nitrogen fixation) and may lead to quite different life stages of the nodule bacteria.Upregulation of fixK and its associated genes was observed in the nifA mutant nodule bacteria. Additional quantitative real-time PCR experiments revealed that the transcript levels of fixLJ were significantly upshifted in the nifA mutant nodule bacteria.Putative NifA binding sites were predicted by a statistical method in the upstream sequences of 13 differentially regulated genes from the nifA- transcriptome.

  15. Metabolic modelling reveals the specialization of secondary replicons for niche adaptation in Sinorhizobium meliloti.

    Science.gov (United States)

    diCenzo, George C; Checcucci, Alice; Bazzicalupo, Marco; Mengoni, Alessio; Viti, Carlo; Dziewit, Lukasz; Finan, Turlough M; Galardini, Marco; Fondi, Marco

    2016-01-01

    The genome of about 10% of bacterial species is divided among two or more large chromosome-sized replicons. The contribution of each replicon to the microbial life cycle (for example, environmental adaptations and/or niche switching) remains unclear. Here we report a genome-scale metabolic model of the legume symbiont Sinorhizobium meliloti that is integrated with carbon utilization data for 1,500 genes with 192 carbon substrates. Growth of S. meliloti is modelled in three ecological niches (bulk soil, rhizosphere and nodule) with a focus on the role of each of its three replicons. We observe clear metabolic differences during growth in the tested ecological niches and an overall reprogramming following niche switching. In silico examination of the inferred fitness of gene deletion mutants suggests that secondary replicons evolved to fulfil a specialized function, particularly host-associated niche adaptation. Thus, genes on secondary replicons might potentially be manipulated to promote or suppress host interactions for biotechnological purposes. PMID:27447951

  16. Crystallization and preliminary crystallographic studies of the recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114

    International Nuclear Information System (INIS)

    The dihydropyrimidinase from S. meliloti CECT4114, with activity towards both hydantoin and dihydrouracil substrates, was crystallized, and diffraction data were collected to 1.85 Å resolution. Dihydropyrimidinases are involved in the reductive pathway of pyrimidine degradation, catalysing the hydrolysis of 5,6-dihydrouracil and 5,6-dihydrothymine to the corresponding N-carbamoyl β-amino acids. This enzyme has often been referred to as hydantoinase owing to its industrial application in the production of optically pure amino acids starting from racemic mixtures of 5-monosubstituted hydantoins. Recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114 (SmelDhp) has been expressed, purified and crystallized. Crystallization was performed using the counter-diffusion method with capillaries of 0.3 mm inner diameter. Crystals of SmelDhp suitable for data collection and structure determination were grown in the presence of agarose at 0.1%(w/v) in order to ensure mass transport controlled by diffusion. X-ray data were collected to a resolution of 1.85 Å. The crystal belongs to the orthorhombic space group C2221, with unit-cell parameters a = 124.89, b = 126.28, c = 196.10 Å and two molecules in the asymmetric unit. A molecular-replacement solution has been determined and refinement is in progress

  17. Spatiotemporal choreography of chromosome and megaplasmids in the Sinorhizobium meliloti cell cycle.

    Science.gov (United States)

    Frage, Benjamin; Döhlemann, Johannes; Robledo, Marta; Lucena, Daniella; Sobetzko, Patrick; Graumann, Peter L; Becker, Anke

    2016-06-01

    A considerable share of bacterial species maintains multipartite genomes. Precise coordination of genome replication and segregation with cell growth and division is vital for proliferation of these bacteria. The α-proteobacterium Sinorhizobium meliloti possesses a tripartite genome composed of one chromosome and the megaplasmids pSymA and pSymB. Here, we investigated the spatiotemporal pattern of segregation of these S. meliloti replicons at single cell level. Duplication of chromosomal and megaplasmid origins of replication occurred spatially and temporally separated, and only once per cell cycle. Tracking of FROS (fluorescent repressor operator system)-labelled origins revealed a strict temporal order of segregation events commencing with the chromosome followed by pSymA and then by pSymB. The repA2B2C2 region derived from pSymA was sufficient to confer the spatiotemporal behaviour of this megaplasmid to a small plasmid. Altering activity of the ubiquitous prokaryotic replication initiator DnaA, either positively or negatively, resulted in an increase in replication initiation events or G1 arrest of the chromosome only. This suggests that interference with DnaA activity does not affect replication initiation control of the megaplasmids. PMID:26853523

  18. Rhizobial galactoglucan determines the predatory pattern of Myxococcus xanthus and protects Sinorhizobium meliloti from predation

    Science.gov (United States)

    Pérez, Juana; Jiménez-Zurdo, José I.; Martínez-Abarca, Francisco; Millán, Vicenta; Shimkets, Lawrence J.; Muñoz-Dorado, José

    2014-01-01

    Summary Myxococcus xanthus is a social bacterium that preys on prokaryotic and eukaryotic microorganisms. Co-culture of M. xanthus with reference laboratory strains and field isolates of the legume symbiont Sinorhizobium meliloti revealed two different predatory patterns that resemble frontal and wolfpack attacks. Use of mutants impaired in the two types of M. xanthus surface motility (A or adventurous and S or social motility) and a csgA mutant, which is unable to form macroscopic travelling waves known as ripples, has demonstrated that both motility systems but not rippling are required for efficient predation. To avoid frontal attack and reduce killing rates, rhizobial cells require a functional expR gene. ExpR regulates expression of genes involved in a variety of functions. The use of S. meliloti mutants impaired in several of these functions revealed that the exopolysaccharide galactoglucan (EPS II) is the major determinant of the M. xanthus predatory pattern. The data also suggest that this biopolymer confers an ecological advantage to rhizobial survival in soil, which may have broad environmental implications. PMID:24707988

  19. Role of oxyR from Sinorhizobium meliloti in Regulating the Expression of Catalases

    Institute of Scientific and Technical Information of China (English)

    Li LUO; Ming-Sheng QI; Shi-Yi YAO; Hai-Ping CHENG; Jia-Bi ZHU; Guan-Qiao YU

    2005-01-01

    The process of symbiotic nitrogen fixation results in the generation of reactive oxygen species such as the superoxide anion (O2-) and hydrogen peroxide (H2O2). The response of rhizobia to these toxic oxygen species is an important factor in nodulation and nitrogen fixation. In Sinorhizobium meliloti, one oxyR homologue and three catalase genes, katA, katB, and katC were detected by sequence analysis. This oxyR gene is located next to and divergently from katA on the chromosome. To investigate the possible roles of oxyR in regulating the expression of catalases at the transcriptional level in S. meliloti, an insertion mutant of this gene was constructed. The mutant was more sensitive and less adaptive to H2O2 than the wild type strain, and total catalase/peroxidase activity was reduced approximately fourfold with the OxyR mutation relative to controls. The activities of KatA and KatB and the expression of katA::lacZ and katB::lacZ promoter fusions were increased in the mutant strain compared with the parental strain grown in the absence of H2O2,indicating that katA and katB are repressed by OxyR. However, when exposed to H2O2, katA expression was also increased in both S. meliloti and Escherichia coli. When exposed to H2O2, OxyR is converted from a reduced to an oxidized form in E. coli. We concluded that the reduced form of OxyR functions as a repressor of katA and katB expression. Thus, in the presence of H2O2, reduced OxyR is converted to the oxidized form of OxyR that then results in increased katA expression. We further showed that oxyR expression is autoregulated via negative feedback.

  20. Systematic insertion mutagenesis of GntR family transcriptional regulator genes in Sinorhizobium meliloti

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    GntR-type transcriptional regulators regulate the most diverse biological processes in bacteria. Although GntR-type transcriptional regulators consist of the second largest family of transcriptional regulators in Sinorhizobium meliloti, little is known about their functions. In this study, we investigated 54 putative genes encoding GntR family of transcriptional regulators in S. meliloti Rm1021. Secondary structure analysis of the C-terminal domain of these putative transcriptional regulators indicated that thirty-seven were members of the FadR subfamily, ten of the HutC subfamily and five of the MocR subfamily. The remaining two did not fall into any specific subfamily category, and may form two new subfamilies. The 54 gntR genes were mutagenized by plasmid insertion mutagenesis to investigate their roles. We found that, of the 54 mutants, only the gtrA1 and gtrB1 mutants had slower growth rates and cell maximal yields on both rich medium and minimal medium, and lower cell motility on swarming plate than wild type Rm1021. All mutants, with the exception of gtrA1 and gtrB1, can establish effective symbioses with alfalfa. Plants inoculated with gtrA1 and gtrB1 mutants grew shorter than those inoculated with wild type, and formed relatively smaller, round and light pink nodules, which were mainly located on lateral roots. And there was an abnormal increase in the number of nodules induced by both mutants. These results suggested that the gtrA1 and gtrB1 mutants were symbiotically deficient. Our work presents a global overview of GntR-like transcriptional regulators involved in symbiosis in S.meliloti, and provides new insight into the functions of GntR-like transcriptional regulators.

  1. Genetic Organization of the Region Encoding Regulation, Biosynthesis, and Transport of Rhizobactin 1021, a Siderophore Produced by Sinorhizobium meliloti

    OpenAIRE

    Lynch, Damien; O'Brien, John; Welch, Timothy; Clarke, Paul; Ó Cuív, Páraic; Crosa, Jorge H.; O'Connell, Michael

    2001-01-01

    Eight genes have been identified that function in the regulation, biosynthesis, and transport of rhizobactin 1021, a hydroxamate siderophore produced under iron stress by Sinorhizobium meliloti. The genes were sequenced, and transposon insertion mutants were constructed for phenotypic analysis. Six of the genes, named rhbABCDEF, function in the biosynthesis of the siderophore and were shown to constitute an operon that is repressed under iron-replete conditions. Another gene in the cluster, n...

  2. Contribution of Individual Chemoreceptors to Sinorhizobium meliloti Chemotaxis Towards Amino Acids of Host and Nonhost Seed Exudates.

    Science.gov (United States)

    Webb, Benjamin A; Helm, Richard F; Scharf, Birgit E

    2016-03-01

    Plant seeds and roots exude a spectrum of molecules into the soil that attract bacteria to the spermosphere and rhizosphere, respectively. The alfalfa symbiont Sinorhizobium meliloti utilizes eight chemoreceptors (McpT to McpZ and IcpA) to mediate chemotaxis. Using a modified hydrogel capillary chemotaxis assay that allows data quantification and larger throughput screening, we defined the role of S. meliloti chemoreceptors in sensing its host, Medicago sativa, and a closely related nonhost, Medicago arabica. S. meliloti wild type and most single-deletion strains displayed comparable chemotaxis responses to host or nonhost seed exudate. However, while the mcpZ mutant responded like wild type to M. sativa exudate, its reaction to M. arabica exudate was reduced by 80%. Even though the amino acid (AA) amounts released by both plant species were similar, synthetic AA mixtures that matched exudate profiles contributed differentially to the S. meliloti wild-type response to M. sativa (23%) and M. arabica (37%) exudates, with McpU identified as the most important chemoreceptor for AA. Our results show that S. meliloti is equally attracted to host and nonhost legumes; however, AA play a greater role in attraction to M. arabica than to M. sativa, with McpZ being specifically important in sensing M. arabica. PMID:26713349

  3. Effects of nutritional and environmental conditions on Sinorhizobium meliloti biofilm formation.

    Science.gov (United States)

    Rinaudi, Luciana; Fujishige, Nancy A; Hirsch, Ann M; Banchio, Erika; Zorreguieta, Angeles; Giordano, Walter

    2006-11-01

    Rhizobia are non-spore-forming soil bacteria that fix atmospheric nitrogen into ammonia in a symbiosis with legume roots. However, in the absence of a legume host, rhizobia manage to survive and hence must have evolved strategies to adapt to diverse environmental conditions. The capacity to respond to variations in nutrient availability enables the persistence of rhizobial species in soil, and consequently improves their ability to colonize and to survive in the host plant. Rhizobia, like many other soil bacteria, persist in nature most likely in sessile communities known as biofilms, which are most often composed of multiple microbial species. We have been employing in vitro assays to study environmental parameters that might influence biofilm formation in the Medicago symbiont Sinorhizobium meliloti. These parameters include carbon source, amount of nitrate, phosphate, calcium and magnesium as well as the effects of osmolarity and pH. The microtiter plate assay facilitates the detection of subtle differences in rhizobial biofilms in response to these parameters, thereby providing insight into how environmental stress or nutritional status influences rhizobial survival. Nutrients such as sucrose, phosphate and calcium enhance biofilm formation as their concentrations increase, whereas extreme temperatures and pH negatively affect biofilm formation. PMID:16887339

  4. Characteristics of the LrhA subfamily of transcriptional regulators from Sinorhizobium meliloti

    Institute of Scientific and Technical Information of China (English)

    Mingsheng Qi; Li Luo; Haiping Cheng; Jiabi Zhu; Guanqiao Yu

    2008-01-01

    In our previous work, we identified 94 putative genes encoding LysR-type transcriptional regulators from Sinorhizobium meliloti. All of these putative lysR genes were mutagenized using plasmid insertions to determine their phenotypes. Six LysR-type regulators, encoded by mutants SMa1979, SMb20715, SMc00820, SMc04163, SMc03975,and SMc04315, showed similar amino acid sequences (30%)and shared the conserved DNA-binding domain with LrhA,HexA, or DgdR. Phenotype analysis of these gene mutants indicated that the regulators control the swimming behaviors of the bacteria, production of quorum-sensing signals, and secretion of extracellular proteins. These characteristics are very similar to those of LrhA, HexA, and DgdR.Thus, we refer to this group as the LrhA subfamily. Sequence analysis showed that a great number of homologous genes of the LrhA subfamily were distributed in the α,β, and γsubdivisions of proteobacteria, and a few in actinobacteria. These findings could provide new clues to the roles of the LysR gene family.

  5. Role of extracellular compounds in Cd-sequestration relative to Cd uptake by bacterium Sinorhizobium meliloti

    Energy Technology Data Exchange (ETDEWEB)

    Slaveykova, Vera I., E-mail: vera.slaveykova@epfl.c [Environmental Biophysical Chemistry, IIE-ENAC, Ecole Polytechnique Federale de Lausanne (EPFL), Station 2, CH-1015 Lausanne (Switzerland); Parthasarathy, Nalini [Department of Inorganic, Analytic and Applied Chemistry, University of Geneva, Sciences II, 30 Quai Ernest Ansermet, 1211 Geneva 4 (Switzerland); Dedieu, Karine; Toescher, Denis [Environmental Biophysical Chemistry, IIE-ENAC, Ecole Polytechnique Federale de Lausanne (EPFL), Station 2, CH-1015 Lausanne (Switzerland)

    2010-08-15

    The role of bacterially derived compounds in Cd(II) complexation and uptake by bacterium Sinorhizobium meliloti wild type (WT) and genetically modified ExoY-mutant, deficient in exopolysaccharide production, was explored combining chemical speciation measurements and assays with living bacteria. Obtained results demonstrated that WT- and ExoY-strains excreted siderophores in comparable amounts, while WT-strain produced much higher amount of exopolysaccharides and less exoproteins. An evaluation of Cd(II) distribution in bacterial suspensions under short term exposure conditions, showed that most of the Cd is bound to bacterial surface envelope, including Cd bound to the cell wall and to the attached extracellular polymeric substances. However, the amount of Cd bound to the dissolved extracellular compounds increases at high Cd(II) concentrations. The implications of these findings to more general understanding of the Cd(II) fate and cycling in the environment is discussed. - Bacterial excreted extracellular compounds play minor role in Cd(II) sequestration relative to bacteria.

  6. Rhizobia from Lanzarote, the Canary Islands, That Nodulate Phaseolus vulgaris Have Characteristics in Common with Sinorhizobium meliloti Isolates from Mainland Spain▿

    OpenAIRE

    Zurdo-Piñeiro, José Luis; García-Fraile, Paula; Rivas, Raúl; Peix, Alvaro; León-Barrios, Milagros; Willems, Anne; Mateos, Pedro Francisco; Martínez-Molina, Eustoquio; Velázquez, Encarna; Van Berkum, Peter

    2009-01-01

    The stable, low-molecular-weight (LMW) RNA fractions of several rhizobial isolates of Phaseolus vulgaris grown in the soil of Lanzarote, an island of the Canary Islands, were identical to a less-common pattern found within Sinorhizobium meliloti (assigned to group II) obtained from nodules of alfalfa and alfalfa-related legumes grown in northern Spain. The P. vulgaris isolates and the group II LMW RNA S. meliloti isolates also were distinguishable in that both had two conserved inserts of 20 ...

  7. Exploring the symbiotic pangenome of the nitrogen-fixing bacterium Sinorhizobium meliloti

    Directory of Open Access Journals (Sweden)

    Daligault Hajnalka

    2011-05-01

    Full Text Available Abstract Background Sinorhizobium meliloti is a model system for the studies of symbiotic nitrogen fixation. An extensive polymorphism at the genetic and phenotypic level is present in natural populations of this species, especially in relation with symbiotic promotion of plant growth. AK83 and BL225C are two nodule-isolated strains with diverse symbiotic phenotypes; BL225C is more efficient in promoting growth of the Medicago sativa plants than strain AK83. In order to investigate the genetic determinants of the phenotypic diversification of S. meliloti strains AK83 and BL225C, we sequenced the complete genomes for these two strains. Results With sizes of 7.14 Mbp and 6.97 Mbp, respectively, the genomes of AK83 and BL225C are larger than the laboratory strain Rm1021. The core genome of Rm1021, AK83, BL225C strains included 5124 orthologous groups, while the accessory genome was composed by 2700 orthologous groups. While Rm1021 and BL225C have only three replicons (Chromosome, pSymA and pSymB, AK83 has also two plasmids, 260 and 70 Kbp long. We found 65 interesting orthologous groups of genes that were present only in the accessory genome, consequently responsible for phenotypic diversity and putatively involved in plant-bacterium interaction. Notably, the symbiosis inefficient AK83 lacked several genes required for microaerophilic growth inside nodules, while several genes for accessory functions related to competition, plant invasion and bacteroid tropism were identified only in AK83 and BL225C strains. Presence and extent of polymorphism in regulons of transcription factors involved in symbiotic interaction were also analyzed. Our results indicate that regulons are flexible, with a large number of accessory genes, suggesting that regulons polymorphism could also be a key determinant in the variability of symbiotic performances among the analyzed strains. Conclusions In conclusions, the extended comparative genomics approach revealed a

  8. Mining the Sinorhizobium meliloti transportome to develop FRET biosensors for sugars, dicarboxylates and cyclic polyols.

    Directory of Open Access Journals (Sweden)

    Alexandre Bourdès

    Full Text Available Förster resonance energy transfer (FRET biosensors are powerful tools to detect biologically important ligands in real time. Currently FRET bisosensors are available for twenty-two compounds distributed in eight classes of chemicals (two pentoses, two hexoses, two disaccharides, four amino acids, one nucleobase, two nucleotides, six ions and three phytoestrogens. To expand the number of available FRET biosensors we used the induction profile of the Sinorhizobium meliloti transportome to systematically screen for new FRET biosensors.Two new vectors were developed for cloning genes for solute-binding proteins (SBPs between those encoding FRET partner fluorescent proteins. In addition to a vector with the widely used cyan and yellow fluorescent protein FRET partners, we developed a vector using orange (mOrange2 and red fluorescent protein (mKate2 FRET partners. From the sixty-nine SBPs tested, seven gave a detectable FRET signal change on binding substrate, resulting in biosensors for D-quinic acid, myo-inositol, L-rhamnose, L-fucose, β-diglucosides (cellobiose and gentiobiose, D-galactose and C4-dicarboxylates (malate, succinate, oxaloacetate and fumarate. To our knowledge, we describe the first two FRET biosensor constructs based on SBPs from Tripartite ATP-independent periplasmic (TRAP transport systems.FRET based on orange (mOrange2 and red fluorescent protein (mKate2 partners allows the use of longer wavelength light, enabling deeper penetration of samples at lower energy and increased resolution with reduced back-ground auto-fluorescence. The FRET biosensors described in this paper for four new classes of compounds; (i cyclic polyols, (ii L-deoxy sugars, (iii β-linked disaccharides and (iv C4-dicarboxylates could be developed to study metabolism in vivo.

  9. Draft Genome Sequence of Sinorhizobium meliloti CCNWSX0020, a Nitrogen-Fixing Symbiont with Copper Tolerance Capability Isolated from Lead-Zinc Mine Tailings

    Science.gov (United States)

    Li, Zhefei; Ma, Zhanqiang; Hao, Xiuli

    2012-01-01

    Sinorhizobium meliloti CCNWSX0020 was isolated from Medicago lupulina plants growing in lead-zinc mine tailings, which can establish a symbiotic relationship with Medicago species. Also, the genome of this bacterium contains a number of protein-coding sequences related to metal tolerance. We anticipate that the genomic sequence provides valuable information to explore environmental bioremediation. PMID:22328762

  10. A glutamine-amidotransferase-like protein modulates FixT anti-kinase activity in Sinorhizobium meliloti

    Directory of Open Access Journals (Sweden)

    Boistard Pierre

    2001-05-01

    Full Text Available Abstract Background Nitrogen fixation gene expression in Sinorhizobium meliloti, the alfalfa symbiont, depends on a cascade of regulation that involves both positive and negative control. On top of the cascade, the two-component regulatory system FixLJ is activated under the microoxic conditions of the nodule. In addition, activity of the FixLJ system is inhibited by a specific anti-kinase protein, FixT. The physiological significance of this negative regulation by FixT was so far unknown. Results We have isolated by random Tn5 mutagenesis a S. meliloti mutant strain that escapes repression by FixT. Complementation test and DNA analysis revealed that inactivation of an asparagine synthetase-like gene was responsible for the phenotype of the mutant. This gene, that was named asnO, encodes a protein homologous to glutamine-dependent asparagine synthetases. The asnO gene did not appear to affect asparagine biosynthesis and may instead serve a regulatory function in S. meliloti. We provide evidence that asnO is active during symbiosis . Conclusions Isolation of the asnO mutant argues for the existence of a physiological regulation associated with fixT and makes it unlikely that fixT serves a mere homeostatic function in S. meliloti. Our data suggest that asnO might control activity of the FixT protein, in a way that remains to be elucidated. A proposed role for asnO might be to couple nitrogen fixation gene expression in S. meliloti to the nitrogen needs of the cells.

  11. Sinorhizobium meliloti nifA gene exerts a pleiotropic effect on nodulation through the enhanced plant defense response

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Sinorhizobium meliloti nifA gene is required for the expression of a bunch of nif and fix genes. Here, we report its pleiotropic effects on the nodule formation. Compared with wild type strain, nifA mutant significantly reduced nodule suppression rate in split-root system. The plants inoculated with mutant strain produced lower amount of daidzein and less necrotic cells on their roots. In addition, the defense genes failed to be evoked by nifA mutant at the early nodulation stage. These findings indicated that host defense response was one of the mechanisms mediated by nifA gene to regulate nodule formation during symbiosis. Even though nifA mutant could increase the number of nodules in host plant, it synthesized lower Nod factors than wild type. This suggested that nifA gene mediated multiple and diverse instances in nodulation formation.

  12. Altered susceptibility to infection by Sinorhizobium meliloti and Nectria haematococca in alfalfa roots with altered cell cycle.

    Science.gov (United States)

    Woo, H-H; Hirsch, A M; Hawes, M C

    2004-07-01

    Most infections of plant roots are initiated in the region of elongation; the mechanism for this tissue-specific localization pattern is unknown. In alfalfa expressing PsUGT1 antisense mRNA under the control of the cauliflower mosaic virus (CaMV) 35S promoter, the cell cycle in roots is completed in 48 h instead of 24 h, and border cell number is decreased by more than 99%. These plants were found to exhibit increased root-tip infection by a fungal pathogen and reduced nodule formation by a bacterial symbiont. Thus, the frequency of infection in the region of elongation by Nectria haematocca was unaffected, but infection of the root tip was increased by more than 90%; early stages of Sinorhizobium meliloti infection and nodule morphology were normal, but the frequency of nodulation was fourfold lower than in wild-type roots. PMID:15042410

  13. Altered susceptibility to infection by Sinorhizobium meliloti and Nectria haematococca in alfalfa roots with altered cell cycle.

    Science.gov (United States)

    Woo, H-H; Hirsch, A M; Hawes, M C

    2004-07-01

    Most infections of plant roots are initiated in the region of elongation; the mechanism for this tissue-specific localization pattern is unknown. In alfalfa expressing PsUGT1 antisense mRNA under the control of the cauliflower mosaic virus (CaMV) 35S promoter, the cell cycle in roots is completed in 48 h instead of 24 h, and border cell number is decreased by more than 99%. These plants were found to exhibit increased root-tip infection by a fungal pathogen and reduced nodule formation by a bacterial symbiont. Thus, the frequency of infection in the region of elongation by Nectria haematocca was unaffected, but infection of the root tip was increased by more than 90%; early stages of Sinorhizobium meliloti infection and nodule morphology were normal, but the frequency of nodulation was fourfold lower than in wild-type roots.

  14. Genome-engineered Sinorhizobium meliloti for the production of poly(lactic-co-3-hydroxybutyric) acid copolymer.

    Science.gov (United States)

    Tran, Tam T; Charles, Trevor C

    2016-02-01

    Economically competitive commercial production of biodegradable bioplastics with desirable properties is an important goal. In this study, we demonstrate the use of chromosome engineering of an alternative bacterial host, Sinorhizobium meliloti, for production of the copolymer, poly(lactate-co-3-hydroxybutyrate). Codon-optimized genes for 2 previously engineered enzymes, Clostridium propionicum propionate CoA transferase (Pct532Cp) and Pseudomonas sp. strain MBEL 6-19 polyhydroxyalkanoate (PHA) synthase 1 (PhaC1400Ps6-19), were introduced into S. meliloti Rm1021 by chromosome integration, replacing the native phbC gene. On the basis of phenotypic analysis and detection of polymer product by gas chromatography analysis, synthesis and accumulation of the copolymer was confirmed. The chromosome integrant strain, with the introduced genes under the control of the native phbC promoter, is able to produce over 15% cell dry mass of poly(lactate-co-3-hydroxybutyrate), containing 30 mol% lactate, from growth on mannitol. We were also able to purify the polymer from the culture and confirm the structure by NMR and GC-MS. To our knowledge, this is the first demonstration of production of this copolymer in the Alphaproteobacteria. Further optimization of this system may eventually yield strains that are able to produce economically viable commercial product. PMID:26639519

  15. Crystallization and preliminary crystallographic studies of an active-site mutant hydantoin racemase from Sinorhizobium meliloti CECT4114

    International Nuclear Information System (INIS)

    Crystals of an active-site mutated hydantoin racemase from S. meliloti have been obtained in the presence and absence of d,l-5-isopropyl-hydantoin and characterized by X-ray diffraction. A recombinant active-site mutant of hydantoin racemase (C76A) from Sinorhizobium meliloti CECT 4114 (SmeHyuA) has been crystallized in the presence and absence of the substrate d,l-5-isopropyl hydantoin. Crystals of the SmeHyuA mutant suitable for data collection and structure determination were grown using the counter-diffusion method. X-ray data were collected to resolutions of 2.17 and 1.85 Å for the free and bound enzymes, respectively. Both crystals belong to space group R3 and contain two molecules of SmeHyuA per asymmetric unit. The crystals of the free and complexed SmeHyuA have unit-cell parameters a = b = 85.43, c = 152.37 Å and a = b = 85.69, c = 154.38 Å, crystal volumes per protein weight (VM) of 1.94 and 1.98 Å3 Da−1 and solvent contents of 36.7 and 37.9%, respectively

  16. Screening of Highly Effective Sinorhizobium meliloti Strains for 'Vector' Alfalfa and Testing of Its Competitive Nodulation Ability in the Field

    Institute of Scientific and Technical Information of China (English)

    ZENG Zhao-Hai; CHEN Wen-Xin; HU Yue-Gao; SUI Xin-Hua; CHEN Dan-Ming

    2007-01-01

    Seventeen Sinorhizobium meliloti strains from seven provinces in China were used to screen highly effective strains for alfalfa cultivar in a greenhouse study and their symbiotic relationship and competitive ability were studied in the field.CCBAU30138 was the most effective strain,as evidenced by increase in dry weights.A field experiment showed that the inoculation of alfalfa with CCBAU30138 resulted in increases of 11.9%and 19.6%of dry matter production and crude protein production,respectively,in forage of monocultured plants.The total dry matter yields of alfalfa and tall fescue in binary culture were increased by 16.3%by inoculation of alfalfa with this strain.These results showed that S.Meliloti strain CCBAU30138 was an effective inoculant both in the greenhouse and in the field.The analysis of randomly amplified polymorphic DNA(RAPD)by polymerase chain reaction(PCR)from nodule extracts showed that the strain CCBAU30138 had high competitiveness in the field.It occupied 47.5%of nodules in alfalfa monoculture and 44.4% of nodules in alfalfa-tall fescue binary culture after 20 weeks of growth.In conclusion,a simple system to select highly effective and competitive symbiotic strains specific to alfalfa was established.Using this system.A strain suitable for the alfalfa cultivar'Vector’grown in Wuqiao County of Hebei Province was obtained.

  17. oriT-Directed Cloning of Defined Large Regions from Bacterial Genomes: Identification of the Sinorhizobium meliloti pExo Megaplasmid Replicator Region

    OpenAIRE

    Patrick S G Chain; Hernandez-Lucas, Ismael; Golding, Brian; Finan, Turlough M.

    2000-01-01

    We have developed a procedure to directly clone large fragments from the genome of the soil bacterium Sinorhizobium meliloti. Specific regions to be cloned are first flanked by parallel copies of an origin of transfer (oriT) together with a plasmid replication origin capable of replicating large clones in Escherichia coli but not in the target organism. Supplying transfer genes in trans specifically transfers the oriT-flanked region, and in this process, site-specific recombination at the ori...

  18. The induction of Sinorhizobium meliloti C4-dicarboxylate transport system(Dct)is regulated by oxygen concentration

    Institute of Scientific and Technical Information of China (English)

    WEN Jin; NAN Beiyan; Fergal O'Gara; WANG Yiping

    2005-01-01

    The Sinorhizobium meliloti C4-dicarboxylate transport (Dct) system is essential for symbiotic nitrogen fixation. The dctA gene, encoding the C4-dicarboxylate permease, is expressed in both free living and symbiotic cells. But in free living cells expression of dctD and dctB is absolutely required for the expression of dctA. In this study, in order to investigate the effect of oxygen concentration on the induction of Dct system, E. coli DH5α strain which carries the plasmid-encoded dctABD operon was used in tube assays. It was found that the specific induction of Dct system occurred only at a certain depth under the surface of M63- 0.6% agar media, suggesting that Dct system could respond to oxygen concentration during succinate-induced expression. Furthermore, when measured at different oxygen concentrations, the highest expression level was observed at oxygen concentration of 2%. Thus, we predict that in addition to dicarboxylates, the induction of Dct system may also regulated by oxygen concentration.

  19. Analysis of the downstream region of nodD3 P1 promoter by deletion and complementation tests in Sinorhizobium meliloti

    Institute of Scientific and Technical Information of China (English)

    陈迪; 刘彦杰; 朱家璧; 沈善炯; 俞冠翘

    2003-01-01

    In Sinorhizobium meliloti, the nodD3 gene is transcriptionally controlled by two promoters, P1 and P2. Under P1, there is a 660 bp sequence including a small open reading frame, ORF2, followed by the nodD3 coding region. Genetic analysis using the different deletions on the 3′ends of P1 downstream sequence showed that the downstream sequence +1-+125nt is essential for P1 expression. Complementation, mutations and nodulation tests demonstrated that the ORF2 auto-represses P1 expression, while the P1 downstream sequence +1-+125nt counteracts it.

  20. The stress-related, rhizobial small RNA RcsR1 destabilizes the autoinducer synthase encoding mRNA sinI in Sinorhizobium meliloti.

    Science.gov (United States)

    Baumgardt, Kathrin; Šmídová, Klára; Rahn, Helen; Lochnit, Günter; Robledo, Marta; Evguenieva-Hackenberg, Elena

    2016-05-01

    Quorum sensing is a cell density-dependent communication system of bacteria relying on autoinducer molecules. During the analysis of the post-transcriptional regulation of quorum sensing in the nitrogen fixing plant symbiont Sinorhizobium meliloti, we predicted and verified a direct interaction between the 5'-UTR of sinI mRNA encoding the autoinducer synthase and a small RNA (sRNA), which we named RcsR1. In vitro, RcsR1 prevented cleavage in the 5'-UTR of sinI by RNase E and impaired sinI translation. In line with low ribosomal occupancy and transcript destabilization upon binding of RcsR1 to sinI, overproduction of RcsR1 in S. meliloti resulted in lower level and shorter half-life of sinI mRNA, and in decreased autoinducer amount. Although RcsR1 can influence quorum sensing via sinI, its level did not vary at different cell densities, but decreased under salt stress and increased at low temperature. We found that RcsR1 and its stress-related expression pattern, but not the interaction with sinI homologs, are conserved in Sinorhizobium, Rhizobium and Agrobacterium. Consistently, overproduction of RcsR1 in S. meliloti and Agrobacterium tumefaciens inhibited growth at high salinity. We identified conserved targets of RcsR1 and showed that most conserved interactions and the effect on growth under salt stress are mediated by the first stem-loop of RcsR1, while its central part is responsible for the species-specific interaction with sinI. We conclude that RcsR1 is an ancient, stress-related riboregulator in rhizobia and propose that it links stress responses to quorum sensing in S. meliloti. PMID:26588798

  1. Nodulation by Sinorhizobium meliloti originated from a mining soil alleviates Cd toxicity and increases Cd-phytoextraction in Medicago sativa L.

    Science.gov (United States)

    Ghnaya, Tahar; Mnassri, Majda; Ghabriche, Rim; Wali, Mariem; Poschenrieder, Charlotte; Lutts, Stanley; Abdelly, Chedly

    2015-01-01

    Besides their role in nitrogen supply to the host plants as a result of symbiotic N fixation, the association between legumes and Rhizobium could be useful for the rehabilitation of metal-contaminated soils by phytoextraction. A major limitation presents the metal-sensitivity of the bacterial strains. The aim of this work was to explore the usefulness of Sinorhizobium meliloti originated from a mining site for Cd phytoextraction by Medicago sativa. Inoculated and non-inoculated plants were cultivated for 60 d on soils containing 50 and/or 100 mg Cd kg−1 soil. The inoculation hindered the occurrence of Cd- induced toxicity symptoms that appeared in the shoots of non-inoculated plants. This positive effect of S. meliloti colonization was accompanied by an increase in biomass production and improved nutrient acquisition comparatively to non-inoculated plants. Nodulation enhanced Cd absorption by the roots and Cd translocation to the shoots. The increase of plant biomass concomitantly with the increase of Cd shoot concentration in inoculated plants led to higher potential of Cd-phytoextraction in these plants. In the presence of 50 mg Cd kg−1 in the soil, the amounts of Cd extracted in the shoots were 58 and 178 μg plant−1 in non-inoculated and inoculated plants, respectively. This study demonstrates that this association M. sativa-S. meliloti may be an efficient biological system to extract Cd from contaminated soils. PMID:26528320

  2. The tep1 gene of Sinorhizobium meliloti coding for a putative transmembrane efflux protein and N-acetyl glucosamine affect nod gene expression and nodulation of alfalfa plants

    Directory of Open Access Journals (Sweden)

    Soto María

    2009-01-01

    Full Text Available Abstract Background Soil bacteria collectively known as Rhizobium, characterized by their ability to establish beneficial symbiosis with legumes, share several common characteristics with pathogenic bacteria when infecting the host plant. Recently, it was demonstrated that a fadD mutant of Sinorhizobium meliloti is altered in the control of swarming, a type of co-ordinated movement previously associated with pathogenicity, and is also impaired in nodulation efficiency on alfalfa roots. In the phytopathogen Xanthomonas campestris, a fadD homolog (rpfB forms part of a cluster of genes involved in the regulation of pathogenicity factors. In this work, we have investigated the role in swarming and symbiosis of SMc02161, a S. meliloti fadD-linked gene. Results The SMc02161 locus in S. meliloti shows similarities with members of the Major Facilitator Superfamily (MFS of transporters. A S. meliloti null-mutant shows increased sensitivity to chloramphenicol. This indication led us to rename the locus tep1 for transmembrane efflux protein. The lack of tep1 does not affect the appearance of swarming motility. Interestingly, nodule formation efficiency on alfalfa plants is improved in the tep1 mutant during the first days of the interaction though nod gene expression is lower than in the wild type strain. Curiously, a nodC mutation or the addition of N-acetyl glucosamine to the wild type strain lead to similar reductions in nod gene expression as in the tep1 mutant. Moreover, aminosugar precursors of Nod factors inhibit nodulation. Conclusion tep1 putatively encodes a transmembrane protein which can confer chloramphenicol resistance in S. meliloti by expelling the antibiotic outside the bacteria. The improved nodulation of alfalfa but reduced nod gene expression observed in the tep1 mutant suggests that Tep1 transports compounds which influence nodulation. In contrast to Bradyrhizobium japonicum, we show that in S. meliloti there is no feedback regulation

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

    Directory of Open Access Journals (Sweden)

    Khatira Anwari

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

  4. A consolidated analysis of the physiologic and molecular responses induced under acid stress in the legume-symbiont model-soil bacterium Sinorhizobium meliloti.

    Science.gov (United States)

    Draghi, W O; Del Papa, M F; Hellweg, C; Watt, S A; Watt, T F; Barsch, A; Lozano, M J; Lagares, A; Salas, M E; López, J L; Albicoro, F J; Nilsson, J F; Torres Tejerizo, G A; Luna, M F; Pistorio, M; Boiardi, J L; Pühler, A; Weidner, S; Niehaus, K; Lagares, A

    2016-01-01

    Abiotic stresses in general and extracellular acidity in particular disturb and limit nitrogen-fixing symbioses between rhizobia and their host legumes. Except for valuable molecular-biological studies on different rhizobia, no consolidated models have been formulated to describe the central physiologic changes that occur in acid-stressed bacteria. We present here an integrated analysis entailing the main cultural, metabolic, and molecular responses of the model bacterium Sinorhizobium meliloti growing under controlled acid stress in a chemostat. A stepwise extracellular acidification of the culture medium had indicated that S. meliloti stopped growing at ca. pH 6.0-6.1. Under such stress the rhizobia increased the O2 consumption per cell by more than 5-fold. This phenotype, together with an increase in the transcripts for several membrane cytochromes, entails a higher aerobic-respiration rate in the acid-stressed rhizobia. Multivariate analysis of global metabolome data served to unequivocally correlate specific-metabolite profiles with the extracellular pH, showing that at low pH the pentose-phosphate pathway exhibited increases in several transcripts, enzymes, and metabolites. Further analyses should be focused on the time course of the observed changes, its associated intracellular signaling, and on the comparison with the changes that operate during the sub lethal acid-adaptive response (ATR) in rhizobia. PMID:27404346

  5. Sinorhizobium meliloti sigma factors RpoE1 and RpoE4 are activated in stationary phase in response to sulfite.

    Directory of Open Access Journals (Sweden)

    Bénédicte Bastiat

    Full Text Available Rhizobia are soil bacteria able to establish a nitrogen-fixing symbiosis with legume plants. Both in soil and in planta, rhizobia spend non-growing periods resembling the stationary phase of in vitro-cultured bacteria. The primary objective of this work was to better characterize gene regulation in this biologically relevant growth stage in Sinorhizobium meliloti. By a tap-tag/mass spectrometry approach, we identified five sigma factors co-purifying with the RNA polymerase in stationary phase: the general stress response regulator RpoE2, the heat shock sigma factor RpoH2, and three extra-cytoplasmic function sigma factors (RpoE1, RpoE3 and RpoE4 belonging to the poorly characterized ECF26 subgroup. We then showed that RpoE1 and RpoE4 i are activated upon metabolism of sulfite-generating compounds (thiosulfate and taurine, ii display overlapping regulatory activities, iii govern a dedicated sulfite response by controlling expression of the sulfite dehydrogenase SorT, iv are activated in stationary phase, likely as a result of endogenous sulfite generation during bacterial growth. We showed that SorT is required for optimal growth of S. meliloti in the presence of sulfite, suggesting that the response governed by RpoE1 and RpoE4 may be advantageous for bacteria in stationary phase either by providing a sulfite detoxification function or by contributing to energy production through sulfite respiration. This paper therefore reports the first characterization of ECF26 sigma factors, the first description of sigma factors involved in control of sulphur metabolism, and the first indication that endogenous sulfite may act as a signal for regulation of gene expression upon entry of bacteria in stationary phase.

  6. Comparative toxicity assessment of CeO{sub 2} and ZnO nanoparticles towards Sinorhizobium meliloti, a symbiotic alfalfa associated bacterium: Use of advanced microscopic and spectroscopic techniques

    Energy Technology Data Exchange (ETDEWEB)

    Bandyopadhyay, Susmita [Environmental Science and Engineering PhD Program, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); University of California Center for Environmental Implications of Nanotechnology (UC CEIN), The University of Texas at El Paso (United States); Peralta-Videa, Jose R. [Department of Chemistry, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); Plascencia-Villa, German; Jose-Yacaman, Miguel [Department of Physics and Astronomy, The University of Texas at San Antonio, One UTSA Circle, San Antonio, TX 78249 (United States); Gardea-Torresdey, Jorge L., E-mail: jgardea@utep.edu [Environmental Science and Engineering PhD Program, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); Department of Chemistry, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX 79968 (United States); University of California Center for Environmental Implications of Nanotechnology (UC CEIN), The University of Texas at El Paso (United States)

    2012-11-30

    Highlights: Black-Right-Pointing-Pointer First cytotoxicity study of CeO{sub 2} and ZnO nanoparticles to Sinorhizobium meliloti. Black-Right-Pointing-Pointer First report upon the mechanisms of CeO{sub 2} and ZnO NPs toxicity to S. meliloti. Black-Right-Pointing-Pointer ZnO NPs were found to be bactericidal in lower concentration. Black-Right-Pointing-Pointer CeO{sub 2} NPs had bacteriostatic effect on S. meliloti. - Abstract: Cerium oxide (CeO{sub 2}) and zinc oxide (ZnO) nanoparticles (NPs) are extensively used in a variety of instruments and consumer goods. These NPs are of great concern because of potential toxicity towards human health and the environment. The present work aimed to assess the toxic effects of 10 nm CeO{sub 2} and ZnO NPs towards the nitrogen fixing bacterium Sinorhizobium meliloti. Toxicological parameters evaluated included UV/Vis measurement of minimum inhibitory concentration, disk diffusion tests, and dynamic growth. Ultra high-resolution scanning transmission electron microscopy (STEM) and infrared spectroscopy (FTIR) were utilized to determine the spatial distribution of NPs and macromolecule changes in bacterial cells, respectively. Results indicate that ZnO NPs were more toxic than CeO{sub 2} NPs in terms of inhibition of dynamic growth and viable cells counts. STEM images revealed that CeO{sub 2} and ZnO NPs were found on bacterial cell surfaces and ZnO NPs were internalized into the periplasmic space of the cells. FTIR spectra showed changes in protein and polysaccharide structures of extra cellular polymeric substances present in bacterial cell walls treated with both NPs. The growth data showed that CeO{sub 2} NPs have a bacteriostatic effect, whereas ZnO NPs is bactericidal to S. meliloti. Overall, ZnO NPs were found to be more toxic than CeO{sub 2} NPs.

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

    Directory of Open Access Journals (Sweden)

    Menck Carlos FM

    2007-03-01

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

  8. La simbiosis fijadora de nitrógeno Sinorhizobium meliloti-alfalfa: aproximaciones ómicas aplicadas a la identificación y caracterización de determinantes genéticos del rizobio asociados a la colonización temprana de la raíz de alfalfa (Medicago sativa)

    OpenAIRE

    Salas, María Eugenia

    2015-01-01

    Sinorhizobium meliloti es una α-proteobacteria capaz de establecer asociaciones simbióticas con plantas de los géneros Medicago, Melilotus y Trigonella. Esta asociación es el resultado de un complejo diálogo molecular entre los simbiontes, que se diferencian a lo largo de la interacción para dar lugar a un nuevo órgano en las raíces de las plantas, el nódulo fijador de nitrógeno. El nicho simbiótico accesible a los rizobios está naturalmente limitado, y resulta ocupado por aquellas cepas que ...

  9. Denitrification by Rhizobium meliloti

    Energy Technology Data Exchange (ETDEWEB)

    Rosen, A.

    1996-10-01

    Rhizobium meliloti strains were investigated for their denitrification activity as free-living cells and in nodules on lucerne (Medicago sativa) roots. They were also investigated for presence of nitrous oxide reductase (nos) activity and for genes using a nosZ probe derived from the Pseudomonas stutzeri. To decide whether R. meliloti strains used as inoculants contribute to the total denitrification activity in a lucerne ley, strains with different denitrifying capacities were used in field and laboratory experiments. The nitrate reduction activity of R. meliloti during anaerobic respiration was compared with that of a strain of Pseudomonas aeruginosa. A great diversity in the denitrification activity was found within strains of R. meliloti, and four of thirteen investigated strains showed an obvious denitrification activity. Two denitrifying bacteria were used as references, one strain each of Bradyrhizobium japonicum and P. aeruginosa. All but one of the R. meliloti strains hybridized to the PstI-fragment of the nosZ-gene from P. stutzeri. Two sizes of the hybridizing fragment, 5 and 7 kb, were noticed. Nos activity was only shown in three R. meliloti strains, and these were all characterized by a high denitrification activity. The potential denitrification activity was about 20, 40, and 80 times higher than the actual denitrification activity for lucerne, fallow, and grass, respectively. The potential denitrification activity was almost the same in lucerne and grass planted soils. Compared with the unplanted soil, the presence of lucerne roots in the soil increased the actual denitrification activity, while roots of both plant species, grass and lucerne, increased the potential denitrification activity in the soil. 32 refs, 7 figs, 1 tab

  10. Sinorhizobium fredii HH103 bacteroids are not terminally differentiated and show altered O-antigen in nodules of the Inverted Repeat-Lacking Clade legume Glycyrrhiza uralensis.

    Science.gov (United States)

    Crespo-Rivas, Juan C; Guefrachi, Ibtissem; Mok, Kenny C; Villaécija-Aguilar, José A; Acosta-Jurado, Sebastián; Pierre, Olivier; Ruiz-Sainz, José E; Taga, Michiko E; Mergaert, Peter; Vinardell, José M

    2016-09-01

    In rhizobial species that nodulate inverted repeat-lacking clade (IRLC) legumes, such as the interaction between Sinorhizobium meliloti and Medicago, bacteroid differentiation is driven by an endoreduplication event that is induced by host nodule-specific cysteine rich (NCR) antimicrobial peptides and requires the participation of the bacterial protein BacA. We have studied bacteroid differentiation of Sinorhizobium fredii HH103 in three host plants: Glycine max, Cajanus cajan and the IRLC legume Glycyrrhiza uralensis. Flow cytometry, microscopy analyses and viability studies of bacteroids as well as confocal microscopy studies carried out in nodules showed that S. fredii HH103 bacteroids, regardless of the host plant, had deoxyribonucleic acid (DNA) contents, cellular sizes and survival rates similar to those of free-living bacteria. Contrary to S. meliloti, S. fredii HH103 showed little or no sensitivity to Medicago NCR247 and NCR335 peptides. Inactivation of S. fredii HH103 bacA neither affected symbiosis with Glycyrrhiza nor increased bacterial sensitivity to Medicago NCRs. Finally, HH103 bacteroids isolated from Glycyrrhiza, but not those isolated from Cajanus or Glycine, showed an altered lipopolysaccharide. Our studies indicate that, in contrast to the S. meliloti-Medicago model symbiosis, bacteroids in the S. fredii HH103-Glycyrrhiza symbiosis do not undergo NCR-induced and bacA-dependent terminal differentiation. PMID:26521863

  11. Three way interactions between Thymus vulgaris, Medicago truncatula and Sinorhizobium meliloti

    DEFF Research Database (Denmark)

    Grøndahl, Eva; Ehlers, Bodil Kirstine

    2012-01-01

    Thymus vulgaris is a dominating component of the Mediterranean garrigue vegetation. It produces aromatic oil, containing monoterpenes, which affects the performance (growth, survival) of other plants, and microorganisms. Annual plant species of the genus Medicago are commonly found in Mediterranean...

  12. Mixed Nodule Infection in Sinorhizobium meliloti–Medicago sativa Symbiosis Suggest the Presence of Cheating Behavior

    Science.gov (United States)

    Checcucci, Alice; Azzarello, Elisa; Bazzicalupo, Marco; Galardini, Marco; Lagomarsino, Alessandra; Mancuso, Stefano; Marti, Lucia; Marzano, Maria C.; Mocali, Stefano; Squartini, Andrea; Zanardo, Marina; Mengoni, Alessio

    2016-01-01

    In the symbiosis between rhizobia and legumes, host plants can form symbiotic root nodules with multiple rhizobial strains, potentially showing different symbiotic performances in nitrogen fixation. Here, we investigated the presence of mixed nodules, containing rhizobia with different degrees of mutualisms, and evaluate their relative fitness in the Sinorhizobium meliloti–Medicago sativa model symbiosis. We used three S. meliloti strains, the mutualist strains Rm1021 and BL225C and the non-mutualist AK83. We performed competition experiments involving both in vitro and in vivo symbiotic assays with M. sativa host plants. We show the occurrence of a high number (from 27 to 100%) of mixed nodules with no negative effect on both nitrogen fixation and plant growth. The estimation of the relative fitness as non-mutualist/mutualist ratios in single nodules shows that in some nodules the non-mutualist strain efficiently colonized root nodules along with the mutualist ones. In conclusion, we can support the hypothesis that in S. meliloti–M. sativa symbiosis mixed nodules are formed and allow non-mutualist or less-mutualist bacterial partners to be less or not sanctioned by the host plant, hence allowing a potential form of cheating behavior to be present in the nitrogen fixing symbiosis. PMID:27379128

  13. A highly conserved protein of unknown function in Sinorhizobium meliloti affects sRNA regulation similar to Hfq

    OpenAIRE

    Pandey, Shree P.; Minesinger, Brenda K.; Kumar, Janesh; Walker, Graham C.

    2011-01-01

    The SMc01113/YbeY protein, belonging to the UPF0054 family, is highly conserved in nearly every bacterium. However, the function of these proteins still remains elusive. Our results show that SMc01113/YbeY proteins share structural similarities with the MID domain of the Argonaute (AGO) proteins, and might similarly bind to a small-RNA (sRNA) seed, making a special interaction with the phosphate on the 5′-side of the seed, suggesting they may form a component of the bacterial sRNA pathway. In...

  14. A FIELD STUDY WITH GENETICALLY ENGINEERED ALFALFA INOCULATED WITH RECOMBINANT SINORHIZOBIUM MELILOTI: EFFECTS ON THE SOIL ECOSYSTEM

    Science.gov (United States)

    The agricultural use of genetically engineered plants and microorganisms has become increasingly common. Because genetically engineered plants and microorganisms can produce compounds foreign to their environment, there is concern that they may become established outside of thei...

  15. Cloning and Identification of Ammonia Monooxygenase Gene of Sinorhizobium sp. NP1%中华根瘤菌NP1氨单加氧酶基因的克隆与功能鉴定

    Institute of Scientific and Technical Information of China (English)

    刘钰莹; 邱枫; 宋琴; 窦鑫; 许雷

    2010-01-01

    以中华根瘤菌NP1(Sinorhizobium sp.NP1)为原始菌株,通过同源克隆与Tail-PCR方法,获得1 089 bp的氨单加氧酶基因(amo)全长序列.该基因编码362个氨基酸,其二级结构与Sinorhizobium meliloti 1021 AMO的二级结构相似,该蛋白有9个跨膜区段.以自杀穿梭质粒pJQ200SK为原始载体,构建NP1 amo基因敲除质粒pJQ200SK-amo-Tc.采用三亲本杂交的方法将该质粒转入原始菌株NP1中,获得amo基因敲除菌株NP1∷amo.通过本贝洛氏(Berthelot)法对氨氮进行测定,发现NP1∷amo的脱氮效率比原始菌株NP1下降约35%.该结果表明,本实验中所克隆的氨单加氧酶基因为脱氮关键酶基因.

  16. Proteome Analysis of Inhibitory Effect of Gadolinium on Sinorhizobium fredii

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The inhibitory effect of gadolinium on Sinorhizobium fredii USDA 205 was studied on a global scale using two-dimensional gel electrophoresis and MALDI-TOF MS. The results indicated that 22 proteins were significantly affected by 1 mmol·L-1 Gd3+ treatment when compared with an untreated control. Among these proteins, nine were up-regulated and thirteen were down-regulated. The differently expressed proteins were classified into 8 functional categories based on their functions, including transporters, proteins for cellular defence, and proteins involved in metabolism.

  17. Alfalfa microsymbionts from different ITS and nodC lineages of Ensifer meliloti and Ensifer medicae symbiovar meliloti establish efficient symbiosis with alfalfa in Spanish acid soils.

    Science.gov (United States)

    Ramírez-Bahena, Martha-Helena; Vargas, Margarita; Martín, María; Tejedor, Carmen; Velázquez, Encarna; Peix, Álvaro

    2015-06-01

    Alfalfa (Medicago sativa L.) is an important crop worldwide whose cropping in acid soils is hampered by the poor nodulation and yield commonly attributed to the sensitivity of its endosymbionts to acid pH. In this work, we isolated several acid-tolerant strains from alfalfa nodules in three acid soils in northwestern Spain. After grouping by RAPD fingerprinting, most strains were identified as Ensifer meliloti and only two strains as Ensifer medicae according to their 16S-23S intergenic spacer (ITS) sequences that allowed the differentiation of two groups within each one of these species. The two ITS groups of E. meliloti and the ITS group I of E. medicae have been previously found in Medicago nodules; however, the group II of E. medicae has been only found to date in Prosopis alba nodules. The analysis of the nodC gene showed that all strains isolated in this study belong to the symbiovar meliloti, grouping with the type strains of E. meliloti or E. medicae, but some harboured nodC gene alleles different from those found to date in alfalfa nodules. The strains of E. medicae belong to the symbiovar meliloti which should be also recognised in this species, although they harboured a nodC allele phylogenetically divergent to those from E. meliloti strains. Microcosm experiments showed that inoculation of alfalfa with selected acid-tolerant strains significantly increased yields in acid soils representing a suitable agricultural practice for alfalfa cropping in these soils. PMID:25586575

  18. Biochemical characterization of a fructokinase mutant of Rhizobium meliloti.

    OpenAIRE

    Gardiol, A; Arias, A.; Cerveñansky, C; Gaggero, C; Martínez-Drets, G

    1980-01-01

    A double mutant strain (UR3) of Rhizobium meliloti L5-30 was isolated from a phosphoglucose isomerase mutant (UR1) on the basis of its resistance to fructose inhibition when grown on fructose-rich medium. UR3 lacked both phosphoglucose isomerase and fructokinase activity. A mutant strain (UR4) lacking only the fructokinase activity was derived from UR3; it grew on the same carbon sources as the parent strain, but not on fructose, mannitol, or sorbitol. A spontaneous revertant (UR5) of normal ...

  19. Bacterial Molecular Signals in the Sinorhizobium fredii-Soybean Symbiosis

    Directory of Open Access Journals (Sweden)

    Francisco J. López-Baena

    2016-05-01

    Full Text Available Sinorhizobium (Ensifer fredii (S. fredii is a rhizobial species exhibiting a remarkably broad nodulation host-range. Thus, S. fredii is able to effectively nodulate dozens of different legumes, including plants forming determinate nodules, such as the important crops soybean and cowpea, and plants forming indeterminate nodules, such as Glycyrrhiza uralensis and pigeon-pea. This capacity of adaptation to different symbioses makes the study of the molecular signals produced by S. fredii strains of increasing interest since it allows the analysis of their symbiotic role in different types of nodule. In this review, we analyze in depth different S. fredii molecules that act as signals in symbiosis, including nodulation factors, different surface polysaccharides (exopolysaccharides, lipopolysaccharides, cyclic glucans, and K-antigen capsular polysaccharides, and effectors delivered to the interior of the host cells through a symbiotic type 3 secretion system.

  20. Genetic regulation of nitrogen fixation in Rhizobium meliloti.

    Science.gov (United States)

    Cebolla, A; Palomares, A J

    1994-12-01

    The soil bacterium Rhizobium meliloti fixes dinitrogen when associated with root nodules formed on its plant host, Medicago sativa (alfalfa). The expression of most of the known genes required for nitrogen fixation (nif and fix genes), including the structural genes for nitrogenase, is induced in response to a decrease in oxygen concentration. Induction of nif and fix gene expression by low oxygen is physiologically relevant because a low-oxygen environment is maintained in root nodules to prevent inactivation of the highly oxygen-sensitive nitrogenase enzyme. The genes responsible for sensing and transducing the low oxygen signal, fixL and fixJ, encode proteins (FixL and FixJ, respectively) that are homologous to a large family of bacterial proteins involved in signal transduction, the two component regulatory system proteins. The two components consist of a sensor protein, to which FixL is homologous, and a response regulator protein, to which FixJ is homologous. The sensor protein respond to an activating signal by autophosphorylating and then transferring the phosphate to its cognate response regulator protein. The phosphorylated response regulator, which is often a transcriptional activator, is then able to activate its target. A cascade model of nif and fix gene regulation in R. meliloti has been proposed, whereby FixL acts as an oxygen sensor as the initial event in the cascade and transmits this information to FixJ. FixJ, which possesses a putative helix-turn-helix DNA-binding motif, then activates transcription of the nifA and fixK genes. The nifA and fixK gene products, are transcriptional activators of at least 14 other nif and fix genes.

  1. Structure and Biological Roles of Sinorhizobium fredii HH103 Exopolysaccharide

    Science.gov (United States)

    Acosta-Jurado, Sebastián; Soto, María J.; Margaret, Isabel; Crespo-Rivas, Juan C.; Sanjuan, Juan; Temprano, Francisco; Gil-Serrano, Antonio; Ruiz-Sainz, José E.; Vinardell, José M.

    2014-01-01

    Here we report that the structure of the Sinorhizobium fredii HH103 exopolysaccharide (EPS) is composed of glucose, galactose, glucuronic acid, pyruvic acid, in the ratios 5∶2∶2∶1 and is partially acetylated. A S. fredii HH103 exoA mutant (SVQ530), unable to produce EPS, not only forms nitrogen fixing nodules with soybean but also shows increased competitive capacity for nodule occupancy. Mutant SVQ530 is, however, less competitive to nodulate Vigna unguiculata. Biofilm formation was reduced in mutant SVQ530 but increased in an EPS overproducing mutant. Mutant SVQ530 was impaired in surface motility and showed higher osmosensitivity compared to its wild type strain in media containing 50 mM NaCl or 5% (w/v) sucrose. Neither S. fredii HH103 nor 41 other S. fredii strains were recognized by soybean lectin (SBL). S. fredii HH103 mutants affected in exopolysaccharides (EPS), lipopolysaccharides (LPS), cyclic glucans (CG) or capsular polysaccharides (KPS) were not significantly impaired in their soybean-root attachment capacity, suggesting that these surface polysaccharides might not be relevant in early attachment to soybean roots. These results also indicate that the molecular mechanisms involved in S. fredii attachment to soybean roots might be different to those operating in Bradyrhizobium japonicum. PMID:25521500

  2. Structure and biological roles of Sinorhizobium fredii HH103 exopolysaccharide.

    Science.gov (United States)

    Rodríguez-Navarro, Dulce N; Rodríguez-Carvajal, Miguel A; Acosta-Jurado, Sebastián; Soto, María J; Margaret, Isabel; Crespo-Rivas, Juan C; Sanjuan, Juan; Temprano, Francisco; Gil-Serrano, Antonio; Ruiz-Sainz, José E; Vinardell, José M

    2014-01-01

    Here we report that the structure of the Sinorhizobium fredii HH103 exopolysaccharide (EPS) is composed of glucose, galactose, glucuronic acid, pyruvic acid, in the ratios 5∶2∶2∶1 and is partially acetylated. A S. fredii HH103 exoA mutant (SVQ530), unable to produce EPS, not only forms nitrogen fixing nodules with soybean but also shows increased competitive capacity for nodule occupancy. Mutant SVQ530 is, however, less competitive to nodulate Vigna unguiculata. Biofilm formation was reduced in mutant SVQ530 but increased in an EPS overproducing mutant. Mutant SVQ530 was impaired in surface motility and showed higher osmosensitivity compared to its wild type strain in media containing 50 mM NaCl or 5% (w/v) sucrose. Neither S. fredii HH103 nor 41 other S. fredii strains were recognized by soybean lectin (SBL). S. fredii HH103 mutants affected in exopolysaccharides (EPS), lipopolysaccharides (LPS), cyclic glucans (CG) or capsular polysaccharides (KPS) were not significantly impaired in their soybean-root attachment capacity, suggesting that these surface polysaccharides might not be relevant in early attachment to soybean roots. These results also indicate that the molecular mechanisms involved in S. fredii attachment to soybean roots might be different to those operating in Bradyrhizobium japonicum. PMID:25521500

  3. A 13C-NMR study of exopolysaccharide synthesis in Rhizobium meliloti Su47 strain

    Science.gov (United States)

    Tavernier, P.; Portais, J.-C.; Besson, I.; Courtois, J.; Courtois, B.; Barbotin, J.-N.

    1998-02-01

    Metabolic pathways implied in the synthesis of succinoglycan produced by the Su47 strain of R. meliloti were evaluated by 13C-NMR spectroscopy after incubation with [1{-}13C] or [2{-}13C] glucose. The biosynthesis of this polymer by R. meliloti from glucose occurred by a direct polymerisation of the introduced glucose and by the pentose phosphate pathway. Les voies métaboliques impliquées dans la synthèse du succinoglycane produit par la souche Su47 de R. meliloti ont été évaluées par la spectroscopie de RMN du carbone 13 après incubation des cellules avec du [1{-}13C] ou [2{-}13C] glucose. La biosynthèse de ce polymère à partir du glucose se produit par polymérisation directe du glucose et par la voie des pentoses phosphate.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-01-01

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

  5. Inhibition of nodule development by multicopy promoters of Rhizobium meliloti nif/fix genes

    Institute of Scientific and Technical Information of China (English)

    吴桐; 朱家璧; 俞冠翘; 沈善炯

    1995-01-01

    Using luc gene as a reporter to study the activation of Rhizobium meliloti nif/fix genes in thedevelopment of symbiosis,the authors observed that nodule development and nitrogen fixation were inhibitedby both multicopy promoters of nifHDK and fixABCX.The phenotype of R.meliloti containing multicopynif/fix promoters appeared exactly like that of nifA mutant.Using lacZ as a reporter,the authors got the same re-sults.By contrast,the rhizobia containing low-copy promoters of nif/fix genes were normal fornodule development and nitrogen fixation.These results substantiate the evidence that the product of nifAgene not only acts as a transcriptional activator of nif/fix genes,but also plays an important role in thedevelopment of root nodules.

  6. Isolation and characterization of Rhizobium meliloti mutants affected in exopolysaccharide production.

    Science.gov (United States)

    Rodríguez-Navarro, D N; Palomares, A J; Casadesús, J

    1991-06-01

    Rhizobium meliloti mutants affected in the production of exopolysaccharide (EPS) were isolated after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. The mutants were classified into three phenotypic classes: (I) Exo-, rough mutants lacking exopolysaccharide; (II) Exos (for "small") which form tiny, compact colonies and synthesize reduced amounts of EPS; and (III) Exoc (for "constitutive"), hypermucoid mutants which overproduce EPS. Hypermucoid strains showed increased resistance to desiccation. All the mutants were able to nodulate, although a significant decrease in infectivity degree and/or competitiveness was found in rough and compact strains. Two mutants proved to be deficient in nitrogen fixation. Complementation analysis with cloned R. meliloti exo genes could not be applied to the study of these Fix- mutants because introduction of plasmids derived from cosmid vector pLAFR1 caused loss of nodulating ability. However, complementation of calcofluor staining and EPS production was observed. Complementation with certain exo genes also caused a marked increase in motility.

  7. Anoxic growth of Ensifer meliloti 1021 by N2O-reduction, a potential mitigation strategy

    OpenAIRE

    Bueno, Emilio; Mania, Daniel; Frostegard, Ǻsa; Bedmar, Eulogio J.; Bakken, Lars R.; Delgado, Maria J.

    2015-01-01

    Denitrification in agricultural soils is a major source of N2O. Legume crops enhance N2O emission by providing N-rich residues, thereby stimulating denitrification, both by free-living denitrifying bacteria and by the symbiont (rhizobium) within the nodules. However, there are limited data concerning N2O production and consumption by endosymbiotic bacteria associated with legume crops. It has been reported that the alfalfa endosymbiont Ensifer meliloti strain 1021, despite possessing and expr...

  8. Functional nodFE genes are present in Sinorhizobium sp. strain MUS10, a symbiont of tropical legume Sesbania rostrata

    Science.gov (United States)

    Sinorhizobium sp. strain MUS10, a rhizobium from the Indian subcontinent, forms nitrogen-fixing nodules on the stems and roots of tropical legume Sesbania rostrata. The structure of Nod factors (NFs) of MUS10 are similar to those of Azorhizobium caulinodans, S. saheli bv sesbaniae and S. terangae bv...

  9. Erwinia herbicola isolates from alfalfa plants may play a role in nodulation of alfalfa by Rhizobium meliloti.

    OpenAIRE

    Handelsman, J; Brill, W J

    1985-01-01

    Erwinia herbicola was isolated from roots of plants derived from surface-sterilized seeds of all alfalfa varieties that were tested. Some of these E. herbicola strains affected nodulation by certain strains of Rhizobium meliloti. In previously published work we presented the isolation of slow-and fast-nodulating variants from a single culture of R. meliloti 102F51. In the absence of E. herbicola, the slow-nodulating variant induced the formation of nodules on alfalfa as rapidly as the faster-...

  10. Anoxic growth of Ensifer meliloti 1021 by N2O-reduction, a potential mitigation strategy

    Science.gov (United States)

    Bueno, Emilio; Mania, Daniel; Frostegard, Ǻsa; Bedmar, Eulogio J.; Bakken, Lars R.; Delgado, Maria J.

    2015-01-01

    Denitrification in agricultural soils is a major source of N2O. Legume crops enhance N2O emission by providing N-rich residues, thereby stimulating denitrification, both by free-living denitrifying bacteria and by the symbiont (rhizobium) within the nodules. However, there are limited data concerning N2O production and consumption by endosymbiotic bacteria associated with legume crops. It has been reported that the alfalfa endosymbiont Ensifer meliloti strain 1021, despite possessing and expressing the complete set of denitrification enzymes, is unable to grow via nitrate respiration under anoxic conditions. In the present study, we have demonstrated by using a robotized incubation system that this bacterium is able to grow through anaerobic respiration of N2O to N2. N2O reductase (N2OR) activity was not dependent on the presence of nitrogen oxyanions or NO, thus the expression could be induced by oxygen depletion alone. When incubated at pH 6, E. meliloti was unable to reduce N2O, corroborating previous observations found in both, extracted soil bacteria and Paracoccus denitrificans pure cultures, where expression of functional N2O reductase is difficult at low pH. Furthermore, the presence in the medium of highly reduced C-substrates, such as butyrate, negatively affected N2OR activity. The emission of N2O from soils can be lowered if legumes plants are inoculated with rhizobial strains overexpressing N2O reductase. This study demonstrates that strains like E. meliloti 1021, which do not produce N2O but are able to reduce the N2O emitted by other organisms, could act as even better N2O sinks. PMID:26074913

  11. Anoxic growth of Ensifer meliloti 1021 by N2O-reduction, a potential mitigation strategy

    Directory of Open Access Journals (Sweden)

    Emilio eBueno

    2015-05-01

    Full Text Available Denitrification in agricultural soils is a major source of N2O. Legume crops enhance N2O emission by providing N-rich residues, thereby stimulating denitrification, both by free-living denitrifying bacteria and by the symbiont (rhizobium within the nodules. However, there are limited data concerning N2O production and consumption by endosymbiotic bacteria associated with legume crops. It has been reported that the alfalfa endosymbiont Ensifer meliloti strain 1021, despite possessing and expressing the complete set of denitrification enzymes, is unable to grow via nitrate respiration under anoxic conditions. In the present study, we have demonstrated by using a robotized incubation system that this bacterium is able to grow through anaerobic respiration of N2O to N2. N2O reductase (N2OR activity was not dependent on the presence of nitrogen oxyanions or NO, thus the expression could be induced by oxygen depletion alone. When incubated at pH 6, E. meliloti was unable to reduce N2O, corroborating previous observations found in both, extracted soil bacteria and Paracoccus denitrificans pure cultures, where expression of functional N2O reductase is difficult at low pH. Furthermore, the presence in the medium of highly reduced C-substrates, such as butyrate, negatively affected N2OR activity. The emission of N2O from soils can be lowered if legumes plants are inoculated with rhizobial strains overexpressing N2O reductase. This study demonstrates that strains like E. meliloti 1021, which do not produce N2O but are able to reduce the N2O emitted by other organisms, could act as even better N2O sinks.

  12. Anoxic growth of Ensifer meliloti 1021 by N2O-reduction, a potential mitigation strategy.

    Science.gov (United States)

    Bueno, Emilio; Mania, Daniel; Frostegard, Ǻsa; Bedmar, Eulogio J; Bakken, Lars R; Delgado, Maria J

    2015-01-01

    Denitrification in agricultural soils is a major source of N2O. Legume crops enhance N2O emission by providing N-rich residues, thereby stimulating denitrification, both by free-living denitrifying bacteria and by the symbiont (rhizobium) within the nodules. However, there are limited data concerning N2O production and consumption by endosymbiotic bacteria associated with legume crops. It has been reported that the alfalfa endosymbiont Ensifer meliloti strain 1021, despite possessing and expressing the complete set of denitrification enzymes, is unable to grow via nitrate respiration under anoxic conditions. In the present study, we have demonstrated by using a robotized incubation system that this bacterium is able to grow through anaerobic respiration of N2O to N2. N2O reductase (N2OR) activity was not dependent on the presence of nitrogen oxyanions or NO, thus the expression could be induced by oxygen depletion alone. When incubated at pH 6, E. meliloti was unable to reduce N2O, corroborating previous observations found in both, extracted soil bacteria and Paracoccus denitrificans pure cultures, where expression of functional N2O reductase is difficult at low pH. Furthermore, the presence in the medium of highly reduced C-substrates, such as butyrate, negatively affected N2OR activity. The emission of N2O from soils can be lowered if legumes plants are inoculated with rhizobial strains overexpressing N2O reductase. This study demonstrates that strains like E. meliloti 1021, which do not produce N2O but are able to reduce the N2O emitted by other organisms, could act as even better N2O sinks. PMID:26074913

  13. Identification of salt-tolerant Sinorhizobium sp. strain BL3 membrane proteins based on proteomics

    DEFF Research Database (Denmark)

    Tanthanuch, Waraporn; Tittabutr, Panlada; Mohammed, Shabaz;

    2010-01-01

    Sinorhizobium sp. BL3 is a salt-tolerant strain that can fix atmospheric nitrogen in symbiosis with leguminous host plants under salt-stress conditions. Since cell membranes are the first barrier to environmental change, it is interesting to explore the membrane proteins within this protective...... functional categories, the two biggest of which were energy production and conversion, and proteins not in clusters of orthologous groups (COGs). In addition, a comparative analysis of membrane proteins between salt-stressed and non-stressed BL3 cells was conducted using a membrane enrichment method and off...... barrier under salt stress. The protein contents of membrane-enriched fractions obtained from BL3 were analyzed by nanoflow liquid chromatography interfaced with electrospray ionization tandem mass spectrometry. A total of 105 membrane proteins were identified. These proteins could be classified into 17...

  14. Regulation of symbiotic nitrogen fixation in root nodules of alfalfa (Medicago sativa) infected with Rhizobium meliloti.

    Science.gov (United States)

    Kamberger, W

    1977-10-24

    Symbiotic nitrogen fixation of Rhizobium meliloti bacteroids in Medicago sativa root nodules was suppressed by several inorganic nitrogen sources. Amino acids like glutamine, glutamic acid and aspartic acid, which can serve as sole nitrogen sources for the unnodulated plant did not influence nitrogenase activity of effective nodules, even at high concetrations. Ammonia and nitrate suppressed symbiotic nitrogen fixation in vivo only at concentrations much higher than those needed for suppression of nitrogenase activity in free living nitrogen fixing bacteria. The kinetics of suppression were slow compared with that of free living nitrogen fixing bacteria. On the other hand, nitrite, which acts as a direct inhibitor of nitrogenase, suppressed very quickly and at low concentrations. Glutamic acid and glutamine enhanced the effect of ammonia dramatically, while the suppression by nitrate was enhanced only slightly.

  15. Genetic and computational identification of a conserved bacterial metabolic module.

    Directory of Open Access Journals (Sweden)

    Cara C Boutte

    2008-12-01

    Full Text Available We have experimentally and computationally defined a set of genes that form a conserved metabolic module in the alpha-proteobacterium Caulobacter crescentus and used this module to illustrate a schema for the propagation of pathway-level annotation across bacterial genera. Applying comprehensive forward and reverse genetic methods and genome-wide transcriptional analysis, we (1 confirmed the presence of genes involved in catabolism of the abundant environmental sugar myo-inositol, (2 defined an operon encoding an ABC-family myo-inositol transmembrane transporter, and (3 identified a novel myo-inositol regulator protein and cis-acting regulatory motif that control expression of genes in this metabolic module. Despite being encoded from non-contiguous loci on the C. crescentus chromosome, these myo-inositol catabolic enzymes and transporter proteins form a tightly linked functional group in a computationally inferred network of protein associations. Primary sequence comparison was not sufficient to confidently extend annotation of all components of this novel metabolic module to related bacterial genera. Consequently, we implemented the Graemlin multiple-network alignment algorithm to generate cross-species predictions of genes involved in myo-inositol transport and catabolism in other alpha-proteobacteria. Although the chromosomal organization of genes in this functional module varied between species, the upstream regions of genes in this aligned network were enriched for the same palindromic cis-regulatory motif identified experimentally in C. crescentus. Transposon disruption of the operon encoding the computationally predicted ABC myo-inositol transporter of Sinorhizobium meliloti abolished growth on myo-inositol as the sole carbon source, confirming our cross-genera functional prediction. Thus, we have defined regulatory, transport, and catabolic genes and a cis-acting regulatory sequence that form a conserved module required for myo

  16. Production of nodulation factors by Rhizobium meliloti: fermentation, purification and characterization of glycolipids.

    Science.gov (United States)

    Kohring, B; Baier, R; Niehaus, K; Pühler, A; Flaschel, E

    1997-12-01

    Lipooligosaccharides, synthesized by soil bacteria of the genera Rhizobium, are known to have multifunctional effects on a wide variety of plants as signal substances in symbiosis initiation, cell response elicitation and growth regulation. These so called nodulation (Nod-) factors represent interesting biotechnological products with respect to fundamental studies of symbiotic interactions as well as for potential applications. Therefore, a batch fermentation process on a scale of 30 l has been developed by means of the Rhizobium meliloti strain R.m. 1021 (pEK327) strongly overexpressing the genes for the synthesis of Nod factors. Induction by the flavone luteolin led to growth associated production of the lipooligosaccharides. Ultrafiltration was used for separating the biomass from the filtrate containing the extracellular Nod factors. Simultaneously, ultrafiltration reduced the amount of lipophilic substances, which would otherwise interfere with processes downstream. The second separation step consisted in adsorption on XAD-2, a nonspecific hydrophobic adsorptive resin. Adsorption of Nod factors was carried out by batch operation of a stirred tank. Desorption was performed by elution with methanol in a fixed bed column. A semi-preparative reversed phase HPLC (Polygoprep 100-30 C18) was chosen as the final purification step. The Nod factors were obtained after evaporation and lyophilization. Thus, about 600 mg of Nod factors were produced from 20 l of fermentation broth. The Nod factors produced by Rhizobium meliloti R.m. 1021 (pEK327) were identified by liquid secondary ion mass spectrometry and by reversed-phase HPLC as fluorescent derivatives of 2-aminobenzamide. The biological activity of the products was demonstrated by means of the root hair deformation (HAD-) assay.

  17. The structure of Sinorhizobium meliloti phage ΦM12, which has a novel T=19l triangulation number and is the founder of a new group of T4-superfamily phages.

    Science.gov (United States)

    Stroupe, M Elizabeth; Brewer, Tess E; Sousa, Duncan R; Jones, Kathryn M

    2014-02-01

    ΦM12 is the first example of a T=19l geometry capsid, encapsulating the recently sequenced genome. Here, we present structures determined by cryo-EM of full and empty capsids. The structure reveals the pattern for assembly of 1140 HK97-like capsid proteins, pointing to interactions at the pseudo 3-fold symmetry axes that hold together the asymmetric unit. The particular smooth surface of the capsid, along with a lack of accessory coat proteins encoded by the genome, suggest that this interface is the primary mechanism for capsid assembly. Two-dimensional averages of the tail, including the neck and baseplate, reveal that ΦM12 has a relatively narrow neck that attaches the tail to the capsid, as well as a three-layer baseplate. When free from DNA, the icosahedral edges expand by about 5nm, while the vertices stay at the same position, forming a similarly smooth, but bowed, T=19l icosahedral capsid.

  18. NCBI nr-aa BLAST: CBRC-DRER-26-0161 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0161 ref|NP_384590.1| acyl-CoA synthase [Sinorhizobium meliloti 1021] emb|CAC29432.2| long...-chain acyl-CoA synthetase [Sinorhizobium meliloti] emb|CAC41921.1| PROBABLE LONG-CHAIN-

  19. NCBI nr-aa BLAST: CBRC-DRER-26-0474 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0474 ref|NP_384172.1| SENSOR HISTIDINE KINASE TRANSMEMBRANE PROTEIN [S...inorhizobium meliloti 1021] emb|CAC41453.1| SENSOR HISTIDINE KINASE TRANSMEMBRANE PROTEIN [Sinorhizobium meliloti] NP_384172.1 1e-159 68% ...

  20. Construction and pilot screening of a signature-tagged mutant library of Sinorhizobium fredii.

    Science.gov (United States)

    Wang, Dan; Wang, Yuan Chun; Wu, Li Juan; Liu, Jian Xin; Zhang, Pan; Jiao, Jian; Yan, Hui; Liu, Tao; Tian, Chang Fu; Chen, Wen Xin

    2016-03-01

    Sinorhizobium fredii is well known for its ability to establish symbiosis with diverse legumes such as Glycine max (soybean, determinate nodules) and Cajanus cajan (pigeon pea, indeterminate nodules). In order to make screening of S. fredii genes related to symbiosis cost-effective, we constructed a large Tn5 insertion mutant library of S. fredii CCBAU45436 using the signature-tagged mutagenesis (STM) technique. This STM library contains a total of 25,500 independent mutants distributed in 17 sublibraries tagged by corresponding distinct DNA bar-code sequences. After the pilot screening of 255 mutants in 15 batches, Tag85-4, Tag4-17, Tag4-11 and Tag10-13 were found to have attenuated competitiveness (0-30 % in nodule occupation) compared to the wild-type strain when inoculated on soybean. Further characterization of these mutants suggests that Tag4-11 (a pyrC mutant) and Tag10-13 (a nrdJ mutant) are defective in establishing symbiosis with soybean. The pyrC mutant induced uninfected pseudonodules while the nrdJ mutant formed significantly more nodules containing bacteroids with poor persistence ability. When these two mutants were tested on pigeon pea, host-specific symbiotic defects were found. These results demonstrated the STM library as a valuable resource for identifying S. fredii genes relevant to symbiosis. PMID:26472206

  1. A proteomic analysis of bacterial strain Sinorhizobium fredii RT19 subjected to salt shock

    Institute of Scientific and Technical Information of China (English)

    QI Suwei; YANG Pingfang; JING Yuxiang; SHEN Shihua; YANG Susheng

    2004-01-01

    Sinorhizobium fredii RT19, a strain of freeliving bacteria, was subjected to salt shock and its protein expression profiles were analyzed by differential display proteome approaches. The results of separation by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) showed that the number of resolved proteins was 481, 465 and 424, corresponding to salt-free control, 5 and 50 min 1 mol/L salt treatment, respectively. Among the resolved proteins, 82 in total had altered expression in response to salt-shock stress. 26 out of the 82 proteins were induced and 23 were completely inhibited, while 12 were up-regulated and 21 down-regulated in response to salt shock. In addition, the appearance of differentially displayed proteins responding to different salt shock periods is also reported. The identity of the 26 induced proteins was revealed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) followed by database searching. Among them, 20 were assigned to proteins with known functions. Their roles in response to salt shock stress are discussed.

  2. Toxic effects of arsenic on Sinorhizobium-Medicago sativa symbiotic interaction

    Energy Technology Data Exchange (ETDEWEB)

    Pajuelo, Eloisa [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain); Rodriguez-Llorente, Ignacio D. [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain)], E-mail: irodri@us.es; Dary, Mohammed; Palomares, Antonio J. [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain)

    2008-07-15

    Recently, the Rhizobium-legume symbiotic interaction has been proposed as an interesting tool in bioremediation. However, little is known about the effect of most common contaminants on this process. The phytotoxic effects of arsenic on nodulation of Medicago sativa have been examined in vitro using the highly arsenic resistant and symbiotically effective Sinorhizobium sp. strain MA11. The bacteria were able to grow on plates containing As concentrations as high as 10 mM. Nevertheless, as little as 25-35 {mu}M arsenite produced a 75% decrease in the total number of nodules, due to a 90% reduction in the number of rhizobial infections, as could be determined using the strain MA11 carrying a lacZ reporter gene. This effect was associated to root hair damage and a shorter infective root zone. However, once nodulation was established nodule development seemed to continue normally, although earlier senescence could be observed in nodules of arsenic-grown plants. - First steps of nodulation of alfalfa, in particular infection thread formation, are more sensitive to As than nitrogen fixation due to plant effects.

  3. Toxic effects of arsenic on Sinorhizobium-Medicago sativa symbiotic interaction

    International Nuclear Information System (INIS)

    Recently, the Rhizobium-legume symbiotic interaction has been proposed as an interesting tool in bioremediation. However, little is known about the effect of most common contaminants on this process. The phytotoxic effects of arsenic on nodulation of Medicago sativa have been examined in vitro using the highly arsenic resistant and symbiotically effective Sinorhizobium sp. strain MA11. The bacteria were able to grow on plates containing As concentrations as high as 10 mM. Nevertheless, as little as 25-35 μM arsenite produced a 75% decrease in the total number of nodules, due to a 90% reduction in the number of rhizobial infections, as could be determined using the strain MA11 carrying a lacZ reporter gene. This effect was associated to root hair damage and a shorter infective root zone. However, once nodulation was established nodule development seemed to continue normally, although earlier senescence could be observed in nodules of arsenic-grown plants. - First steps of nodulation of alfalfa, in particular infection thread formation, are more sensitive to As than nitrogen fixation due to plant effects

  4. Mutualistic co-evolution of type III effector genes in Sinorhizobium fredii and Bradyrhizobium japonicum.

    Directory of Open Access Journals (Sweden)

    Jeffrey A Kimbrel

    2013-02-01

    Full Text Available Two diametric paradigms have been proposed to model the molecular co-evolution of microbial mutualists and their eukaryotic hosts. In one, mutualist and host exhibit an antagonistic arms race and each partner evolves rapidly to maximize their own fitness from the interaction at potential expense of the other. In the opposing model, conflicts between mutualist and host are largely resolved and the interaction is characterized by evolutionary stasis. We tested these opposing frameworks in two lineages of mutualistic rhizobia, Sinorhizobium fredii and Bradyrhizobium japonicum. To examine genes demonstrably important for host-interactions we coupled the mining of genome sequences to a comprehensive functional screen for type III effector genes, which are necessary for many Gram-negative pathogens to infect their hosts. We demonstrate that the rhizobial type III effector genes exhibit a surprisingly high degree of conservation in content and sequence that is in contrast to those of a well characterized plant pathogenic species. This type III effector gene conservation is particularly striking in the context of the relatively high genome-wide diversity of rhizobia. The evolution of rhizobial type III effectors is inconsistent with the molecular arms race paradigm. Instead, our results reveal that these loci are relatively static in rhizobial lineages and suggest that fitness conflicts between rhizobia mutualists and their host plants have been largely resolved.

  5. Escherichia coli BdcA controls biofilm dispersal in Pseudomonas aeruginosa and Rhizobium meliloti

    Directory of Open Access Journals (Sweden)

    Wood Thomas K

    2011-10-01

    Full Text Available Abstract Background Previously we showed that BdcA controls Escherichia coli biofilm dispersal by binding the ubiquitous bacterial signal cyclic diguanylate (c-di-GMP; upon reducing the concentration of c-di-GMP, the cell shifts to the planktonic state by increasing motility, decreasing aggregation, and decreasing production of biofilm adhesins. Findings Here we report that BdcA also increases biofilm dispersal in other Gram-negative bacteria including Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhizobium meliloti. BdcA binds c-di-GMP in these strains and thereby reduces the effective c-di-GMP concentrations as demonstrated by increases in swimming motility and swarming motility as well as by a reduction in extracellular polysaccharide production. We also develop a method to displace existing biofilms by adding BdcA via conjugation from E. coli in mixed-species biofilms. Conclusion Since BdcA shows the ability to control biofilm dispersal in diverse bacteria, BdcA has the potential to be used as a tool to disperse biofilms for engineering and medical applications.

  6. Performance of fenugreek bioinoculated with Rhizobium meliloti strains under semi-arid condition.

    Science.gov (United States)

    Singh, N K; Patel, D B

    2016-01-01

    Rhizobium meliloti strains were isolated from the fields of S.D. Agricultural University (Gujarat, India) and were maintained in the Congo Red Yeast Extract Mannitol Agar medium. These strains were tested for their effectiveness for fenugreek crop grown under semi-arid condition. Among the six Rhizobium strains, FRS-7 strain showed best plant growth parameters like shoot length, shoot dry weight, shoot total nitrogen, root length, root dry weight, root total nitrogen, seed yield, 1000 grain weight, number of root nodules, and nodules fresh and dry weight. The performance of this strain was better as compared to 20 kgN ha(-1) treatment through urea and was even far better over control plot. Seed yields obtained with FRS-7 during two years were 10.14 and 9.66 q ha(-1); which was about 36.8% and 45.9% high over control. This strain resulted in saving of about 20 kgN ha(-1) accompanied with better crop yield and soil health. Results of the present experiments can be utilized in integrated nutrient management for cultivation of fenugreek in semi-arid areas to provide sustainability to agricultural productivity in such regions. PMID:26930857

  7. Genetic and structural analysis of the Rhizobium meliloti fixA, fixB, fixC, and fixX genes.

    OpenAIRE

    Earl, C D; Ronson, C W; Ausubel, F M

    1987-01-01

    The fixA, fixB, fixC, and fixX genes of Rhizobium meliloti 1021 constitute an operon and are required for nitrogen fixation in alfalfa nodules. DNA homologous to the R. meliloti fixABC genes is present in all other Rhizobium and Bradyrhizobium species examined, but fixABC-homologous sequences were found in only one free-living diazotroph, Azotobacter vinelandii. To determine whether the fixABCX genes share sequence homology with any of the 17 Klebsiella pneumoniae nif genes, we determined the...

  8. Ensifer meliloti overexpressing Escherichia coli phytase gene ( appA) improves phosphorus (P) acquisition in maize plants

    Science.gov (United States)

    Sharma, Vikas; Kumar, Ajit; Archana, G.; Kumar, G. Naresh

    2016-10-01

    The Escherichia coli phytase gene appA encoding enzyme AppA was cloned in a broad host range plasmid pBBR1MCS2 ( lac promoter), termed pVA1, and transformed into the Ensifer meliloti 1020. Transformation of pVA1 in Ensifer meliloti { E. m (pVA1)} increased its phosphatase and phytase activity by ˜9- and ˜50-fold, respectively, compared to the transformants containing empty plasmid as control { E. m (pBBR1MCS2)}. The western blot experiments using rabbit anti-AppA antibody showed that AppA is translocated into the periplasm of the host after its expression. Ensifer meliloti harboring AppA protein { E. m (pVA1)} and { E. m (pBBR1MCS2)} could acidify the unbuffered phytate minimal media (pH 8.0) containing Ca-phytate or Na-phytate as sole organic P (Po) source to below pH 5.0 and released P. However, both { E. m (pVA1)} and { E. m (pBBR1MCS2)} neither dropped pH of the medium nor released P when the medium was buffered at pH 8.0 using Tris-Cl, indicating that acidification of medium was important for the enzymatic hydrolysis of phytate. Further experiments proved that maize plants inoculated with { E. m. (pVA1)} showed increase in growth under sterile semi solid agar (SSA) medium containing Na-phytate as sole P source. The present study could be helpful in generating better transgenic bioinoculants harboring phosphate mineralization properties that ultimately promote plant growth.

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

    Directory of Open Access Journals (Sweden)

    Erik Machado-Ferreira

    2012-01-01

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

  10. Functional analysis of the cysteine motifs in the ferredoxin-like protein FdxN of Rhizobium meliloti involved in symbiotic nitrogen fixation.

    Science.gov (United States)

    Masepohl, B; Kutsche, M; Riedel, K U; Schmehl, M; Klipp, W; Pühler, A

    1992-05-01

    The Rhizobium meliloti fdxN gene, which is part of the nifA-nifB-fdxN operon, is absolutely required for symbiotic nitrogen fixation. The deduced sequence of the FdxN protein is characterized by two cysteine motifs typical of bacterial-type ferredoxins. The Fix-phenotype of an R. meliloti fdxN::[Tc] mutant could be rescued by the R. leguminosarum fdxN gene, whereas no complementation was observed with nif-associated genes encoding ferredoxins from Bradyrhizobium japonicum, Azotobacter vinelandii, A. chroococcum and Rhodobacter capsulatus. In addition to these heterologous genes, several R. meliloti fdxN mutant genes constructed by site-directed mutagenesis were analyzed. Not only a cysteine residue within the second cysteine motif (position 42), which is known to coordinate the Fe-S cluster in homologous proteins, but also a cysteine located down-stream of this motif (position 61), was found to be essential for the activity of the R. meliloti FdxN protein. Changing the amino acid residue proline in position 56 into methionine resulted in a FdxN mutant protein with decreased activity, whereas changes in positions 35 (Asp35Glu) and 45 (Gly45Glu) had no significant effect on the function of the FdxN mutant proteins. In contrast to bacterial-type ferredoxins, which contain two identical cysteine motifs of the form C-X2-C-X2-C-X3-C, nif-associated ferredoxins, including R. meliloti FdxN, are characterized by two different cysteine motifs. Six "additional" amino acids separate the second (Cys42) and the third cysteine (Cys51) in the C-terminal motif (C-X2-C-X8-C-X3-C).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1603075

  11. Regulatory role of the sequences downstream from nodD3 P1 promoter of Rhizobium meliloti

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The 660 bp region between nodD3 P1 promoter and the following coding region of Rhizobium meliloti has been studied.This region is designated "downstream sequences".It consists of two potential open reading frames,ORF1 and ORF2.Studies on the role of the downstream sequences on the activity of nodD3 P1 with nod D3(P1)-lacZ fusion show that deletion of the sequences containing ORF2 causes the increase of the activity of the fusion; on the contrary,addition of extra copies of ORF2 markedly decreases the activity of the fusion.These results indicate that the product of ORF2 plays a negative role in the expression of nod D3.

  12. N2-Fixation in Alfalfa (Medicago sativa L.) Seedlings and Rhizobium meliloti L. Grown in Vitro Under Salt and Drought Stresses

    OpenAIRE

    Mohamad, Ramzi Muhiddin

    1987-01-01

    Alfalfa, Medicago sativa L., cultivars, breeding lines and germplasm releases (populations) and Rhizobium meliloti L. strains that exhibit nitrogen fixation efficiency and tolerance to salinity and drought stresses should enhance seedling establishment, increase yields, reduce nitrogen dependency on petroleum-based nitrogen fertilizers, and allow wider use of irrigated lands in semiarid and a rid regions. In vitro experiments were conducted to determine relative salt (sodium chloride -- NaCl)...

  13. The 32-kilobase exp gene cluster of Rhizobium meliloti directing the biosynthesis of galactoglucan: genetic organization and properties of the encoded gene products.

    OpenAIRE

    Becker, A.; Rüberg, S; Küster, H.; Roxlau, A A; Keller, M; Ivashina, T; H.P. Cheng; Walker, G C; Pühler, A

    1997-01-01

    Proteins directing the biosynthesis of galactoglucan (exopolysaccharide II) in Rhizobium meliloti Rm2011 are encoded by the exp genes. Sequence analysis of a 32-kb DNA fragment of megaplasmid 2 containing the exp gene cluster identified previously (J. Glazebrook and G. C. Walker, Cell 56:661-672, 1989) revealed the presence of 25 open reading frames. Homologies of the deduced exp gene products to proteins of known function suggested that the exp genes encoded four proteins involved in the bio...

  14. The Sinorhizobium fredii HH103 Genome: A Comparative Analysis With S. fredii Strains Differing in Their Symbiotic Behavior With Soybean.

    Science.gov (United States)

    Vinardell, José-María; Acosta-Jurado, Sebastián; Zehner, Susanne; Göttfert, Michael; Becker, Anke; Baena, Irene; Blom, Jochem; Crespo-Rivas, Juan Carlos; Goesmann, Alexander; Jaenicke, Sebastian; Krol, Elizaveta; McIntosh, Matthew; Margaret, Isabel; Pérez-Montaño, Francisco; Schneiker-Bekel, Susanne; Serranía, Javier; Szczepanowski, Rafael; Buendía, Ana-María; Lloret, Javier; Bonilla, Ildefonso; Pühler, Alfred; Ruiz-Sainz, José-Enrique; Weidner, Stefan

    2015-07-01

    Sinorhizobium fredii HH103 is a fast-growing rhizobial strain infecting a broad range of legumes including both American and Asiatic soybeans. In this work, we present the sequencing and annotation of the HH103 genome (7.25 Mb), consisting of one chromosome and six plasmids and representing the structurally most complex sinorhizobial genome sequenced so far. Comparative genomic analyses of S. fredii HH103 with strains USDA257 and NGR234 showed that the core genome of these three strains contains 4,212 genes (61.7% of the HH103 genes). Synteny plot analysis revealed that the much larger chromosome of USDA257 (6.48 Mb) is colinear to the HH103 (4.3 Mb) and NGR324 chromosomes (3.9 Mb). An additional region of the USDA257 chromosome of about 2 Mb displays similarity to plasmid pSfHH103e. Remarkable differences exist between HH103 and NGR234 concerning nod genes, flavonoid effect on surface polysaccharide production, and quorum-sensing systems. Furthermore a number of protein secretion systems have been found. Two genes coding for putative type III-secreted effectors not previously described in S. fredii, nopI and gunA, have been located on the HH103 genome. These differences could be important to understand the different symbiotic behavior of S. fredii strains HH103, USDA257, and NGR234 with soybean.

  15. The Sinorhizobium (Ensifer) fredii HH103 Type 3 Secretion System Suppresses Early Defense Responses to Effectively Nodulate Soybean.

    Science.gov (United States)

    Jiménez-Guerrero, Irene; Pérez-Montaño, Francisco; Monreal, José Antonio; Preston, Gail M; Fones, Helen; Vioque, Blanca; Ollero, Francisco Javier; López-Baena, Francisco Javier

    2015-07-01

    Plants that interact with pathogenic bacteria in their natural environments have developed barriers to block or contain the infection. Phytopathogenic bacteria have evolved mechanisms to subvert these defenses and promote infection. Thus, the type 3 secretion system (T3SS) delivers bacterial effectors directly into the plant cells to alter host signaling and suppress defenses, providing an appropriate environment for bacterial multiplication. Some rhizobial strains possess a symbiotic T3SS that seems to be involved in the suppression of host defenses to promote nodulation and determine the host range. In this work, we show that the inactivation of the Sinorhizobium (Ensifer) fredii HH103 T3SS negatively affects soybean nodulation in the early stages of the symbiotic process, which is associated with a reduction of the expression of early nodulation genes. This symbiotic phenotype could be the consequence of the bacterial triggering of soybean defense responses associated with the production of salicylic acid (SA) and the impairment of the T3SS mutant to suppress these responses. Interestingly, the early induction of the transcription of GmMPK4, which negatively regulates SA accumulation and defense responses in soybean via WRKY33, could be associated with the differential defense responses induced by the parental and the T3SS mutant strain.

  16. Identification of Sinorhizobium (Ensifer) medicae based on a specific genomic sequence unveiled by M13-PCR fingerprinting.

    Science.gov (United States)

    Dourado, Ana Catarina; Alves, Paula I L; Tenreiro, Tania; Ferreira, Eugénio M; Tenreiro, Rogério; Fareleira, Paula; Crespo, M Teresa Barreto

    2009-12-01

    A collection of nodule isolates from Medicago polymorpha obtained from southern and central Portugal was evaluated by M13-PCR fingerprinting and hierarchical cluster analysis. Several genomic clusters were obtained which, by 16S rRNA gene sequencing of selected representatives, were shown to be associated with particular taxonomic groups of rhizobia and other soil bacteria. The method provided a clear separation between rhizobia and co-isolated non-symbiotic soil contaminants. Ten M13-PCR groups were assigned to Sinorhizobium (Ensifer) medicae and included all isolates responsible for the formation of nitrogen-fixing nodules upon re-inoculation of M. polymorpha test-plants. In addition, enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting indicated a high genomic heterogeneity within the major M13- PCR clusters of S. medicae isolates. Based on nucleotide sequence data of an M13-PCR amplicon of ca. 1500 bp, observed only in S. medicae isolates and spanning locus Smed_3707 to Smed_3709 from the pSMED01 plasmid sequence of S. medicae WSM419 genome's sequence, a pair of PCR primers was designed and used for direct PCR amplification of a 1399-bp sequence within this fragment. Additional in silico and in vitro experiments, as well as phylogenetic analysis, confirmed the specificity of this primer combination and therefore the reliability of this approach in the prompt identification of S. medicae isolates and their distinction from other soil bacteria. PMID:20112226

  17. Stable isotope labelling reveals that NaCl stress decreases the production of Ensifer (Sinorhizobium) arboris lipochitooligosaccharide signalling molecules.

    Science.gov (United States)

    Penttinen, Petri; Räsänen, Leena A; Lortet, Gilles; Lindström, Kristina

    2013-12-01

    Ensifer (Sinorhizobium) arboris is a symbiont of salt-tolerant leguminous trees in the genera Acacia and Prosopis that are utilized in the prevention of soil erosion and desertification and in phytoremediation of salinized soil. Signalling between the plant and the rhizobia is essential for the formation of effective symbiosis that increases the success of reclaiming saline sites. We assessed the effect of salt stress on the growth and the production of lipochitooligosaccharide signalling molecules (LCOs) of S. arboris HAMBI 2361, an LCO-overproducing derivative of the S. arboris type strain HAMBI 1552. The strain tolerated NaCl up to 750 mM. To obtain both qualitative and quantitative information on the LCO production under salt stress, we devised a method where LCOs were differentially labelled by stable isotopes of nitrogen, (14)N and (15)N, and analysed by mass spectrometry. Under control conditions, the strain produced altogether 27 structural LCO variants. In 380 mM NaCl, 13 LCO variants were produced in detectable amounts, and six of these were reliably quantified, ranging from one-tenth to one-third of the non-stressed one.

  18. Isolation and characterization of a Sinorhizobium fredii mutant that cannot utilize proline as the sole carbon and nitrogen source

    Institute of Scientific and Technical Information of China (English)

    HUANG Sheng; BAI Xueliang; MA Qingsheng; TANG Xianlai; WU Bo

    2004-01-01

    Sinorhizobium fredii strain HN01 can use proline as the sole carbon and nitrogen source. A mutant strain GXHN100 unable to catabolize proline was screened from 6000 Tn5gusA5 random insertional mutants of S.fredii strain HN01. Sequencing analysis showed that an open reading frame, named pmrA (proline metabolic relative), was inserted by the Tn5gusA5. A positive clone, named pGXHN100 which containing 3.3kb foreign DNA fragment of S.fredii strain HN01, was isolated from a partial gene library of S.fredii HN01 by colony in situ hybridization. Sequence analysis showed that pGXHN100 contained the entire pmrA gene. The 3.3kb DNA fragment of pGXHN100 was cloned into a broad-host-range cosmid vector pLAFR3 to form plasmid pGXHN200 which was subsequently introduced into GXHN100 to form a complemented strain GXHN200. Plant test showed that GXHN100 was effective and no obvious changes in nitrogenase activity comparing with parental strain. But GXHN100 nodulated 2 days later on soybean and its nodulation efficiency and competitiveness were decreased. The complemented strain GXHN200 restored the nodulation efficiency and competitiveness of GXHN100 to the wild type.

  19. Three phylogenetic groups of nodA and nifH genes in Sinorhizobium and Mesorhizobium isolates from leguminous trees growing in Africa and Latin America.

    Science.gov (United States)

    Haukka, K; Lindström, K; Young, J P

    1998-02-01

    The diversity and phylogeny of nodA and nifH genes were studied by using 52 rhizobial isolates from Acacia senegal, Prosopis chilensis, and related leguminous trees growing in Africa and Latin America. All of the strains had similar host ranges and belonged to the genera Sinorhizobium and Mesorhizobium, as previously determined by 16S rRNA gene sequence analysis. The restriction patterns and a sequence analysis of the nodA and nifH genes divided the strains into the following three distinct groups: sinorhizobia from Africa, sinorhizobia from Latin America, and mesorhizobia from both regions. In a phylogenetic tree also containing previously published sequences, the nodA genes of our rhizobia formed a branch of their own, but within the branch no correlation between symbiotic genes and host trees was apparent. Within the large group of African sinorhizobia, similar symbiotic gene types were found in different chromosomal backgrounds, suggesting that transfer of symbiotic genes has occurred across species boundaries. Most strains had plasmids, and the presence of plasmid-borne nifH was demonstrated by hybridization for some examples. The nodA and nifH genes of Sinorhizobium teranga ORS1009T grouped with the nodA and nifH genes of the other African sinorhizobia, but Sinorhizobium saheli ORS609T had a totally different nodA sequence, although it was closely related based on the 16S rRNA gene and nifH data. This might be because this S. saheli strain was originally isolated from Sesbania sp., which belongs to a different cross-nodulation group than Acacia and Prosopis spp. The factors that appear to have influenced the evolution of rhizobial symbiotic genes vary in importance at different taxonomic levels.

  20. Tolerância ao sal e às altas temperaturas de estirpes de Sinorhizobium provenientes de zonas secas do Alentejo Salt and temperature tolerance of Sinorhizobium strains isolated from dry environments in Alentejo

    Directory of Open Access Journals (Sweden)

    P. Fareleira

    2007-07-01

    Full Text Available O trabalho teve como principal objectivo a obtenção de estirpes de rizóbio adequadas à nodulação de luzernas anuais e adaptadas a sobreviver nas condições ambientais susceptíveis de ocorrer em solos degradados. Dado que as populações de rizóbio apresentam variabilidade considerável no que respeita à tolerância a factores ambientais, os estudos focaram-se na pesquisa e na selecção de estirpes resistentes a condições adversas. Efectuaram-se colheitas de solos em diversas zonas no sul do País, seleccionando-se locais afectados por secura, temperaturas elevadas e, pontualmente, salinidade. A partir destes solos, isolaram-se estirpes de rizóbio, usando como planta hospedeira a luzerna anual Medicago polymorpha. Estudaram-se os efeitos de condições de stresse ambiental, como a salinidade e as altas temperaturas, no crescimento das estirpes isoladas. Dos 41 isolamentos analisados, 11 apresentaram crescimento em meio con-tendo 1,4 M de cloreto de sódio e suplementado com 10% de extracto de terra, e 22 cresceram quando incubadas a 45 ºC em meio sem aditivos. Três estirpes mostraram ter capacidade para crescer sob os efeitos conjuntos da salinidade e da alta temperatura. A análise de extractos etanólicos de estirpes tolerantes à salinidade revelou, na maior parte dos casos, a acumulação, induzida pelo sal, dos solutos compatíveis de Sinorhizobium: o dipéptido N - acetilglutaminilglutamina amida, vários tipos de betaínas, trealose, glutamato e prolina. A observação, por NMR de 31P in vivo, de uma estirpe tolerante ao sal, proveniente de um solo xistoso de baixo teor em fósforo, mostrou a presença de níveis elevados de reservas intracelulares de fosfato inorgânico (polifosfato, sugerindo um bom potencial para utilização em solos onde os riscos de salinização se conjuguem com deficiências em fósforo assimilável.The main objective of this work was to obtain rhizobial strains able to nodulate annual medics and

  1. DNA同源性分析最终确定新疆中华根瘤菌 (Sinorhizobium xinjiangensis) 为一独立种%SINORHIZOBIUM XINJIANGENSIS AS AN INDEPENDENT SPECIES CONFIRMED BY DNA HOMOLOGY ANALYSIS

    Institute of Scientific and Technical Information of China (English)

    彭桂香; 谭志远; 陈文新

    2000-01-01

    针对S. xinjiangensis分类地位的争议,从新疆再次分离获得34株快生大豆根瘤菌,在16S rDNA PCR-RFLP分析和SDS全细胞蛋白电泳分群的基础上,进行了16S rDNA 全序列相似性和DNA同源性分析.所测4个菌株和S. fredii模式菌株USDA 205的16S rDNA全序列有很高的相似性.而DNA同源性分析说明新分离的菌株代表与原定的S. xinjiangensis为一个DNA同源群.其模式菌株CCBAU 110 与S. fredii的2 株参比菌株USDA 194、2048之间的DNA同源性分别为31.5%和28.7%.S. fredii的模式菌株USDA 205与新分离的菌株代表及原定的S. xinjiangensis的模式菌株和2个参比菌株之间的DNA同源性为20.8%~39%,远低于70%.属于种水平上的差异.按照国际细菌分类委员会以DNA同源性≥70%且△Tm≤5 ℃作为定种的标准,S. xinjiangensis是Sinorhizobium属中不同于S. fredii的一个独立的种. 表3 参20%In this study, based on the results from 16S rDNA PCR-RFLP of 34 fast-growing soybean rhizobia strains isolated from Xinjiang in comparison with several type and reference strains of related species, and SDS-PAGE whole-cell protein patterns of new representative isolates and other strains in Sinorhizobium, new representative isolates were further analyzed by 16S rDNA full sequencing, G+C mol% tests and DNA-DNA hybridization. 4 strains were analyzed by 16S rDNA full sequencing, indicating that type strain CCBAU 110 of S. xinjiangensis and type strain USDA 205 of S. fredii shared high similarity. The DNA homologies indicated that new representative isolates and S. xinjiangensis were of the same DNA homology groups. The DNA homologies between CCBAU 110 and the reference strains of S. fredii: USDA 194, 2048 were 31.5% and 28.7%, respectively. The DNA-DNA hybridization values between USDA 205 and CCBAU 110, 2 reference strains of S. xinjiangensis, representative of new isolates were 20.8%~39%, which confirmed that S. xinjiangensis and S. fredii belonged to the

  2. NopC Is a Rhizobium-Specific Type 3 Secretion System Effector Secreted by Sinorhizobium (Ensifer) fredii HH103

    Science.gov (United States)

    Medina, Carlos; Ollero, Francisco Javier; López-Baena, Francisco Javier

    2015-01-01

    Sinorhizobium (Ensifer) fredii HH103 is a broad host-range nitrogen-fixing bacterium able to nodulate many legumes, including soybean. In several rhizobia, root nodulation is influenced by proteins secreted through the type 3 secretion system (T3SS). This specialized secretion apparatus is a common virulence mechanism of many plant and animal pathogenic bacteria that delivers proteins, called effectors, directly into the eukaryotic host cells where they interfere with signal transduction pathways and promote infection by suppressing host defenses. In rhizobia, secreted proteins, called nodulation outer proteins (Nops), are involved in host-range determination and symbiotic efficiency. S. fredii HH103 secretes at least eight Nops through the T3SS. Interestingly, there are Rhizobium-specific Nops, such as NopC, which do not have homologues in pathogenic bacteria. In this work we studied the S. fredii HH103 nopC gene and confirmed that its expression was regulated in a flavonoid-, NodD1- and TtsI-dependent manner. Besides, in vivo bioluminescent studies indicated that the S. fredii HH103 T3SS was expressed in young soybean nodules and adenylate cyclase assays confirmed that NopC was delivered directly into soybean root cells by means of the T3SS machinery. Finally, nodulation assays showed that NopC exerted a positive effect on symbiosis with Glycine max cv. Williams 82 and Vigna unguiculata. All these results indicate that NopC can be considered a Rhizobium-specific effector secreted by S. fredii HH103. PMID:26569401

  3. NopC Is a Rhizobium-Specific Type 3 Secretion System Effector Secreted by Sinorhizobium (Ensifer) fredii HH103.

    Science.gov (United States)

    Jiménez-Guerrero, Irene; Pérez-Montaño, Francisco; Medina, Carlos; Ollero, Francisco Javier; López-Baena, Francisco Javier

    2015-01-01

    Sinorhizobium (Ensifer) fredii HH103 is a broad host-range nitrogen-fixing bacterium able to nodulate many legumes, including soybean. In several rhizobia, root nodulation is influenced by proteins secreted through the type 3 secretion system (T3SS). This specialized secretion apparatus is a common virulence mechanism of many plant and animal pathogenic bacteria that delivers proteins, called effectors, directly into the eukaryotic host cells where they interfere with signal transduction pathways and promote infection by suppressing host defenses. In rhizobia, secreted proteins, called nodulation outer proteins (Nops), are involved in host-range determination and symbiotic efficiency. S. fredii HH103 secretes at least eight Nops through the T3SS. Interestingly, there are Rhizobium-specific Nops, such as NopC, which do not have homologues in pathogenic bacteria. In this work we studied the S. fredii HH103 nopC gene and confirmed that its expression was regulated in a flavonoid-, NodD1- and TtsI-dependent manner. Besides, in vivo bioluminescent studies indicated that the S. fredii HH103 T3SS was expressed in young soybean nodules and adenylate cyclase assays confirmed that NopC was delivered directly into soybean root cells by means of the T3SS machinery. Finally, nodulation assays showed that NopC exerted a positive effect on symbiosis with Glycine max cv. Williams 82 and Vigna unguiculata. All these results indicate that NopC can be considered a Rhizobium-specific effector secreted by S. fredii HH103. PMID:26569401

  4. NCBI nr-aa BLAST: CBRC-CREM-01-1341 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available m meliloti 1021] sp|P20672|DCTA_RHIME C4-dicarboxylate transport protein gb|AAA26253.1| C4-dicarboxylate car... protein [Sinorhizobium meliloti] gb|AAA26248.1| dctA protein emb|CAC49923.1| C4-

  5. Colonization and nitrogenase activity of Triticum aestivum (cv. Baccross and Mahdavi) to the dual inoculation with Azospirillum brasilense and Rhizobium meliloti plus 2,4-D.

    Science.gov (United States)

    Mehry, Askary; Akbar, Mostajeran; Giti, Emtiazi

    2008-06-15

    The potential enhancement of root colonization and nitrogenase activity of wheat cultivars (Baccross and Mahdavi) was studied with application of two Azospirillum brasilense strains (native and Sp7) co-inoculated with two Rhizobium meliloti strains (native and DSMZ 30135). The results indicated that the colonization was different due to the strains and cultivars of wheat were used. Native A. brasilense colonized wheat root better than Sp7 strain. However, Baccross cv. reacted better with native Azospirillum compared to Mahdavi cv. which reacted better with Sp7. When plants inoculated with dual inoculants (SP7 with standard Rhizobium), the colonization of Azospirillum were increased significantly (from 1.67 x 10(5) to 22 x 10(5) cfu g(-1) FW for Baccras cv. and 3.67 x 10(5) to 26 x 10(5) cfu g(-1) FW for Mahdavi cultivar). When the standard Rhizobium as co-inoculants changed to the native Rhizobium, the colonization of Azospirillum was higher when compared to the single inoculants but was almost the same when compared to the standard Rhizobium. When the standard or native strains of Rhizobium used as single inoculation of wheat roots, the number of Rhizobium in the wheat roots were not changed significantly. However, when plants co-inoculated with Rhizobium and Azospirillum, the colonization of Rhizobium was increased. Co-inoculation of standard strain of R. melilot with A. brasilense Sp7 showed that the colonization of Rhizobium were increased from 0.67 x 10(5) to 21 x 10(5) cfu g(-1) FW for Baccross cv. and 0.33 x 10(5) to 18 x 10(5) cfu g(-1) FW for Mahdavi cv. This behavior was the same when inoculation of Rhizobium was happened with the native one. In dual inoculation, the highest nitrogenase activity was measured in combination of the local strains (native A. brasilense with the native R. meliloti) and the lower one belongs to the combination of standard strains (Sp7 with standard R. meliloti). The difference in nirtogenase activity for different cultivars of

  6. The development of plasmid-free strains of Agrobacterium tumefaciens by using incompatibility with a Rhizobium meliloti plasmid to eliminate pAtC58.

    Science.gov (United States)

    Hynes, M F; Simon, R; Pühler, A

    1985-03-01

    Agrobacterium tumefaciens strains LBA275 and LBA290 were cured of their cryptic plasmid pAtC58 by the introduction of the Rhizobium meliloti plasmid pRme41a, which is incompatible with pAtC58. pRme41a and pTiC58, the resident Ti plasmid of LBA275, were subsequently eliminated by growth at supraoptimal temperature (40 degrees C). The resulting plasmid-free Agrobacterium strains, UBAPF1 and UBAPF2, have proved extremely useful for the study of Rhizobium plasmids. The loss of the cryptic plasmid pAtC58 has no effect on the tumor-forming ability of the Agrobacterium strains; when the Ti plasmid is present, normal tumors are formed on Kalanchoe daigremontiana. PMID:4001194

  7. 亚砷酸氧化菌Sinorhizobium sp.GW3的鉴定与亚砷酸氧化酶基因的分离%Identification of A Novel Arsenite-Oxidizing Bacterium Sinorhizobium sp.GW3 and Isolation of Arsenite Oxidase Gene

    Institute of Scientific and Technical Information of China (English)

    汪耀; 涂书新; 王革娇

    2010-01-01

    亚砷酸氧化细菌能够将毒性大的As(Ⅲ)氧化成毒性小的As(V),在生物修复砷污染方面具有应用价值.对一株新型亚砷酸氧化菌Sinorhizobium sp.GW3进行了较全面的鉴定,并分离及分析了亚砷酸氧化酶基因aoxAB.形态学、生理生化鉴定和16S rRNA基因等分析结果表明该菌为Sinorhizobium属.该菌对As(Ⅲ)抗性的MIC(Minimum inhibitory concentration)为9 mmol/L.首次在Sinorhizobium属中分离了包括编码小亚基aoxA和大亚基aoxB在内的亚砷酸氧化酶基因,其编码的大小亚基与已发现的Agrobacterium tumefaciens(ABB51929)的大小亚基在氨基酸水平上分别有86%、80%的同源性.

  8. Molecular Cloning and Characterization of cgt, the Brucella abortus Cyclic β-1,2-Glucan Transporter Gene, and Its Role in Virulence

    OpenAIRE

    Roset, Mara S.; Ciocchini, Andrés E.; Ugalde, Rodolfo A.; Iñón de Iannino, Nora

    2004-01-01

    The animal pathogen Brucella abortus contains a gene cgt, which complemented Sinorhizobium meliloti nodule development (ndvA) and Agrobacterium tumefaciens chromosomal virulence (chvA) mutants. Complemented strains recovered the presence of anionic cyclic β-1,2-glucan, motility, tumor induction in A. tumefaciens, and nodule occupancy in S. meliloti, all traits strictly associated with the presence of cyclic β-1,2-glucan in the periplasm. Nucleotide sequencing revealed that B. abortus cgt cont...

  9. Final report for DOE grant FG02-06ER15805

    Energy Technology Data Exchange (ETDEWEB)

    Gage, Daniel

    2012-05-31

    DOE funding was used to investigate the role of the phosphotransferase system (PTS) in the symbiotic, nodulating bacterium Sinorhizobium meliloti. This system is well studied in several bacterial species. However, it's organization and function in S. meliloti is substantially different than in the those other, well-studied bacteria. The S. meliloti PTS, through our DOE-funded work, has become a model for how this important signal transduction system works in the a-proteobacteria. We have found that the PTS is relatively simple, used for only signal transduction and not transport, and is involved in regulation of carbon metabolism in response to carbon availability and nitrogen availability.

  10. GenBank blastx search result: AK061948 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061948 001-042-E07 U17226.1 Sinorhizobium meliloti carbon flux regulator (aniA), beta-ketothiolase (phb...A), and acetoacetyl CoA reductase (phbB) genes, complete cds.|BCT BCT 1e-54 +3 ...

  11. GenBank blastx search result: AK059654 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059654 001-031-C10 U17226.1 Sinorhizobium meliloti carbon flux regulator (aniA), beta-ketothiolase (phb...A), and acetoacetyl CoA reductase (phbB) genes, complete cds.|BCT BCT 2e-69 +1 ...

  12. Acacia senegal and Prosopis chilensis-nodulating rhizobia Sinorhizobium arboris HAMBI 2361 and S. kostiense HAMBI 2362 produce tetra- and pentameric LCOs that are N-methylated, O-6-carbamoylated and partially sulfated.

    Science.gov (United States)

    Nowak, Petri; Soupas, Laura; Thomas-Oates, Jane; Lindström, Kristina

    2004-04-28

    Sinorhizobium arboris and S. kostiense are rhizobia that nodulate the tropical leguminous trees Acacia senegal and Prosopis chilensis. The lipochito-oligosaccharidic signalling molecules (LCOs) of S. arboris HAMBI 2361 and S. kostiense HAMBI 2362 were analyzed by mass spectrometry. The major LCOs produced by the strains were shown to be pentameric, acylated with common fatty acids, N-methylated, O-6-carbamoylated and partially sulfated, as are the LCOs characterized to date for other Acacia-nodulating rhizobia. Besides the major LCOs the two strains produced (i) tetrameric LCOs, (ii) LCOs acylated with fatty acids other than those commonly found, (iii) LCOs with only an acyl substituent and (iv) noncarbamoylated LCOs. Production of LCOs (i) to (iii) are novel among Acacia-nodulating rhizobia. The roles of the different structural characteristics of LCOs in the rhizobium-A. senegal symbiosis are discussed. Specific structural features of the LCOs are proposed to be important in the selection of effective nitrogen-fixing rhizobia by A. senegal.

  13. Subcloning and Sequencing of DNA Fragment Related to Salt Tolerance in Sinorhizobium fredii RT19%费氏中华根瘤菌与耐盐有关的DNA片段的亚克隆和测序

    Institute of Scientific and Technical Information of China (English)

    卞学琳; 葛世超; 杨苏声

    2000-01-01

    将费氏中华根瘤菌(Sinorhizobium fredii) RT19与耐盐有关的23kb DNA片段用BamH酶切成大小不同的长度,分别与质粒pML122连接,然后转化大肠杆菌(Escherichia coli) S17-1,筛选出3个转化子。以这些转化子为供体, RT19的盐敏感突变株RC3-3为受体,分别 进行二亲本杂交,筛选到接合子BR2,得到4.4kb与耐盐有关的DNA片段。根据其物理图谱,酶切成6个DNA片段,并分别连接到质粒pUC18进行测序。测序分析表明,该4.4kb DNA片段含有fixO、fixN基因和3个开放阅读框(ORF)。%A 23kb DNA fragment related to salt tolerance was obtained from the gene library of S. fredii strain RT19. In this study, BamH was selected to digest 23kb DNA fragment into different length of DNA fragments. The resulting fragments were ligated with plasmid pML122, then the recombinant plasmids were transformed to competent cells of E. coli S17-1 on selective medium and three transformants TR were obtained. Two-Parental mating experiments were carried out with these transformants as donor and salt sensitive S. fredii strain RC3-3 as recipient, and the transconjugant BR2 was selected on FY plates containing gentamycin and 0.4mol/L NaCl. Thus, a 4.4kb DNA fragment related to salt tolerance was obtained. Based on its physical map, six restriction fragments were subcloned into plasmid pUC18 for DNA sequencing. Subsequently, sequencing and analysis of 4.4kb DNA fragment showed that fixO, fixN genes and three ORFs were obtained.

  14. Genetic characterization of fast-growing rhizobia able to nodulate Prosopis alba in North Spain.

    Science.gov (United States)

    Iglesias, Olga; Rivas, Raúl; García-Fraile, Paula; Abril, Adriana; Mateos, Pedro F; Martinez-Molina, Eustoquio; Velázquez, Encarna

    2007-12-01

    Prosopis is a Mimosaceae legume tree indigenous to South America and not naturalized in Europe. In this work 18 rhizobial strains nodulating Prosopis alba roots were isolated from a soil in North Spain that belong to eight different randomly amplified polymorphic DNA groups phylogenetically related to Sinorhizobium medicae, Sinorhizobium meliloti and Rhizobium giardinii according to their intergenic spacer and 16S rRNA gene sequences. The nodC genes of isolates close to S. medicae and S. meliloti were identical to those of S. medicae USDA 1,037(T) and S. meliloti LMG 6,133(T) and accordingly all these strains were able to nodulate both alfalfa and Prosopis. These nodC genes were phylogenetically divergent from those of the isolates close to R. giardinii that were identical to that of R. giardinii H152(T) and therefore all these strains formed nodules in common beans and Prosopis. The nodC genes of the strains isolated in Spain were phylogenetically divergent from that carried by Mesorhizobium chacoense Pr-5(T) and Sinorhizobium arboris LMG 1,4919(T) nodulating Prosopis in America and Africa, respectively. Therefore, Prosopis is a promiscuous host which can establish symbiosis with strains carrying very divergent nodC genes and this promiscuity may be an important advantage for this legume tree to be used in reforestation.

  15. Analysis of Simple Sequence Repeats in Genomes of Rhizobia

    Institute of Scientific and Technical Information of China (English)

    GAO Ya-mei; HAN Yi-qiang; TANG Hui; SUN Dong-mei; WANG Yan-jie; WANG Wei-dong

    2008-01-01

    Simple sequence repeats (SSRs) or microsatellites, as genetic markers, are ubiquitous in genomes of various organisms. The analysis of SSR in rhizobia genome provides useful information for a variety of applications in population genetics of rhizobia. We analyzed the occurrences, relative abundance, and relative density of SSRs, the most common in Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti genomes se-quenced in the microorganisms tandem repeats database, and SSRs in the three species genomes were compared with each other. The result showed that there were 1 410, 859, and 638 SSRs in B. japonicum, M. loti, and 5. meliloti genomes, respectively. In the genomes of B. japonicum, M. loti, and 5. meliloti, tetranucleotide, pentanucleotide, and hexanucleotide repeats were more abundant and indicated higher mutation rates in these species. The least abundance was mononucleotide repeat. The SSRs type and distribution were similar among these species.

  16. Methods for the isolation of genes encoding novel PHB cycle enzymes from complex microbial communities.

    Science.gov (United States)

    Nordeste, Ricardo F; Trainer, Maria A; Charles, Trevor C

    2010-01-01

    Development of different PHAs as alternatives to petrochemically derived plastics can be facilitated by mining metagenomic libraries for diverse PHA cycle genes that might be useful for synthesis of bioplastics. The specific phenotypes associated with mutations of the PHA synthesis pathway genes in Sinorhizobium meliloti allows for the use of powerful selection and screening tools to identify complementing novel PHA synthesis genes. Identification of novel genes through their function rather than sequence facilitates finding functional proteins that may otherwise have been excluded through sequence-only screening methodology. We present here methods that we have developed for the isolation of clones expressing novel PHA metabolism genes from metagenomic libraries. PMID:20830568

  17. ProMEX – a mass spectral reference database for Plant Proteomics

    Directory of Open Access Journals (Sweden)

    Stefanie eWienkoop

    2012-06-01

    Full Text Available The ProMEX database is one of the main collection of annotated tryptic peptides in plant proteomics. The main objective of the ProMEX Database is to provide experimental MS/MS-based information for cell type-specific or subcellular proteomes in Arabidopsis thaliana, Medicago truncatula, Chlamydomonas reinhardtii, Lotus japonicus, Lotus corniculatus, Phaseolus vulgaris, Lycopersicon esculentum, Solanum tuberosum, Nicotiana tabacum, Glycine max, Zea mays, Bradyrhizobium japonicum and Sinorhizobium meliloti. Direct links at the protein level to the most relevant databases are present in ProMEX. Furthermore, the spectral sequence information are linked to their respective pathways and can be viewed in pathway maps.

  18. Molecular cloning and characterization of cgt, the Brucella abortus cyclic beta-1,2-glucan transporter gene, and its role in virulence.

    Science.gov (United States)

    Roset, Mara S; Ciocchini, Andrés E; Ugalde, Rodolfo A; Iñón de Iannino, Nora

    2004-04-01

    The animal pathogen Brucella abortus contains a gene cgt, which complemented Sinorhizobium meliloti nodule development (ndvA) and Agrobacterium tumefaciens chromosomal virulence (chvA) mutants. Complemented strains recovered the presence of anionic cyclic beta-1,2-glucan, motility, tumor induction in A. tumefaciens, and nodule occupancy in S. meliloti, all traits strictly associated with the presence of cyclic beta-1,2-glucan in the periplasm. Nucleotide sequencing revealed that B. abortus cgt contains a 1,797-bp open reading frame coding for a predicted membrane protein of 599 amino acids (65.9 kDa) that is 58.5 and 59.9% identical to S. meliloti NdvA and A. tumefaciens ChvA, respectively. Additionally, B. abortus cgt, like S. meliloti ndvA and A. tumefaciens chvA possesses ATP-binding motifs and the ABC signature domain features of a typical ABC transporter. Characterization of Cgt was carried out by the construction of null mutants in B. abortus 2308 and S19 backgrounds. Both mutants do not transport cyclic beta-1,2-glucan to the periplasm, as shown by the absence of anionic cyclic glucan, and they display reduced virulence in mice and defective intracellular multiplication in HeLa cells. These results suggest that cyclic beta-1,2-glucan must be transported into the periplasmatic space to exert its action as a virulence factor. PMID:15039351

  19. Present status and development on biological nitrogen fixation research in China

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    This presentation introduces the advances in biological nitrogen fixation research abroad, in particular, describes the great progress and achievements on its research in China as follows: collection of rhizobial resources and establishment of the largest database of Rhizobium in China, correction and development of Rhizobium taxonomy in international; discovery of a couple of nif genes, identification and unification of linkage among the nif gene operons of Klebsiella pneumoniae, finding of regulative mechanism of positive regulation nif gene and its sensitivity to oxygen, temperature; finding of the activity of nodulation gene nodD3 product in Sinorhizobium meliloti which is not controlled by flavonoid produced from its host alfalfa; finding of the association between expression of genes coding the products for carbon utilization and nitrogen metabolism and their regulations; chemical synthesis of nodulation factor of Sinorhizobium meliloti; constructions of engineered nitrogen fixers and utilization in practice based on the research of gene expression and regulation; chemical simulation of the structure and function of nitrogenase and bringing forward the model of nitrogenase active center for the first time in international and synthesis of model compounds which were paid attention by colleagues abroad. Finally, the development of nitrogen fixation research in China in future has been put forward, suggesting that the nif gene regulation and its role in providing crops with nitrogen element, signal transduction and molecular interactions between Rhizobium and legume, coupling between carbon and nitrogen metabolisms, nitrogen fixation and photosynthesis, and functional genomics of nitrogen-fixing nodule symbiosis, etc., would be actively worked on.

  20. Multifaceted Investigation of Metabolites During Nitrogen Fixation in Medicago via High Resolution MALDI-MS Imaging and ESI-MS

    Science.gov (United States)

    Gemperline, Erin; Jayaraman, Dhileepkumar; Maeda, Junko; Ané, Jean-Michel; Li, Lingjun

    2015-01-01

    Legumes have developed the unique ability to establish a symbiotic relationship with soil bacteria known as rhizobia. This interaction results in the formation of root nodules in which rhizobia thrive and reduce atmospheric dinitrogen into plant-usable ammonium through biological nitrogen fixation (BNF). Owing to the availability of genetic information for both of the symbiotic partners, the Medicago truncatula- Sinorhizobium meliloti association is an excellent model for examining the BNF process. Although metabolites are important in this symbiotic association, few studies have investigated the array of metabolites that influence this process. Of these studies, most target only a few specific metabolites, the roles of which are either well known or are part of a well-characterized metabolic pathway. Here, we used a multifaceted mass spectrometric (MS) approach to detect and identify the key metabolites that are present during BNF using the Medicago truncatula- Sinorhizobium meliloti association as the model system. High mass accuracy and high resolution matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) Orbitrap instruments were used in this study and provide complementary results for more in-depth characterization of the nitrogen-fixation process. We used well-characterized plant and bacterial mutants to highlight differences between the metabolites that are present in functional versus nonfunctional nodules. Our study highlights the benefits of using a combination of mass spectrometric techniques to detect differences in metabolite composition and the distributions of these metabolites in plant biology.

  1. Roles of Extracellular Polysaccharides and Biofilm Formation in Heavy Metal Resistance of Rhizobia

    Directory of Open Access Journals (Sweden)

    Natalia Nocelli

    2016-05-01

    Full Text Available Bacterial surface components and extracellular compounds, particularly flagella, lipopolysaccharides (LPSs, and exopolysaccharides (EPSs, in combination with environmental signals and quorum-sensing signals, play crucial roles in bacterial autoaggregation, biofilm development, survival, and host colonization. The nitrogen-fixing species Sinorhizobium meliloti (S. meliloti produces two symbiosis-promoting EPSs: succinoglycan (or EPS I and galactoglucan (or EPS II. Studies of the S. meliloti/alfalfa symbiosis model system have revealed numerous biological functions of EPSs, including host specificity, participation in early stages of host plant infection, signaling molecule during plant development, and (most importantly protection from environmental stresses. We evaluated functions of EPSs in bacterial resistance to heavy metals and metalloids, which are known to affect various biological processes. Heavy metal resistance, biofilm production, and co-culture were tested in the context of previous studies by our group. A range of mercury (Hg II and arsenic (As III concentrations were applied to S. meliloti wild type strain and to mutant strains defective in EPS I and EPS II. The EPS production mutants were generally most sensitive to the metals. Our findings suggest that EPSs are necessary for the protection of bacteria from either Hg (II or As (III stress. Previous studies have described a pump in S. meliloti that causes efflux of arsenic from cells to surrounding culture medium, thereby protecting them from this type of chemical stress. The presence of heavy metals or metalloids in culture medium had no apparent effect on formation of biofilm, in contrast to previous reports that biofilm formation helps protect various microorganism species from adverse environmental conditions. In co-culture experiments, EPS-producing heavy metal resistant strains exerted a protective effect on AEPS-non-producing, heavy metal-sensitive strains; a phenomenon

  2. Diversity of endophytic bacteria associated with nodules of two indigenous legumes at different altitudes of the Qilian Mountains in China.

    Science.gov (United States)

    Xu, Lin; Zhang, Yong; Wang, Li; Chen, Weimin; Wei, Gehong

    2014-09-01

    A total of 201 endophytic root nodule-associated bacteria collected from two legumes indigenous to different Qilian Mountain altitudes (Hexi Corridor) were characterized through 16S rDNA polymerase chain reaction (PCR)-restriction fragment length polymorphism, 16S rRNA gene sequence analysis, and enterobacterial repetitive intergenic consensus-PCR clustering. The isolates phylogenetically belonged to 35 species in the Phyllobacterium, Ensifer, Rhizobium, Microvirga, Sphingomonas, Paracoccus, Mycobacterium, Paenibacillus, Cohnella, Sporosarcina, Bacillus, Staphylococcus, Brevibacterium, Xenophilus, Erwinia, Leclercia, Acinetobacter, and Pseudomonas genera. Phylogenetic nodA sequence analysis showed higher similarity to Sinorhizobium meliloti with strains related to the Rhizobium, Sinorhizobium, and Acinetobacter genera. Sequence analysis of the nifH gene revealed that the strains belonging to Xenophilus, Acinetobacter, Phyllobacterium, and Rhizobium had genes similar to those of Mesorhizobium and Sinorhizobium. The results indicated that horizontal gene transfer could have occurred between rhizobia and non-rhizobial endophytes. Canonical correspondence analysis revealed that altitude and host plant species contributed more to the bacterial endosymbiont separation than other ecological factors. This study provided valuable information on the interactions between symbiotic bacteria, non-symbiotic bacteria and their habitats, and thus provided knowledge on their genetic diversity and ecology. PMID:24985194

  3. Molecular Signals Controlling the Inhibition of Nodulation by Nitrate in Medicago truncatula.

    Science.gov (United States)

    van Noorden, Giel E; Verbeek, Rob; Dinh, Quy Dung; Jin, Jian; Green, Alexandra; Ng, Jason Liang Pin; Mathesius, Ulrike

    2016-01-01

    The presence of nitrogen inhibits legume nodule formation, but the mechanism of this inhibition is poorly understood. We found that 2.5 mM nitrate and above significantly inhibited nodule initiation but not root hair curling in Medicago trunatula. We analyzed protein abundance in M. truncatula roots after treatment with either 0 or 2.5 mM nitrate in the presence or absence of its symbiont Sinorhizobium meliloti after 1, 2 and 5 days following inoculation. Two-dimensional gel electrophoresis combined with mass spectrometry was used to identify 106 differentially accumulated proteins responding to nitrate addition, inoculation or time point. While flavonoid-related proteins were less abundant in the presence of nitrate, addition of Nod gene-inducing flavonoids to the Sinorhizobium culture did not rescue nodulation. Accumulation of auxin in response to rhizobia, which is also controlled by flavonoids, still occurred in the presence of nitrate, but did not localize to a nodule initiation site. Several of the changes included defense- and redox-related proteins, and visualization of reactive oxygen species indicated that their induction in root hairs following Sinorhizobium inoculation was inhibited by nitrate. In summary, the presence of nitrate appears to inhibit nodulation via multiple pathways, including changes to flavonoid metabolism, defense responses and redox changes. PMID:27384556

  4. The Effects of Clinorotation on the Host Plant, Medicago truncatula, and Its Microbial Symbionts

    Directory of Open Access Journals (Sweden)

    Ariel J.C. Dauzart

    2016-02-01

    Full Text Available Understanding the outcome of the plant-microbe symbiosis in altered gravity is vital to developing life support systems for long-distance space travel and colonization of other planets. Thus, the aim of this research was to understand mutualistic relationships between plants and endophytic microbes under the influence of altered gravity. This project utilized the model tripartite relationship among Medicago truncatula ¬– Sinorhizobium meliloti – Rhizophagus irregularis. Plants were inoculated with rhizobial bacteria (S. meliloti, arbuscular mycorrhizal fungi (R. irregularis, or both microbes, and placed on a rotating clinostat. Vertical and horizontal static controls were also performed. Clinorotation significantly reduced M. truncatula dry mass and fresh mass compared to the static controls. The addition of rhizobia treatments under clinorotation also altered total root length and root-to-shoot fresh mass ratio. Nodule size decreased under rhizobia + clinorotation treatment, and nodule density was significantly decreased compared to the vertical treatment. However, inoculation with arbuscular mycorrhizal fungi was shown to increase biomass accumulation and nodule size. Thus, clinorotation significantly affected M. truncatula and its symbiotic relationships with S. meliloti and R. irregularis. In the long term, the results observed in this clinostat study on the changes of plant-microbe mutualism need to be investigated in spaceflight experiments. Thus, careful consideration of the symbiotic microbes of plants should be included in the design of bioregenerative life support systems needed for space travel.

  5. The Effects of Clinorotation on the Host Plant, Medicago truncatula, and Its Microbial Symbionts

    Science.gov (United States)

    Dauzart, Ariel; Vandenbrink, Joshua; Kiss, John

    2016-02-01

    Understanding the outcome of the plant-microbe symbiosis in altered gravity is vital to developing life support systems for long-distance space travel and colonization of other planets. Thus, the aim of this research was to understand mutualistic relationships between plants and endophytic microbes under the influence of altered gravity. This project utilized the model tripartite relationship among Medicago truncatula ¬- Sinorhizobium meliloti - Rhizophagus irregularis. Plants were inoculated with rhizobial bacteria (S. meliloti), arbuscular mycorrhizal fungi (R. irregularis), or both microbes, and placed on a rotating clinostat. Vertical and horizontal static controls were also performed. Clinorotation significantly reduced M. truncatula dry mass and fresh mass compared to the static controls. The addition of rhizobia treatments under clinorotation also altered total root length and root-to-shoot fresh mass ratio. Nodule size decreased under rhizobia + clinorotation treatment, and nodule density was significantly decreased compared to the vertical treatment. However, inoculation with arbuscular mycorrhizal fungi was shown to increase biomass accumulation and nodule size. Thus, clinorotation significantly affected M. truncatula and its symbiotic relationships with S. meliloti and R. irregularis. In the long term, the results observed in this clinostat study on the changes of plant-microbe mutualism need to be investigated in spaceflight experiments. Thus, careful consideration of the symbiotic microbes of plants should be included in the design of bioregenerative life support systems needed for space travel.

  6. Homoserine Lactones Influence the Reaction of Plants to Rhizobia

    Directory of Open Access Journals (Sweden)

    Karl-Heinz Kogel

    2013-08-01

    Full Text Available Bacterial quorum sensing molecules not only grant the communication within bacterial communities, but also influence eukaryotic hosts. N-acyl-homoserine lactones (AHLs produced by pathogenic or beneficial bacteria were shown to induce diverse reactions in animals and plants. In plants, the reaction to AHLs depends on the length of the lipid side chain. Here we investigated the impact of two bacteria on Arabidopsis thaliana, which usually enter a close symbiosis with plants from the Fabaceae (legumes family and produce a long-chain AHL (Sinorhizobium meliloti or a short-chain AHL (Rhizobium etli. We demonstrate that, similarly to the reaction to pure AHL molecules, the impact, which the inoculation with rhizosphere bacteria has on plants, depends on the type of the produced AHL. The inoculation with oxo-C14-HSL-producing S. meliloti strains enhanced plant resistance towards pathogenic bacteria, whereas the inoculation with an AttM lactonase-expressing S. meliloti strain did not. Inoculation with the oxo-C8-HSL-producing R. etli had no impact on the resistance, which is in agreement with our previous hypothesis. In addition, plants seem to influence the availability of AHLs in the rhizosphere. Taken together, this report provides new insights in the role of N-acyl-homoserine lactones in the inter-kingdom communication at the root surface.

  7. Occurrence and Potential Diagnostic Applications of Serological Cross-Reactivities between Brucella and Other Alpha-Proteobacteria

    Science.gov (United States)

    Delpino, M. Victoria; Fossati, Carlos A.; Baldi, Pablo C.

    2004-01-01

    Agrobacterium, Sinorhizobium, and Ochrobactrum are genera closely related to Brucella but, in contrast to the latter, are not pathogenic for humans and animals. We studied by an indirect enzyme-linked immunosorbent assay (ELISA) the reactivities of brucellosis sera against cytosolic (CYT) and membrane (MA) antigens from these nonpathogenic bacteria, and we evaluated the potential usefulness of these cross-reactions for the diagnosis of brucellosis in humans, sheep, cows, and dogs. Canine infection by Brucella canis was detected with high specificity by CYT antigen-based ELISAs (96% for Agrobacterium, 96% for Sinorhizobium, and 91% for Ochrobactrum), while sensitivity was variable (58% for Agrobacterium, 88% for Sinorhizobium, and 84% for Ochrobactrum). In addition, it was possible to diagnose canine disease shortly after exposure to the pathogen (15 days). Similar results for canine brucellosis were obtained with MA antigens. In contrast, normal sera from humans, sheep, and cattle reacted strongly with all the antigens (CYT and MA antigens from the three bacteria), producing high cutoff values and, consequently, low sensitivities. While for some host species the reactivity patterns of normal sera by Western blotting were similar to those produced with sera from infected individuals, the reactivity pattern of bovine sera against Sinorhizobium meliloti antigens exhibited some differential bands for the two groups of sera. These results show that crude fractions from nonpathogenic alpha-proteobacteria can be used to diagnose canine brucellosis but may need to be further separated into simpler fractions to have diagnostic usefulness in ovine, bovine, or human infection. By reducing the biosafety requirements, the use of antigens derived from these nonpathogenic bacteria would simplify the production of diagnostic kits for brucellosis, especially in settings where biosafety level-3 facilities are scarce or absent. PMID:15358645

  8. Harvesting of novel polyhydroxyalkanaote (PHA) synthase encoding genes from a soil metagenome library using phenotypic screening.

    Science.gov (United States)

    Schallmey, Marcus; Ly, Anh; Wang, Chunxia; Meglei, Gabriela; Voget, Sonja; Streit, Wolfgang R; Driscoll, Brian T; Charles, Trevor C

    2011-08-01

    We previously reported the construction of metagenomic libraries in the IncP cosmid vector pRK7813, enabling heterologous expression of these broad-host-range libraries in multiple bacterial hosts. Expressing these libraries in Sinorhizobium meliloti, we have successfully complemented associated phenotypes of polyhydroxyalkanoate synthesis mutants. DNA sequence analysis of three clones indicates that the complementing genes are homologous to, but substantially different from, known polyhydroxyalkanaote synthase-encoding genes. Thus we have demonstrated the ability to isolate diverse genes for polyhydroxyalkanaote synthesis by functional complementation of defined mutants. Such genes might be of use in the engineering of more efficient systems for the industrial production of bioplastics. The use of functional complementation will also provide a vehicle to probe the genetics of polyhydroxyalkanaote metabolism and its relation to carbon availability in complex microbial assemblages. PMID:21631577

  9. Optical disassembly of cellular clusters by tunable tug-of-war tweezers

    CERN Document Server

    Bezryadina, Anna; Chen, Joseph C; Chen, Zhigang

    2016-01-01

    Bacterial biofilms underlie many persistent infections, posing major hurdles in antibiotic treatment. Here, we design and demonstrate tug-of-war optical tweezers that can facilitate assessment of cell-cell adhesion - a key contributing factor to biofilm development, thanks to the combined actions of optical scattering and gradient forces. With a customized optical landscape distinct from that of conventional tweezers, not only can such tug-of-war tweezers stably trap and stretch a rod-shaped bacterium in the observing plane, but, more importantly, they can also impose a tunable lateral force that pulls apart cellular clusters without any tethering or mechanical movement. As a proof of principle, we examined a Sinorhizobium meliloti strain that forms robust biofilms and found that the strength of intercellular adhesion depends on the growth medium. This technique may herald new photonic tools for optical manipulation and biofilm study, as well as other biological applications.

  10. Optical disassembly of cellular clusters by tunable ‘tug-of-war’ tweezers

    Science.gov (United States)

    Bezryadina, Anna S; Preece, Daryl C; Chen, Joseph C; Chen, Zhigang

    2016-01-01

    Bacterial biofilms underlie many persistent infections, posing major hurdles in antibiotic treatment. Here we design and demonstrate ‘tug-of-war’ optical tweezers that can facilitate the assessment of cell–cell adhesion—a key contributing factor to biofilm development, thanks to the combined actions of optical scattering and gradient forces. With a customized optical landscape distinct from that of conventional tweezers, not only can such ‘tug-of-war’ tweezers stably trap and stretch a rod-shaped bacterium in the observing plane, but, more importantly, they can also impose a tunable lateral force that pulls apart cellular clusters without any tethering or mechanical movement. As a proof of principle, we examined a Sinorhizobium meliloti strain that forms robust biofilms and found that the strength of intercellular adhesion depends on the growth medium. This technique may herald new photonic tools for optical manipulation and biofilm study, as well as other biological applications.

  11. Novel associations between rhizobial populations and legume species within the genera Lathyrus and Oxytropis grown in the temperate region of China

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Fifty rhizobial isolates of Lathyrus and Oxytropis collected from northern regions of China were studied in their genotypic characterization based upon analyses of ARDRA, 16S-23S IGS PCR-RFLP, TP-RAPD, MLEE, sequences of 16S rDNA gene and housekeeping genes of atpD, recA and glnII. The results demonstrated that most of the Lathyrus rhizobia belonged to Rhizobium and most of the Oxytropis rhizobia belonged to Sinorhizobium. A novel group of Rhizobium sp. I and S. meliloti were identified as the main microsymbionts respectively associated with Lathyrus and Oxytropis species in the collection area, which were new associations between rhizobia and the mentioned hosts. This study also provides new evidence for biogeography of rhizobia.

  12. Dicty_cDB: Contig-U13420-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 7 2e-58 AL939112_11( AL939112 |pid:none) Streptomyces coelicolor A3(2) com... 179... tab... 92 4e-20 4 ( BM160833 ) EST563356 PyBS Plasmodium yoelii yoelii cDNA clon... 66 4e-20 5 ( AE014831 ) Plasmodi...e bacterium HF10_... 166 4e-51 CP000614_1902( CP000614 |pid:none) Burkholderia vie...tnamiensis G4 c... 162 6e-51 AE017197_777( AE017197 |pid:none) Rickettsia typhi str. Wilmin...:none) Rickettsia peacockii str. Rustic... 170 8e-50 AL591688_1732( AL591688 |pid:none) Sinorhizobium meliloti 1021 co

  13. Dicty_cDB: Contig-U16505-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 12D chromos... 166 2e-54 CP000614_746( CP000614 |pid:none) Burkholderia vietnamiensis G4 ch... 159 3e-54 C...:none) Enterococcus faecium response regu... 121 7e-26 AL939117_51( AL939117 |pid:none) Streptomyces coelicolor A3(2) co...:none) Mus musculus C-terminal binding pr... 87 6e-18 AL591688_2739( AL591688 |pid:none) Sinorhizobium meliloti 1021 co...:none) Streptomyces coelicolor A3(2) co... 157 8e-47 CP001083_3404( CP001083 |pid...3 4 ( FH512937 ) CHO_OF4160xi05f1.ab1 CHO_OF4 Nicotiana tabacum ge... 50 0.30 1 ( AC115599 ) Dictyostelium discoid

  14. Dicty_cDB: Contig-U15660-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 91688_2070( AL591688 |pid:none) Sinorhizobium meliloti 1021 com... 63 3e-08 CP000615_1487( CP000615 |pid:none) Burkholderia vietnam...P000614_2409( CP000614 |pid:none) Burkholderia vietnamiensis G4 c... 56 4e-15 FM242711_2089( FM242711 |pid:n...18 |pid:none) Beutenbergia cavernae DSM 12333... 53 1e-14 CP000617_63( CP000617 |pid:none) Burkholderia vietnam...( CP000615 |pid:none) Burkholderia vietnamiensis G4 c... 48 1e-09 CP001026_1668( ... CP000615_1365( CP000615 |pid:none) Burkholderia vietnamiensis G4 c... 50 7e-06 AM902716_4053( AM902716 |pid

  15. The transcriptional activator LdtR from 'Candidatus Liberibacter asiaticus' mediates osmotic stress tolerance.

    Directory of Open Access Journals (Sweden)

    Fernando A Pagliai

    2014-04-01

    Full Text Available The causal agent of Huanglongbing disease, 'Candidatus Liberibacter asiaticus', is a non-culturable, gram negative, phloem-limited α-proteobacterium. Current methods to control the spread of this disease are still limited to the removal and destruction of infected trees. In this study, we identified and characterized a regulon from 'Ca. L. asiaticus' involved in cell wall remodeling, that contains a member of the MarR family of transcriptional regulators (ldtR, and a predicted L,D-transpeptidase (ldtP. In Sinorhizobium meliloti, mutation of ldtR resulted in morphological changes (shortened rod-type phenotype and reduced tolerance to osmotic stress. A biochemical approach was taken to identify small molecules that modulate LdtR activity. The LdtR ligands identified by thermal shift assays were validated using DNA binding methods. The biological impact of LdtR inactivation by the small molecules was then examined in Sinorhizobium meliloti and Liberibacter crescens, where a shortened-rod phenotype was induced by growth in presence of the ligands. A new method was also developed to examine the effects of small molecules on the viability of 'Ca. Liberibacter asiaticus', using shoots from HLB-infected orange trees. Decreased expression of ldtRLas and ldtPLas was observed in samples taken from HLB-infected shoots after 6 h of incubation with the LdtR ligands. These results provide strong proof of concept for the use of small molecules that target LdtR, as a potential treatment option for Huanglongbing disease.

  16. Efecto de la inoculación con bacterias rizosféricas en dos variedades de trigo. Fase I: condiciones controladas The effect of innoculation with rhizospheric bacteria on two varities of wheat. Phase1: controlled conditions

    Directory of Open Access Journals (Sweden)

    Carlos José Bécquer Granados

    Full Text Available Se llevó a cabo un experimento bajo condiciones controladas para determinar la respuesta de dos variedades de trigo a la inoculación simple y combinada realizada con Sinorhizobium y Azospirillum. Se utilizó una cepa de A. zeae, y dos cepas pertenecientes a S. meliloti. Los materiales y métodos aplicados correspondieron a lo descrito en las metodologías prestablecidas en este campo de estudio. Diseño experimental: completamente aleatorizado, con 20 tratamientos y 4 réplicas. No se utilizó tratamiento fertilizado. Se evaluaron diferentes variables agronómicas, relacionadas con la arquitectura radical y la biomasa aérea. Análisis de varianza bifactorial. En caso de aparecer diferencias, se aplicó análisis de varianza de un factor a la interacción en primera instancia, o a los factores probados. Diferencias entre medias por LSD de Fisher. Se transformaron datos de conteo de dígitos por √x. Se calculó correlación y regresión múltiple entre variables. Se concluye que la inoculación combinada de Sinorhizobium con Azospirillum, así como la inoculación simple con Sinorhizobium, resultaron de alta importancia en las alternativas de inoculación que se realizaron en el experimento. Existió una alta diferenciación entre las dos variedades de trigo en determinadas variables agronómicas, lo que indica una influencia marcada de las características varietales de las plantas. Se observó una fuerte relación estadística entre las variables peso seco aéreo y variables de la raíz para los tratamientos inoculados con A2 y A2+N7, respectivamente.The experiment was carried out under controlled conditions to determine the response of the two wheat varieties to the simple inoculation and the combined inoculation which was carried out with Sinorhizobium, and Azospirillum. A strain of A. zeae and two strains belonging to S. meliloti were used. The materials and methods that were applied corresponded to what was described in the already

  17. Conservation and Occurrence of Trans-Encoded sRNAs in the Rhizobiales

    Directory of Open Access Journals (Sweden)

    Jan Reinkensmeier

    2011-11-01

    Full Text Available Post-transcriptional regulation by trans-encoded sRNAs, for example via base-pairing with target mRNAs, is a common feature in bacteria and influences various cell processes, e.g., response to stress factors. Several studies based on computational and RNA-seq approaches identified approximately 180 trans-encoded sRNAs in Sinorhizobium meliloti. The initial point of this report is a set of 52 trans-encoded sRNAs derived from the former studies. Sequence homology combined with structural conservation analyses were applied to elucidate the occurrence and distribution of conserved trans-encoded sRNAs in the order of Rhizobiales. This approach resulted in 39 RNA family models (RFMs which showed various taxonomic distribution patterns. Whereas the majority of RFMs was restricted to Sinorhizobium species or the Rhizobiaceae, members of a few RFMs were more widely distributed in the Rhizobiales. Access to this data is provided via the RhizoGATE portal [1,2].

  18. LDSS-P: an advanced algorithm to extract functional short motifs associated with coordinated gene expression

    Science.gov (United States)

    Ichida, Hiroyuki; Long, Sharon R.

    2016-01-01

    Identifying functional elements in promoter sequences is a major goal in computational and experimental genome biology. Here, we describe an algorithm, Local Distribution of Short Sequences for Prokaryotes (LDSS-P), to identify conserved short motifs located at specific positions in the promoters of co-expressed prokaryotic genes. As a test case, we applied this algorithm to a symbiotic nitrogen-fixing bacterium, Sinorhizobium meliloti. The LDSS-P profiles that overlap with the 5′ section of the extracytoplasmic function RNA polymerase sigma factor RpoE2 consensus sequences displayed a sharp peak between -34 and -32 from TSS positions. The corresponding genes overlap significantly with RpoE2 targets identified from previous experiments. We further identified several groups of genes that are co-regulated with characterized marker genes. Our data indicate that in S. meliloti, and possibly in other Rhizobiaceae species, the master cell cycle regulator CtrA may recognize an expanded motif (AACCAT), which is positionally shifted from the previously reported CtrA consensus sequence in Caulobacter crescentus. Bacterial one-hybrid experiments showed that base substitution in the expanded motif either increase or decrease the binding by CtrA. These results show the effectiveness of LDSS-P as a method to delineate functional promoter elements. PMID:27190233

  19. Microgravity Effects on the Early Events of Biological Nitrogen Fixation in Medicago Truncatula: Results from the SyNRGE Experiment

    Science.gov (United States)

    Stutte, Gary W.; Roberts, Michael S.

    2013-02-01

    SyNRGE (Symbiotic Nodulation in a Reduced Gravity Environment) was a sortie mission on STS-135 in the Biological Research in Canisters (BRIC) hardware to study the effect of μg on a plant-microbe symbiosis resulting in biological nitrogen fixation. Medicago truncatula, a model species for the legume family, was inoculated with its bacterial symbiont, Sinorhizobium meliloti, to observe early biomolecular events associated with infection and nodulation in Petri Dish Fixation Units (PDFU’s). Two sets of experiments were conducted in orbit and in 24-hour delayed ground controls. Experiments were designed to determine if S. meliloti would infect M. truncatula and initiate biomolecular changes associated with nodule formation and if the μg environment altered the host plant and/or bacteria to induce nodule formation upon return to 1g. Initial analysis results demonstrate that the legumes and bacteria cultivated in μg have potential to develop a symbiotic interaction, but suggest that μg alters their ability to form nodules upon return to 1g. (Research supported by NASA ESMD/ Advance Capabilities Division grant NNX10AR09A)

  20. Diversity and numbers of root-nodule bacteria (rhizobia in Polish soils

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    Stefan Martyniuk

    2011-04-01

    Full Text Available Using a sand pouch-plant infection method, populations of several species of root-nodule bacteria (rhizobia were enumerated in eighty soils collected throughout Poland. Rhizobium leguminosarum bv. viciae (symbionts of pea, faba bean, vetch and R. leguminosarum bv. trifolii (symbionts of clover were detected in 77 and 76 soils, respectively. Most of these soils contained moderate and high numbers of these species of the rhizobia. Symbionts of beans, R. leguminosarum bv. phaseoli, were assessed in 76 soils; of this number 15 soils had no detectable populations of bean rhizobia and in 40 soils high or moderate numbers of these bacteria were found. Bradyrhizobium sp. (Lupinus, root-nodule bacteria of lupine and serradella, were absent in 19 soils, out of 80 tested, and 34 soils were colonised by high or moderate populations of bradyrhizobia. Sinorhizobium meliloti, rhizobia nodulating alfalfa, were sparse in the examined soils; with 56 soil containing no detectable numbers of S. meliloti and only 6 soils harbouring high or moderate populations of this species. The estimated numbers of the rhizobia in the studied soils were also related to some physical and chemical properties of these soils.

  1. NPR1 protein regulates pathogenic and symbiotic interactions between Rhizobium and legumes and non-legumes.

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    Smadar Peleg-Grossman

    Full Text Available BACKGROUND: Legumes are unique in their ability to establish symbiotic interaction with rhizobacteria from Rhizobium genus, which provide them with available nitrogen. Nodulation factors (NFs produced by Rhizobium initiate legume root hair deformation and curling that entrap the bacteria, and allow it to grow inside the plant. In contrast, legumes and non-legumes activate defense responses when inoculated with pathogenic bacteria. One major defense pathway is mediated by salicylic acid (SA. SA is sensed and transduced to downstream defense components by a redox-regulated protein called NPR1. METHODOLOGY/PRINCIPAL FINDINGS: We used Arabidopsis mutants in SA defense pathway to test the role of NPR1 in symbiotic interactions. Inoculation of Sinorhizobium meliloti or purified NF on Medicago truncatula or nim1/npr1 A. thaliana mutants induced root hair deformation and transcription of early and late nodulins. Application of S. meliloti or NF on M. truncatula or A. thaliana roots also induced a strong oxidative burst that lasted much longer than in plants inoculated with pathogenic or mutualistic bacteria. Transient overexpression of NPR1 in M. truncatula suppressed root hair curling, while inhibition of NPR1 expression by RNAi accelerated curling. CONCLUSIONS/SIGNIFICANCE: We show that, while NPR1 has a positive effect on pathogen resistance, it has a negative effect on symbiotic interactions, by inhibiting root hair deformation and nodulin expression. Our results also show that basic plant responses to Rhizobium inoculation are conserved in legumes and non-legumes.

  2. FadD Is Required for Utilization of Endogenous Fatty Acids Released from Membrane Lipids ▿ †

    Science.gov (United States)

    Pech-Canul, Ángel; Nogales, Joaquina; Miranda-Molina, Alfonso; Álvarez, Laura; Geiger, Otto; Soto, María José; López-Lara, Isabel M.

    2011-01-01

    FadD is an acyl coenzyme A (CoA) synthetase responsible for the activation of exogenous long-chain fatty acids (LCFA) into acyl-CoAs. Mutation of fadD in the symbiotic nitrogen-fixing bacterium Sinorhizobium meliloti promotes swarming motility and leads to defects in nodulation of alfalfa plants. In this study, we found that S. meliloti fadD mutants accumulated a mixture of free fatty acids during the stationary phase of growth. The composition of the free fatty acid pool and the results obtained after specific labeling of esterified fatty acids with a Δ5-desaturase (Δ5-Des) were in agreement with membrane phospholipids being the origin of the released fatty acids. Escherichia coli fadD mutants also accumulated free fatty acids released from membrane lipids in the stationary phase. This phenomenon did not occur in a mutant of E. coli with a deficient FadL fatty acid transporter, suggesting that the accumulation of fatty acids in fadD mutants occurs inside the cell. Our results indicate that, besides the activation of exogenous LCFA, in bacteria FadD plays a major role in the activation of endogenous fatty acids released from membrane lipids. Furthermore, expression analysis performed with S. meliloti revealed that a functional FadD is required for the upregulation of genes involved in fatty acid degradation and suggested that in the wild-type strain, the fatty acids released from membrane lipids are degraded by β-oxidation in the stationary phase of growth. PMID:21926226

  3. FadD is required for utilization of endogenous fatty acids released from membrane lipids.

    Science.gov (United States)

    Pech-Canul, Ángel; Nogales, Joaquina; Miranda-Molina, Alfonso; Álvarez, Laura; Geiger, Otto; Soto, María José; López-Lara, Isabel M

    2011-11-01

    FadD is an acyl coenzyme A (CoA) synthetase responsible for the activation of exogenous long-chain fatty acids (LCFA) into acyl-CoAs. Mutation of fadD in the symbiotic nitrogen-fixing bacterium Sinorhizobium meliloti promotes swarming motility and leads to defects in nodulation of alfalfa plants. In this study, we found that S. meliloti fadD mutants accumulated a mixture of free fatty acids during the stationary phase of growth. The composition of the free fatty acid pool and the results obtained after specific labeling of esterified fatty acids with a Δ5-desaturase (Δ5-Des) were in agreement with membrane phospholipids being the origin of the released fatty acids. Escherichia coli fadD mutants also accumulated free fatty acids released from membrane lipids in the stationary phase. This phenomenon did not occur in a mutant of E. coli with a deficient FadL fatty acid transporter, suggesting that the accumulation of fatty acids in fadD mutants occurs inside the cell. Our results indicate that, besides the activation of exogenous LCFA, in bacteria FadD plays a major role in the activation of endogenous fatty acids released from membrane lipids. Furthermore, expression analysis performed with S. meliloti revealed that a functional FadD is required for the upregulation of genes involved in fatty acid degradation and suggested that in the wild-type strain, the fatty acids released from membrane lipids are degraded by β-oxidation in the stationary phase of growth.

  4. Effectivenes of inoculation in alfalfa breeding in ecological conditions of the Bjelovar and Bilogora county

    Directory of Open Access Journals (Sweden)

    Darko Uher

    2012-09-01

    Full Text Available Development and basic existence of animal production as well as production of high quality milk depends upon possibility of sufficient production of quality and protein sufficient forage. Forage crop that satisfies these demands is alfalfa which is one of the most important perennial forage crop legumes. The aim of this study was to enhance alfalfa production on acid soil by liming and alfalfa seed inoculation with efficient Sinorhizobium meliloti strains in order to reduce the use of mineral nitrogen fertilization and enable qualitative and cost effective production of forage on the dairy farms. Field trial was established at family farm in the area of Bjelovar and Bilogora county. During two years experimental period statistically significant influence of inoculation and liming on forage and dry matteryield was determined. Significantly the lowest yields were determined on untreated plots without liming material. In all untreated plots, significantly lower yields were determined, but significant differences in yields were also obtained by inoculation with different S. meliloti strains, emphasizing the importance of strains selection used for alfalfa inoculation. In both experimental years total forage yield were ranging from 34 t/ha (untreated plots without liming up to 60 t/ha on plots inoculated with strain 2011 and without liming. Values of total dry matter yield for both experimental years ranged from 6.5 t/ha (untreated plots without liming up to 15,7 t/ha on plots inoculated with strain 2011 without liming. Results of this study showed that application of liming materials for acidity removal had positive effect on alfalfa yields in both experimental years and significantly improved alfalfa production on acid soils. The results of this study clearly showed that inoculation with selected S. meliloti strains may improve alfalfa production on acid soils and may contribute to more efficient forage production for dairy farms under particular

  5. Diverse role of fast growing rhizobia in growth promotion and enhancement of psoralen content in Psoralea corylifolia L

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    Chandra Prabha

    2013-01-01

    Full Text Available Background: Psoralea corylifolia (Bakuchi, a weed, which possesses a highly potent and medicinally important compound psoralen. P. corylifolia has been widely exploited since ages for its biological potential. Materials and Methods: Fifteen root nodulating bacteria as pure culture collection (PCC were isolated from P. corylifolia in India. Further, these strains were evaluated for their effect on the psoralen content in P. corylifolia. High performance liquid chromatography (HPLC method was used for the estimation of psoralen in P. corylifolia seed extracts. The effectiveness of these rhizobial strains was assessed on the basis of screening of various plant growth promoting attributes. Results: The 16S ribosomal RNA sequencing analysis revealed the identity of two most effective rhizobial isolates PCC2 and PCC7 as Rhizobium leguminosarum and Sinorhizobium meliloti, respectively. The R. leguminosarum PCC2 (JN546144 and Ensifer meliloti PCC7 (JN546145 strains showed solubilization of insoluble inorganic phosphate, secreted indole acetic acid (IAA, produced siderophore, showed ACC deaminase activity, and were positive for nodulation and nitrogen fixing genes. Seeds of P. corylifolia were bacterized with combination of R. leguminosarum PCC2 and Ensifer meliloti PCC7 along with their individual application that resulted in enhancement of various early vegetative and late reproduction parameters of plants in two consecutive field trials in the year 2009 and 2010. The psoralen content in the seeds of P. corylifolia was observed to be increased in the field trials where the combination of rhizobial strains PCC2 and PCC7 was used (2.79% compared to control (1.91%. Conclusion: These findings indicate that rhizobial strains PCC2 and PCC7 showing good plant growth promoting attributes can be effective for increasing the psoralen content in the seeds of P. corylifolia to a certain level.

  6. From Endosymbiont to Host-Controlled Organelle: The Hijacking of Mitochondrial Protein Synthesis and Metabolism

    NARCIS (Netherlands)

    Gabaldon, T.; Huynen, M.A.

    2007-01-01

    Mitochondria are eukaryotic organelles that originated from the endosymbiosis of an alpha-proteobacterium. To gain insight into the evolution of the mitochondrial proteome as it proceeded through the transition from a free-living cell to a specialized organelle, we compared a reconstructed ancestral

  7. From endosymbiont to host-controlled organelle: the hijacking of mitochondrial protein synthesis and metabolism.

    NARCIS (Netherlands)

    Gabaldon, T.; Huynen, M.A.

    2007-01-01

    Mitochondria are eukaryotic organelles that originated from the endosymbiosis of an alpha-proteobacterium. To gain insight into the evolution of the mitochondrial proteome as it proceeded through the transition from a free-living cell to a specialized organelle, we compared a reconstructed ancestral

  8. Efecto de la inoculación con bacterias rizosféricas en dos variedades de trigo. Fase II: invernadero Effect of inoculation with rihizospheric bacteria in two varieties of wheat. Phase II: greenhouse

    Directory of Open Access Journals (Sweden)

    Carlos José Bécquer Granados

    Full Text Available Se llevó a cabo un experimento de invernadero para evaluar la influencia de la inoculación simple y combinada, efectuada con las bacterias rizosféricas Sinorhizobium y Azospirillum, en dos variedades de trigo. Materiales y métodos según lo descrito en las metodologías convencionales para este campo de estudio. El diseño experimental fue completamente aleatorizado, con 4 réplicas y 10 tratamientos. Análisis estadístico varianza bifactorial. Se utilizó tratamiento fertilizado con NH4NO3 (150 ppm/kg suelo. Se evaluó contenido de clorofila foliar, peso seco aéreo, peso seco radical, longitud del tallo y germinación. En caso de aparecer diferencias, se determinaron mediante la prueba de Duncan, y las diferencias entre las variedades con t-Student. Se concluye que la inoculación combinada de la cepaA2 (Sinorhizobium meliloti con la cepaN7 (Azospirillum zeae, fue la de mayor influencia positiva en el contenido de clorofila de las plantas. Por otra parte, existió una alta diferenciación entre las dos variedades de trigo en la longitud del tallo, peso seco aéreo y peso seco radical. Los resultados en peso seco aéreo y peso seco radical, al combinarse los dos factores estudiados, dependieron notablemente de las características varietales de la planta y del efecto significativo de la población autóctona de rizobacterias. La germinación de las plantas no estuvo vinculada a ninguno de los factores aplicados en el experimento.The experiment was carried out in a greenhouse so that the inoculation, both simple and combined (with the rhizospheric bacteria- Sinorhizobium andAzospirllium, en two varieties of Wheat. Materials and methodologies were used in accordance with previously prescribed conventional methodologies for this study. The experimental design was completely randomized, with 4 replicas and 10 treatments. A statistical analysis using two-way variance was done. Fertilized treatment was applied with NH4NO3 (150 ppm/kg soil. The

  9. Growth and Nutrient Uptake of Orchardgrass (Dactylis glomerata L. and Meadow Fescue (Festuca pratensis Huds. as Affected by Rhizobacteria

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    Olivera STAJKOVIĆ-SRBINOVIĆ

    2016-06-01

    Full Text Available A diverse group of soil bacteria found in the rhizosphere which can colonize plant roots and improve plant growth are designated as plant growth promoting rhizobacteria. The aim of this study was isolation and screening of different rhizobacterial strains for plant growth promoting characteristics and their ability to improve growth of two grass species, orchardgrass (Dactylis glomerata L. and meadow fescue (Festuca pratensis Huds.. The strains investigated, belonging to the genera Azotobacter, Bacillus, Pseudomonas and rhizobial bacteria, showed various plant growth promoting traits, such as phosphate solubilisation, siderophore production, and indole-3-acetic acid (IAA production. Co-inoculation of meadow fescue with Azotobacter chroococcum A2 and Sinorhizobium meliloti or Pseudomonas sp., and A. chroococcum A5 with S. meliloti, significantly increased shoot dry weight (SDW(25-33%, as well as total N (26-33%, P (24-31% and K (26-28% contents in plants (mg pot-1, compared to uninoculated control. In addition, inoculation of orchardgrass with A. chroococcum strain A1, as well as co-inoculation with B. megaterium and A. chroococcum A1 or A31, significantly increased SDW (51-59% and total N (54-59%, P (51-74% and K (49-55% contents, compared to uninoculated control. Nitrogen percentage in SDW was slightly higher than sufficiency ranges, while K percentage was optimal in all treatments in both species. Phosphorous percentage was lower than sufficiency ranges as a consequence of very low soil P content. The results emphasize the potential of particular rhizobacteria to improve the growth of forage grasses.

  10. Sulphoacetaldehyde acetyltransferase yields acetyl phosphate: purification from Alcaligenes defragrans and gene clusters in taurine degradation.

    Science.gov (United States)

    Ruff, Jürgen; Denger, Karin; Cook, Alasdair M

    2003-01-15

    The facultatively anaerobic bacterium Alcaligenes defragrans NKNTAU was found to oxidize taurine (2-aminoethanesulphonate) with nitrate as the terminal electron acceptor. Taurine was transaminated to 2-sulphoacetaldehyde. This was not converted into sulphite and acetate by a "sulphoacetaldehyde sulpho-lyase" (EC 4.4.1.12), but into sulphite and acetyl phosphate, which was identified by three methods. The enzyme, which required the addition of phosphate, thiamin diphosphate and Mg(2+) ions for activity, was renamed sulphoacetaldehyde acetyltransferase (Xsc; EC 2.3.1.-). Inducible Xsc was expressed at high levels, and a three-step 11-fold purification yielded an essentially homogeneous soluble protein, which was a homotetramer in its native form; the molecular mass of the subunit was found to be between about 63 kDa (SDS/PAGE) and 65.3 kDa (matrix-assisted laser-desorption ionization-time-of-flight MS). The N-terminal and two internal amino acid sequences were determined, and PCR primers were generated. The xsc gene was amplified and sequenced; the derived molecular mass of the processed protein was 65.0 kDa. The downstream gene presumably encoded the inducible phosphate acetyltransferase (Pta) found in crude extracts. The desulphonative enzymes ("EC 4.4.1.12") from Achromobacter xylosoxidans NCIMB 10751 and Desulfonispora thiosulfatigenes GKNTAU were shown to be Xscs. We detected at least three subclasses of xsc in Proteobacteria and in Gram-positive bacteria, and they comprised a distinct group within the acetohydroxyacid synthase supergene family. Genome sequencing data revealed xsc genes in Burkholderia fungorum (80% sequence identity) and Sinorhizobium meliloti (61%) with closely linked pta genes. Different patterns of regulation for the transport and dissimilation of taurine were hypothesized for S. meliloti and B. fungorum. PMID:12358600

  11. Dibenzofuran degradation by Sphingomonas wittichii RW1 under environmental stresses

    OpenAIRE

    Coronado E.

    2013-01-01

    Sphingomonas wittichii is a gram-negative Alpha-proteobacterium, capable of degrading xenobiotic compounds such as dibenzofuran (DBF), dibenzo-p-dioxin, carbazole, 2-hydroxybiphenyl or nitro diphenyl ether herbicides. The metabolism of strain RW1 has been the subject of previous studies and a number of genes involved in DBF degradation have been characterized. It is known that RW1 posseses a unique initial DBF dioxygenase (encoded by the dxnAl gene) that catalyzes the first step in the degrad...

  12. A Transposable Partitioning Locus Used To Stabilize Plasmid-Borne Hydrogen Oxidation and Trifolitoxin Production Genes in a Sinorhizobium Strain

    OpenAIRE

    Kent, Angela D.; Wojtasiak, Michelle L.; Robleto, Eduardo A.; Eric W Triplett

    1998-01-01

    Improved nitrogen-fixing inoculum strains for leguminous crops must be able to effectively compete with indigenous strains for nodulation, enhance legume productivity compared to the productivity obtained with indigenous strains, and maintain stable expression of any added genes in the absence of selection pressure. We constructed a transposable element containing the tfx region for expression of increased nodulation competitiveness and the par locus for plasmid stability. The transposon was ...

  13. Different biochemical mechanisms ensure network-wide balancing of reducing equivalents in microbial metabolism.

    Science.gov (United States)

    Fuhrer, Tobias; Sauer, Uwe

    2009-04-01

    To sustain growth, the catabolic formation of the redox equivalent NADPH must be balanced with the anabolic demand. The mechanisms that ensure such network-wide balancing, however, are presently not understood. Based on 13C-detected intracellular fluxes, metabolite concentrations, and cofactor specificities for all relevant central metabolic enzymes, we have quantified catabolic NADPH production in Agrobacterium tumefaciens, Bacillus subtilis, Escherichia coli, Paracoccus versutus, Pseudomonas fluorescens, Rhodobacter sphaeroides, Sinorhizobium meliloti, and Zymomonas mobilis. For six species, the estimated NADPH production from glucose catabolism exceeded the requirements for biomass synthesis. Exceptions were P. fluorescens, with balanced rates, and E. coli, with insufficient catabolic production, in which about one-third of the NADPH is supplied via the membrane-bound transhydrogenase PntAB. P. versutus and B. subtilis were the only species that appear to rely on transhydrogenases for balancing NADPH overproduction during growth on glucose. In the other four species, the main but not exclusive redox-balancing mechanism appears to be the dual cofactor specificities of several catabolic enzymes and/or the existence of isoenzymes with distinct cofactor specificities, in particular glucose 6-phosphate dehydrogenase. An unexpected key finding for all species, except E. coli and B. subtilis, was the lack of cofactor specificity in the oxidative pentose phosphate pathway, which contrasts with the textbook view of the pentose phosphate pathway dehydrogenases as being NADP+ dependent.

  14. Choline uptake in Agrobacterium tumefaciens by the high-affinity ChoXWV transporter.

    Science.gov (United States)

    Aktas, Meriyem; Jost, Kathinka A; Fritz, Christiane; Narberhaus, Franz

    2011-10-01

    Agrobacterium tumefaciens is a facultative phytopathogen that causes crown gall disease. For successful plant transformation A. tumefaciens requires the membrane lipid phosphatidylcholine (PC), which is produced via the methylation and the PC synthase (Pcs) pathways. The latter route is dependent on choline. Although choline uptake has been demonstrated in A. tumefaciens, the responsible transporter(s) remained elusive. In this study, we identified the first choline transport system in A. tumefaciens. The ABC-type choline transporter is encoded by the chromosomally located choXWV operon (ChoX, binding protein; ChoW, permease; and ChoV, ATPase). The Cho system is not critical for growth and PC synthesis. However, [14C]choline uptake is severely reduced in A. tumefaciens choX mutants. Recombinant ChoX is able to bind choline with high affinity (equilibrium dissociation constant [KD] of ≈2 μM). Since other quaternary amines are bound by ChoX with much lower affinities (acetylcholine, KD of ≈80 μM; betaine, KD of ≈470 μM), the ChoXWV system functions as a high-affinity transporter with a preference for choline. Two tryptophan residues (W40 and W87) located in the predicted ligand-binding pocket are essential for choline binding. The structural model of ChoX built on Sinorhizobium meliloti ChoX resembles the typical structure of substrate binding proteins with a so-called "Venus flytrap mechanism" of substrate binding. PMID:21803998

  15. Exploring root symbiotic programs in the model legume Medicago truncatula using EST analysis

    Science.gov (United States)

    Journet, Etienne-Pascal; van Tuinen, Diederik; Gouzy, Jérome; Crespeau, Hervé; Carreau, Véronique; Farmer, Mary-Jo; Niebel, Andreas; Schiex, Thomas; Jaillon, Olivier; Chatagnier, Odile; Godiard, Laurence; Micheli, Fabienne; Kahn, Daniel; Gianinazzi-Pearson, Vivienne; Gamas, Pascal

    2002-01-01

    We report on a large-scale expressed sequence tag (EST) sequencing and analysis program aimed at characterizing the sets of genes expressed in roots of the model legume Medicago truncatula during interactions with either of two microsymbionts, the nitrogen-fixing bacterium Sinorhizobium meliloti or the arbuscular mycorrhizal fungus Glomus intraradices. We have designed specific tools for in silico analysis of EST data, in relation to chimeric cDNA detection, EST clustering, encoded protein prediction, and detection of differential expression. Our 21 473 5′- and 3′-ESTs could be grouped into 6359 EST clusters, corresponding to distinct virtual genes, along with 52 498 other M.truncatula ESTs available in the dbEST (NCBI) database that were recruited in the process. These clusters were manually annotated, using a specifically developed annotation interface. Analysis of EST cluster distribution in various M.truncatula cDNA libraries, supported by a refined R test to evaluate statistical significance and by ‘electronic northern’ representation, enabled us to identify a large number of novel genes predicted to be up- or down-regulated during either symbiotic root interaction. These in silico analyses provide a first global view of the genetic programs for root symbioses in M.truncatula. A searchable database has been built and can be accessed through a public interface. PMID:12490726

  16. Photosynthetic and Molecular Markers of CO2-mediated Photosynthetic Downregulation in Nodulated Alfalfa

    Institute of Scientific and Technical Information of China (English)

    (A)lvaro Sanz-Sáez; Gorka Erice; Iker Aranjuelo; Ricardo Aroca; Juan Manuel Ruíz-Lozano; Jone Aguirreolea; Juan José Irigoyen

    2013-01-01

    Elevated CO2 leads to a decrease in potential net photosynthesis in long-term experiments and thus to a reduction in potential growth.This process is known as photosynthetic downregulation.There is no agreement on the definition of which parameters are the most sensitive for detecting CO2 acclimation.In order to investigate the most sensitive photosynthetic and molecular markers of CO2 acclimation,the effects of elevated CO2,and associated elevated temperature were analyzed in alfalfa plants inoculated with different Sinorhizobium meliloti strains.Plants (Medicago sativa L.cv.Aragón) were grown in summer or autumn in temperature gradient greenhouses (TGG).At the end of the experiment,all plants showed acclimation in both seasons,especially under elevated summer temperatures.This was probably due to the lower nitrogen (N) availability caused by decreased N2-fixation under higher temperatures.Photosynthesis measured at growth CO2 concentration,rubisco in vitro activity and maximum rate of carboxylation were the most sensitive parameters for detecting downregulation.Severe acclimation was also related with decreases in leaf nitrogen content associated with declines in rubisco content (large and small subunits) and activity that resulted in a drop in photosynthesis.Despite the sensitivity of rubisco content as a marker of acclimation,it was not coordinated with gene expression,possibly due to a lag between gene transcription and protein translation.

  17. Azide resistance in Rhizobium ciceri linked with superior symbiotic nitrogen fixation.

    Science.gov (United States)

    Bhaskar, V Vijay

    2004-12-01

    Isolated azide resistant (AzR) native R. ciceri strain 18-7 was resistant to sodium azide at 10 microg/ml. To find if nif-reiteration is responsible for azide resistance and linked to superior symbiotic nitrogen fixation, transposon (Tn5) induced azide sensitive mutants were generated. Using 4 kb nif-reiterated Sinorhizobium meliloti DNA, a clone C4 that complemented azide sensitivity was isolated by DNA hybridization from genomic library of chickpea Rhizobium strain Rcd301. EcoRI restriction mapping revealed the presence of 7 recognition sites with a total insert size of 19.17 kb. Restriction analysis of C4 clone and nif-reiterated DNA (pRK 290.7) with EcoRI and XhoI revealed similar banding pattern. Wild type strain 18-7, mutant M126 and complemented mutant M126(C4) were characterized for symbiotic properties (viz., acetylene reduction assay, total nitrogen content, nodule number and fresh and dry weight of the infected plants) and explanta nitrogenase activity. Our results suggested that azide resistance, nif-reiteration, and superior symbiotic effectiveness were interlinked with no correlation between ex-planta nitrogenase activity and azide resistance in R. ciceri.

  18. The destructive citrus pathogen, 'Candidatus Liberibacter asiaticus' encodes a functional flagellin characteristic of a pathogen-associated molecular pattern.

    Directory of Open Access Journals (Sweden)

    Huasong Zou

    Full Text Available Huanglongbing (HLB is presently the most devastating citrus disease worldwide. As an intracellular plant pathogen and insect symbiont, the HLB bacterium, 'Candidatus Liberibacter asiaticus' (Las, retains the entire flagellum-encoding gene cluster in its significantly reduced genome. Las encodes a flagellin and hook-associated protein (Fla of 452 amino acids that contains a conserved 22 amino acid domain (flg22 at positions 29 to 50 in the N-terminus. The phenotypic alteration in motility of a Sinorhizobium meliloti mutant lacking the fla genes was partially restored by constitutive expression of Fla(Las. Agrobacterium-mediated transient expression in planta revealed that Fla(Las induced cell death and callose deposition in Nicotiana benthamiana, and that the transcription of BAK1 and SGT1, which are associated with plant innate immunity, was upregulated. Amino acid substitution experiments revealed that residues 38 (serine and 39 (aspartate of Fla(Las were essential for callose induction. The synthetic flg22(Las peptide could not induce plant cell death but retained the ability to induce callose deposition at a concentration of 20 µM or above. This demonstrated that the pathogen-associated molecular pattern (PAMP activity of flg22 in Las was weaker than those in other well-studied plant pathogenic bacteria. These results indicate that Fla(Las acts as a PAMP and may play an important role in triggering host plant resistance to the HLB bacteria.

  19. Biodegradation pathway and detoxification of the diazo dye Reactive Black 5 by Phanerochaete chrysosporium.

    Science.gov (United States)

    Enayatizamir, Naeimeh; Tabandeh, Fatemeh; Rodríguez-Couto, Susana; Yakhchali, Bagher; Alikhani, Hossein A; Mohammadi, Leila

    2011-11-01

    The in vivo biodegradation of the diazo dye Reactive Black 5 (RB5) by Phanerochaete chrysosporium immobilised on cubes of nylon sponge and on sunflower-seed shells (SS) in laboratory-scale bioreactors was investigated. The SS cultivation led to the best results with a decolouration percentage of 90.3% in 72 h for an initial RB5 concentration of 100 mg/L. It was found that the addition of 0.4 mM veratryl alcohol (VA) into the medium considerably increased the decolouration rate in SS cultivation. However, the addition of VA had no effect in the nylon cultivation. Thin layer chromatography (TLC) revealed that RB5 was transformed into one metabolite after 24 h. UV-vis spectroscopy and Fourier Transform Infrared (FT-IR) also confirmed the biodegradation of RB5. Toxicity of RB5 solutions before and after fungal treatment was assayed using Sinorhizobium meliloti as a sensitive soil microorganism. P. chrysosporium transformed the toxic dye RB5 into a non-toxic product.

  20. Genomic analyses of metal resistance genes in three plant growth promoting bacteria of legume plants in Northwest mine tailings, China

    Institute of Scientific and Technical Information of China (English)

    Pin Xie; Xiuli Hao; Martin Herzberg; Yantao Luo; Dietrich H.Nies; Gehong Wei

    2015-01-01

    To better understand the diversity of metal resistance genetic determinant from microbes that survived at metal tailings in northwest of China,a highly elevated level of heavy metal containing region,genomic analyses was conducted using genome sequence of three native metal-resistant plant growth promoting bacteria (PGPB).It shows that:Mesorhizobium amorphae CCNWGS0123 contains metal ~nsporters from P-type ATPase,CDF (Cation Diffusion Facilitator),HupE/UreJ and CHR (chromate ion transporter) family involved in copper,zinc,nickel as well as chromate resistance and homeostasis.Meanwhile,the putative CopA/CueO system is expected to mediate copper resistance in Sinorhizobium meliloti CCNWSX0020 while ZntA transporter,assisted with putative CzcD,determines zinc tolerance in Agrobacterium tumefaciens CCNWGS0286.The greenhouse experiment provides the consistent evidence of the plant growth promoting effects of these microbes on their hosts by nitrogen fixation and/or indoleacetic acid (IAA) secretion,indicating a potential in-site phytoremediation usage in the mining tailing regions of China.

  1. Enhanced resistance to nanoparticle toxicity is conferred by overproduction of extracellular polymeric substances

    Energy Technology Data Exchange (ETDEWEB)

    Joshi, Nimisha, E-mail: joshi.nimisha@gmail.com [School of GeoSciences, Microbial Geochemistry Laboratory, University of Edinburgh, West Mains Road, Edinburgh EH9 3JW (United Kingdom); Ngwenya, Bryne T. [School of GeoSciences, Microbial Geochemistry Laboratory, University of Edinburgh, West Mains Road, Edinburgh EH9 3JW (United Kingdom); French, Christopher E. [School of Biological Sciences, Institute of Cell Biology, Darwin Building, University of Edinburgh, Mayfield Road, Edinburgh EH9 3JR (United Kingdom)

    2012-11-30

    Highlights: Black-Right-Pointing-Pointer Demonstration that bacteria engineered for EPS overproduction have better survival against Ag nanotoxicity. Black-Right-Pointing-Pointer EPS destabilises Ag nanoparticles and promotes their aggregation. Black-Right-Pointing-Pointer TEM demonstration that EPS traps the Ag nanoparticles outside the cell. Black-Right-Pointing-Pointer EPS from overexpressing strains offers protection to non-EPS strains of bacteria. Black-Right-Pointing-Pointer EPS polymer analogues such as xanthan also produce a similar response. - Abstract: The increasing production and use of engineered nanoparticles, coupled with their demonstrated toxicity to different organisms, demands the development of a systematic understanding of how nanoparticle toxicity depends on important environmental parameters as well as surface properties of both cells and nanomaterials. We demonstrate that production of the extracellular polymeric substance (EPS), colanic acid by engineered Escherichia coli protects the bacteria against silver nanoparticle toxicity. Moreover, exogenous addition of EPS to a control strain results in an increase in cell viability, as does the addition of commercial EPS polymer analogue xanthan. Furthermore, we have found that an EPS producing strain of Sinorhizobium meliloti shows higher survival upon exposure to silver nanoparticles than the parent strain. Transmission electron microscopy (TEM) observations showed that EPS traps the nanoparticles outside the cells and reduces the exposed surface area of cells to incoming nanoparticles by inducing cell aggregation. Nanoparticle size characterization in the presence of EPS and xanthan indicated a marked tendency towards aggregation. Both are likely effective mechanisms for reducing nanoparticle toxicity in the natural environment.

  2. Enhanced resistance to nanoparticle toxicity is conferred by overproduction of extracellular polymeric substances

    International Nuclear Information System (INIS)

    Highlights: ► Demonstration that bacteria engineered for EPS overproduction have better survival against Ag nanotoxicity. ► EPS destabilises Ag nanoparticles and promotes their aggregation. ► TEM demonstration that EPS traps the Ag nanoparticles outside the cell. ► EPS from overexpressing strains offers protection to non-EPS strains of bacteria. ► EPS polymer analogues such as xanthan also produce a similar response. - Abstract: The increasing production and use of engineered nanoparticles, coupled with their demonstrated toxicity to different organisms, demands the development of a systematic understanding of how nanoparticle toxicity depends on important environmental parameters as well as surface properties of both cells and nanomaterials. We demonstrate that production of the extracellular polymeric substance (EPS), colanic acid by engineered Escherichia coli protects the bacteria against silver nanoparticle toxicity. Moreover, exogenous addition of EPS to a control strain results in an increase in cell viability, as does the addition of commercial EPS polymer analogue xanthan. Furthermore, we have found that an EPS producing strain of Sinorhizobium meliloti shows higher survival upon exposure to silver nanoparticles than the parent strain. Transmission electron microscopy (TEM) observations showed that EPS traps the nanoparticles outside the cells and reduces the exposed surface area of cells to incoming nanoparticles by inducing cell aggregation. Nanoparticle size characterization in the presence of EPS and xanthan indicated a marked tendency towards aggregation. Both are likely effective mechanisms for reducing nanoparticle toxicity in the natural environment.

  3. Improvement of Nitrogen Fixation Efficiency and Plasmid Stability in Bradyrhizobium japonicum by the Introduction of dctABD and parCBA/DE Genes%导入dctABD和parCBA/DE基因提高大豆慢生根瘤菌固氮效率和稳定性的研究

    Institute of Scientific and Technical Information of China (English)

    李友国; 李杰; 刘墨青; 周俊初

    2000-01-01

    以pLAFR3为载体构建重组质粒pHN207,携带有来自苜蓿根瘤菌(Sinorhizobium meliloti)的四碳二羧酸转移酶基因dctABD、来自pTR102的parCBA/DE基因和标记发光酶基因luxAB。利用2亲本杂交法,将重组质粒pHN207导入大豆慢生根瘤菌(Bradyrhizobium japonicum)TA11和CB1809,分别考察了转移接合子中外源重组质粒在人工培养条件和共生条件下的稳定性,结果表明par基因的引入明显提高pLAFR3在TA11和CB1809中的稳定性。dctABD基因可显著提高TA11和CB1809在大豆黑龙33、宁镇一号和渝豆一号上的共生固氮能力,使结瘤植物的地上部分干重(生物量)和总氮量等指标较对照组有显著提高。%ArecombinantplasmidpHN207containingC4-dicarboxylicacidtransport genes(dctABD) from Sinorhizobium meliloti, parCBA/DE genes from pTR102 and reporter genes luxAB from pDB30 was constructed by using pLAFR3 as the vector. The pHN207 was then introduced into the Bradyrhizobium japonicum TA11 and CB1809 by bi-parental mating. It was confirmed that parCBA/DE genes could increase the stability of pLAFR3 in the transconjugants under both free-living and symbiotic condition. The results of plant pot experiment indicated that the introduction of dctABD genes could significantly improve the symbiotic nitrogen fixation efficiency of TA11 and CB1809 with soybean varieties of Heilong 33, Ningzhen No.1 and Yudou No.1. Compared with the control, the shoot dry weight (biomass) and total nitrogen content of the plants tested were significantly increased.

  4. The non-specific lipid transfer protein N5 of Medicago truncatula is implicated in epidermal stages of rhizobium-host interaction

    Directory of Open Access Journals (Sweden)

    Pii Youry

    2012-12-01

    Full Text Available Abstract Background The symbiotic interaction between leguminous plants and rhizobia involves two processes: bacterial infection, resulting in the penetration of bacteria in epidermal and cortical cells, and root nodule organogenesis. Root nodule symbiosis is activated by rhizobial signalling molecules, called Nodulation factors (NFs. NF perception induces the expression of several genes called early nodulins. The early nodulin N5 of Medicago truncatula is a lipid transfer protein that has been shown to positively regulate nodulation although it displays in vitro inhibitory activity against Sinorhizobium meliloti. The purpose of this work was to investigate the role of MtN5 by studying its spatial and temporal pattern of expression during the symbiotic interaction, also in relation to known components of the symbiotic signalling pathway, and by analysing the phenotypic alterations displayed by rhizobia-inoculated MtN5-silenced roots. Results We show here that MtN5 is a NF-responsive gene expressed at a very early phase of symbiosis in epidermal cells and root hairs. MtN5 expression is induced in vitro by rhizobial effector molecules and by auxin and cytokinin, phytohormones involved in nodule organogenesis. Furthermore, lipid signaling is implicated in the response of MtN5 to rhizobia, since the activity of phospholipase D is required for MtN5 induction in S. meliloti-inoculated roots. MtN5-silenced roots inoculated with rhizobia display an increased root hair curling and a reduced number of invaded primordia compared to that in wild type roots, but with no impairment to nodule primordia formation. This phenotype is associated with the stimulation of ENOD11 expression, an early marker of infection, and with the down-regulation of Flotillin 4 (FLOT4, a protein involved in rhizobial entry. Conclusions These data indicate that MtN5 acts downstream of NF perception and upstream of FLOT4 in regulating pre-infection events. The positive effect of MtN5

  5. Fixação do nitrogênio em alfafa nodulada sob supressão e ressuprimento de fósforo Nitrogen fixation in alfalfa nodulated under phosphorus supression and resupply

    Directory of Open Access Journals (Sweden)

    Fernando Teixeira Gomes

    2002-12-01

    Full Text Available Estudaram-se os efeitos da supressão e do ressuprimento de fósforo (Pi sobre a fixação biológica do nitrogênio atmosférico (N2 em plantas de alfafa cv. Flórida 77 inoculadas com Sinorhizobium meliloti, em diferentes estádios do desenvolvimento vegetativo (V3 e V4 e reprodutivo (R6 e R8. O ensaio foi conduzido em casa de vegetação e as plantas cultivadas em solução nutritiva. O período de supressão de Pi por dez dias reduziu os teores de Pi nas folhas e nos nódulos em todos os estádios do desenvolvimento, enquanto nas raízes essa redução somente foi observada nos estádios vegetativos. Após o ressuprimento de Pi à solução nutritiva por igual período, dez dias, foi observada a recuperação nos teores de Pi nos estádios R6 e R8 para folhas, V3 e V4 para raízes e V3 para os nódulos. A supressão de Pi alterou o processo de fixação do N2, estimado pela concentração de aminoácidos totais nas folhas e nas raízes. Os teores de aminoácidos nas folhas e nas raízes foram significativamente menores nas plantas sob supressão de Pi, em relação aos das plantas do tratamento controle. Após o ressuprimento os teores de aminoácidos totais nas raízes, em todos os estádios do desenvolvimento, alcançaram valores similares àqueles das plantas do tratamento controle, enquanto nas folhas isso só ocorreu nos estádios vegetativos. A supressão de Pi não influenciou a proporção de aminoácidos livres na seiva do xilema.Phosphorus (Pi suppression and resupply effects were studied on nitrogen biological fixation (N2 in alfalfa cv. Florida 77 inoculated with Sinorhizobium meliloti in different vegetative (V3 and V4 and reproductive (R6 and R8 stages. The experiment was carried in greenhouse and the plants cultivated in nutritive solution. The inorganic phosphorus (Pi ten days suppression period decreased Pi levels in leaves and nodules in all growth stages, whereas in the roots this decrease was observed only in the

  6. Development of a Microemulsion Formulation for Antimicrobial SecA Inhibitors

    Science.gov (United States)

    Wang, Nian

    2016-01-01

    In our previous study, we have identified five antimicrobial small molecules via structure based design, which inhibit SecA of Candidatus Liberibacter asiaticus (Las). SecA is a critical protein translocase ATPase subunit and is involved in pre-protein translocation across and integration into the cellular membrane in bacteria. In this study, eleven compounds were identified using similarity search method based on the five lead SecA inhibitors identified previously. The identified SecA inhibitors have poor aqueous solubility. Thus a microemulsion master mix (MMX) was developed to address the solubility issue and for application of the antimicrobials. MMX consists of N-methyl-2-pyrrolidone and dimethyl sulfoxide as solvent and co-solvent, as well as polyoxyethylated castor oil, polyalkylene glycol, and polyoxyethylene tridecyl ether phosphate as surfactants. MMX has significantly improved the solubility of SecA inhibitors and has no or little phytotoxic effects at concentrations less than 5.0% (v/v). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the SecA inhibitors and streptomycin against eight bacteria including Agrobacterium tumefaciens, Liberibacter crescens, Rhizobium etli, Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti phylogenetically related to Las were determined using the broth microdilution method. MIC and MBC results showed that the 16 SecA inhibitors have antibacterial activities comparable to that of streptomycin. Overall, we have identified 11 potent SecA inhibitors using similarity search method. We have developed a microemulsion formulation for SecA inhibitors which improved the antimicrobial activities of SecA inhibitors. PMID:26963811

  7. Characterization of rhizobia from legumes of agronomic interest grown in semi-arid areas of Central Spain relates genetic differences to soil properties.

    Science.gov (United States)

    Ruiz-Díez, Beatriz; Fajardo, Susana; Felipe, María del Rosario de; Fernández-Pascual, Mercedes

    2012-02-01

    A study of symbiotic bacteria from traditional agricultural legumes from Central Spain was performed to create a collection of rhizobia from soils differing in physicochemical, analytical and/or agroecological properties which could be well-adapted to the environmental conditions of this region, and be used for sustainable agricultural practices. Thirty-six isolates were obtained from root-nodules of fifteen legume species (including Cicer arietinum, Lathyrus sativus, Lens culinaris, Lupinus spp., Medicago sativa, Phaseolus vulgaris, Pisum sativum, and Vicia spp.) from three agriculture areas with soils of different pHs and from a forest area with undisturbed soils. Phenotypical characterization revealed uniformity across the thirty-six isolates, with important exceptions in terms of environmental tolerance (three isolates survived at high temperatures, three at high salinity and three at acid pH). The molecular analysis of 16S rRNA gene showed a close relationship of twenty-nine isolates to Rhizobium leguminosarum, one to Rhizobium gallicum, one to Mesorhizobium ciceri, two to Sinorhizobium (Ensifer) meliloti and three to Bradyrhizobium canariense. The sequence analysis of a symbiosis-specific gene, nod C, showed a correlation with the plant host and grouped twenty-six isolates with Rhizobium leguminosarum bv. viciae, establishing the diversity in relation to legume-host. The 16S-23S rRNA intergenic spacer (IGS) region allowed for intraspecific differentiation, so that strains with equal 16S rRNA were grouped by means of their soil origin. These results indicated that phenotypical and genetically related strains may be widely distributed in this region and that soil abiotic characteristics could have a substantial bearing on the selection of the strains living in each environment. PMID:21953333

  8. utilization of bio fertilizers and organic sources in arable soils under saline conditions using tracer technique

    International Nuclear Information System (INIS)

    Recently, more attention has been paid to conserve and save surrounding environment via minimizing the excessive use of chemical fertilizers and, in general, the agrochemicals applied in heavy quantities in agricultural agroecosystems. Therefore, the attention of most of agronomists was turned towards the use of so called clean agriculture or organic farming. Many of organic systems was pointed out such as the recycling of farm wastes i.e. crop residues, animal manure, organic conditioners for reclamation of soil and in the same time enhancement of plant growth and improving yield quality. The application of organic wastes combined with or without microbial inoculants to plant media are considered as a good management practice in any agricultural production system because it improves, plant quality and soil fertility. Therefore, we have the opportunity to conduct some experiments for achieving the clean agriculture approach, combating the adverse effects of salinity and avoiding the environmental pollution. Series of laboratory and greenhouse experiments were carried out to evaluate the impact of (1) potent isolated fungi (Aspergillus oryzae and Aspergillus terreus) on degrading plant residues (Leucaena and Acacia green parts), and (2) biofertilizers (Sinorhizobium meliloti, Azospirillum brasilense, and Pseudomonas aeruginosa) in assessing barley and spinach plants to combat salinity of soil and irrigation water.15N-tracer technique that considered unique and more reliable technique may benefits in clarifying the responsible mechanisms related to plant growth and gave us the opportunity to quantify the exact amounts of N derived from the different sources of nitrogen available to spinach and barley plants grown on sandy saline soil and irrigated with saline water.

  9. Molybdate uptake by Agrobacterium tumefaciens correlates with the cellular molybdenum cofactor status.

    Science.gov (United States)

    Hoffmann, Marie-Christine; Ali, Koral; Sonnenschein, Marleen; Robrahn, Laura; Strauss, Daria; Narberhaus, Franz; Masepohl, Bernd

    2016-09-01

    Many enzymes require the molybdenum cofactor, Moco. Under Mo-limiting conditions, the high-affinity ABC transporter ModABC permits molybdate uptake and Moco biosynthesis in bacteria. Under Mo-replete conditions, Escherichia coli represses modABC transcription by the one-component regulator, ModE, consisting of a DNA-binding and a molybdate-sensing domain. Instead of a full-length ModE protein, many bacteria have a shorter ModE protein, ModE(S) , consisting of a DNA-binding domain only. Here, we asked how such proteins sense the intracellular molybdenum status. We show that the Agrobacterium tumefaciens ModE(S) protein Atu2564 is essential for modABC repression. ModE(S) binds two Mo-boxes in the modA promoter as shown by electrophoretic mobility shift assays. Northern analysis revealed cotranscription of modE(S) with the upstream gene, atu2565, which was dispensable for ModE(S) activity. To identify genes controlling ModE(S) function, we performed transposon mutagenesis. Tn5 insertions resulting in derepressed modA transcription mapped to the atu2565-modE(S) operon and several Moco biosynthesis genes. We conclude that A. tumefaciens ModE(S) activity responds to Moco availability rather than to molybdate concentration directly, as is the case for E. coli ModE. Similar results in Sinorhizobium meliloti suggest that Moco dependence is a common feature of ModE(S) regulators. PMID:27196733

  10. Response of Root Properties to Tripartite Symbiosis between Lucerne (Medicago sativa L., Rhizobia and Mycorrhiza Under Dry Organic Farming Conditions

    Directory of Open Access Journals (Sweden)

    M. R. Ardakani

    2009-01-01

    Full Text Available Problem statement: It is generally considered that root turnover is a major contributor to organic matter and mineral nutrient cycles in organic managed agroecosystems. Approach: This study designed to investigate whether microbial activity could affect on root properties of Lucerne in an organically managed field under dry weather conditions. The trial was laid out as a factorial experiment in the fields of the University of Natural Resources and Applied Life Sciences, Vienna- Austria at Raasdorf in 2007. The experimental factors of Rhizobium (Sinorhizobium meliloti and Arbuscular Mycorrhiza (AM including Glomus etunicatum, G. intraradices and G. claroideum and irrigation levels were tested. Results: Results showed that increasing water deficit affected root dry weigh, specific root mass and root length significantly at 1% level and co-inoculation of rhizobium and mycorrhiza with irrigation increased all root parameters. Datas of variance analysis for mycorrhizal colonization showed that main effect of using mycorrhiza had significant effects on root parameters at 5 and 1% probability level at first and second harvest, respectively. Results of mean comparisons by Duncans Multiple Range Test showed that mycorrhizal colonization was higher in the inoculated treatments by rhizobium, mycorrhiza and irrigated plots in both harvests. Double interaction of mycorrhiza and irrigation was higher in both harvests (37.05 and 65.73%, respectively. Conclusion: It can be suggested that the tripartite symbiosis of Rhizobium, AM and Lucerne can improve the performance of Lucerne in organic farming and under dry conditions. Such traits could be incorporated into breeding programs to improve drought tolerance especially in organic fields.

  11. Molybdate uptake by Agrobacterium tumefaciens correlates with the cellular molybdenum cofactor status.

    Science.gov (United States)

    Hoffmann, Marie-Christine; Ali, Koral; Sonnenschein, Marleen; Robrahn, Laura; Strauss, Daria; Narberhaus, Franz; Masepohl, Bernd

    2016-09-01

    Many enzymes require the molybdenum cofactor, Moco. Under Mo-limiting conditions, the high-affinity ABC transporter ModABC permits molybdate uptake and Moco biosynthesis in bacteria. Under Mo-replete conditions, Escherichia coli represses modABC transcription by the one-component regulator, ModE, consisting of a DNA-binding and a molybdate-sensing domain. Instead of a full-length ModE protein, many bacteria have a shorter ModE protein, ModE(S) , consisting of a DNA-binding domain only. Here, we asked how such proteins sense the intracellular molybdenum status. We show that the Agrobacterium tumefaciens ModE(S) protein Atu2564 is essential for modABC repression. ModE(S) binds two Mo-boxes in the modA promoter as shown by electrophoretic mobility shift assays. Northern analysis revealed cotranscription of modE(S) with the upstream gene, atu2565, which was dispensable for ModE(S) activity. To identify genes controlling ModE(S) function, we performed transposon mutagenesis. Tn5 insertions resulting in derepressed modA transcription mapped to the atu2565-modE(S) operon and several Moco biosynthesis genes. We conclude that A. tumefaciens ModE(S) activity responds to Moco availability rather than to molybdate concentration directly, as is the case for E. coli ModE. Similar results in Sinorhizobium meliloti suggest that Moco dependence is a common feature of ModE(S) regulators.

  12. Toxicogenomic Responses of the Model Legume Medicago truncatula to Aged Biosolids Containing a Mixture of Nanomaterials (TiO₂, Ag, and ZnO) from a Pilot Wastewater Treatment Plant.

    Science.gov (United States)

    Chen, Chun; Unrine, Jason M; Judy, Jonathan D; Lewis, Ricky W; Guo, Jing; McNear, David H; Tsyusko, Olga V

    2015-07-21

    Toxicogenomic responses in Medicago truncatula A17 were monitored following exposure to biosolids-amended soils. Treatments included biosolids produced using a pilot wastewater treatment plant with either no metal introduced into the influent (control); bulk/ionic TiO2, ZnO, and AgNO3 added to influent (bulk/dissolved treatment); or Ag, ZnO, and TiO2 engineered nanomaterials added to influent (ENM treatment) and then added to soil, which was aged in the field for 6 months. In our companion study, we found inhibition of nodulation in the ENM but not in the bulk/dissolved treatment. Gene expression profiling revealed highly distinct profiles with more than 10-fold down-regulation in 239 genes in M. truncatula roots from the ENM treatment, while gene expression patterns were similar between bulk/dissolved and control treatments. In response to ENM exposure, many of the identified biological pathways, gene ontologies, and individual genes are associated with nitrogen metabolism, nodulation, metal homeostasis, and stress responses. Expression levels of nine genes were independently confirmed with qRT-PCR. Exposure to ENMs induced unique shifts in expression profiles and biological pathways compared with bulk/dissolved treatment, despite the lack of difference in bioavailable metal fractions, metal oxidation state, and coordination environment between ENM and bulk/dissolved biosolids. As populations of Sinorhizobium meliloti Rm2011 were similar in bulk/dissolved and ENM treatments, our results suggest that inhibition of nodulation in the ENM treatment was primarily due to phytotoxicity, likely caused by enhanced bioavailability of Zn ions.

  13. Multifunctionality and diversity of culturable bacterial communities strictly associated with spores of the plant beneficial symbiont Rhizophagus intraradices.

    Science.gov (United States)

    Battini, Fabio; Cristani, Caterina; Giovannetti, Manuela; Agnolucci, Monica

    2016-02-01

    Arbuscular Mycorrhizal Fungi (AMF) live in symbiosis with most crop plants and represent essential elements of soil fertility and plant nutrition and productivity, facilitating soil mineral nutrient uptake and protecting plants from biotic and abiotic stresses. These beneficial services may be mediated by the dense and active spore-associated bacterial communities, which sustain diverse functions, such as the promotion of mycorrhizal activity, biological control of soilborne diseases, nitrogen fixation, and the supply of nutrients and growth factors. In this work, we utilised culture-dependent methods to isolate and functionally characterize the microbiota strictly associated to Rhizophagus intraradices spores, and molecularly identified the strains with best potential plant growth promoting (PGP) activities by 16S rDNA sequence analysis. We isolated in pure culture 374 bacterial strains belonging to different functional groups-actinobacteria, spore-forming, chitinolytic and N2-fixing bacteria-and screened 122 strains for their potential PGP activities. The most common PGP trait was represented by P solubilization from phytate (69.7%), followed by siderophore production (65.6%), mineral P solubilization (49.2%) and IAA production (42.6%). About 76% of actinobacteria and 65% of chitinolytic bacteria displayed multiple PGP activities. Nineteen strains with best potential PGP activities, assigned to Sinorhizobium meliloti, Streptomyces spp., Arthrobacter phenanthrenivorans, Nocardiodes albus, Bacillus sp. pumilus group, Fictibacillus barbaricus and Lysinibacillus fusiformis, showed the ability to produce IAA and siderophores and to solubilize P from mineral phosphate and phytate, representing suitable candidates as biocontrol agents, biofertilisers and bioenhancers, in the perspective of targeted management of beneficial symbionts and their associated bacteria in sustainable food production systems. PMID:26805620

  14. Characterization of BKC-1 class A carbapenemase from Klebsiella pneumoniae clinical isolates in Brazil.

    Science.gov (United States)

    Nicoletti, Adriana Giannini; Marcondes, Marcelo F M; Martins, Willames M B S; Almeida, Luiz G P; Nicolás, Marisa F; Vasconcelos, Ana T R; Oliveira, Vitor; Gales, Ana Cristina

    2015-09-01

    Three Klebsiella pneumoniae clinical isolates demonstrating carbapenem resistance were recovered from different patients hospitalized at two medical centers in São Paulo, Brazil. Resistance to all β-lactams, quinolones, and some aminoglycosides was observed for these isolates that were susceptible to polymyxin B. Carbapenem hydrolysis, which was inhibited by clavulanic acid, was observed for all K. pneumoniae isolates that belonged to the same pulsed-field gel electrophoresis (PFGE) type and a novel sequence type (ST), ST1781 (clonal complex 442 [CC442]). A 10-kb nonconjugative incompatibility group Q (IncQ) plasmid, denominated p60136, was transferred to Escherichia coli strain TOP10 cells by electroporation. The full sequencing of p60136 showed that it was composed of a mobilization system, ISKpn23, the phosphotransferase aph3A-VI, and a 941-bp open reading frame (ORF) that codified a 313-amino acid protein. This ORF was named bla BKC-1. Brazilian Klebsiella carbapenemase-1 (BKC-1) showed a pI of 6.0 and possessed the highest identity (63%) with a β-lactamase of Sinorhizobium meliloti, an environmental bacterium. Hydrolysis studies demonstrated that purified BKC-1 not only hydrolyzed carbapenems but also penicillins, cephalosporins, and monobactams. However, the carbapenems were less efficiently hydrolyzed due to their very low kcat values (0.0016 to 0.031 s(-1)). In fact, oxacillin was the best substrate for BKC-1 (kcat /Km , 53,522.6 mM(-1) s(-1)). Here, we report a new class A carbapenemase, confirming the diversity and rapid evolution of β-lactamases in K. pneumoniae clinical isolates. PMID:26055384

  15. Application of targeted metagenomics to explore abundance and diversity of CO₂-fixing bacterial community using cbbL gene from the rhizosphere of Arachis hypogaea.

    Science.gov (United States)

    Yousuf, Basit; Keshri, Jitendra; Mishra, Avinash; Jha, Bhavanath

    2012-09-10

    Sequestration of CO(2) by autotrophic bacteria is a key process of biogeochemical carbon cycling in soil ecosystem. Rhizosphere is a rich niche of microbial activity and diversity, influenced by change in atmospheric CO(2). Structural changes in rhizosphere composition influence microbial communities and the nutrient cycling. In the present study, the bacterial diversity and population dynamics were established using cbbL and 16S rRNA gene targeted metagenomics approach from the rhizosphere of Arachis hypogaea. A total of 108 cbbL clones were obtained from the rhizospheric soil which revealed predominance of cbbL sequences affiliated to Rhizobium leguminosarum, Bradyrhizobium sp., Sinorhizobium meliloti, Ochrobactrum anthropi and a variety of uncultured cbbL harboring bacteria. The 16S rRNA gene clone library exhibited the dominance of Firmicutes (34.4%), Proteobacteria (18.3%), Actinobacteria (17.2%) and Bacteroidetes (16.1%). About 43% nucleotide sequences of 16S rRNA gene clone library were novel genera which showed <95% homology with published sequences. Gene copy number of cbbL and 16S rRNA genes, determined by quantitative real-time PCR (qRT PCR), was 9.38 ± 0.75 × 10(7) and 5.43 ± 0.79 × 10(8) (per g dry soil), respectively. The results exhibited bacterial community structure with high bacterial diversity and abundance of CO(2)-fixing bacteria, which can be explored further for their role in carbon cycling, sustainable agriculture and environment management. PMID:22766402

  16. Development of a Microemulsion Formulation for Antimicrobial SecA Inhibitors.

    Directory of Open Access Journals (Sweden)

    Jiahuai Hu

    Full Text Available In our previous study, we have identified five antimicrobial small molecules via structure based design, which inhibit SecA of Candidatus Liberibacter asiaticus (Las. SecA is a critical protein translocase ATPase subunit and is involved in pre-protein translocation across and integration into the cellular membrane in bacteria. In this study, eleven compounds were identified using similarity search method based on the five lead SecA inhibitors identified previously. The identified SecA inhibitors have poor aqueous solubility. Thus a microemulsion master mix (MMX was developed to address the solubility issue and for application of the antimicrobials. MMX consists of N-methyl-2-pyrrolidone and dimethyl sulfoxide as solvent and co-solvent, as well as polyoxyethylated castor oil, polyalkylene glycol, and polyoxyethylene tridecyl ether phosphate as surfactants. MMX has significantly improved the solubility of SecA inhibitors and has no or little phytotoxic effects at concentrations less than 5.0% (v/v. The minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC of the SecA inhibitors and streptomycin against eight bacteria including Agrobacterium tumefaciens, Liberibacter crescens, Rhizobium etli, Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti phylogenetically related to Las were determined using the broth microdilution method. MIC and MBC results showed that the 16 SecA inhibitors have antibacterial activities comparable to that of streptomycin. Overall, we have identified 11 potent SecA inhibitors using similarity search method. We have developed a microemulsion formulation for SecA inhibitors which improved the antimicrobial activities of SecA inhibitors.

  17. Development of a Microemulsion Formulation for Antimicrobial SecA Inhibitors.

    Science.gov (United States)

    Hu, Jiahuai; Akula, Nagaraju; Wang, Nian

    2016-01-01

    In our previous study, we have identified five antimicrobial small molecules via structure based design, which inhibit SecA of Candidatus Liberibacter asiaticus (Las). SecA is a critical protein translocase ATPase subunit and is involved in pre-protein translocation across and integration into the cellular membrane in bacteria. In this study, eleven compounds were identified using similarity search method based on the five lead SecA inhibitors identified previously. The identified SecA inhibitors have poor aqueous solubility. Thus a microemulsion master mix (MMX) was developed to address the solubility issue and for application of the antimicrobials. MMX consists of N-methyl-2-pyrrolidone and dimethyl sulfoxide as solvent and co-solvent, as well as polyoxyethylated castor oil, polyalkylene glycol, and polyoxyethylene tridecyl ether phosphate as surfactants. MMX has significantly improved the solubility of SecA inhibitors and has no or little phytotoxic effects at concentrations less than 5.0% (v/v). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the SecA inhibitors and streptomycin against eight bacteria including Agrobacterium tumefaciens, Liberibacter crescens, Rhizobium etli, Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti phylogenetically related to Las were determined using the broth microdilution method. MIC and MBC results showed that the 16 SecA inhibitors have antibacterial activities comparable to that of streptomycin. Overall, we have identified 11 potent SecA inhibitors using similarity search method. We have developed a microemulsion formulation for SecA inhibitors which improved the antimicrobial activities of SecA inhibitors. PMID:26963811

  18. Partial Protection against Brucella Infection in Mice by Immunization with Nonpathogenic Alphaproteobacteria▿

    Science.gov (United States)

    Delpino, M. Victoria; Estein, Silvia M.; Fossati, Carlos A.; Baldi, Pablo C.

    2007-01-01

    Previous findings indicate that Brucella antigens and those from nonpathogenic alphaproteobacteria (NPAP) are cross-recognized by the immune system. We hypothesized that immunization with NPAP would protect mice from Brucella infection. Mice were immunized subcutaneously with heat-killed Ochrobactrum anthropi, Sinorhizobium meliloti, Mesorhizobium loti, Agrobacterium tumefaciens, or Brucella melitensis H38 (standard positive control) before intravenous challenge with Brucella abortus 2308. Cross-reacting serum antibodies against Brucella antigens were detected at the moment of challenge in all NPAP-immunized mice. Thirty days after B. abortus challenge, splenic CFU counts were significantly lower in mice immunized with O. anthropi, M. loti, and B. melitensis H38 than in the phosphate-buffered saline controls (protection levels were 0.80, 0.66, and 1.99 log units, respectively). In mice immunized intraperitoneally with cytosoluble extracts from NPAP or Brucella abortus, protection levels were 1.58 for the latter, 0.63 for O. anthropi, and 0.40 for M. loti. To test whether the use of live NPAP would increase protection further, mice were both immunized and challenged by the oral route. Immunization with NPAP induced a significant increase in serum immunoglobulin G (IgG), but not serum or fecal IgA, against Brucella antigens. After challenge, anti-Brucella IgA increased significantly in the sera and feces of mice orally immunized with O. anthropi. For all NPAP, protection levels were higher than those obtained with systemic immunizations but were lower than those obtained by oral immunization with heat-killed B. abortus. These results show that immunization with NPAP, especially O. anthropi, confers partial protection against Brucella challenge. However, such protection is lower than that conferred by immunization with whole Brucella or its cytosoluble fraction. PMID:17715332

  19. On the Unusual Homeoviscous Adaptation of the Membrane Fatty Acyl Components against the Thermal Stress in Rhi{Zeta}obium meliloti

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Seb Yung; Jung, Seun Ho [Konkuk University, Seoul (Korea, Republic of); Choi, Yong Hoon; Yang, Chul Hak [Seoul National University, Seoul (Korea, Republic of); Kim, Hyun Won [Yonsei University Wonju College of Medicine, Wonju (Korea, Republic of)

    1999-06-15

    In order to maintain the optimal fluidity in membrane, microorganism genetically regulates the ratio of the unsaturated fatty acids (Ufos) to saturated ones of its biological membrane in response to external perturbing condition such as the change of temperature. The remodelling of fatty acyl chain composition is the most frequently observed response to altered growth temperature. It is reflected in the elevated proportions of unsaturated fatty acid (UFAs) at low temperature. Because cis double bonds, normally positioned at the middle of fatty acyl chains, introduce a kink of approximately 30 .deg. into acyl chain, UFAs pack less compactly and exhibit lower melting points than their saturated homologues. Thus, enrichment of membranes with UFAs offsets, to a significant degree, the increase in membrane order caused by a drop in temperature. This is so called homeoviscous adaptation of the membrane fatty acyl chains against thermal stress. Membrane maintains the optimal viscosity using homeoviscous adaptation.

  20. Identification and Characterization of a Brucella abortus ATP-Binding Cassette Transporter Homolog to Rhizobium meliloti ExsA and Its Role in Virulence and Protection in Mice

    OpenAIRE

    G.M.S. Rosinha; Freitas, Daniela A.; Miyoshi, Anderson; Azevedo, Vasco; Campos, Eleonora; Cravero, Silvio L; Rossetti, Osvaldo; Splitter, Gary; S.C. Oliveira

    2002-01-01

    Brucella abortus is a facultative intracellular bacterial pathogen that causes abortion in domestic animals and undulant fever in humans. The mechanism of virulence of Brucella spp. is not fully understood yet. Furthermore, genes that allow Brucella to reach the intracellular niche and to interact with host cells need to be identified. Using the genomic survey sequence (GSS) approach, we identified the gene encoding an ATP-binding cassette (ABC) transporter of B. abortus strain S2308. The ded...

  1. Exploring mitochondrial evolution and metabolism organization principles by comparative analysis of metabolic networks.

    Science.gov (United States)

    Chang, Xiao; Wang, Zhuo; Hao, Pei; Li, Yuan-Yuan; Li, Yi-Xue

    2010-06-01

    The endosymbiotic theory proposed that mitochondrial genomes are derived from an alpha-proteobacterium-like endosymbiont, which was concluded from sequence analysis. We rebuilt the metabolic networks of mitochondria and 22 relative species, and studied the evolution of mitochondrial metabolism at the level of enzyme content and network topology. Our phylogenetic results based on network alignment and motif identification supported the endosymbiotic theory from the point of view of systems biology for the first time. It was found that the mitochondrial metabolic network were much more compact than the relative species, probably related to the higher efficiency of oxidative phosphorylation of the specialized organelle, and the network is highly clustered around the TCA cycle. Moreover, the mitochondrial metabolic network exhibited high functional specificity to the modules. This work provided insight to the understanding of mitochondria evolution, and the organization principle of mitochondrial metabolic network at the network level.

  2. New criteria for selecting the origin of DNA replication in Wolbachia and closely related bacteria

    DEFF Research Database (Denmark)

    Ioannidis, Panagiotis; Dunning Hotopp, Julie C; Sapountzis, Panagiotis;

    2007-01-01

    as their patterns of sequence evolution will aid studies of cell replication and cell density, as well as the potential genetic manipulation of these widespread intracellular bacteria. RESULTS: Using features that have been previously experimentally verified in the alpha-Proteobacterium Caulobacter crescentus......BACKGROUND: The annotated genomes of two closely related strains of the intracellular bacterium Wolbachia pipientis have been reported without the identifications of the putative origin of replication (ori). Identifying the ori of these bacteria and related alpha-Proteobacteria as well......, the origin of DNA replication (ori) regions were identified in silico for Wolbachia strains and eleven other related bacteria belonging to Ehrlichia, Anaplasma, and Rickettsia genera. These features include DnaA-, CtrA- and IHF-binding sites as well as the flanking genes in C. crescentus. The Wolbachia ori...

  3. Wolbachia infections in natural Anopheles populations affect egg laying and negatively correlate with Plasmodium development

    Science.gov (United States)

    Shaw, W. Robert; Marcenac, Perrine; Childs, Lauren M.; Buckee, Caroline O.; Baldini, Francesco; Sawadogo, Simon P.; Dabiré, Roch K.; Diabaté, Abdoulaye; Catteruccia, Flaminia

    2016-01-01

    The maternally inherited alpha-proteobacterium Wolbachia has been proposed as a tool to block transmission of devastating mosquito-borne infectious diseases like dengue and malaria. Here we study the reproductive manipulations induced by a recently identified Wolbachia strain that stably infects natural mosquito populations of a major malaria vector, Anopheles coluzzii, in Burkina Faso. We determine that these infections significantly accelerate egg laying but do not induce cytoplasmic incompatibility or sex-ratio distortion, two parasitic reproductive phenotypes that facilitate the spread of other Wolbachia strains within insect hosts. Analysis of 221 blood-fed A. coluzzii females collected from houses shows a negative correlation between the presence of Plasmodium parasites and Wolbachia infection. A mathematical model incorporating these results predicts that infection with these endosymbionts may reduce malaria prevalence in human populations. These data suggest that Wolbachia may be an important player in malaria transmission dynamics in Sub-Saharan Africa. PMID:27243367

  4. Frequency of infection with A and B supergroup Wolbachia in insects and pests associated with mulberry and silkworm

    Indian Academy of Sciences (India)

    B M Prakash; H P Puttaraju

    2007-06-01

    Wolbachia is a ubiquitous, Gram-negative, vertically transmitted, alpha-proteobacterium that causes an array of reproductive abnormalities including cytoplasmic incompatibility, feminization of genetic males, parthenogenesis in a number of insect species, among others. Wolbachia is now being exploited as an agent for pest and vector control. Previous surveys indicated that it is commonly seen in 16–76% of arthropods. In this paper, using polymerase chain reaction assay based on specific amplification of the ftsZ-A and -B supergroup Wolbachia gene fragments, we found that 30% of insects and pests screened were positive for Wolbachia. Among them 66.7% harbour double Wolbachia infection, while 33.3% harbour single Wolbachia infection. These results indicate widespread infection with both double and single Wolbachia, and provide a wealth of information to exploit this endobacterium for the management of pests and vectors.

  5. Genetic Diversity Affects the Daily Transcriptional Oscillations of Marine Microbial Populations.

    Science.gov (United States)

    Shilova, Irina N; Robidart, Julie C; DeLong, Edward F; Zehr, Jonathan P

    2016-01-01

    Marine microbial communities are genetically diverse but have robust synchronized daily transcriptional patterns at the genus level that are similar across a wide variety of oceanic regions. We developed a microarray-inspired gene-centric approach to resolve transcription of closely-related but distinct strains/ecotypes in high-throughput sequence data. Applying this approach to the existing metatranscriptomics datasets collected from two different oceanic regions, we found unique and variable patterns of transcription by individual taxa within the abundant picocyanobacteria Prochlorococcus and Synechococcus, the alpha Proteobacterium Pelagibacter and the eukaryotic picophytoplankton Ostreococcus. The results demonstrate that marine microbial taxa respond differentially to variability in space and time in the ocean. These intra-genus individual transcriptional patterns underlie whole microbial community responses, and the approach developed here facilitates deeper insights into microbial population dynamics.

  6. 费氏中华根瘤菌HN01的putA基因克隆%Cloning of the putA gene from Sinorhizobium fredii HN01

    Institute of Scientific and Technical Information of China (English)

    唐美琼; 申佩弘; 许兢; 蒋承建; 陈钢; 武波

    2010-01-01

    从快生型大豆根瘤菌费氏中华根瘤菌HN01的基因组文库中克隆到一个putA(脯氨酸脱氢酶)基因,该基因与已报道的苜蓿中华根瘤菌1021中putA基因在核苷酸和氨基酸水平上分别有82%和86%相似性.利用自杀性质粒pK18mob构建含有putA基因部分片段的重组质粒,通过三亲本结合导入出发菌株HN01中,获得正向插入的极性突变体GXHNPA和反向插入非极性突变体GXHNPB.将putA基因完整的ORF连接到广宿主载体pLAFRJ上,获得用于互补的质粒pGXHN37,通过三亲接合,将pLAFRJ导入GXHNPA和GXHNPB获得互补菌株GXHNPHA和GXHNPHB.对出发菌株、突变菌株、互补菌株进一步的研究发现,以脯氨酸为唯一C、N源的MM培养基中培养时,突变体均不能生长,互补菌株和野生型HN01没有差异,而在完全培养基YMB中培养时,突变菌株生长不受影响.植株实验发现,突变体均能有效结瘤,但是固氮酶活与出发菌株相比有所下降,结瘤时间延迟1 d,竞争结瘤能力下降,而互补菌株与野生型HN01没有明显的差异.

  7. NCBI nr-aa BLAST: CBRC-CBRE-01-0167 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRE-01-0167 ref|YP_001327006.1| phage tape measure protein [Sinorhizobium med...icae WSM419] gb|ABR60171.1| phage tape measure protein [Sinorhizobium medicae WSM419] YP_001327006.1 2.9 26% ...

  8. Effects of Effective Microbial Inoculants on Alfalfa Growth Character%苜蓿根际有益菌接种剂对苜蓿生长特性影响的研究

    Institute of Scientific and Technical Information of China (English)

    韩华雯; 孙丽娜; 姚拓; 张英; 王国基

    2013-01-01

    为探讨微生物肥源替代或部分替代化肥的应用潜力,利用前期从苜蓿(Medicago sativa L.)和小麦(Triticum aestivum L.)根际分离的3株溶磷菌(Bacillus sp.,Pseudomonas sp.和Azotobacter sp.)和1株根瘤菌(Sinorhizobium meliloti)研制苜蓿根际新型专用接种剂,并进行田间随机区组试验,测定其对苜蓿生长特性的影响.结果表明:单一菌种接种剂+半量磷肥对苜蓿的促生效果不及复合菌种接种剂+半量磷肥,与对照(全量磷肥)相比,复合菌种接种剂+半量磷肥处理对苜蓿的各项生长指标均有明显的促生效应,其中以复合接种剂+半量磷肥处理的效果最佳:苜蓿株高、叶绿素含量、叶茎比、干鲜比及产量分别较对照增加9.00%,51.98%,13.79%,19.57%,11.98%(第1茬)和8.26%,48.08%,16.87%,20.07%,20.95%(第2茬);单一菌种接种剂+半量磷肥处理的效果不及全量磷肥处理,但处理根瘤接种剂+半量磷肥效果较好.因此,推荐根瘤接种剂和Jm170+Jm92+ Lx191溶磷菌+根瘤菌复合接种剂为适用于苜蓿的最佳单一及复合菌株接种剂,研制的各接种剂质量达到农业部微生物肥料行业标准(NY227-94)的要求.%Maintenance of soil fertility is one of the more important requirements for sustainable agriculture in China because increasing chemical fertilizer use and highly productive systems has created environmental problems and resource overexploitation. In recent years, bio-fertilizers have emerged as an important component of the integrated nutrient supply system and show great promise to improve crop yields. The objective of this paper is to survey the possibility of applying bio-fertilizers to replace chemical fertilizers. Three phosphate-solubilizing bacteria strains (Bacillus sp. , Pseudomonas sp. and Azotobacter sp. ) and one rhi-zobium (Sinorhizobium meliloti) , isolated from alfalfa and wheat rhizosphere, were used to produce single and compound inoculants. A field

  9. 系列混合碳源条件下颗粒化EBPR系统茵群结构变化规律研究%A study of microbial diversity of granule-based enhanced biological phosphorus removal systems cultivated with ratiometric propionate and acetate as mixed carbon sources

    Institute of Scientific and Technical Information of China (English)

    蒋涛; 孙培德; 金均

    2012-01-01

    A series of mixed carbon sources with different ratios of propionate and acetate was applied in granule-based enhanced biological phosphorus removal (EBPR) sludge in SBR reactor. Microbial diversity change during the granular process and functional bacteria competition under different carbon sources were studied. Significant microbial diversity change in EBPR system was exhibited during granulation. Uncultured bacteria previously dominated in the system disappeared rapidly, while uncultured rhodocyclaceae bacterium and portions of candidatus competibacter phosphatis, denitrifying bacterium, acinetobacter and uncultured alpha proteobacterium were gradually washed out. Uncultured chlorobi bacterium and uncultured alpha proteobacterium were the primary phosphorus removal bacteria in developed granular EBPR system. The change of bacteria population ofcandidatus competibacterphosphatis and uncultured chlorobi bacterium was evidenced as a result of microbial diversity under different ratios of mixed carbon sources. The population of candidatus competibacter phosphatis increased monotonically with acetate concentration, decreaseing the system phosphorus removal efficiency. Meanwhile, the population of uncultured chlorobi bacterium had a positive correlation with propionate concentration, which maintained good phosphorus removal efficiency of the EBPR system.%在SBR反应器中接种富含聚磷菌的活性污泥,采用一系列不同丙酸/乙酸比例混合的碳源进行EBPR系统污泥的颗粒化培养,并考察了颗粒化进程中的系统菌群结构变化,以及不同混合碳源条件对系统功能菌种竞争的影响.结果表明,污泥颗粒化过程对EBPR系统菌群结构产生了较大的筛选作用.原本在系统中占优势的一类Uncultured bacterium被迅速淘汰;Uncultured Rhodocyclaceae bacterium、部分Candidatus Competibacter phosphatis、部分Denitrifying bacterium、Acinetobacter及部分Uncultured alpha proteobacterium分别逐渐被淘

  10. Genetic diversity of rhizobial populations recovered from three Lotus species cultivated in the infra-arid Tunisian soils

    Institute of Scientific and Technical Information of China (English)

    Mokhtar Rejili; Maria José Lorite; Mosbah Mahdhi; Juan Sanjuan Pinilla; Ali Ferchichi; Mohamed Mars

    2009-01-01

    gments from each strain based on the UPGMA algorithm from the combined patterns showed that Lotus isolates are very diverse and that they were affiliated to Sinorhizobium,Rhizobium,and Mesorhizobium genera.

  11. Bacterial study of Vostok drilling fluid: the tool to make ice core finding confident

    Science.gov (United States)

    Alekhina, I. A.; Petit, J. R.; Lukin, V. V.; Bulat, S. A.

    2003-04-01

    Decontamination of Vostok ice core is a critical issue in molecular biology studies. Core surface contains a film of hardly removable 'dirty' drilling fluid representing a mixture of polyhydrocarbons (PHC) including polyaromatic hydrocarbons (PAH) and freon. To make ice microbial finding more confident the original Vostok drilling fluid sampled from different depths (110m - 3600m) was analyzed for bacterial content by ribosomal DNA sequencing. Total, 33 clones of 16S ribosomal DNA were recovered from four samples of drilling fluid at 110, 2750, 3400, and 3600m leading to identification of 8 bacterial species. No overlapping was observed even for neighboring samples (3400m and 3600m). At present four major bacteria with the titer more than 103-104 cells per ml (as estimated from PCR results) are identified. Among them we found: unknown representative of Desulfobacteraceae which are able to oxidize sulphates and degrade benzenes (110m); PAH-degrading alpha-proteobacterium Sphingomonas natatoria (3400m); alpha-proteobacterium representing closely-related group of Sphingomonas sp. (e.g., S. aurantiaca) which are able to degrade PAH as well, and human pathogen closely related to Haloanella gallinarum of CFB group (3600m). Four additional species were revealed as single clones and showed relatedness to human pathogens and saprophytes as well as soil bacteria. These bacteria may represent drilling fluid contaminants introduced during its sampling or DNA extraction procedure. Of four major bacteria revealed, one species, Sphingomonas natatoria, has been met by us in the Vostok core from 3607 m depth (AF532054) whereas another Sphingomonas sp. which we refer to as S. aurantiaca was found in Antarctic microbial endolithic community (AF548567), hydrocarbon-containing soil near Scott Base in Antarctica (AF184221) and even isolated from 3593m Vostok accretion ice (AF324199) and Taylor Dome core (AF395031). The source for major human pathogen-related bacteria is rather uncertain

  12. The Genome of the Obligately Intracellular Bacterium Ehrlichia canis Reveals Themes of Complex Membrane Structure and Immune Evasion Strategies

    Energy Technology Data Exchange (ETDEWEB)

    Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Doyle, C Kuyler [Center for Biodenfense and Emerging Infectious Diseases; Lykidis, A [U.S. Department of Energy, Joint Genome Institute; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Francino, M P [U.S. Department of Energy, Joint Genome Institute; Chain, Patrick S [ORNL; Shin, M [U.S. Department of Energy, Joint Genome Institute; Malfatti, Stephanie [Lawrence Livermore National Laboratory (LLNL); Larimer, Frank W [ORNL; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Detter, J C [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Richardson, P M [U.S. Department of Energy, Joint Genome Institute; Yu, X J [Center for Biodenfense and Emerging Infectious Diseases; Walker, D H [Center for Biodenfense and Emerging Infectious Diseases; McBride, J W [Center for Biodenfense and Emerging Infectious Diseases; Kyripides, N C [U.S. Department of Energy, Joint Genome Institute

    2006-01-01

    Ehrlichia canis, a small obligately intracellular, tick-transmitted, gram-negative, {alpha}-proteobacterium, is the primary etiologic agent of globally distributed canine monocytic ehrlichiosis. Complete genome sequencing revealed that the E. canis genome consists of a single circular chromosome of 1,315,030 bp predicted to encode 925 proteins, 40 stable RNA species, 17 putative pseudogenes, and a substantial proportion of noncoding sequence (27%). Interesting genome features include a large set of proteins with transmembrane helices and/or signal sequences and a unique serine-threonine bias associated with the potential for O glycosylation that was prominent in proteins associated with pathogen-host interactions. Furthermore, two paralogous protein families associated with immune evasion were identified, one of which contains poly(G-C) tracts, suggesting that they may play a role in phase variation and facilitation of persistent infections. Genes associated with pathogen-host interactions were identified, including a small group encoding proteins (n = 12) with tandem repeats and another group encoding proteins with eukaryote-like ankyrin domains (n = 7).

  13. Regulation of chromosomal replication in Caulobacter crescentus.

    Science.gov (United States)

    Collier, Justine

    2012-03-01

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

  14. Genomic Sequencing and Biological Characteristics of a Novel Escherichia Coli Bacteriophage 9g, a Putative Representative of a New Siphoviridae Genus

    Directory of Open Access Journals (Sweden)

    Eugene E. Kulikov

    2014-12-01

    Full Text Available Bacteriophage 9g was isolated from horse feces using Escherichia coli C600 as a host strain. Phage 9g has a slightly elongated capsid 62 × 76 nm in diameter and a non-contractile tail about 185 nm long. The complete genome sequence of this bacteriophage consists of 56,703 bp encoding 70 predicted open reading frames. The closest relative of phage 9g is phage PhiJL001 infecting marine alpha-proteobacterium associated with Ircinia strobilina sponge, sharing with phage 9g 51% of amino acid identity in the main capsid protein sequence. The DNA of 9g is resistant to most restriction endonucleases tested, indicating the presence of hypermodified bases. The gene cluster encoding a biosynthesis pathway similar to biosynthesis of the unusual nucleoside queuosine was detected in the phage 9g genome. The genomic map organization is somewhat similar to the typical temperate phage gene layout but no integrase gene was detected. Phage 9g efficiently forms stable associations with its host that continues to produce the phage over multiple passages, but the phage can be easily eliminated via viricide treatment indicating that no true lysogens are formed. Since the sequence, genomic organization and biological properties of bacteriophage 9g are clearly distinct from other known Enterobacteriaceae phages, we propose to consider it as the representative of a novel genus of the Siphoviridae family.

  15. A novel thermoalkalostable esterase from Acidicaldus sp. strain USBA-GBX-499 with enantioselectivity isolated from an acidic hot springs of Colombian Andes.

    Science.gov (United States)

    López, Gina; Chow, Jennifer; Bongen, Patrick; Lauinger, Benjamin; Pietruszka, Jörg; Streit, Wolfgang R; Baena, Sandra

    2014-10-01

    Several thermo- and mesoacidophilic bacterial strains that revealed high lipolytic activity were isolated from water samples derived from acidic hot springs in Los Nevados National Natural Park (Colombia). A novel lipolytic enzyme named 499EST was obtained from the thermoacidophilic alpha-Proteobacterium Acidicaldus USBA-GBX-499. The gene estA encoded a 313-amino-acid protein named 499EST. The deduced amino acid sequence showed the highest identity (58 %) with a putative α/β hydrolase from Acidiphilium sp. (ZP_08632277.1). Sequence alignments and phylogenetic analysis indicated that 499EST is a new member of the bacterial esterase/lipase family IV. The esterase reveals its optimum catalytic activity at 55 °C and pH 9.0. Kinetic studies showed that 499EST preferentially hydrolyzed middle-length acyl chains (C6-C8), especially p-nitrophenyl (p-NP) caproate (C6). Its thermostability and activity were strongly enhanced by adding 6 mM FeCl3. High stability in the presence of water-miscible solvents such as dimethyl sulfoxide and glycerol was observed. This enzyme also exhibits stability under harsh environmental conditions and enantioselectivity towards naproxen and ibuprofen esters, yielding the medically relevant (S)-enantiomers. In conclusion, according to our knowledge, 499EST is the first thermoalkalostable esterase derived from a Gram-negative thermoacidophilic bacterium.

  16. Phase Preference by Active, Acetate-Utilizing Bacteria at the Rifle, CO Integrated Field Research Challenge Site

    Energy Technology Data Exchange (ETDEWEB)

    Kerkhof, L.; Williams, K.H.; Long, P.E.; McGuinness, L.

    2011-02-21

    Previous experiments at the Rifle, Colorado Integrated Field Research Challenge (IFRC) site demonstrated that field-scale addition of acetate to groundwater reduced the ambient soluble uranium concentration. In this report, sediment samples collected before and after acetate field addition were used to assess the active microbes via {sup 13}C acetate stable isotope probing on 3 phases [coarse sand, fines (8-approximately 150 {micro}m), groundwater (0.2-8 {micro}m)] over a 24-day time frame. TRFLP results generally indicated a stronger signal in {sup 13}C-DNA in the 'fines' fraction compared to the sand and groundwater. Before the field-scale acetate addition, a Geobacter-like group primarily synthesized {sup 13}C-DNA in the groundwater phase, an alpha Proteobacterium primarily grew on the fines/sands, and an Acinetobacter sp. and Decholoromonas-like OTU utilized much of the {sup 13}C acetate in both groundwater and particle-associated phases. At the termination of the field-scale acetate addition, the Geobacter-like species was active on the solid phases rather than the groundwater, while the other bacterial groups had very reduced newly synthesized DNA signal. These findings will help to delineate the acetate utilization patterns of bacteria in the field and can lead to improved methods for stimulating distinct microbial populations in situ.

  17. Bacteria associated with the bleached and cave coral Oculina patagonica.

    Science.gov (United States)

    Koren, Omry; Rosenberg, Eugene

    2008-04-01

    The relative abundance of bacteria in the mucus and tissues of Oculina patagonica taken from bleached and cave (azooxanthellae) corals was determined by analyses of the 16S rRNA genes from cloned libraries of extracted DNA and from isolated colonies. The results were compared to previously published data on healthy O. patagonica. The bacterial community of bleached, cave, and healthy corals were completely different from each other. A tight cluster (>99.5% identity) of bacteria, showing 100% identity to Acinetobacter species, dominated bleached corals, comprising 25% of the 316 clones sequenced. The dominant bacterial cluster found in cave corals, representing 29% of the 97 clones sequenced, showed 98% identity to an uncultured bacterium from the Great Barrier Reef. Vibrio splendidus was the most dominant species in healthy O. patagonica. The culturable bacteria represented 0.1-1.0% of the total bacteria (SYBR Gold staining) of the corals. The most abundant culturable bacteria in bleached, cave, and healthy corals were clusters that most closely matched Microbulbifer sp., an alpha-proteobacterium previously isolated from healthy corals and an alpha-protobacterium (AB026194), respectively. Three generalizations emerge from this study on O. patagonica: (1) More bacteria are associated with coral tissue than mucus; (2) tissue and mucus populations are different; (3) bacterial populations associated with corals change dramatically when corals lack their symbiotic zooxanthellae, either as a result of the bleaching disease or when growing in the absence of light.

  18. Role of the Irr protein in the regulation of iron metabolism in Rhodobacter sphaeroides.

    Directory of Open Access Journals (Sweden)

    Verena Peuser

    Full Text Available In Rhizobia the Irr protein is an important regulator for iron-dependent gene expression. We studied the role of the Irr homolog RSP_3179 in the photosynthetic alpha-proteobacterium Rhodobacter sphaeroides. While Irr had little effect on growth under iron-limiting or non-limiting conditions its deletion resulted in increased resistance to hydrogen peroxide and singlet oxygen. This correlates with an elevated expression of katE for catalase in the Irr mutant compared to the wild type under non-stress conditions. Transcriptome studies revealed that Irr affects the expression of genes for iron metabolism, but also has some influence on genes involved in stress response, citric acid cycle, oxidative phosphorylation, transport, and photosynthesis. Most genes showed higher expression levels in the wild type than in the mutant under normal growth conditions indicating an activator function of Irr. Irr was however not required to activate genes of the iron metabolism in response to iron limitation, which showed even stronger induction in the absence of Irr. This was also true for genes mbfA and ccpA, which were verified as direct targets for Irr. Our results suggest that in R. sphaeroides Irr diminishes the strong induction of genes for iron metabolism under iron starvation.

  19. Detection of S-nitrosothiol and nitrosylated proteins in Arachis hypogaea functional nodule: response of the nitrogen fixing symbiont.

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    Debasis Maiti

    Full Text Available To detect the presence of NO, ROS and RNS in nodules of crack entry legumes, we used Arachis hypogaea functional nodule. The response of two cognate partner rhizobia was compared towards NO and GSNO using S. meliloti and Bradyrhizobium sp NC921001. ROS, NO, nitrosothiol and bacteroids were detected by fluorescence microscopy. Redox enzymes and thiol pools were detected biochemically. Nitrosothiols were found to be present but ROS and NO were absent in A. hypogaea nodule. A number of S-nitrosylated proteins were also detected. The total thiol pool and most of the redox enzymes were low in nodule cytosolic extract but these were found to be high in the partner microorganisms indicating partner rhizobia could protect the nodule environment against the nitrosothiols. Both S. meliloti and Bradyrhizobium sp NC921001 were found to contain GSNO reductase. Interestingly, there was a marked difference in growth pattern between S. meliloti and Bradyrhizobium sp in presence of sodium nitroprusside (SNP and S-nitrosoglutathione (GSNO. Bradyrhizobium sp was found to be much more tolerant to NO donor compounds than the S. meliloti. In contrast, S. meliloti showed resistance to GSNO but was sensitive to SNP. Together our data indicate that nodule environment of crack entry legumes is different than the nodules of infection mode entry in terms of NO, ROS and RNS. Based on our biochemical characterization, we propose that exchange of redox molecules and reactive chemical species is possible between the bacteroid and nodule compartment.

  20. Characterization of root-nodulating bacteria on Retama raetam in arid Tunisian soils

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The aim of this study is to investigate the diversity of Retama raetam root-nodule bacteria isolated from arid regions of Tunisia.Twelve isolates, chosen as representative for different 16S rRNA gene patterns, were characterized by 16S rRNA gene sequencing and phenotypic analysis. Isolates were assigned to Sinorhizobium, Rhizobium and Agrobacterium. Symbiotic properties of Sinorhizobium and Rhizobium isolates showed a large diversity in their capacity to infect their host plant and fix atmospheric nitrogen. Strain RK 22 identified as Rhizobium was the most effective isolate.

  1. Biological nitrogen fixation by lucerne (Medicago sativa L.) in acid soils.

    OpenAIRE

    Pijnenborg, J.W.M.

    1990-01-01

    Growth of lucerne( Medicago sativa L.) is poor in soils with values of pH-H2O below 6. This is often due to nitrogen deficiency, resulting from a hampered performance of the symbiosis withRhizobium meliloti. This thesis deals with the factors affecting biological nitrogen fixation by lucerne in acid soils.In a field experiment, lucerne seeds were either inoculated withR.meliloti only,or inoculated and pelleted with lime, before sowing in a sandy soil of pH 5.2. Lime-pelleting significantly im...

  2. New criteria for selecting the origin of DNA replication in Wolbachia and closely related bacteria

    Directory of Open Access Journals (Sweden)

    Baldo Laura

    2007-06-01

    Full Text Available Abstract Background The annotated genomes of two closely related strains of the intracellular bacterium Wolbachia pipientis have been reported without the identifications of the putative origin of replication (ori. Identifying the ori of these bacteria and related alpha-Proteobacteria as well as their patterns of sequence evolution will aid studies of cell replication and cell density, as well as the potential genetic manipulation of these widespread intracellular bacteria. Results Using features that have been previously experimentally verified in the alpha-Proteobacterium Caulobacter crescentus, the origin of DNA replication (ori regions were identified in silico for Wolbachia strains and eleven other related bacteria belonging to Ehrlichia, Anaplasma, and Rickettsia genera. These features include DnaA-, CtrA- and IHF-binding sites as well as the flanking genes in C. crescentus. The Wolbachia ori boundary genes were found to be hemE and COG1253 protein (CBS domain protein. Comparisons of the putative ori region among related Wolbachia strains showed higher conservation of bases within binding sites. Conclusion The sequences of the ori regions described here are only similar among closely related bacteria while fundamental characteristics like presence of DnaA and IHF binding sites as well as the boundary genes are more widely conserved. The relative paucity of CtrA binding sites in the ori regions, as well as the absence of key enzymes associated with DNA replication in the respective genomes, suggest that several of these obligate intracellular bacteria may have altered replication mechanisms. Based on these analyses, criteria are set forth for identifying the ori region in genome sequencing projects.

  3. Quantification of ploidy in proteobacteria revealed the existence of monoploid, (mero-oligoploid and polyploid species.

    Directory of Open Access Journals (Sweden)

    Vito Pecoraro

    Full Text Available Bacteria are generally assumed to be monoploid (haploid. This assumption is mainly based on generalization of the results obtained with the most intensely studied model bacterium, Escherichia coli (a gamma-proteobacterium, which is monoploid during very slow growth. However, several species of proteobacteria are oligo- or polyploid, respectively. To get a better overview of the distribution of ploidy levels, genome copy numbers were quantified in four species of three different groups of proteobacteria. A recently developed Real Time PCR approach, which had been used to determine the ploidy levels of halophilic archaea, was optimized for the quantification of genome copy numbers of bacteria. Slow-growing (doubling time 103 minutes and fast-growing (doubling time 25 minutes E. coli cultures were used as a positive control. The copy numbers of the origin and terminus region of the chromosome were determined and the results were in excellent agreement with published data. The approach was also used to determine the ploidy levels of Caulobacter crescentus (an alpha-proteobacterium and Wolinella succinogenes (an epsilon-proteobacterium, both of which are monoploid. In contrast, Pseudomonas putida (a gamma-proteobacterium contains 20 genome copies and is thus polyploid. A survey of the proteobacteria with experimentally-determined genome copy numbers revealed that only three to four of 11 species are monoploid and thus monoploidy is not typical for proteobacteria. The ploidy level is not conserved within the groups of proteobacteria, and there are no obvious correlations between the ploidy levels with other parameters like genome size, optimal growth temperature or mode of life.

  4. Molecular characterization of a mosaic locus in the genome of 'Candidatus Liberibacter asiaticus'

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    Wang Xuefeng

    2012-01-01

    Full Text Available Abstract Background Huanglongbing (HLB is a highly destructive disease of citrus production worldwide. 'Candidatus Liberibacter asiaticus', an unculturable alpha proteobacterium, is a putative pathogen of HLB. Information about the biology and strain diversity of 'Ca. L. asiaticus' is currently limited, inhibiting the scope of HLB research and control. Results A genomic region (CLIBASIA_05640 to CLIBASIA_05650 of 'Ca. L. asiaticus' showing hyper-sequence variation or locus mosaicism was identified and investigated using 262 bacterial strains (188 from China and 74 from Florida. Based on the characteristic electrophoretic profiles of PCR amplicons generated by a specific primer set, eight electrophoretic types (E-types were identified, six E-types (A, B, C, D, E, and F in China and four E-types (A, C, G, and H in Florida. The 'Ca. L. asiaticus' strains from China consisted predominately of E-type A (71.3% and E-type B (19.7%. In contrast, the 'Ca. L. asiaticus' strains from Florida was predominated by E-type G (82.4%. Diversity of 'Ca. L. asiaticus' in China was also evidenced. Strains from the high altitude Yunnan Province consisted of five E-types with E-type B being the majority (62.8%, whereas strains from the low altitude coastal Guangdong Province consisted of only two E-types with E-type A as the majority (97.0%. Sequence analyses revealed that variation of DNA amplicons was due to insertion/deletion events at CLIBASIA_05650 and the downstream intergenic region. Conclusions This study demonstrated the genomic mosaicism of 'Ca. L. asiaticus' resulted from active DNA insertion/deletion activities. Analyses of strain variation depicted the significant inter- and intra-continent diversity of 'Ca. L. asiaticus'.

  5. The complete genome sequence of 'Candidatus Liberibacter solanacearum', the bacterium associated with potato zebra chip disease.

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    Hong Lin

    Full Text Available Zebra Chip (ZC is an emerging plant disease that causes aboveground decline of potato shoots and generally results in unusable tubers. This disease has led to multi-million dollar losses for growers in the central and western United States over the past decade and impacts the livelihood of potato farmers in Mexico and New Zealand. ZC is associated with 'Candidatus Liberibacter solanacearum', a fastidious alpha-proteobacterium that is transmitted by a phloem-feeding psyllid vector, Bactericera cockerelli Sulc. Research on this disease has been hampered by a lack of robust culture methods and paucity of genome sequence information for 'Ca. L. solanacearum'. Here we present the sequence of the 1.26 Mbp metagenome of 'Ca. L. solanacearum', based on DNA isolated from potato psyllids. The coding inventory of the 'Ca. L. solanacearum' genome was analyzed and compared to related Rhizobiaceae to better understand 'Ca. L. solanacearum' physiology and identify potential targets to develop improved treatment strategies. This analysis revealed a number of unique transporters and pathways, all potentially contributing to ZC pathogenesis. Some of these factors may have been acquired through horizontal gene transfer. Taxonomically, 'Ca. L. solanacearum' is related to 'Ca. L. asiaticus', a suspected causative agent of citrus huanglongbing, yet many genome rearrangements and several gene gains/losses are evident when comparing these two Liberibacter. species. Relative to 'Ca. L. asiaticus', 'Ca. L. solanacearum' probably has reduced capacity for nucleic acid modification, increased amino acid and vitamin biosynthesis functionalities, and gained a high-affinity iron transport system characteristic of several pathogenic microbes.

  6. Screening a wide host-range, waste-water metagenomic library in tryptophan auxotrophs of Rhizobium leguminosarum and of Escherichia coli reveals different classes of cloned trp genes.

    Science.gov (United States)

    Li, Youguo; Wexler, Margaret; Richardson, David J; Bond, Philip L; Johnston, Andrew W B

    2005-12-01

    A metagenomic cosmid library was constructed, in which the insert DNA was derived from bacteria in a waste-water treatment plant and the vector was the wide host-range cosmid pLAFR3. The library was screened for clones that could correct defined tryptophan auxotrophs of the alpha-proteobacterium Rhizobium leguminosarum and of Escherichia coli. A total of 26 different cosmids that corrected at least one trp mutant in one or both of these species were obtained. Several cosmids corrected the auxotrophy of one or more R. leguminosarum trp mutants, but not the corresponding mutants in E. coli. Conversely, one cosmid corrected trpA, B, C, D and E mutants of E. coli but none of the trp mutants of R. leguminosarum. Two of the Trp+ cosmids were examined in more detail. One contained a trp operon that resembled that of the pathogen Chlamydophila caviae, containing the unusual kynU gene, which specifies kynureninase. The other, whose trp genes functioned in R. leguminosarum but not in E. coli, contained trpDCFBA in an operon that is likely co-transcribed with five other genes, most of which had no known link with tryptophan synthesis. The sequences of these TRP proteins, and the products of nine other genes encoded by this cosmid, failed to affiliate them with any known bacterial lineage. For one metagenomic cosmid, lac reporter fusions confirmed that its cloned trp genes were transcribed in R. leguminosarum, but not in E. coli. Thus, rhizobia, with their many sigma-factors, may be well-suited hosts for metagenomic libraries, cloned in wide host-range vectors. PMID:16309391

  7. Biological nitrogen fixation by lucerne (Medicago sativa L.) in acid soils.

    NARCIS (Netherlands)

    Pijnenborg, J.W.M.

    1990-01-01

    Growth of lucerne( Medicago sativa L.) is poor in soils with values of pH-H2O below 6. This is often due to nitrogen deficiency, resulting from a hampered performance of the symbiosis withRhizobium meliloti. This thesis deals with the factors affecting biological nitrogen fixat

  8. Characterization of Rhizobial Isolates of Phaseolus vulgaris by Staircase Electrophoresis of Low-Molecular-Weight RNA

    OpenAIRE

    Velázquez, Encarna; Martínez-Romero, Esperanza; Rodríguez-Navarro, Dulce Nombre; Trujillo, Martha E.; Daza, Antonio; Mateos, Pedro F.; Martínez-Molina, Eustoquio; Van Berkum, Peter

    2001-01-01

    Low-molecular-weight (LMW) RNA molecules were analyzed to characterize rhizobial isolates that nodulate the common bean growing in Spain. Since LMW RNA profiles, determined by staircase electrophoresis, varied across the rhizobial species nodulating beans, we demonstrated that bean isolates recovered from Spanish soils presumptively could be characterized as Rhizobium etli, Rhizobium gallicum, Rhizobium giardinii, Rhizobium leguminosarum bv. viciae and bv. trifolii, and Sinorhizobium fredii.

  9. The effect of inoculation of an indigenous bacteria on the early growth of Acacia farnesiana in a degraded area

    OpenAIRE

    Eliane Ceccon; Anayeli Almazo-Rogel; Esperanza Martínez-Romero; Ivonne Toledo

    2012-01-01

    Restoration of native vegetation and fuelwood production are important environmental pending goals for Mexico, where years of wrong management practices resulted in ecosystemic degradation and fuelwood scarcity. In degraded areas, native rhizobial strains are often undetectable, therefore, the restoration of natural vegetation associated with an effective nodulation of the leguminous trees is mostly appropriate. Sinorhizobium americanum is a native nitrogen-fixing bacteria isolated from nodul...

  10. Conservation of Plasmid-Encoded Traits among Bean-Nodulating Rhizobium Species

    OpenAIRE

    Brom, Susana; Girard, Lourdes; García-de los Santos, Alejandro; Sanjuan-Pinilla, Julio M.; Olivares, José; Sanjuan, Juan

    2002-01-01

    Rhizobium etli type strain CFN42 contains six plasmids. We analyzed the distribution of genetic markers from some of these plasmids in bean-nodulating strains belonging to different species (Rhizobium etli, Rhizobium gallicum, Rhizobium giardinii, Rhizobium leguminosarum, and Sinorhizobium fredii). Our results indicate that independent of geographic origin, R. etli strains usually share not only the pSym plasmid but also other plasmids containing symbiosis-related genes, with a similar organi...

  11. Symbiotic and taxonomic diversity of rhizobia isolated from Acacia tortilis subsp. raddiana in Africa.

    Science.gov (United States)

    Ba, Salif; Willems, Anne; de Lajudie, Philippe; Roche, Philippe; Jeder, Habib; Quatrini, Paola; Neyra, Marc; Ferro, Myriam; Promé, Jean-Claude; Gillis, Monique; Boivin-Masson, Catherine; Lorquin, Jean

    2002-04-01

    A collection of rhizobia isolated from Acacia tortilis subsp. raddiana from various sites in the North and South of Sahara was analyzed for their diversity at both taxonomic and symbiotic levels. On the basis of whole cell protein (SDS-PAGE) and 16S rDNA sequence analysis, most of the strains were found to belong to the Sinorhizobium and Mesorhizobium genera where they may represent several different genospecies. Despite their chromosomal diversity, most A. tortilis Mesorhizobium and Sinorhizobium symbionts exhibited very similar symbiotic characters. Nodulation tests showed that the strains belong to the Acacia-Leucaena-Prosopis nodulation group, although mainly forming non-fixing nodules on species other than A. tortilis. Most of the strains tested responded similarly to flavonoid nod gene inducers, as estimated by using heterologous nodA-lacZ fusions. Thin layer chromatography analysis of the Nod factors synthesized by overproducing strains showed that most of the strains exhibited similar profiles. The structures of Nod factors produced by four different Sinorhizobium sp. strains were determined and found to be similar to other Acacia-Prosopis-Leucaena nodulating rhizobia of the Sinorhizobium-Mesorhizobium-Rhizobium branch. They are chitopentamers, N-methylated and N-acylated by common fatty acids at the terminal non reducing sugar. The molecules can also be 6-O sulfated at the reducing end and carbamoylated at the non reducing end. The phylogenetic analysis of available NodA sequences, including new sequences from A. tortilis strains, confirmed the clustering of the NodA sequences of members of the Acacia-Prosopis-Leucaena nodulation group.

  12. Identification of a bacteria-like ferrochelatase in Strongyloides venezuelensis, an animal parasitic nematode.

    Directory of Open Access Journals (Sweden)

    Eiji Nagayasu

    Full Text Available Heme is an essential molecule for vast majority of organisms serving as a prosthetic group for various hemoproteins. Although most organisms synthesize heme from 5-aminolevulinic acid through a conserved heme biosynthetic pathway composed of seven consecutive enzymatic reactions, nematodes are known to be natural heme auxotrophs. The completely sequenced Caenorhabditis elegans genome, for example, lacks all seven genes for heme biosynthesis. However, genome/transcriptome sequencing of Strongyloides venezuelensis, an important model nematode species for studying human strongyloidiasis, indicated the presence of a gene for ferrochelatase (FeCH, which catalyzes the terminal step of heme biosynthesis, whereas the other six heme biosynthesis genes are apparently missing. Phylogenetic analyses indicated that nematode FeCH genes, including that of S. venezuelensis (SvFeCH have a fundamentally different evolutionally origin from the FeCH genes of non-nematode metazoa. Although all non-nematode metazoan FeCH genes appear to be inherited vertically from an ancestral opisthokont, nematode FeCH may have been acquired from an alpha-proteobacterium, horizontally. The identified SvFeCH sequence was found to function as FeCH as expected based on both in vitro chelatase assays using recombinant SvFeCH and in vivo complementation experiments using an FeCH-deficient strain of Escherichia coli. Messenger RNA expression levels during the S. venezuelensis lifecycle were examined by real-time RT-PCR. SvFeCH mRNA was expressed at all the stages examined with a marked reduction at the infective third-stage larvae. Our study demonstrates the presence of a bacteria-like FeCH gene in the S. venezuelensis genome. It appeared that S. venezuelensis and some other animal parasitic nematodes reacquired the once-lost FeCH gene. Although the underlying evolutionary pressures that necessitated this reacquisition remain to be investigated, it is interesting that the presence of Fe

  13. Greenhouse evaluation of rhizobia as biocontrol agent of root-infecting fungi in okra

    Directory of Open Access Journals (Sweden)

    I. A. Siddiqui

    2013-12-01

    Full Text Available Nine rhizobial strains isolated from the root nodules of Cicer arietinum, Vigna radiata, V. mungo, Samania saman, Sesbania sesban, Leucinia sp., Prosopis cineraria and Medicago sativa were used to study their effects on root-infecting fungi viz., Macrophomina phaseolina, Fusarium solani and Rhizoctonia solani. In dual culture plate assay, strains of Bradyrhizobium sp., and R. meliloti were found to inhibit radial growth of M. phaseolina, F. solani and R. solani producing zones of inhibition. Bradyrhizobium sp., and R. meliloti either used as seed dressing or as soil drench significantly suppressed root-rot infection caused by M. phaseolina, F. solani and R. solani in okra, a non-1eguminous crop under greenhouse conditions. Biomass of plants was also higher in the presence of rhizobia.

  14. Greenhouse evaluation of rhizobia as biocontrol agent of root-infecting fungi in okra

    OpenAIRE

    Siddiqui, I. A.; S. Ehteshamul-Haque; Zaki, M. J.; Ghaffar, A.

    2013-01-01

    Nine rhizobial strains isolated from the root nodules of Cicer arietinum, Vigna radiata, V. mungo, Samania saman, Sesbania sesban, Leucinia sp., Prosopis cineraria and Medicago sativa were used to study their effects on root-infecting fungi viz., Macrophomina phaseolina, Fusarium solani and Rhizoctonia solani. In dual culture plate assay, strains of Bradyrhizobium sp., and R. meliloti were found to inhibit radial growth of M. phaseolina, F. solani and R. solani producing zones of inhibition. ...

  15. Degradation of the Herbicide Glyphosate by Members of the Family Rhizobiaceae

    OpenAIRE

    Liu, C.-M.; McLean, P. A.; Sookdeo, C. C.; Cannon, F. C.

    1991-01-01

    Several strains of the family Rhizobiaceae were tested for their ability to degrade the phosphonate herbicide glyphosate (isopropylamine salt of N-phosphonomethylglycine). All organisms tested (seven Rhizobium meliloti strains, Rhizobium leguminosarum, Rhizobium galega, Rhizobium trifolii, Agrobacterium rhizogenes, and Agrobacterium tumefaciens) were able to grow on glyphosate as the sole source of phosphorus in the presence of the aromatic amino acids, although growth on glyphosate was not a...

  16. Isolation and characterization of insertion sequence elements from gram-negative bacteria by using new broad-host-range, positive selection vectors.

    Science.gov (United States)

    Simon, R; Hötte, B; Klauke, B; Kosier, B

    1991-01-01

    On the basis of an RSF1010-derived broad-host-range vector, three different systems which enable positive detection and isolation of insertion sequence (IS) elements from gram-negative bacteria were constructed. Vectors pSUP104-pheS, pSUP104-rpsL, and pSUP104-sac were used successfully in a number of Rhizobium strains and in Xanthomonas campestris. More than 20 different IS elements were isolated and characterized. The 16 IS elements from Rhizobium meliloti were further used to characterize various R. meliloti strains by hybridization. The resulting hybridization patterns were different for every strain and gave a clear and definite IS fingerprint of each strain. These IS fingerprints can be used to identify and characterize R. meliloti strains rapidly and unequivocally, as they proved to be relatively stable. Some of the IS elements were found to be identical when the IS fingerprints from a given strain were compared. This method of IS fingerprinting can also establish whether IS elements are the same, related, or different. Images PMID:1847366

  17. Isolation and characterization of insertion sequence elements from gram-negative bacteria by using new broad-host-range, positive selection vectors.

    Science.gov (United States)

    Simon, R; Hötte, B; Klauke, B; Kosier, B

    1991-02-01

    On the basis of an RSF1010-derived broad-host-range vector, three different systems which enable positive detection and isolation of insertion sequence (IS) elements from gram-negative bacteria were constructed. Vectors pSUP104-pheS, pSUP104-rpsL, and pSUP104-sac were used successfully in a number of Rhizobium strains and in Xanthomonas campestris. More than 20 different IS elements were isolated and characterized. The 16 IS elements from Rhizobium meliloti were further used to characterize various R. meliloti strains by hybridization. The resulting hybridization patterns were different for every strain and gave a clear and definite IS fingerprint of each strain. These IS fingerprints can be used to identify and characterize R. meliloti strains rapidly and unequivocally, as they proved to be relatively stable. Some of the IS elements were found to be identical when the IS fingerprints from a given strain were compared. This method of IS fingerprinting can also establish whether IS elements are the same, related, or different.

  18. Variation of clonal, mesquite-associated rhizobial and bradyrhizobial populations from surface and deep soils by symbiotic gene region restriction fragment length polymorphism and plasmid profile analysis.

    Science.gov (United States)

    Thomas, P M; Golly, K F; Zyskind, J W; Virginia, R A

    1994-04-01

    Genetic characteristics of 14 Rhizobium and 9 Bradyrhizobium mesquite (Prosopis glandulosa)-nodulating strains isolated from surface (0- to 0.5-m) and deep (4- to 6-m) rooting zones were determined in order to examine the hypothesis that surface- and deep-soil symbiont populations were related but had become genetically distinct during adaptation to contrasting soil conditions. To examine genetic diversity, Southern blots of PstI-digested genomic DNA were sequentially hybridized with the nodDABC region of Rhizobium meliloti, the Klebsiella pneumoniae nifHDK region encoding nitrogenase structural genes, and the chromosome-localized ndvB region of R. meliloti. Plasmid profile and host plant nodulation assays were also made. Isolates from mesquite nodulated beans and cowpeas but not alfalfa, clover, or soybeans. Mesquite was nodulated by diverse species of symbionts (R. meliloti, Rhizobium leguminosarum bv. phaseoli, and Parasponia bradyrhizobia). There were no differences within the groups of mesquite-associated rhizobia or bradyrhizobia in cross-inoculation response. The ndvB hybridization results showed the greatest genetic diversity among rhizobial strains. The pattern of ndvB-hybridizing fragments suggested that surface and deep strains were clonally related, but groups of related strains from each soil depth could be distinguished. Less variation was found with nifHDK and nodDABC probes. Large plasmids (>1,500 kb) were observed in all rhizobia and some bradyrhizobia. Profiles of plasmids of less than 1,000 kb were related to the soil depth and the genus of the symbiont. We suggest that interacting selection pressures for symbiotic competence and free-living survival, coupled with soil conditions that restrict genetic exchange between surface and deep-soil populations, led to the observed patterns of genetic diversity. PMID:16349226

  19. Variation of clonal, mesquite-associated rhizobial and bradyrhizobial populations from surface and deep soils by symbiotic gene region restriction fragment length polymorphism and plasmid profile analysis.

    Science.gov (United States)

    Thomas, P M; Golly, K F; Zyskind, J W; Virginia, R A

    1994-04-01

    Genetic characteristics of 14 Rhizobium and 9 Bradyrhizobium mesquite (Prosopis glandulosa)-nodulating strains isolated from surface (0- to 0.5-m) and deep (4- to 6-m) rooting zones were determined in order to examine the hypothesis that surface- and deep-soil symbiont populations were related but had become genetically distinct during adaptation to contrasting soil conditions. To examine genetic diversity, Southern blots of PstI-digested genomic DNA were sequentially hybridized with the nodDABC region of Rhizobium meliloti, the Klebsiella pneumoniae nifHDK region encoding nitrogenase structural genes, and the chromosome-localized ndvB region of R. meliloti. Plasmid profile and host plant nodulation assays were also made. Isolates from mesquite nodulated beans and cowpeas but not alfalfa, clover, or soybeans. Mesquite was nodulated by diverse species of symbionts (R. meliloti, Rhizobium leguminosarum bv. phaseoli, and Parasponia bradyrhizobia). There were no differences within the groups of mesquite-associated rhizobia or bradyrhizobia in cross-inoculation response. The ndvB hybridization results showed the greatest genetic diversity among rhizobial strains. The pattern of ndvB-hybridizing fragments suggested that surface and deep strains were clonally related, but groups of related strains from each soil depth could be distinguished. Less variation was found with nifHDK and nodDABC probes. Large plasmids (>1,500 kb) were observed in all rhizobia and some bradyrhizobia. Profiles of plasmids of less than 1,000 kb were related to the soil depth and the genus of the symbiont. We suggest that interacting selection pressures for symbiotic competence and free-living survival, coupled with soil conditions that restrict genetic exchange between surface and deep-soil populations, led to the observed patterns of genetic diversity.

  20. Underexpression of Ap from R-Plasmids in Fast-Growing Rhizobium Species

    Science.gov (United States)

    Sikka, Virendra K.; Kumar, Sushil

    1984-01-01

    The presence of the plasmid RP1 in the cells of Rhizobium leguminosarum strains Rld1, 300, and 248, R. phaseoli 1233, R. trifolii strains T1 and 6661, and R. meliloti 4013 was found to appreciably increase bacterial resistance toward kanamycin and tetracycline but not toward ampicillin. The presence of 16 other R-plasmids in R. leguminosarum was also found to either not increase or only marginally increase bacterial resistance toward ampicillin. It appears now that underexpression of the plasmid-specified ampicillin function is common to most fast- and slow-growing rhizobia. PMID:16346686

  1. Low Earth orbit journey and ground simulations studies point out metabolic changes in the ESA life support organism Rhodospirillum rubrum

    Science.gov (United States)

    Mastroleo, Felice; Leys, Natalie; Benotmane, Rafi; Vanhavere, Filip; Janssen, Ann; Hendrickx, Larissa; Wattiez, Ruddy; Mergeay, Max

    MELiSSA (Micro-Ecological Life Support System Alternative) is a project of closed regenerative life support system for future space flights developed by the European Space Agency. It consists of interconnected processes (i.e. bioreactors, higher plant compartments, filtration units,..) targeting the total recycling of organic waste into oxygen, water and food. Within the MELiSSA loop, the purple non-sulfur alpha-proteobacterium R. rubrum ATCC25903 is used to convert fatty acids released from the upstream raw waste digesting reactor to CO2 and biomass, and to complete the mineralization of aminoacids into NH4+ that will be forwarded to the nitrifying compartment. Among the numerous challenges of the project, the functional stability of the bioreactors in long term and under space flight conditions is of paramount importance for the efficiency of the life support system and consequently the crew safety. Therefore, the physiological and metabolic changes induced by space flight were investigated for R. rubrum. The bacterium grown on solid medium during 2 different 10-day space flights to the ISS (MES- SAGE2, BASE-A experiments) were compared to cells grown on Earth 1 g gravity or modeled microgravity and normal Earth radiation or simulated space flight radiation conditions in order to relate each single stress to its respective cellular response. For simulating the radiation environment, pure gamma and neutron sources were combined, while simulation of changes in gravity where performed using the Random Positioning Machine technology. Transcriptome analysis using R. rubrum total genome DNA-chip showed up-regulation of genes involved in oxidative stress response after a 10-day mission inside the ISS, without loss of viability. As an example, alkyl hydroperoxide reductase, thioredoxin reductase and bacterioferritin genes are least 2 fold induced although the radiation dose experienced by the bacterium (4 mSv) is very low compared to its radiotolerance (D10 = 100 Sv

  2. Bacterial diversity analysis of Huanglongbing pathogen-infected citrus, using PhyloChip and 16S rRNA gene clone library sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Shankar Sagaram, U.; DeAngelis, K.M.; Trivedi, P.; Andersen, G.L.; Lu, S.-E.; Wang, N.

    2009-03-01

    The bacterial diversity associated with citrus leaf midribs was characterized 1 from citrus groves that contained the Huanglongbing (HLB) pathogen, which has yet to be cultivated in vitro. We employed a combination of high-density phylogenetic 16S rDNA microarray and 16S rDNA clone library sequencing to determine the microbial community composition of symptomatic and asymptomatic citrus midribs. Our results revealed that citrus leaf midribs can support a diversity of microbes. PhyloChip analysis indicated that 47 orders of bacteria from 15 phyla were present in the citrus leaf midribs while 20 orders from phyla were observed with the cloning and sequencing method. PhyloChip arrays indicated that nine taxa were significantly more abundant in symptomatic midribs compared to asymptomatic midribs. Candidatus Liberibacter asiaticus (Las) was detected at a very low level in asymptomatic plants, but was over 200 times more abundant in symptomatic plants. The PhyloChip analysis was further verified by sequencing 16S rDNA clone libraries, which indicated the dominance of Las in symptomatic leaves. These data implicate Las as the pathogen responsible for HLB disease. Citrus is the most important commercial fruit crop in Florida. In recent years, citrus Huanglongbing (HLB), also called citrus greening, has severely affected Florida's citrus production and hence has drawn an enormous amount of attention. HLB is one of the most devastating diseases of citrus (6,13), characterized by blotchy mottling with green islands on leaves, as well as stunting, fruit decline, and small, lopsided fruits with poor coloration. The disease tends to be associated with a phloem-limited fastidious {alpha}-proteobacterium given a provisional Candidatus status (Candidatus Liberobacter spp. later changed to Candidatus Liberibacter spp.) in nomenclature (18,25,34). Previous studies indicate that HLB infection causes disorder in the phloem and severely impairs the translocation of assimilates in

  3. 非抗虫转基因棉花对土壤细菌群落多样性的影响%Effects of Insect Non-resistant Transgenic Cottons on Bacterial Community Diversity in Soil

    Institute of Scientific and Technical Information of China (English)

    赵云丽; 李刚; 修伟明; 多立安; 曹璇; 雒珺瑜; 崔金杰; 杨殿林; 赵建宁

    2015-01-01

    There are increasing public concerns over the ecological risks of transgenic plants. Under field conditions, the diversity and com-position of bacterial community in soils grown with three insect non-resistant transgenic cottons(high-yield transgenic cotton expressing the RNA recognition motif 2 gene, disease-resistant transgenic cotton expressing the gastrodia antifungal protein and high-quality transgenic cotton expressing the [1-aminocyclopropane-1-carboxylate(ACC)oxidase])and one conventional cotton CCRI 12(as control)were evalu-ated at the boll-opening stage by denaturing gradient gel electrophoresis(DGGE). The results showed that planting three transgenic cottons did not show significant effects on Shannon-wiener index(H), evenness(EH)and richness(S)of soil bacteria. High degree of similarity in community structure was observed between transgenic and conventional cottons, indicating no influence of transgenic cottons on bacterial community diversity in the short term. High-yield RRM2 transgenic cotton, disease-resistant GAFP transgenic cotton, high-quality ACO2 transgenic cotton and conventional cotton had 67%similarity in community levels and were thus regarded as one group. According to the se-quence analysis of DGGE dominant bands, microorganisms which presented the highest homology belonged to families of Flavobacteria, Bacteriovorax, Segetibacter, alpha proteobacterium, Geobacte, Paenisporosarcina, and Acidobacterium, respectively, and all of them were not cultivatable.%田间试验条件下,为了探究非抗虫转基因棉花对土壤细菌群落多样性的影响,应用PCR-DGGE技术对转RRM2基因高产棉、转GAFP基因抗病棉、转ACO2基因优质棉及非转基因常规棉(中棉所12)种植后在吐絮期的土壤细菌群落多样性进行分析。结果表明,与常规棉相比,3种转基因棉的种植均未对土壤细菌香农-威纳指数(H)、均匀度(EH)和丰富度(S)造成显著影

  4. Biodegradation of geosmin in drinking water by novel bacteria isolated from biologically active carbon.

    Science.gov (United States)

    Zhou, Beihai; Yuan, Rongfang; Shi, Chunhong; Yu, Liying; Gu, Junnong; Zhang, Chunlei

    2011-01-01

    Three strains of Gram-negative bacteria capable of removing geosmin from drinking water were isolated from biologically active carbon and identified to be Chryseobacterium sp., Sinorhizobium sp. and Stenotrophomonas sp. based on physio-biochemistry analysis and 16S rRNA gene sequence analysis. Removal efficiencies of 2 mg/L geosmin in mineral salts medium were 84.0%, 80.2% and 74.4% for Chryseobacterium sp., Sinorhizobium sp. and Stenotrophomonas sp., respectively, while removal efficiencies of 560 ng/L geosmin in filter influent were 84.8%, 82.3% and 82.5%, respectively. The biodegradation of geosmin was determined to be a pseudo first-order reaction, with rate constants at 2 mg/L and 560 ng/L being 0.097 and 0.086 day(-1), 0.089 and 0.084 day(-1), 0.074 and 0.098 day(-1) for the above mentioned degraders, respectively. The biomass of culture in the presence of geosmin was much higher than that in the absence of geosmin.

  5. Biodegradation of geosmin in drinking water by novel bacteria isolated from biologically active carbon

    Institute of Scientific and Technical Information of China (English)

    Beihai Zhou; Rongfang Yuan; Chunhong Shi; Liying Yu; Junnong Gu; Chunlei Zhang

    2011-01-01

    Three strains of Gram-negative bacteria capable of removing geosmin from drinking water were isolated from biologically active carbon and identified to be Chryseobacterium sp., Sinorhizobium sp.and Stenotrophomonas sp.based on physio-biochemistry analysis and 16S rRNA gene sequence analysis.Removal efficiencies of 2 mg/L geosmin in mineral salts medium were 84.0%, 80.2% and 74.4% for Chryseobacterium sp., Sinorhizobium sp.and Stenotrophomonas sp., respectively, while removal efficiencies of 560 ng/L geosmin in filter influent were 84.8%, 82.3% and 82.5%, respectively.The biodegradation of geosmin was determined to be a pseudo first-order reaction, with rate constants at 2 mg/L and 560 ng/L being 0.097 and 0.086 day-1, 0.089 and 0.084 day-1, 0.074 and 0.098 day-1 for the above mentioned degraders, respectively.The biomass of culture in the presence of geosmin was much higher than that in the absence of geosmin.

  6. Genetic Diversity of Fast-growing Rhizobia Associated with Glycine max in Subtropical Regions of China%我国亚热带地区快生型大豆根瘤菌遗传多样性研究

    Institute of Scientific and Technical Information of China (English)

    杨成运; 周俊初; 杨江科; 段广才

    2011-01-01

    Genetic diversity of 31 isolates from effective nodules of soybean in different geographical regions of China was studied using 16S rDNA gene RELP patterns, 16S rDNA gene sequencing, 16S— 23S rDN A IGS region RE LP patterns and 16S — 23S rDN A IGS sequencing assays. The isolates were clustered into one genospecies with Sinorhizobium fredii USDA205 on the basis of their 16S rDNA gene PCR - RFLP patterns. 16S rDNA gene sequence of strains indicated that the isolates were very closely related (identities higher than 99.5%) to Sinorhizobium fredii USDA205. The analysis of the 16S—23S intergenic spacer (IGS) divided the isolates into 10 genotypes and four groups. The 16S—23S rDNA IGS sequencing assays divided the isolates into two groups. Group Ⅰ was clustered with Sinorhizobium fredii USDA205. Group Ⅱ was less similarity to Sinorhizobium fredii USDA205 than Sinorhizobium xinjiangense. The isolates from the subtropical regions of China had higher genetic similarity.%利用16S rDNA PCR-RELP、16S rDNA基因序列分析以及16S-23S rDNA IGS PCRRELP技术,对分离自我国江苏盐城、浙江温州、湖北仙桃及重庆等亚热带地区的31株大豆快生型根瘤茵(fast-growing rhizobia)进行了群体遗传多样性研究.16S rDNA PCR-RELP分析结果表明,所有供试大豆根瘤菌都与费氏中华根瘤菌USDA205聚成一类.供试代表菌株YcS2的16S rDNA基因序列与费氏中华根瘤菌USDA205的该序列相似性超过99.5%.16S-23S rDNA IGS PCR-RFLP研究结果表明:所有供试菌株分为10个基因型,在87%的相似性上分为4个类群.在此基础上筛选出10个代表菌株进行16S-23S rDNA基因序列分析,结果表明,10个供试菌株分为2个群,群Ⅱ(YcS3、ZzS1、YcS14)与新疆中华根瘤茵CCBAU110聚在一起,群I(其余7个菌株)与费氏中华根瘤茵USDA205聚在一起.中国亚热带地区的快生型大豆根瘤茵具有高度的相似性.

  7. Characterization of root-nodulating bacteria associated to Prosopis farcta growing in the arid regions of Tunisia.

    Science.gov (United States)

    Fterich, A; Mahdhi, M; Caviedes, M A; Pajuelo, E; Rivas, R; Rodriguez-Llorente, I D; Mars, M

    2011-06-01

    Diversity of 50 bacterial isolates recovered from root nodules of Prosopis farcta grown in different arid soils in Tunisia, was investigated. Characterization of isolates was assessed using a polyphasic approach including phenotypic characteristics, 16S rRNA gene PCR--RFLP and sequencing, nodA gene sequencing and MLSA. It was found that most of isolates are tolerant to high temperature (40°C) and salinity (3%). Genetic characterization emphasizes that isolates were assigned to the genus Ensifer (80%), Mesorhizobium (4%) and non-nodulating endophytic bacteria (16%). Forty isolates belonging to the genus Ensifer were affiliated to Ensifer meliloti, Ensifer xinjiangense/Ensifer fredii and Ensifer numidicus species. Two isolates belonged to the genus Mesorhizobium. Eight isolates failing to renodulate their host plant were endophytic bacteria and belonged to Bacillus, Paenibacillus and Acinetobacter genera. Symbiotic properties of nodulating isolates showed a diversity in their capacity to infect their host plant and fix atmospheric nitrogen. Isolate PG29 identified as Ensifer meliloti was the most effective one. Ability of Prosopis farcta to establish symbiosis with rhizobial species confers an important advantage for this species to be used in reforestation programs. This study offered the first systematic information about the diversity of microsymbionts nodulating Prosopis farcta in the arid regions of Tunisia. PMID:21359955

  8. Multiplication and Viability of some Rhizobium Strains to be used as Inoculants for Agricultural Biomass Production

    Directory of Open Access Journals (Sweden)

    Simina Neo

    2012-05-01

    Full Text Available Rhizobia are well known for their capacity to establish a symbiosis with legumes. They inhabit root nodules, where they reduce atmospheric nitrogen and make it available to the plant. Biological nitrogen fixation is an important component of sustainable agriculture, and rhizobial inoculants have been applied frequently as biofertilizers. In this review we approach the subject of legumes inoculation in order to improve the nitrogen fixing capacity. In the first part of the experiment, the Rhizobium strains were cultivated on media indicated in the literature as optimal for bacterial growing and development in laboratory conditions. Afterwards, the Rhizobium strains that have grown and accumulate biomass were tested in different conditions of pH and salinity. The biomass accumulation was determinate by spectrophotometer. The obtained values shown that the Rhizobium strains tested can be used to inoculate the legumes cultivated on acid, basic and alkaline soils. Finally, the stability in real time of two strains of Rhizobium (Rhizobium meliloti and Rhizobium japonicum mixed with different supports was evaluated during a 6- months period. The supports studied were: peat, peat and calcium carbonate, zeolite, and ceramic. The highest number of viable cells at the end of the experiment was obtained in ceramic with Rhizobium japonicum (8x105 cells/gram, and the lowest number of viable cells was obtained in zeolite with Rhizobium meliloti (1,1x103 cells/gram.

  9. STABILITY IN REAL TIME OF SOME CRYOPRESERVED MICROBIAL STRAINS WITH REFERENCE TO GENETICALLY MODIFIED MICROORGANISMS

    Directory of Open Access Journals (Sweden)

    DANIELA VINTILĂ

    2013-12-01

    Full Text Available The aim of this work is to analyze the viability of microorganisms from Collection of Industrial Microorganisms from Faculty of Animal Science and Biotechnology – Timisoara, during freezing and thawing as part of cryopreservation technique. The stability in real time of 19 strains cryopreserved in 16% glycerol was evaluated during a 6-months period. The strains studied were: Escherichia coli, Lactobacillus acidophilus, Rhizobium meliloti, Saccharomyces cerevisiae, Aspergillus oryzae, Aspergillus niger, Trichoderma viride, Bacillus globigii, Bacillus licheniformis, and 9 strains of Bacillus subtilis. The strains cryopreserved at -20oC and -70oC were activated using the fast thawing protocol. A better cell recovery was achieved with the -70oC protocol reaching an average viability for E. coli of 86,3%, comparing with 78,6% in -20oC protocol. The cell recovery percentages for the other strains were: 92,4% for L. acidophilus, 93,9% for A.niger, 89% for A. oryzae, 86,7% for T. viride, 94,2% for R. meliloti, 82,1% for S. cerevisiae, 89,9% for B. licheniformis. Regarding the viability of genetically modified microorganisms, the values shows a good recovering after freezing and thawing, even after 180 days of cryopreservation. With the -20oC protocol lower viability was observed due probably to the formation of eutectic mixtures and recrystalization processes.

  10. Characterization of root-nodulating bacteria associated to Prosopis farcta growing in the arid regions of Tunisia.

    Science.gov (United States)

    Fterich, A; Mahdhi, M; Caviedes, M A; Pajuelo, E; Rivas, R; Rodriguez-Llorente, I D; Mars, M

    2011-06-01

    Diversity of 50 bacterial isolates recovered from root nodules of Prosopis farcta grown in different arid soils in Tunisia, was investigated. Characterization of isolates was assessed using a polyphasic approach including phenotypic characteristics, 16S rRNA gene PCR--RFLP and sequencing, nodA gene sequencing and MLSA. It was found that most of isolates are tolerant to high temperature (40°C) and salinity (3%). Genetic characterization emphasizes that isolates were assigned to the genus Ensifer (80%), Mesorhizobium (4%) and non-nodulating endophytic bacteria (16%). Forty isolates belonging to the genus Ensifer were affiliated to Ensifer meliloti, Ensifer xinjiangense/Ensifer fredii and Ensifer numidicus species. Two isolates belonged to the genus Mesorhizobium. Eight isolates failing to renodulate their host plant were endophytic bacteria and belonged to Bacillus, Paenibacillus and Acinetobacter genera. Symbiotic properties of nodulating isolates showed a diversity in their capacity to infect their host plant and fix atmospheric nitrogen. Isolate PG29 identified as Ensifer meliloti was the most effective one. Ability of Prosopis farcta to establish symbiosis with rhizobial species confers an important advantage for this species to be used in reforestation programs. This study offered the first systematic information about the diversity of microsymbionts nodulating Prosopis farcta in the arid regions of Tunisia.

  11. CONDITIONING MICROBIAL PRODUCTS CONTAINING NITROGEN FIXING BACTERIA WITH DIFFERENT SOLID EXCIPIENTS

    Directory of Open Access Journals (Sweden)

    VINTILĂ T.

    2007-01-01

    Full Text Available The stability in real time of two strains of Rhizobium (Rhizobium meliloti andRhizobium japonicum mixed with different excipients was evaluated during a6-months period. The excipients studied were: peat, peat and calciumcarbonate, zeolite, and ceramic. Liquid cultures and excipients mixtures weredried (12-14% humidity, sealed in plastic bags and preserved at +4oC. Thecells were activated periodically by suspending aliquots from dry products in0.9% saline solution. The viability of Rhizobium cells was evaluated bycultivation of diluted suspensions in YMA plates. The number of viable cells isdecreasing during drying in all cases, increase in the first month of storage,and remains constant or decrease very slowly during storage for all obtaineddry products containing rhizobia mixed with solid dry excipients. The highestnumber of viable cells at the end of the experiment was obtained in ceramicwith Rhizobium japonicum (8x105 cells/gram, and the lowest number ofviable cells was obtained in zeolite with Rhizobium meliloti (1,1x103cells/gram.

  12. Practical use of CMC-amended rhizobial inoculant for Mucuna pruriens cultivation to enhance the growth and protection against Macrophomina phaseolina.

    Science.gov (United States)

    Aeron, Abhinav; Khare, Ekta; Kumar Arora, Naveen; Kumar Maheshwari, Dinesh

    2012-01-01

    In many parts of the world Mucuna pruriens is used as an important medicinal, forage and green manure crop. In the present investigation the effect of the addition of CMC in carrier during development of bioformulation on shelflife, plant growth promotive and biocontrol activity against Macrophomina phaseolina was screened taking M. pruriens as a test crop. Ensifer meliloti RMP6(Ery+Kan+) and Bradyrhizobium sp. BMP7(Tet+Kan+) (kanamycin resistance engineered by Tn5 transposon mutagenesis) used in the study showed production of siderophore, IAA, solubilizing phosphate and biocontrol of M. phaseolina. RMP6(Ery+Kan+) also showed ACC deaminase activity. The survival of both the strains in sawdust-based bioformulation was enhanced with an increase in the concentration of CMC from 0 to 1%. At 0% CMC Bradyrhizobium sp. BMP7(Tet+Kan+) showed more increase in nodule number/plant (500.00%) than E. meliloti RMP6(Ery+Kan+) (52.38%), over the control in M. phaseolina-infested soil. There was 185.94% and 59.52% enhancement in nodule number/plant by RMP6(Ery+Kan+) and BMP7(Tet+Kan+) with an increase in the concentration of CMC from 0% to 1% in the bioformulations. However further increase in concentration of CMC did not result in enhancement in survival of either the strains or nodule number/plant. PMID:22688243

  13. Efficacy of wild plant in combination with microbial antagonists for the control of root rot fungi on mungbean and cowpea

    International Nuclear Information System (INIS)

    Present work was carried out to investigate the efficacy of Aerva javanica in combination with different microbial antagonists namely Rhizobium meliloti, Pseudomonas aeruginosa, Trichoderma harzianum and Aspergillus niger. Soil amended with A. javanica stem, leaves, flower powder at the rate1% w/w and seeds of cowpea (Vigna unguiculata L.) and mungbean (Vigna radiata L.) were coated with microbial antagonists for the control of root infecting fungi like Macrophomina phaseolina (Tassi) Goid, Fusarium spp. and Rhizoctonia solani Kiihn. Infection of M. phaseolina and R. solani were completely suppressed when seeds were coated with P. aeruginosa, T. harzianum, A. niger, R. meliloti and A. javanica leaves powder mixed in soil at the rate 1% w/w. All antagonists showed reduction in combination with A. javanica leaves powder at the rate1% but T. harzianum and P. aeruginosa in combination with A. javanica leaves showed promising results in complete reduction of R. solani and M. phaseolina on both crops. All growth parameters were maximum when soil was amended with A. javanica leaves powder at the rate 1% w/w and seeds were coated with T. harzianum and P. aeruginosa. (author)

  14. Tolerância de rizóbios de diferentes procedências ao zinco, cobre e cádmio Tolerance of rhizobia genera from different origins to zinc, copper and cadmium

    Directory of Open Access Journals (Sweden)

    Alexandre Matsuda

    2002-03-01

    Full Text Available Sessenta estirpes/isolados dos gêneros Bradyrhizobium, Rhizobium, Sinorhizobium, Mesorhizobium e Azorhizobium, procedentes de diferentes locais (Mata Atlântica, Amazônia, culturas agrícolas e experimentos com metais pesados e de espécies hospedeiras pertencentes às subfamílias Papilionoideae, Mimosoideae e Caesalpinoideae, foram avaliadas quanto à tolerância a Zn, Cu e Cd em meio YMA modificado pela adição de tampões biológicos (HEPES e MES e suplementados com Cu (0 a 60 mg L-1, Cd (0 a 60 mg L-1 e Zn (0 a 1.000 mg L-1. Mediante padrões de crescimento atribuídos às culturas nas diferentes concentrações dos metais, avaliaram-se as concentrações máximas toleradas e as doses tóxicas destes metais para redução de crescimento em 25% (DT25 e 50% (DT50. Não houve influência da procedência na concentração máxima de metal tolerada. A ordem de sensibilidade aos metais, considerando-se as concentrações máximas toleradas, foi Azorhizobium > Rhizobium = Mesorhizobium = Sinorhizobium > Bradyrhizobium. A DT25 e a DT50 foram úteis para diferenciarem estirpes/isolados de um mesmo gênero, que atingiram a mesma concentração máxima tolerada a Zn, Cu e Cd. A ordem de toxicidade dos metais estudados foi Cu > Cd > Zn.Sixty strains/isolates of the genera Bradyrhizobium, Rhizobium, Sinorhizobium, Mesorhizobium and Azorhizobium, isolated from different hosts (legume subfamilies: Papilionoideae, Mimosoideae and Caesalpinoideae and location (Atlantic Forest, Amazon region, crop plantings and heavy metal experiments, were evaluated for Zn, Cu and Cd tolerance in YMA medium modified by the addition of biological buffers (HEPES and MES and supplemented with Cu (0 to 60 mg L-1, Cd (0 to 60 mg L-1, and Zn (0 to 1,000 mg L-1sulphates. Growth standards were applied to evaluate rhizobia cultures growth at different metal concentrations, allowing evaluation of highest tolerated concentrations of Zn, Cu, and Cd and the toxic doses

  15. Detection and organization of atrazine-degrading genetic potential of seventeen bacterial isolates belonging to divergent taxa indicate a recent common origin of their catabolic functions

    DEFF Research Database (Denmark)

    El Azhari, Najoi

    2007-01-01

    A collection of 17 atrazine-degrading bacteria isolated from soils was studied to determine the composition of the atrazine-degrading genetic potential (i.e. trzN, trzD and atz) and the presence of IS1071. The characterization of seven new atrazine-degrading bacteria revealed for the first time...... the trzN-atzBC gene composition in Gram-negative bacteria such as Sinorhizobium sp. or Polaromonas sp. Three main atrazine-degrading gene combinations (i) trzN–atzBC, (ii) atzABC–trzD and (iii) atzABCDEF were observed. The atz and trz genes were often located on plasmids, suggesting that plasmid...... the atrazine-degrading genes....

  16. Symbiotic plasmid is required for NolR to fully repress nodulation genes in Rhizobium leguminosarum A34

    Institute of Scientific and Technical Information of China (English)

    Fengqing Li; Bihe Hou; Guofan Hong

    2008-01-01

    NolR is a reguiator of noduiation genes present in Rhizobium and Sinorhizobium. However, the mechanism by which NolR participates in the inducible transcription ofnoduiation genes remains unclear. To investigate whether there are other factors regulating the function of NoIR, an insertion mutant of NolR in Rhizobium leguminosarum strain 8401, which lacks the symbiotic plasmid, was constructed by homologous recombination. We investigated the effects of NolR inactivation on the expression of nodulation genes. Three inducible nodulation genes (nodA, nodF and nodM) were expressed constitutively in NoiR-mutant, MRl14. Our results suggested that the symbiotic plasmid is required for NolR to fully repress nodulation genes in Rhizobium ieguminosarum A34. In addition, MRl14 has provided a useful tool for further study of molecular interactions between NolR and other factors.

  17. Genetic diversity of nodulating and non-nodulating rhizobia associated with wild soybean (Glycine soja Sieb. & Zucc.) in different ecoregions of China.

    Science.gov (United States)

    Wu, Li Juan; Wang, Hai Qing; Wang, En Tao; Chen, Wen Xin; Tian, Chang Fu

    2011-06-01

    A total of 99 bacterial isolates that originated from root nodules of Glycine soja were characterized with restriction analyses of amplified 16S ribosomal DNA and 16S-23S rDNA intergenic spacers (ITS), and sequence analyses of 16S rRNA, rpoB, atpD, recA and nodC genes. When tested for nodulation of G. soja, 72 of the isolates were effective symbionts, and these belonged to five species: Bradyrhizobium japonicum, Bradyrhizobium elkanii, Bradyrhizobium yuanmingense, Bradyrhizobium liaoningense and Sinorhizobium fredii. All of these, except some B. yuanmingense strains, also formed effective nodules on the domesticated soybean Glycine max. The remaining 27 isolates did not nodulate either host, but were identified as Rhizobium. Phylogeny nodC in the G. soja symbionts suggested that this symbiosis gene was mainly maintained by vertical gene transfer. Different nodC sublineages and rrs-ITS clusters reflected the geographic origins of isolates in this study.

  18. Seleção de bactérias endofíticas de tomateiro como potenciais agentes de biocontrole e de promoção de crescimento Screening of endophytic bacteria isolated from tomato plants as potencial biocontrol agents and growth promotion

    Directory of Open Access Journals (Sweden)

    Patrícia Baston Barretti

    2009-01-01

    Full Text Available Quarenta isolados bacterianos endofíticos de plantas sadias de tomateiro foram avaliados quanto à sua potencialidade como agentes de biocontrole de doenças do tomateiro. Foi realizada, em casa de vegetação, uma seleção massal utilizando-se Pseudomonas syringae pv. tomato e Alternaria solani, como patógenos desafiantes. Com base na média do número de lesões por planta, quatro isolados foram selecionados como potenciais agentes de biocontrole dessas enfermidades fúngica e bacteriana do tomateiro. Esses isolados foram identificados, por meio do sequenciamento do gene 16S do DNA ribossômico, como Acinetobacter johnsonii (UFV-E05, Serratia marcescens (UFV-E13, Sinorhizobium sp. (UFV-E25 e Bacillus megaterium (UFV-E26. Os mesmos isolados selecionados para o biocontrole também foram avaliados quanto à sua capacidade de promover o crescimento em plantas e somente S. marcescens (UFV-E13 proporcionou aumento na altura das plantas.Forty isolates of endophytic bacteria obtained from healthy tomato plants were tested for their potential as biocontrol agents of tomato diseases. A massal screening was performed at greenhouse using Pseudomonas syringae pv. tomato and Alternaria solani as challenging pathogens. Based on the average number of lesions per plant, four isolates were selected as potential agents of biocontrol of these tomato diseases caused by fungi and bacteria. These isolates were identified by 16S ribosomal DNA sequence analysis as Acinetobacter johnsonii (UFV-E05, Serratia marcescens (UFV-E13, Sinorhizobium sp. (UFV-E25 and Bacillus megaterium (UFV-E26. The four endophytes selected for biocontrol were also evaluated for their ability of promoting plant growth and only S. marcescens (UFV-E13 presented increase in the height of the plants.

  19. The effect of inoculation of an indigenous bacteria on the early growth of Acacia farnesiana in a degraded area

    Directory of Open Access Journals (Sweden)

    Eliane Ceccon

    2012-03-01

    Full Text Available Restoration of native vegetation and fuelwood production are important environmental pending goals for Mexico, where years of wrong management practices resulted in ecosystemic degradation and fuelwood scarcity. In degraded areas, native rhizobial strains are often undetectable, therefore, the restoration of natural vegetation associated with an effective nodulation of the leguminous trees is mostly appropriate. Sinorhizobium americanum is a native nitrogen-fixing bacteria isolated from nodules of the native Acacia species in the region. Acacia farnesiana is a multipurpose leguminous shrub from Mexican seasonally dry tropical forests (SDTF. In this study we analyzed the effect of inoculation with S. americanum on A. farnesiana growth in a greenhouse and in a very degraded area and compared with non-inoculated seedlings. In a greenhouse, we measured the biomass dry weight of different parts of the plant, using destructive sampling after 15, 20, 30, 45 and 120 days of growth. We also calculated the relative growth rate (RGR and the resources allocation (root/shoot weight ratio and root length/root dry weight of seedlings. In a degraded area we measured the seedling length and survival and calculated the RGR. In the greenhouse and in the degraded area, the inoculation positively affected the growth of seedlings. However in the greenhouse, the inoculation did not have effect on resource allocation patterns. Therefore, the inoculation with Sinorhizobium americanum could improve the A. farnesiana growth and the re-establishment of important plant-soil interactions in degraded areas, being a recommendable technique for land restoration and the improvement of fuelwood production.

  20. CARACTERIZACIÓN FENOTÍPICA DE AISLADOS DE RIZOBIOS PROCEDENTES DE LA LEGUMINOSA FORRAJERA Canavalia ensiformis

    Directory of Open Access Journals (Sweden)

    Ionel Hernández Forte

    2012-01-01

    Full Text Available El objetivo del presente trabajo fue caracterizar fenotípicamente aislados de rizobios procedentes de la leguminosa forrajera Canavalia ensiformis. Se caracterizaron doce aislados, cinco posibles miembros del género Rhizobium/ Sinorhizobium, siete posibles integrantes del género Bradyrhizobium y dos cepas comerciales, BR 2001 y BR 2003a. Para la caracterización fenotípica de los aislados bacterianos se estudió su capacidad de utilizar seis fuentes de carbono, la tolerancia a condiciones de acidez y a diferentes niveles de cloruro de sodio (NaCl, la resistencia a seis antibióticos así como su crecimiento en tres temperaturas de incubación. Se determinó además la capacidad de estos aislados de producir polihidroxibutiratos (PHB. Todos los aislados estudiados utilizaron la lactosa, glucosa y manitol como fuentes de carbono. Uno de los aislados de rápido crecimiento resultó el más tolerante a pH 4.5. Los aislados posibles miembros del género Rhizobium/ Sinorhizobium toleraron una mayor concentración de NaCl que los posibles integrantes del género Bradyrhizobium. Tres de los aislados de rápido crecimiento fueron resistentes a los seis antibióticos utilizados. Cuando los aislados fueron incubados a 370C todos, con excepción del el aislado CP3 y la cepa comercial BR 2001, presentaron un crecimiento similar al control. El aislado CP10, de lento crecimiento, tuvo la capacidad de producir las mayores concentraciones de PHB (0.023 g.L-1.

  1. NMR Structure of the hypothetical protein encoded by the YjbJ gene from Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Pineda-Lucena, Antonio; Liao, Jack; Wu, Bin; Yee, Adelinda; Cort, John R.; Kennedy, Michael A.; Edwards, Aled M.; Arrowsmith, Cheryl H.

    2002-06-01

    Here we describe the solution structure of YjbJ (gil418541) as part of a structural proteomics project on the feasibility of the high-throughput generation of samples from Escherichia coli for structural studies. YjbJ is a hypothetical protein from Escherichia coli protein of unknown function. It is conserved, showing significant sequence identity to four predicted prokaryotic proteins, also of unknown function (Figure 1A). These include gil16762921 from Salmonella enterica (S. typhi), gil17938413 from Agrobacterium tumefaciens, gil16265654 from Sinorizhobium meliloti, and gil15599932 from Pseudomona aeruginosa. The structure of YjbJ reveals a new variation of a common motif (four-helix bundle) that could not be predicted from the protein sequence. Although the biochemical function is unknown, the existence of patterns of conserved residues on the protein surface suggest that the fold and function of all these proteins could be similar.

  2. Azospirillum IV

    Energy Technology Data Exchange (ETDEWEB)

    Klingmuller, W.

    1988-01-01

    This book's contents include: Advances in the genetics of Azospirillum brasilense Sp7: Use of Tn5 mutagenesis for gene mapping and identification; Characterization of DNA segments adjacent to the nifHDK genes of Azospirillum brasilense by Sp7 Tn5 site-directed mutagenesis; Selection at the chemostat of Azospirillum brasilense Cd N/sub 2/-fixing at high O/sub 2/ pressure. Root hair deformation induced on maize and medicago by an Azospirillum transconjugant containing a Rhizobium meliloti nodulation region. Azospirilla are bacteria that live in association with the roots of many grain crops. Since these bacteria bind molecular nitrogen from the air and excrete plant growth substances, interest has focussed on their potential to increase crop yields.

  3. Pleiotropic effect of fluoranthene on anthocyanin synthesis and nodulation of Medicago sativa is reversed by the plant flavone luteolin

    Energy Technology Data Exchange (ETDEWEB)

    Wetzel, A.; Parniske, M.; Werner, D. [Univ. of Marburg (Germany)

    1995-05-01

    The symbiosis between leguminous plants and soil bacteria of the genus Rhizobium is of considerable agronominal importance. Recently it has been found, that polycyclic aromatic hydrocarbons (PAHs; e.g. anthracene, phenanthrene, fluoranthene), occurring as ubiquitous environmental contaminants can inhibit nodulation of Medicago sativa. Fluoranthene is one of the dominant PAHs found in urban particulate matter, sewage sludge or beside motorways. Several organisms have been shown to be able to metabolize and mineralize fluoranthene but the uptake of fluoranthene is limited due to low solubility of fluoranthene in water and strong adsorption to humic substances in soil. Rhizobium meliloti cannot degrade fluoranthene. Toxic effects of fluoranthene on bacterial growth have never been observed. In contrast to their rhizobial symbiotic partners, alfalfa plants grown on a solidified fluoranthene-containing medium, exhibited symptoms of toxicity. They showed a dose-responsive decrease in shoot length and, if inoculated with R. meliloti, inhibition of nodule formation. Growth retardation is accompanied by a decrease in anthocyanin pigmentation of shoots, and an atypical accumulation of anthocyanins in roots. Plant flavonoids are known to play a central role in the signal exchange of the Legume-Rhizobium symbiosis. Phenylpropane derived compounds and flavonoids have been implicated in nodule development. Since fluoranthene impairs nodulation and induces the production of anthocyanins, it is possible that these events are causally linked via phenylpropanoid metabolism. These experiments attempt to overcome the inhibitory effects of fluoranthene by exogeneous application of the flavonoid luteolin. This paper demonstrates that luteolin antagonizes the fluoranthene mediated inhibition of nodule formation and prevents the accumulation of anthocyanins in roots. 29 refs., 4 figs., 1 tab.

  4. Rhizobium leguminosarum symbiovar trifolii, Ensifer numidicus and Mesorhizobium amorphae symbiovar ciceri (or Mesorhizobium loti) are new endosymbiotic bacteria of Lens culinaris Medik.

    Science.gov (United States)

    Sami, Dhaoui; Mokhtar, Rejili; Peter, Mergaert; Mohamed, Mars

    2016-08-01

    A total of 142 rhizobial bacteria were isolated from root nodules of Lens culinaris Medik endemic to Tunisia and they belonged to the species Rhizobium leguminosarum, and for the first time to Ensifer and Mesorhizobium, genera never previously described as microsymbionts of lentil. Phenotypically, our results indicate that L. culinaris Medik strains showed heterogenic responses to the different phenotypic features and they effectively nodulated their original host. Based on the concatenation of the 16S rRNA with relevant housekeeping genes (glnA, recA, dnaK), rhizobia that nodulate lentil belonged almost exclusively to the known R. leguminosarum sv. viciae. Interestingly, R. leguminosarum sv. trifolii, Ensifer numidicus (10 isolates) and Mesorhizobium amorphae (or M. loti) (9 isolates) isolates species, not considered, up to now, as a natural symbiont of lentil are reported. The E. numidicus and M. amorphae (or M. loti) strains induced fixing nodules on Medicago sativa and Cicer arietinum host plants, respectively. Symbiotic gene phylogenies showed that the E. numidicus, new symbiont of lentil, markedly diverged from strains of R. leguminosarum, the usual symbionts of lentil, and converged to the symbiovar meliloti so far described within E. meliloti Indeed, the nodC and nodA genes from the M. amorphae showed more than 99% similarity with respect to those from M. mediterraneum, the common chickpea nodulating species, and would be included in the new infrasubspecific division named M. amorphae symbiovar ciceri, or to M. loti, related to the strains able to effectively nodulate C. arietinum host plant. On the basis of these data, R. leguminosarum sv. trifolii (type strain LBg3 (T)), M. loti or M. amorphae sv. ciceri (type strain LB4 (T)) and E. numidicus (type strain LBi2 (T)) are proposed as new symbionts of L. culinaris Medik. PMID:27267929

  5. [MEDICAGO SATIVA L: IMPROVEMENT AND NEW APPROACHES OF ITS NUTRITIONAL AND FUNCTIONAL VALUE BY BACTERIAL CO-INOCULATION].

    Science.gov (United States)

    Martínez, Rosario; Nebot, Elena; Porres, Jesús María; Kapravelou, Garyfallia; Del Moral, Ana; Talbi, Chouhra; Bedmar, Eulogio Jose; López-Jurado, María

    2015-12-01

    Objetivo: estudiar el efecto de la inoculación con Ensifer meliloti y Halomonas maura sobre el crecimiento y el valor nutricional y funcional de la leguminosa Medicago sativa L., cultivada bajo condiciones de salinidad. Método: las plantas de M. sativa se cultivaron con una solución de mezcla de sales CaSO4, MgCl, NaCl and Na- HCO3 y se coinocularon con su rizobio específico y la bacteria H. maura. Se determinaron los parámetros fisiológicos de las plantas, así como el contenido en nitrógeno y minerales, y se llevó a cabo un proceso de digestibilidad in vitro. Resultados: la salinidad ejerció un efecto negativo sobre las plantas; sin embargo, la coinoculación de las mismas incrementó su productividad, el contenido en nitrógeno, minerales totales, Ca y Mg. Además, los parámetros fisiológicos de potencial hídrico y concentración de leghemoglobina se incrementaron. Tanto la salinidad como la coinoculación de las plantas aumentaron la capacidad antioxidante de la leguminosa en los dializados y retenidos obtenidos tras someter a la planta a un proceso de digestibilidad in vitro. Conclusión: la coinoculación con E. meliloti y H. maura podría mejorar el cultivo de la alfalfa bajo condiciones específicas de salinidad, aumentando su composición nutricional y funcional, pudiendo considerarse en la formulación de suplementos nutricionales para el consumo humano.

  6. Bacterial diversity analysis of Huanglongbing pathogen-infected citrus, using PhyloChip and 16S rRNA gene clone library sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Shankar Sagaram, U.; DeAngelis, K.M.; Trivedi, P.; Andersen, G.L.; Lu, S.-E.; Wang, N.

    2009-03-01

    The bacterial diversity associated with citrus leaf midribs was characterized 1 from citrus groves that contained the Huanglongbing (HLB) pathogen, which has yet to be cultivated in vitro. We employed a combination of high-density phylogenetic 16S rDNA microarray and 16S rDNA clone library sequencing to determine the microbial community composition of symptomatic and asymptomatic citrus midribs. Our results revealed that citrus leaf midribs can support a diversity of microbes. PhyloChip analysis indicated that 47 orders of bacteria from 15 phyla were present in the citrus leaf midribs while 20 orders from phyla were observed with the cloning and sequencing method. PhyloChip arrays indicated that nine taxa were significantly more abundant in symptomatic midribs compared to asymptomatic midribs. Candidatus Liberibacter asiaticus (Las) was detected at a very low level in asymptomatic plants, but was over 200 times more abundant in symptomatic plants. The PhyloChip analysis was further verified by sequencing 16S rDNA clone libraries, which indicated the dominance of Las in symptomatic leaves. These data implicate Las as the pathogen responsible for HLB disease. Citrus is the most important commercial fruit crop in Florida. In recent years, citrus Huanglongbing (HLB), also called citrus greening, has severely affected Florida's citrus production and hence has drawn an enormous amount of attention. HLB is one of the most devastating diseases of citrus (6,13), characterized by blotchy mottling with green islands on leaves, as well as stunting, fruit decline, and small, lopsided fruits with poor coloration. The disease tends to be associated with a phloem-limited fastidious {alpha}-proteobacterium given a provisional Candidatus status (Candidatus Liberobacter spp. later changed to Candidatus Liberibacter spp.) in nomenclature (18,25,34). Previous studies indicate that HLB infection causes disorder in the phloem and severely impairs the translocation of assimilates in

  7. Effects of biotic and abiotic constraints on the symbiosis between rhizobia and the tropical leguminous trees Acacia and Prosopis.

    Science.gov (United States)

    Räsänen, Leena A; Lindström, Kristina

    2003-10-01

    N2-fixing, drought tolerant and multipurpose Acacia and Prosopis species are appropriate trees for reforestation of degraded areas in arid and semiarid regions of the tropics and subtropics. Acacia and Prosopis trees form N2-fixing nodules with a wide range of rhizobia, for example African acacias mainly with Sinorhizobium sp. and Mesorhizobium sp., and Australian acacias with Bradyrhizobium sp. Although dry and hot seasons restrict formation of N2-fixing nodules on Acacia and Prosopis spp., fully grown trees and their symbiotic partners are well adapted to survive in harsh growth conditions. This review on one hand deals with major constraints of arid and semiarid soils, i.e. drought, salinity and high soil temperature, which affect growth of trees and rhizobia, and on the other hand with adaptation mechanisms by which both organisms survive through unfavourable periods. In addition, defects in infection and nodulation processes due to various abiotic and biotic constraints are reviewed. This knowledge is important when Acacia and Prosopis seedlings are used for forestation of degraded areas in arid and semiarid tropics. PMID:15242281

  8. Construction of the recombinant broad-host-range plasmids providing their bacterial hosts arsenic resistance and arsenite oxidation ability.

    Science.gov (United States)

    Drewniak, Lukasz; Ciezkowska, Martyna; Radlinska, Monika; Sklodowska, Aleksandra

    2015-02-20

    The plasmid pSinA of Sinorhizobium sp. M14 was used as a source of functional phenotypic modules, encoding proteins involved in arsenite oxidation and arsenic resistance, to obtain recombinant broad-host-range plasmids providing their bacterial hosts arsenic resistance and arsenite oxidative ability. An arsenite oxidation module was cloned into pBBR1MCS-2 vector yielding plasmid vector pAIO1, while an arsenic resistance module was cloned into pCM62 vector yielding plasmid pARS1. Both plasmid constructs were introduced (separately and together) into the cells of phylogenetically distant (representing Alpha-, Beta-, and Gammaproteobacteria) and physiologically diversified (unable to oxidize arsenite and susceptible/resistant to arsenite and arsenate) bacteria. Functional analysis of the modified strains showed that: (i) the plasmid pARS1 can be used for the construction of strains with an increased resistance to arsenite [up to 20mM of As(III), (ii) the presence of the plasmid pAIO1 in bacteria previously unable to oxidize As(III) to As(V), contributes to the acquisition of arsenite oxidation abilities by these cells, (iii) the highest arsenite utilization rate are observed in the culture of strains harbouring both the plasmids pAIO1 and pARS1, (iv) the strains harbouring the plasmid pAIO1 were able to grow on arsenic-contaminated mine waters (∼ 3.0 mg As L(-1)) without any supplementation. PMID:25617684

  9. Fast induction of biosynthetic polysaccharide genes lpxA, lpxE, and rkpI of Rhizobium sp. strain PRF 81 by common bean seed exudates is indicative of a key role in symbiosis.

    Science.gov (United States)

    Oliveira, Luciana Ruano; Rodrigues, Elisete Pains; Marcelino-Guimarães, Francismar Corrêa; Oliveira, André Luiz Martinez; Hungria, Mariangela

    2013-06-01

    Rhizobial surface polysaccharides (SPS) are, together with nodulation (Nod) factors, recognized as key molecules for establishment of rhizobia-legume symbiosis. In Rhizobium tropici, an important nitrogen-fixing symbiont of common bean (Phaseolus vulgaris L.), molecular structures and symbiotic roles of the SPS are poorly understood. In this study, Rhizobium sp. strain PRF 81 genes, belonging to the R. tropici group, were investigated: lpxA and lpxE, involved in biosynthesis and modification of the lipid-A anchor of lipopolysaccharide (LPS), and rkpI, involved in synthesis of a lipid carrier required for production of capsular polysaccharides (KPS). Reverse transcription quantitative PCR (RT-qPCR) analysis revealed, for the first time, that inducers released from common bean seeds strongly stimulated expression of all three SPS genes. When PRF 81 cells were grown for 48 h in the presence of seed exudates, twofold increases (p PRF81 and of the type strain of R. tropici CIAT899(T)clustered with orthologous Rhizobium radiobacter and were more related to R. etli and Rhizobium leguminosarum, while rkpI was closer to the Sinorhizobium sp. group. Upregulation of lpxE, lpxA, and rkpI genes suggests that seed exudates can modulate production of SPS of Rhizobium sp. PRF81, leading to cell wall changes necessary for symbiosis establishment.

  10. Analysis of Genetic Diversity and Phylogenesis of Rhizobia from Ammopiptanthus mongolicus%沙冬青根瘤菌遗传多样性和系统发育分析

    Institute of Scientific and Technical Information of China (English)

    毕江涛; 贺达汉; 陈卫民; 谢瑞梅; 韦革宏

    2009-01-01

    利用16S rDNA RFLP、16S-23S rDNA RFLP和16S rDNA序列分析方法,对分离自宁夏和内蒙古阿拉善地区的沙冬青根瘤菌进行了遗传多样性和系统发育分析.结果表明,分离自不同地区沙冬青根瘤菌的44株测试菌株分别归属于中慢生根瘤菌属(Mesorhizobium)、叶杆菌属 (Phylobacterium)、中华根瘤菌属(Sinorhizobium)、根瘤菌属(Rhizobium)、土壤杆菌属(Agrobacterium)5个属种.受寄主和地理环境因素的影响,沙冬青根瘤菌具有丰富的遗传多样性.

  11. Improvement in nitrogen fixation capacity could be part of the domestication process in soybean.

    Science.gov (United States)

    Muñoz, N; Qi, X; Li, M-W; Xie, M; Gao, Y; Cheung, M-Y; Wong, F-L; Lam, H-M

    2016-08-01

    Biological nitrogen fixation (BNF) in soybeans is a complex process involving the interplay between the plant host and the symbiotic rhizobia. As nitrogen supply has a crucial role in growth and development, higher nitrogen fixation capacity would be important to achieve bigger plants and larger seeds, which were important selection criteria during plant domestication by humans. To test this hypothesis, we monitored the nitrogen fixation-related performance in 31 cultivated and 17 wild soybeans after inoculation with the slow-growing Bradyrhizobium diazoefficiens sp. nov. USDA110 and the fast-growing Sinorhizobium (Ensifer) fredii CCBAU45436. Our results showed that, in general, cultivated soybeans gave better performance in BNF. Electron microscopic studies indicated that there was an exceptionally high accumulation of poly-β-hydroxybutyrate bodies in bacteroids in the nodules of all wild soybeans tested, suggesting that the C/N balance in wild soybeans may not be optimized for nitrogen fixation. Furthermore, we identified new quantitative trait loci (QTLs) for total ureides and total nodule fresh weight by employing a recombinant inbred population composed of descendants from a cross between a cultivated and a wild parent. Using nucleotide diversity (θπ), divergence index (Fst) and distribution of fixed single-nucleotide polymorphisms as parameters, we found that some regions in the total ureides QTL on chromosome 17 and the total nodule fresh weight QTL on chromosome 12 exhibited very low diversity among cultivated soybeans, suggesting that these were traits specially selected during the domestication and breeding process.

  12. Bacterial Communities Associated with Different Anthurium andraeanum L. Plant Tissues

    Science.gov (United States)

    Sarria-Guzmán, Yohanna; Chávez-Romero, Yosef; Gómez-Acata, Selene; Montes-Molina, Joaquín Adolfo; Morales-Salazar, Eleacin; Dendooven, Luc; Navarro-Noya, Yendi E.

    2016-01-01

    Plant-associated microbes have specific beneficial functions and are considered key drivers for plant health. The bacterial community structure of healthy Anthurium andraeanum L. plants was studied by 16S rRNA gene pyrosequencing associated with different plant parts and the rhizosphere. A limited number of bacterial taxa, i.e., Sinorhizobium, Fimbriimonadales, and Gammaproteobacteria HTCC2089 were enriched in the A. andraeanum rhizosphere. Endophytes were more diverse in the roots than in the shoots, whereas all shoot endophytes were found in the roots. Streptomyces, Flavobacterium succinicans, and Asteroleplasma were only found in the roots, Variovorax paradoxus only in the stem, and Fimbriimonas 97%-OTUs only in the spathe, i.e., considered specialists, while Brevibacillus, Lachnospiraceae, Pseudomonas, and Pseudomonas pseudoalcaligenes were generalist and colonized all plant parts. The anaerobic diazotrophic bacteria Lachnospiraceae, Clostridium sp., and Clostridium bifermentans colonized the shoot system. Phylotypes belonging to Pseudomonas were detected in the rhizosphere and in the substrate (an equiproportional mixture of soil, cow manure, and peat), and dominated the endosphere. Pseudomonas included nine 97%-OTUs with different patterns of distribution and phylogenetic affiliations with different species. P. pseudoalcaligenes and P. putida dominated the shoots, but were also found in the roots and rhizosphere. P. fluorescens was present in all plant parts, while P. resinovorans, P. denitrificans, P. aeruginosa, and P. stutzeri were only detected in the substrate and rhizosphere. The composition of plant-associated bacterial communities is generally considered to be suitable as an indicator of plant health. PMID:27524305

  13. Endophytic bacteria from Piper tuberculatum Jacq.: isolation, molecular characterization, and in vitro screening for the control of Fusarium solani f. sp piperis, the causal agent of root rot disease in black pepper (Piper nigrum L.).

    Science.gov (United States)

    Nascimento, S B; Lima, A M; Borges, B N; de Souza, C R B

    2015-01-01

    Endophytic bacteria have been found to colonize internal tissues in many different plants, where they can have several beneficial effects, including defense against pathogens. In this study, we aimed to identify endophytic bacteria associated with roots of the tropical piperaceae Piper tuberculatum, which is known for its resistance to infection by Fusarium solani f. sp piperis, the causal agent of black pepper (Piper nigrum) root rot disease in the Amazon region. Based on 16S rRNA gene sequence analysis, we isolated endophytes belonging to 13 genera: Bacillus, Paenibacillus, Pseudomonas, Enterobacter, Rhizobium, Sinorhizobium, Agrobacterium, Ralstonia, Serratia, Cupriavidus, Mitsuaria, Pantoea, and Staphylococcus. The results showed that 56.52% of isolates were associated with the phylum Proteobacteria, which comprised α, β, and γ classes. Other bacteria were related to the phylum Firmicutes, including Bacillus, which was the most abundant genus among all isolates. Antagonistic assays revealed that Pt12 and Pt13 isolates, identified as Pseudomonas putida and Pseudomonas sp, respectively, were able to inhibit F. solani f. sp piperis growth in vitro. We describe, for the first time, the molecular identification of 23 endophytic bacteria from P. tuberculatum, among which two Pseudomonas species have the potential to control the pathogen responsible for root rot disease in black pepper in the Amazon region. PMID:26214435

  14. Robust markers reflecting phylogeny and taxonomy of rhizobia.

    Directory of Open Access Journals (Sweden)

    Yan Ming Zhang

    Full Text Available Genomic ANI (Average Nucleotide Identity has been found to be able to replace DNA-DNA hybridization in prokaryote taxonomy. The ANI of each of the core genes that has a phylogeny congruent with the reference species tree of rhizobia was compared to the genomic ANI. This allowed us to identify three housekeeping genes (SMc00019-truA-thrA whose ANI reflected the intraspecies and interspecies genomic ANI among rhizobial strains, revealing an ANI gap (≥2% between the inter- and intra-species comparisons. The intraspecies (96% and interspecies (94% ANI boundaries calculated from three genes (SMc00019-truA-thrA provided a criterion for bacterial species definition and confirmed 621/629 of known interspecies relationships within Bradyrhizobium, Mesorhizobium, Sinorhizobium and Rhizobium. Some widely studied strains should be renamed. The SMc00019-truA-thrA ANI also correlates well with the genomic ANI of strains in Agrobacterium, Methylobacterium, Ralstonia, Rhodopseudomonas, Cupriavidus and Burkholderia, suggesting their wide applicability in other bacteria.

  15. Characterization of a symbiotically effective Rhizobium resistant to arsenic: Isolated from the root nodules of Vigna mungo (L.) Hepper grown in an arsenic-contaminated field.

    Science.gov (United States)

    Mandal, Santi M; Pati, Bikas R; Das, Amit K; Ghosh, Ananta K

    2008-04-01

    Bacteria were isolated from the root nodules of Vigna mungo (L.) Hepper, grown in an arsenic-contaminated field and the strain was selected by its nodulation ability as well as better arsenic tolerant capacity compared to others. The selected strain was identified as Rhizobium by 16S rDNA sequencing and designated as VMA301. Phylogenetic analysis of the gene sequences showed its close relatedness with Sinorhizobium fredii. LC(50) value of arsenate for the bacteria as determined by flow cytometry was found to be 2.8 mM and arsenic uptake was measured by atomic absorption spectrometry as 0.048 mg g(-1) biomass. The high amount of arsenic was toxic to the cell, which changed the morphology of the bacteria to an elongated shape. Presence of a transcriptional regulatory gene (ArsR) of the ars genetic system was confirmed by amplification and sequencing. The symbiotic property of the isolate was also confirmed by amplification and sequencing of the NodC gene. These results indicate that the isolated Rhizobium bacteria may exert dual roles in the environment, arsenic bioremediation from the soil as well as increase of soil fertility through nitrogen fixation.

  16. Simultaneous arsenite oxidation and nitrate reduction at the electrodes of bioelectrochemical systems.

    Science.gov (United States)

    Nguyen, Van Khanh; Park, Younghyun; Yu, Jaecheul; Lee, Taeho

    2016-10-01

    Arsenic and nitrate contaminations in the soil and groundwater have urged the scientific community to explore suitable technologies for treatment of both contaminants. This study reports, for the first time, a novel application of bioelectrochemical systems for coupling As detoxification at the anode and denitrification at the cathode. A similar As(III) oxidation efficiency was achieved when anode potential was controlled by a potentiostat or a direct current (DC) power supply. However, a slightly lower nitrate reduction rate was obtained in reactors using DC power supply during simultaneous operation of nitrate reduction and As(III) oxidation. Microbial community analysis by denaturing gradient gel electrophoresis indicated the presence of some autotrophic As(III)-oxidizing bacteria, including Achromobacter spp., Ensifer spp., and Sinorhizobium spp., that can flexibly switch their original metabolism of using oxygen as sole electron acceptor to a new metabolism mode of using solid-state anode as sole electron acceptor driving for As(III) oxidation under anaerobic conditions. Although further research is required for validating their applicability, bioelectrochemical systems represent a brilliant technology for remediation of groundwater contaminated with nitrate and/or arsenite. PMID:27438874

  17. In Silico Identification for α-Amino-ε-Caprolactam Racemases by Using Information on the Structure and Function Relationship.

    Science.gov (United States)

    Payoungkiattikun, Wisarut; Okazaki, Seiji; Nakano, Shogo; Ina, Atsutoshi; H-Kittikun, Aran; Asano, Yasuhisa

    2015-07-01

    In silico identification for enzymes having desired functions is attractive because there is a possibility that numerous desirable enzymes have been deposited in databases. In this study, α-amino-ε-caprolactam (ACL) racemases were searched from the NCBI protein database. Four hundred thirteen fold-type I pyridoxal 5'-phosphate-dependent enzymes which are considered to contain sequences of ACL racemase were firstly obtained by submitting the sequence of ACL racemase from Achromobacter obae to the database. By identifying Lys241 as a key amino acid residue, 13 candidates for ACL racemase were selected. Then, putative ACL racemase genes were synthesized as codon-optimized sequences for expression in Escherichia coli. They were subcloned and expressed in E. coli BL21 and underwent His-tag purification. ACL and amino acid amide racemizing activities were detected among ten of the candidates. The locus tags Oant_4493, Smed_5339, and CSE45_2055 derived from Ochrobactrum anthropi ATCC49188, Sinorhizobium medicae WSM 419, and Citreicella sp. SE45, respectively, showed higher racemization activity against D- and L-ACLs rather than that of ACL racemase from A. obae. Our results demonstrate that the newly discovered ACL racemases were unique from ACL racemase from A. obae and might be useful for applications in dynamic kinetic resolution for D- or L-amino acid production. PMID:26206345

  18. Characterization of tropical tree rhizobia and description of Mesorhizobium plurifarium sp. nov.

    Science.gov (United States)

    de Lajudie, P; Willems, A; Nick, G; Moreira, F; Molouba, F; Hoste, B; Torck, U; Neyra, M; Collins, M D; Lindström, K; Dreyfus, B; Gillis, M

    1998-04-01

    A collection of strains isolated from root nodules of Acacia species in Senegal was analysed previously by electrophoresis of total cell protein, auxanographic tests, rRNA-DNA hydridization, 16S rRNA gene sequencing, DNA base composition and DNA-DNA hybridization [de Lajudie, P., Willems, A., Pot, B. & 7 other authors (1994). Int J Syst Bacteriol 44, 715-733]. Strains from Acacia were shown to belong to two groups, Sinorhizobium terangae, and a so-called gel electrophoretic cluster U, which also included some reference strains from Brazil. Further taxonomic characterization of this group using the same techniques plus repetitive extragenic palindromic-PCR and nodulation tests is presented in this paper. Reference strains from Sudan and a number of new rhizobia isolated from nodules of Acacia senegal, Acacia tortilis subsp. raddiana and Prosopis juliflora in Senegal were included. As a result of this polyphasic approach, the creation of a new species, Mesorhizobium plurifarium, is proposed for a genotypically and phenotypically distinct group corresponding to the former cluster U and containing strains isolated from Acacia, Leucaena, Prosopis and Chamaecrista in West Africa (Senegal), East Africa (Sudan) and South America (Brazil). The type strain of Mesorhizobium plurifarium ORS 1032 has been deposited in the LMG collection as LMG 11892.

  19. Effects of biotic and abiotic constraints on the symbiosis between rhizobia and the tropical leguminous trees Acacia and Prosopis.

    Science.gov (United States)

    Räsänen, Leena A; Lindström, Kristina

    2003-10-01

    N2-fixing, drought tolerant and multipurpose Acacia and Prosopis species are appropriate trees for reforestation of degraded areas in arid and semiarid regions of the tropics and subtropics. Acacia and Prosopis trees form N2-fixing nodules with a wide range of rhizobia, for example African acacias mainly with Sinorhizobium sp. and Mesorhizobium sp., and Australian acacias with Bradyrhizobium sp. Although dry and hot seasons restrict formation of N2-fixing nodules on Acacia and Prosopis spp., fully grown trees and their symbiotic partners are well adapted to survive in harsh growth conditions. This review on one hand deals with major constraints of arid and semiarid soils, i.e. drought, salinity and high soil temperature, which affect growth of trees and rhizobia, and on the other hand with adaptation mechanisms by which both organisms survive through unfavourable periods. In addition, defects in infection and nodulation processes due to various abiotic and biotic constraints are reviewed. This knowledge is important when Acacia and Prosopis seedlings are used for forestation of degraded areas in arid and semiarid tropics.

  20. Potential Emergence of Multi-quorum Sensing Inhibitor Resistant (MQSIR) Bacteria.

    Science.gov (United States)

    Koul, Shikha; Prakash, Jyotsana; Mishra, Anjali; Kalia, Vipin Chandra

    2016-03-01

    Expression of certain bacterial genes only at a high bacterial cell density is termed as quorum-sensing (QS). Here bacteria use signaling molecules to communicate among themselves. QS mediated genes are generally involved in the expression of phenotypes such as bioluminescence, biofilm formation, competence, nodulation, and virulence. QS systems (QSS) vary from a single in Vibrio spp. to multiple in Pseudomonas and Sinorhizobium species. The complexity of QSS is further enhanced by the multiplicity of signals: (1) peptides, (2) acyl-homoserine lactones, (3) diketopiperazines. To counteract this pathogenic behaviour, a wide range of bioactive molecules acting as QS inhibitors (QSIs) have been elucidated. Unlike antibiotics, QSIs don't kill bacteria and act at much lower concentration than those of antibiotics. Bacterial ability to evolve resistance against multiple drugs has cautioned researchers to develop QSIs which may not generate undue pressure on bacteria to develop resistance against them. In this paper, we have discussed the implications of the diversity and multiplicity of QSS, in acting as an arsenal to withstand attack from QSIs and may use these as reservoirs to develop multi-QSI resistance. PMID:26843692

  1. Endophytic bacteria from Piper tuberculatum Jacq.: isolation, molecular characterization, and in vitro screening for the control of Fusarium solani f. sp piperis, the causal agent of root rot disease in black pepper (Piper nigrum L.).

    Science.gov (United States)

    Nascimento, S B; Lima, A M; Borges, B N; de Souza, C R B

    2015-07-06

    Endophytic bacteria have been found to colonize internal tissues in many different plants, where they can have several beneficial effects, including defense against pathogens. In this study, we aimed to identify endophytic bacteria associated with roots of the tropical piperaceae Piper tuberculatum, which is known for its resistance to infection by Fusarium solani f. sp piperis, the causal agent of black pepper (Piper nigrum) root rot disease in the Amazon region. Based on 16S rRNA gene sequence analysis, we isolated endophytes belonging to 13 genera: Bacillus, Paenibacillus, Pseudomonas, Enterobacter, Rhizobium, Sinorhizobium, Agrobacterium, Ralstonia, Serratia, Cupriavidus, Mitsuaria, Pantoea, and Staphylococcus. The results showed that 56.52% of isolates were associated with the phylum Proteobacteria, which comprised α, β, and γ classes. Other bacteria were related to the phylum Firmicutes, including Bacillus, which was the most abundant genus among all isolates. Antagonistic assays revealed that Pt12 and Pt13 isolates, identified as Pseudomonas putida and Pseudomonas sp, respectively, were able to inhibit F. solani f. sp piperis growth in vitro. We describe, for the first time, the molecular identification of 23 endophytic bacteria from P. tuberculatum, among which two Pseudomonas species have the potential to control the pathogen responsible for root rot disease in black pepper in the Amazon region.

  2. Diversity of root nodule bacteria from leguminous crops

    Directory of Open Access Journals (Sweden)

    Agrawal Pooja

    2016-01-01

    Full Text Available In the present study, a total of 353 nodule-associated bacteria were isolated from 220 legume plant samples belonging to Cicer arietinum (85, Glycine max (74, Vigna radiata (21 and Cajanus cajan (40. A total of 224 bacteria were identified as fast-growing Rhizobium spp. on the basis of differential staining (Gram staining and carbol fuchsin staining and biochemical tests. All the isolates were tested for indole acetic acid production (IAA, phosphate solubilization and siderophore production on plate assay. To examine the effect of volatile organic metabolites (VOM and water soluble soil components (WSSC on nodule bacteria, culture conditions were optimized by observing the effects of various parameters such as pH, salt content and temperatures on the growth of bacteria. Selected rhizobia were subjected to random amplified polymorphic DNA (RAPD and amplified ribosomal DNA restriction analysis (ARDRA analysis to identify their species. On the basis of RAPD and ARDRA, 10 isolates were identified as Rhizobium meliloti. In this study, Rhizobium GO4, G16, G20, G77, S43, S81, M07, M37, A15 and A55 were observed as the best candidates among the tested bacteria and can be further used as potent bioinoculants.

  3. Phylogenetic Diversity of Ammopiptanthus Rhizobia and Distribution of Rhizobia Associated with Ammopiptanthus mongolicus in Diverse Regions of Northwest China.

    Science.gov (United States)

    Zhao, Liang; Wang, Xinye; Huo, Haibo; Yuan, Guiji; Sun, Yali; Zhang, Dehui; Cao, Ying; Xu, Lin; Wei, Gehong

    2016-07-01

    Aiming to investigate the diversity and distribution of rhizobia associated with Ammopiptanthus, an endangered evergreen legume widely distributed in deserts, we characterized a total of 219 nodule isolates from nine sampling sites in Northwest China with different soil characteristics based upon restriction fragment length polymorphism (RFLP) analysis of 16S ribosomal RNA (rRNA) and symbiotic genes (nodC and nifH). Ten isolates representing different 16S rRNA-RFLP types were selected for further sequence analyses of 16S rRNA and four housekeeping genes. As results, nine genospecies belonging to the genera Ensifer, Neorhizobium, Agrobacterium, Pararhizobium, and Rhizobium could be defined among the isolates. The nodC and nifH phylogenies of 14 isolates representing different symbiotic-RFLP types revealed five lineages linked to Ensifer fredii, Ensifer meliloti, Rhizobium leguminosarum, Mesorhizobium amorphae, and Rhizobium gallicum, which demonstrated the various origins and lateral transfers of symbiotic genes between different genera and species. The rhizobial diversities of Ammopiptanthus mongolicus varied among regions, and the community compositions of rhizobia associated with A. mongolicus were significantly different in wild and cultured fields. Constrained correspondence analysis showed that the distribution of A. mongolicus rhizobia could be explained by available potassium content and that the assembly of symbiotic types was mainly affected by available phosphorus content and carbon-nitrogen ratio. PMID:27079453

  4. Induction of microbial genes for pathogenesis and symbiosis by chemicals from root border cells.

    Science.gov (United States)

    Zhu, Y; Pierson, L S; Hawes, M C

    1997-12-01

    Reporter strains of soil-borne bacteria were used to test the hypothesis that chemicals released by root border cells can influence the expression of bacterial genes required for the establishment of plant-microbe associations. Promoters from genes known to be activated by plant factors included virE, required for Agrobacterium tumefaciens pathogenesis, and common nod genes from Rhizobium leguminosarum bv viciae and Rhizobium meliloti, required for nodulation of pea (Pisum sativum) and alfalfa (Medicago sativum), respectively. Also included was phzB, an autoinducible gene encoding the biosynthesis of antibiotics by Pseudomonas aureofaciens. The virE and nod genes were activated to different degrees, depending on the source of border cells, whereas phzB activity remained unaffected. The homologous interaction between R. leguminosarum bv viciae and its host, pea, was examined in detail. Nod gene induction by border cells was dosage dependent and responsive to environmental signals. The highest levels of gene induction by pea (but not alfalfa) border cells occurred at low temperatures, when little or no bacterial growth was detected. Detached border cells cultured in distilled water exhibited increased nod gene induction (ini) in response to signals from R. leguminosarum bv viciae.

  5. Sequence and structural organization of a nif A-like gene and part of a nifB-like gene of Herbaspirillum seropedicae strain Z78.

    Science.gov (United States)

    Souza, E M; Funayama, S; Rigo, L U; Yates, M G; Pedrosa, F O

    1991-07-01

    The deduced amino acid sequence derived from the sequence of a fragment of DNA from the free-living diazotroph Herbaspirillum seropedicae was aligned to the homologous protein sequences encoded by the nifA genes from Azorhizobium caulinodans, Rhizobium leguminosarum, Rhizobium meliloti and Klebsiella pneumoniae. High similarity was found in the central domain and in the C-terminal region. The H. seropedicae putative NifA sequence was also found to contain an interdomain linker similar to that conserved among rhizobial NifA proteins, but not K. pneumoniae or Azotobacter vinelandii. Analysis of the regulatory sequences found 5' from nifA indicated that the expression of this gene in H. seropedicae is likely to be controlled by NifA, NtrC and RpoN, as judged by the presence of specific NifA- and NtrC-binding sites and characteristic -24/-12 promoters. Possible additional regulatory features included an 'anaerobox' and a site for integration host factor. The N-terminus of another open reading frame was found 3' from nifA and tentatively identified as nifB by amino acid sequence comparison. The putative nifB promoter sequence suggests that expression of H. seropedicae nifB may be activated by NifA and dependent on RpoN. PMID:1840608

  6. Nucleotide sequence of the fixABC region of Azorhizobium caulinodans ORS571: similarity of the fixB product with eukaryotic flavoproteins, characterization of fixX, and identification of nifW.

    Science.gov (United States)

    Arigoni, F; Kaminski, P A; Hennecke, H; Elmerich, C

    1991-03-01

    The nucleotide sequence of a 4.1 kb DNA fragment containing the fixABC region of Azorhizobium caulinodans was established. The three gene products were very similar to the corresponding polypeptides of Rhizobium meliloti. The C-terminal domains of both fixB products displayed a high degree of similarity with the alpha-subunits of rat and human electron transfer flavoproteins, suggesting a role for the FixB protein in a redox reaction. Two open reading frames (ORF) were found downstream of fixC. The first ORF was identified as fixX on the basis of sequence homology with fixX from several Rhizobium and Bradyrhizobium strains. The second ORF potentially encoded a 69 amino acid product and was found to be homologous to a DNA region in the Rhodobacter capsulatus nif cluster I. Insertion mutagenesis of the A. caulinodans fixX gene conferred a Nif- phenotype to bacteria growth in the free-living state and a Fix- phenotype in symbiotic association with the host plant Sesbania rostrata. A crude extract from the fixX mutant had no nitrogenase activity. Furthermore, data presented in this paper also indicate that the previously identified nifO gene located upstream of fixA was probably a homologue of the nifW gene of Klebsiella pneumoniae and Azotobacter vinelandii. PMID:1850088

  7. Optimization of Biodegradability and Toxicity Testing of Degradation Product from Linear Alkyl BenzeneSulfonate (LAS Surfactant as Cleaning Detergent Agent

    Directory of Open Access Journals (Sweden)

    Neera Khairani

    2009-11-01

    Full Text Available Optimization of Biodegradability and Toxicity Testing of Degradation Product from Linear Alkyl BenzeneSulfonate (LAS Surfactant as Cleaning Detergent Agent. Linear Alkyl Benzene Sulfonate (LAS is a surfactantused in laundry detergent as cleaning agent and toxic to aquatic organisms. Results shows, with the LAS concentrationused (20 ppm in medium, adaptation time and Acinetobacter sp. growth has shown better biodegradation ability thanthree other bacteria used Pseudomonas putida, Pseudomonas fluorescence, Bacillaria spp. Thus, Acinetobacter sp isused further for biodegradation process of LAS. Based on its biodegradation half-life using Acinetobacter sp, and withmixed culture (± 52.32% and ± 46.82% respectively could be achieved in 4 (four days, LAS could be categorized as abiodegradable compound. The toxicity assay is based on tetrazolium dye reduction with Rhizobium meliloti as indicatororganism. LAS is more toxic than its intermediate product from biodegradation, with IC50 = 34.35 ppm, and theintermediate product, Ac and Cm, has IC50 = 446.19 ppm and 111.28 ppm respectively. Identification of intermediateproducts using IR and LC-MS analysis shows that the degradation product contains chemicals compounds withfunctional group as follows: benzene, benzoic acid, hydroxyl, and aliphatic carbons with large molecule weight. Untilits half-time degradation time, LAS biodegradation process only occurs at the aliphatic carbon chain, and have not yetreached the stage of aromatic ring opening.

  8. The center for plant and microbial complex carbohydrates at the University of Georgia Complex Carbohydrate Research Center. Annual report, September 15, 1990--December 31, 1991

    Energy Technology Data Exchange (ETDEWEB)

    Albersheim, P.; Darvill, A.

    1991-08-01

    Research from the Complex Carbohydrates Research Center at the University of Georgia is presented. Topics include: Structural determination of soybean isoflavones which specifically induce Bradyrhizobium japonicum nodD1 but not the nodYABCSUIJ operon; structural analysis of the lipopolysaccharides (LPSs) from symbiotic mutants of Bradyrhizobium japonicum; structural characterization of lipooligosaccharides from Bradyrhizobium japonicum that are required for the specific nodulation of soybean; structural characterization of the LPSs from R. Leguminosarum biovar phaseoli, the symbiont of bean; characterization of bacteroid-specific LPS epitopes in R. leguminosarum biovar viciae; analysis of the surface polysaccharides of Rhizobium meliloti mutants whose lipopolysaccharides and extracellular polysaccharides can have the same function in symbiosis; characterization of a polysaccharide produced by certain Bradyrhizobium japonicum strains within soybean nodules; structural analysis of a streptococcal adhesin polysaccharide receptor; conformational studies of xyloglucan, the role of the fucosylated side chain in surface-specific cellulose-xyloglucan interactions; the structure of an acylated glucosamine oligosaccharide signal molecule (nod factor) involved in the symbiosis of Rhizobium leguminosarum biovar viciae with its host Vicia sativa; investigating membrane responses induced by oligogalacturonides in cultured cells; the polygalacturonase inhibitor protein; characterization of the self-incompatability glycoproteins from Petunia hybrida; investigation of the cell wall polysaccharide structures of Arabidopsis thaliana; and the glucan inhibition of virus infection of tabacco.

  9. The center for plant and microbial complex carbohydrates at the University of Georgia Complex Carbohydrate Research Center

    Energy Technology Data Exchange (ETDEWEB)

    Albersheim, P.; Darvill, A.

    1991-08-01

    Research from the Complex Carbohydrates Research Center at the University of Georgia is presented. Topics include: Structural determination of soybean isoflavones which specifically induce Bradyrhizobium japonicum nodD1 but not the nodYABCSUIJ operon; structural analysis of the lipopolysaccharides (LPSs) from symbiotic mutants of Bradyrhizobium japonicum; structural characterization of lipooligosaccharides from Bradyrhizobium japonicum that are required for the specific nodulation of soybean; structural characterization of the LPSs from R. Leguminosarum biovar phaseoli, the symbiont of bean; characterization of bacteroid-specific LPS epitopes in R. leguminosarum biovar viciae; analysis of the surface polysaccharides of Rhizobium meliloti mutants whose lipopolysaccharides and extracellular polysaccharides can have the same function in symbiosis; characterization of a polysaccharide produced by certain Bradyrhizobium japonicum strains within soybean nodules; structural analysis of a streptococcal adhesin polysaccharide receptor; conformational studies of xyloglucan, the role of the fucosylated side chain in surface-specific cellulose-xyloglucan interactions; the structure of an acylated glucosamine oligosaccharide signal molecule (nod factor) involved in the symbiosis of Rhizobium leguminosarum biovar viciae with its host Vicia sativa; investigating membrane responses induced by oligogalacturonides in cultured cells; the polygalacturonase inhibitor protein; characterization of the self-incompatability glycoproteins from Petunia hybrida; investigation of the cell wall polysaccharide structures of Arabidopsis thaliana; and the glucan inhibition of virus infection of tabacco.

  10. A gene cluster for amylovoran synthesis in Erwinia amylovora: characterization and relationship to cps genes in Erwinia stewartii.

    Science.gov (United States)

    Bernhard, F; Coplin, D L; Geider, K

    1993-05-01

    A large ams gene cluster required for production of the acidic extracellular polysaccharide (EPS) amylovoran by the fire blight pathogen Erwinia amylovora was cloned. Tn5 mutagenesis and gene replacement were used to construct chromosomal ams mutants. Five complementation groups, essential for amylovoran synthesis and virulence in E. amylovora, were identified and designated ams A-E. The ams gene cluster is about 7 kb in size and functionally equivalent to the cps gene cluster involved in EPS synthesis by the related pathogen Erwinia stewartii. Mucoidy and virulence were restored to E. stewartii mutants in four cps complementation groups by the cloned E. amylovora ams genes. Conversely, the E. stewartii cps gene cluster was able to complement mutations in E. amylovora ams genes. Correspondence was found between the amsA-E complementation groups and the cpsB-D region, but the arrangement of the genes appears to be different. EPS production and virulence were also restored to E. amylovora amsE and E. stewartii cpsD mutants by clones containing the Rhizobium meliloti exo A gene.

  11. Ensifer glycinis sp. nov., a rhizobial species associated with species of the genus Glycine.

    Science.gov (United States)

    Yan, Hui; Yan, Jun; Sui, Xin Hua; Wang, En Tao; Chen, Wen Xin; Zhang, Xiao Xia; Chen, Wen Feng

    2016-09-01

    Rhizobial strains from root nodules of Astragalus mongholicus and soybean (Glycine max) were characterized phylogenetically as members of the genus Ensifer (formerly named Sinorhizobium), based on 16S rRNA gene sequence comparisons. Results based upon concatenated sequence analysis of three housekeeping genes (recA, atpD and glnII, ≤ 93.8 % similarities to known species) and average nucleotide identity (ANI) values of whole genome sequence comparisons (ranging from 89.6 % to 83.4 % to Ensifer fredii and Ensifer saheli, respectively) indicated the distinct positions of these novel strains within the genus Ensifer. Phylogeny of symbiotic genes (nodC and nifH) of three novel strains clustered them with rhizobial species Ensifer fredii and Ensifer sojae, both isolated from nodules of Glycine max. Cross-nodulation tests showed that the representative strain CCBAU 23380T could form root nodules with nitrogen fixation capability on Glycine soja, Albizia julibrissin, Vigna unguiculata and Cajanus cajan, but failed to nodulate Astragalus mongholicus, its original host legume. Strain CCBAU 23380T formed inefficient nodules on G. max, and it did not contain 18 : 0, 18 : 1ω7c 11-methyl or summed feature 1 fatty acids, which differed from other related strains. Failure to utilize malonic acid as a carbon source distinguished strain CCBAU 23380T from the type strains of related species. The genome size of CCBAU 23380T was 6.0 Mbp, comprising 5624 predicted genes with DNA G+C content of 62.4 mol%. Based on the results above, a novel species, Ensifer glycinis sp. nov., is proposed, with CCBAU 23380T (=LMG 29231T =HAMBI 3645T) as the type strain. PMID:27125987

  12. Ensifer glycinis sp. nov., a rhizobial species associated with species of the genus Glycine.

    Science.gov (United States)

    Yan, Hui; Yan, Jun; Sui, Xin Hua; Wang, En Tao; Chen, Wen Xin; Zhang, Xiao Xia; Chen, Wen Feng

    2016-09-01

    Rhizobial strains from root nodules of Astragalus mongholicus and soybean (Glycine max) were characterized phylogenetically as members of the genus Ensifer (formerly named Sinorhizobium), based on 16S rRNA gene sequence comparisons. Results based upon concatenated sequence analysis of three housekeeping genes (recA, atpD and glnII, ≤ 93.8 % similarities to known species) and average nucleotide identity (ANI) values of whole genome sequence comparisons (ranging from 89.6 % to 83.4 % to Ensifer fredii and Ensifer saheli, respectively) indicated the distinct positions of these novel strains within the genus Ensifer. Phylogeny of symbiotic genes (nodC and nifH) of three novel strains clustered them with rhizobial species Ensifer fredii and Ensifer sojae, both isolated from nodules of Glycine max. Cross-nodulation tests showed that the representative strain CCBAU 23380T could form root nodules with nitrogen fixation capability on Glycine soja, Albizia julibrissin, Vigna unguiculata and Cajanus cajan, but failed to nodulate Astragalus mongholicus, its original host legume. Strain CCBAU 23380T formed inefficient nodules on G. max, and it did not contain 18 : 0, 18 : 1ω7c 11-methyl or summed feature 1 fatty acids, which differed from other related strains. Failure to utilize malonic acid as a carbon source distinguished strain CCBAU 23380T from the type strains of related species. The genome size of CCBAU 23380T was 6.0 Mbp, comprising 5624 predicted genes with DNA G+C content of 62.4 mol%. Based on the results above, a novel species, Ensifer glycinis sp. nov., is proposed, with CCBAU 23380T (=LMG 29231T =HAMBI 3645T) as the type strain.

  13. CARACTERISATION PRELIMINAIRE DES RHIZOBIA ISOLEES A PARTIR DES NODOSITES DE CICER ARIETINUM L. CULTIVE DANS LE NORD-EST ALGERIEN

    Directory of Open Access Journals (Sweden)

    D. CHEKIREB

    2011-10-01

    Full Text Available Dans cette étude nous nous sommes intéressés au rhizobia nodulant Cicer arietinum L. (FLIP 90-13 cultivé dans la wilaya de Guelma (Algérie. Des quarante cinq isolats obtenus à partir de nodosités effectives de Cicer arietinum L, neuf ont nodulé leur plante hôte et deux de ces souches ont nodulé Medicago sp. Les dendrogrammes obtenus après analyse numérique des résultats des tests API 20NE et API 50CH montrent une diversité de ces rhizobia et permet d’apparenter la souche CAO08 au genre Sinorhizobium. D’autres caractères biochimiques comme la production d’acide ou d’alkali et la production du 3-céto-lactose ainsi que la résistance au stress salin, aux antibiotiques et à la variation de la température ont été testés. Les résultats obtenus indiquent que la quasi-totalité des souches acidifie le milieu YEMA en présence de BTB et qu’aucune d’entre elles ne produit le 3-céto-lactose. Toutes les souches testées peuvent croitre en présence de 2% de NaCl et sont capables de pousser à une température de 42°C. D’autre part, ces souches montrent une résistance à des concentrations de kanamycine (100µg/ml et de pénicilline G (60µg/ml.

  14. Cloning and Sequencing of the DNA Fragment Related to Salt Tolerance of Halomonas sp.C6%盐单胞菌C6与耐盐有关DNA片段的克隆和序列分析

    Institute of Scientific and Technical Information of China (English)

    王海洪; 樊振川; 曾静; 杨苏声

    1999-01-01

    提取盐单胞菌(Halomonas)C6的总DNA,用Sau3AⅠ部分酶切,回收15~30 kb的DNA片段,以pLAFR3为载体在大肠杆菌(E.coli)中构建了该菌的基因文库.以C6的基因文库为供体,以费氏中华根瘤菌(Sinorhizobium fredii)盐敏感菌株RC3-3r为受体,在辅助质粒pRK2013的协助下进行三亲本杂交,在选择培养基上筛选到接合子RWH15和RWH17.质粒检测表明,这两个接合子分别含有重组质粒pWH15和pWH17.通过三亲本杂交和功能互补的方法,对质粒pWH15进行两次亚克隆,最终得到3.2 kb的DNA片段.经检测,该片段能使RC3-3r恢复耐盐性.通过DNA测序及开放阅读框架(ORFs)分析,发现该片段上有4个开放阅读框架,其功能有待于进一步研究.

  15. Effect of plant growth promoting rhizobia on seed germination and seedling traits in Acacia senegal

    Directory of Open Access Journals (Sweden)

    S.K. Singh

    2011-11-01

    Full Text Available Among arid zone tree species, Acacia senegal and Prosopis cineraria are the most important dryland resources of Western Rajasthan desert ecosystem. Due to ecological, biological and molecular similarities, they are often studied together. The climatic conditions in this region restrict the build-up of soil organic matter and soils are generally deficient in nitrogen. Studies were carried out to isolate and molecularly characterize the diverse group of plant growth promoting rhizobacteria from root nodules of native A. senegal and P. cineraria and their effect on seed germination and seedling traits in two genotypes of A. senegal. The direct sequencing of 16S rDNA region resulted in molecular identification of plant growth promoting rhizobacteria as Bacillus licheniformis, Sinorhizobium saheli isolated from root nodules of A. senegal and S. kostiense and S. saheli isolated from root nodules of P. cineraria. The partial sequences of 16S rDNA were assigned Gen accession numbers HQ738496, HQ738499, HQ738506 and HQ738508. Scarification treatment with sulphuric acid (98% for 15 minutes was able to break the exogenous seed dormancy and enhanced germination percentage in control treatment to 90% and 92.5% in A. senegal in genotypes CAZRI 113AS and CAZRI 35AS, respectively. The treatments with Bacillus licheniformis or S. kostiense, either inoculated individually or as coinoculants, had positive effect on phenotypic traits of germination. Two A. senegal genotypes exhibited significant differences with regard to all the phenotypic traits. On the other hand, treatments with S. saheli isolated from either A. senegal or P. cineraria had negative effects on germination and related phenotypic traits. Values of the coeffivient of determination (R2 over 80% for root length versus shoot length, root/shoot ratio and seedling weight respectively validate that the observed attributes are inter-dependable and linear progression trend can be predicted.

  16. A Study on Co-Metabolic Biodegradation of Phenol and TCE by Mixed Culture PCL%高效降解菌剂PCL对苯酚及TCE的共代谢降解性能研究

    Institute of Scientific and Technical Information of China (English)

    张施阳

    2016-01-01

    从长期受氯代烃和酚类污染的土壤中驯化筛选得到能以苯酚为共代谢基质降解TCE的混合菌群PCL,其主要由博代氏杆菌(Bordetella sp.)和中华根瘤菌(Sinorhizobium sp.)组成.采用摇瓶振荡培养方法,研究了不同环境条件对混合菌群PCL共代谢苯酚和TCE效能的影响.结果表明,在30℃、pH 7.5条件下,200 mg/L苯酚能被PCL在4d内代谢完全,并且50 mg/L TCE在8d内的降解率能达到85.6%,以8mmol/L H2O2作为氧源,能使TCE降解率提高到91.18%.混合菌群PCL对苯酚和TCE的降解动力学可用Haldane方程来描述,利用非线性拟合求得各自对应的动力学参数k、km、ki分别为0.749/d、46.67 mg/L、386.16 mg/L和0.1332/d、31.15mg/L、0.044 5 mg/L,且拟合效果很好.

  17. Effect of plant growth promoting micro organisms on increasing water use efficiency of alfalfa in water-stress conditions

    Directory of Open Access Journals (Sweden)

    M. Zafari

    2015-11-01

    Full Text Available In order to study the effect of bacterial growth on water use efficiency of alfalfa, a greenhouse experiment, as factorial based on completely randomized blocks design with three replications, was conducted at Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran, in 2012. Treatments consisted of 3 levels of water stress (75, 55 and 35% of field capacity and seed inoculation at 4 levels (no inoculation (control, inoculation with mycorhhiza G. mosseae, inoculation with rhyzobium S. meliloti, and inoculation with combination of mycorhhiza and rhyzobium. Results showed that water stress and seed inoculation have significant effect (P&le0.01 on leaf nutrients content. Water stress reduced absorption of phosphorus (23%, potassium (8%, iron (4% and increased sodium absorption (14% in non-inoculated seeds. Inoculation of seeds reduced stress effects and combined inoculation had the highest effect. Stomatal conductance and water use efficiency were affected (P&le0.01 by inoculation and water stress. Stomatal conductance was decreased during the stress period and seed inoculation with mycorhhiza G. mosseae was most effective on increasing stomatal conductance (47% at the highest level of stress. Water use efficiency increased as a result of water stress and inoculation. The highest value of water use efficiency (0.166 mg/kg was obtained in the combined inoculation with 35% field capacity treatment. Results of regression equations showed that during the inoculation, contribution of phosphorus and potassium in regulation of stomatal conductance was increased and contribution of sodium was decreased.  However, during the stress period, the share of potassium and sodium was increased in stomatal conductance and the share of phosphorus was reduced. Also, stress increased the role of stomatal conductance in water use efficiency. However, inoculation reduced the role of stomatal conductance in water use efficiency.

  18. Auxins upregulate nif and fix genes.

    Science.gov (United States)

    Bianco, Carmen; Defez, Roberto

    2010-10-01

    In a recent publication we analyzed the global effects triggered by IAA overproduction in S. meliloti RD64 under free-living conditions by comparing the gene expression pattern of wild type 1021 with that of RD64 and 1021 treated with IAA and other four chemically or functionally related molecules. Among the genes differentially expressed in RD64 and IAA-treated 1021 cells we found two genes of pho operon, phoT and phoC. Based on this finding we examined the mechanisms for mineral P solubilization in RD64 and the potential ability of this strain to improve Medicago growth under P-starved conditions. Here, we further analyze the expression profiles obtained in microarray analysis and evaluate the specificity and the extent of overlap between all treatments. Venn diagrams indicated that IAA- and 2,4-D-regulated genes were closely related. Furthermore, most differentially expressed genes from pSymA were induced in 1021 cells treated with 2,4-D, ICA, IND and Trp as compared to the untreated 1021 cells. RT-PCR analysis was employed to analyze the differential expression patterns of nitrogen fixation genes under free-living and symbiotic conditions. Under symbiotic condition, the relative expression levels of nif and fix genes were significantly induced in Mt- RD64 plants and in Mt-1021 plants treated with IAA and 2,4-D whereas they were unchanged or repressed in Mt-1021 plants treated with the other selected compounds when compared to the untreated Mt-1021 plants.

  19. 3-Oxoacyl-[ACP] reductase from oilseed rape (Brassica napus).

    Science.gov (United States)

    Sheldon, P S; Kekwick, R G; Smith, C G; Sidebottom, C; Slabas, A R

    1992-04-01

    3-Oxoacyl-[ACP] reductase (E.C. 1.1.1.100, alternatively known as beta-ketoacyl-[ACP] reductase), a component of fatty acid synthetase has been purified from seeds of rape by ammonium sulphate fractionation, Procion Red H-E3B chromatography, FPLC gel filtration and high performance hydroxyapatite chromatography. The purified enzyme appears on SDS-PAGE as a number of 20-30 kDa components and has a strong tendency to exist in a dimeric form, particularly when dithiothreitol is not present to reduce disulphide bonds. Cleveland mapping and cross-reactivity with antiserum raised against avocado 3-oxoacyl-[ACP] reductase both indicate that the multiple components have similar primary structures. On gel filtration the enzyme appears to have a molecular mass of 120 kDa suggesting that the native structure is tetrameric. The enzyme has a strong preference for the acetoacetyl ester of acyl carrier protein (Km = 3 microM) over the corresponding esters of the model substrates N-acetyl cysteamine (Km = 35 mM) and CoA (Km = 261 microM). It is inactivated by dilution but this can be partly prevented by the inclusion of NADPH. Using an antiserum prepared against avocado 3-oxoacyl-[ACP] reductase, the enzyme has been visualised inside the plastids of rape embryo and leaf tissues by immunoelectron microscopy. Amino acid sequencing of two peptides prepared by digestion of the purified enzyme with trypsin showed strong similarities with 3-oxoacyl-[ACP] reductase from avocado pear and the Nod G gene product from Rhizobium meliloti.

  20. Root Exudates of Various Host Plants of Rhizobium leguminosarum Contain Different Sets of Inducers of Rhizobium Nodulation Genes.

    Science.gov (United States)

    Zaat, S A; Wijffelman, C A; Mulders, I H; van Brussel, A A; Lugtenberg, B J

    1988-04-01

    Rhizobium promoters involved in the formation of root nodules on leguminous plants are activated by flavonoids in plant root exudate. A series of Rhizobium strains which all contain the inducible Rhizobium leguminosarum nodA promoter fused to the Escherichia coli lacZ gene, and which differ only in the source of the regulatory nodD gene, were recently used to show that the regulatory nodD gene determines which flavonoids are able to activate the nodA promoter (HP Spaink, CA Wijffelman, E Pees, RJH Okker, BJJ Lugtenberg 1987 Nature 328: 337-340). Since these strains therefore are able to discriminate between various flavonoids, they were used to determine whether or not plants that are nodulated by R. leguminosarum produce different inducers. After chromatographic separation of root exudate constituents from Vicia sativa L. subsp. nigra (L.), V. hirsuta (L.) S.F. Gray, Pisum sativum L. cv Rondo, and Trifolium subterraneum L., the fractions were tested with a set of strains containing a nodD gene of R. leguminosarum, R. trifolii, or Rhizobium meliloti, respectively. It appeared that the source of nodD determined whether, and to what extent, the R. leguminosarum nodA promoter was induced. Lack of induction could not be attributed to the presence of inhibitors. Most of the inducers were able to activate the nodA promoter in the presence of one particular nodD gene only. The inducers that were active in the presence of the R. leguminosarum nodD gene were different in each root exudate.

  1. Sequence and molecular analysis of the nifL gene of Azotobacter vinelandii.

    Science.gov (United States)

    Blanco, G; Drummond, M; Woodley, P; Kennedy, C

    1993-08-01

    In both Klebsiella pneumoniae and Azotobacter vinelandii the nifL gene, which encodes a negative regulator of nitrogen fixation, lies immediately upstream of nifA. We have sequenced the A. vinelandii nifL gene and found that it is more homologous in its C-terminal domain to the histidine protein kinases (HPKs) than is K. pneumoniae NifL. In particular A. vinelandii NifL contains a conserved histidine at a position shown to be phosphorylated in other systems. Both NifL proteins are homologous in their N-termini to a part of the Halobacterium halobium bat gene product; Bat is involved in regulation of bacterio-opsin, the expression of which is oxygen sensitive. The same region showed homology to the haem-binding N-terminal domain of the Rhizobium meliloti fixL gene product, an oxygen-sensing protein. Like K. pneumoniae NifL, A. vinelandii NifL is shown here to prevent expression of nif genes in the presence of NH+4 or oxygen. The sequences found homologous in the C-terminal regions of NifL, FixL and Bat might therefore be involved in oxygen binding or sensing. An in-frame deletion mutation in the nifL coding region resulted in loss of repression by NH+4 and the mutant excreted high amounts of ammonia during nitrogen fixation, thus confirming a phenotype reported earlier for an insertion mutation. In addition, nifLA are cotranscribed in A. vinelandii as in K. pneumoniae, but expression from the A. vinelandii promoter requires neither RpoN nor NtrC. PMID:8231815

  2. Auxins upregulate nif and fix genes.

    Science.gov (United States)

    Bianco, Carmen; Defez, Roberto

    2010-10-01

    In a recent publication we analyzed the global effects triggered by IAA overproduction in S. meliloti RD64 under free-living conditions by comparing the gene expression pattern of wild type 1021 with that of RD64 and 1021 treated with IAA and other four chemically or functionally related molecules. Among the genes differentially expressed in RD64 and IAA-treated 1021 cells we found two genes of pho operon, phoT and phoC. Based on this finding we examined the mechanisms for mineral P solubilization in RD64 and the potential ability of this strain to improve Medicago growth under P-starved conditions. Here, we further analyze the expression profiles obtained in microarray analysis and evaluate the specificity and the extent of overlap between all treatments. Venn diagrams indicated that IAA- and 2,4-D-regulated genes were closely related. Furthermore, most differentially expressed genes from pSymA were induced in 1021 cells treated with 2,4-D, ICA, IND and Trp as compared to the untreated 1021 cells. RT-PCR analysis was employed to analyze the differential expression patterns of nitrogen fixation genes under free-living and symbiotic conditions. Under symbiotic condition, the relative expression levels of nif and fix genes were significantly induced in Mt- RD64 plants and in Mt-1021 plants treated with IAA and 2,4-D whereas they were unchanged or repressed in Mt-1021 plants treated with the other selected compounds when compared to the untreated Mt-1021 plants. PMID:20930554

  3. Microvirga lupini sp. nov., Microvirga lotononidis sp. nov. and Microvirga zambiensis sp. nov. are alphaproteobacterial root-nodule bacteria that specifically nodulate and fix nitrogen with geographically and taxonomically separate legume hosts.

    Science.gov (United States)

    Ardley, Julie K; Parker, Matthew A; De Meyer, Sofie E; Trengove, Robert D; O'Hara, Graham W; Reeve, Wayne G; Yates, Ron J; Dilworth, Michael J; Willems, Anne; Howieson, John G

    2012-11-01

    Strains of Gram-negative, rod-shaped, non-spore-forming bacteria were isolated from nitrogen-fixing nodules of the native legumes Listia angolensis (from Zambia) and Lupinus texensis (from Texas, USA). Phylogenetic analysis of the 16S rRNA gene showed that the novel strains belong to the genus Microvirga, with ≥ 96.1% sequence similarity with type strains of this genus. The closest relative of the representative strains Lut6(T) and WSM3557(T) was Microvirga flocculans TFB(T), with 97.6-98.0% similarity, while WSM3693(T) was most closely related to Microvirga aerilata 5420S-16(T), with 98.8% similarity. Analysis of the concatenated sequences of four housekeeping gene loci (dnaK, gyrB, recA and rpoB) and cellular fatty acid profiles confirmed the placement of Lut6(T), WSM3557(T) and WSM3693(T) within the genus Microvirga. DNA-DNA relatedness values, and physiological and biochemical tests allowed genotypic and phenotypic differentiation of Lut6(T), WSM3557(T) and WSM3693(T) from each other and from other Microvirga species with validly published names. The nodA sequence of Lut6(T) was placed in a clade that contained strains of Rhizobium, Mesorhizobium and Sinorhizobium, while the 100% identical nodA sequences of WSM3557(T) and WSM3693(T) clustered with Bradyrhizobium, Burkholderia and Methylobacterium strains. Concatenated sequences for nifD and nifH show that the sequences of Lut6(T), WSM3557(T) and WSM3693(T) were most closely related to that of Rhizobium etli CFN42(T) nifDH. On the basis of genotypic, phenotypic and DNA relatedness data, three novel species of Microvirga are proposed: Microvirga lupini sp. nov. (type strain Lut6(T) =LMG 26460(T) =HAMBI 3236(T)), Microvirga lotononidis sp. nov. (type strain WSM3557(T) =LMG 26455(T) =HAMBI 3237(T)) and Microvirga zambiensis sp. nov. (type strain WSM3693(T) =LMG 26454(T) =HAMBI 3238(T)). PMID:22199210

  4. Effect of plant growth promoting rhizobia on seed germination and seedling traits in Acacia senegal

    Directory of Open Access Journals (Sweden)

    Sunil Kumar Singh

    2013-12-01

    Full Text Available Among arid zone tree species, Acacia senegal and Prosopis cineraria are the most important dryland resources of Western Rajasthan desert ecosystem. Due to ecological, biological and molecular similarities, they are often studied together. The climatic conditions in this region restrict the build-up of soil organic matter and soils are generally deficient in nitrogen. Studies were carried out to isolate and molecularly characterize the diverse group of plant growth promoting rhizobacteria from root nodules of native A. senegal and P. cineraria and their effect on seed germination and seedling traits in two genotypes of A. senegal. The direct sequencing of 16S rDNA region resulted in molecular identification of plant growth promoting rhizobacteria as Bacillus licheniformis, Sinorhizobium saheli isolated from root nodules of A. senegal and S. kostiense and S. saheli isolated from root nodules of P. cineraria. The partial sequences of 16S rDNA were assigned Gen accession numbers HQ738496, HQ738499, HQ738506 and HQ738508. Scarification treatment with sulphuric acid (98% for 15 minutes was able to break the exogenous seed dormancy and enhanced germination percentage in control treatment to 90% and 92.5% in A. senegal in genotypes CAZRI 11113AS and CAZRI 35AS, respectively. The treatments with Bacillus licheniformis or S. kostiense, either inoculated individually or as coinoculants, had positive effect on phenotypic traits of germination. Two A. senegal genotypes exhibited significant differences with regard to all the phenotypic traits. On the other hand, treatments with S. saheli isolated from either A. senegal or P. cineraria had negative effects on germination and related phenotypic traits. Values of the coeffivient of determination (R2 over 80% for root length versus shoot length, root/shoot ratio and seedling weight respectively validate that the observed attributes are inter-dependable and linear progression trend can be

  5. Analysis of stress resistance and phylogenesis of rhizobia isolated from Caragsana spp.%荒漠植物柠条根瘤菌的抗逆性及其系统发育分析

    Institute of Scientific and Technical Information of China (English)

    代金霞; 王玉炯; 郭晶静; 武雪娟; 张晓玲

    2011-01-01

    对分离自宁夏白芨滩国家级自然保护区的62株柠条根瘤菌的耐盐性、耐酸碱性、生长温度范围和抗生素抗性进行了测定,并采用16S rDNA PCR-RFLP和16S rDNA全序列分析方法对其进行了遗传多样性及系统发育分析.结果表明:柠条根瘤菌具有较强的耐受高温和耐盐、耐酸碱能力,抗逆性较强,但仍存在菌株间差异.70.8%的菌株可耐受4%的NaCl,全部菌株对100 μg/mL的青霉素具有抗性,均可在pH5.0~11.0的培养基中生长.经16S rDNA序列测定鉴定为豌豆根瘤菌的CNXE4、CNXLW24和中华根瘤菌的CNXC3,这3株菌在pH4.0~12.0的范围内都可生长,可以耐受6%的NaCl、50μg/mL浓度的卡那霉素和链霉素、300 μg/mL的青霉素,在60℃热激后可生长,表现出极强的抗逆性.16S rDNA PCR-RFLP将62株菌分为7种图谱类型,16S rDNA全序列分析的结果表明,它们分别归属于根瘤菌属、中华根瘤菌属、土壤杆菌属、叶杆菌属和慢生根瘤菌属,遗传多样性较为丰富.%Several biochemical characteristics of 62 rhizobia isolated from Caragsana spp . In Baijitan National Na-ture Reserve in Ninaxia were detected , including resistance to salt , acid-alkali, temperature variation and intrinsic an tibiotics . The results indicated that 70 .8% strains could tolerate NaCl stress at 4% concentration . All of the strains could tolerate Amp stress at 100 μg/mL as well as could grow at a range of pH from 5~11. Strains CNXE4, CNXLW24 and CNXC3 had high resistance , they could grow at pH from 4~ 12 ,could tolerate 6% concentration NaCl, 50 μg/mL KM and SM , 300 μg/mL AMP , and could grow after being exposed to 60°C for 10 min . The methods of 16S rDNA PCR RFLP and 16S rDNA sequencing were used to analysis the diversity and phylogenesis of 62 rhizobia strains . The results revealed that all the tested strains belonged to the genus of Rhizobium , Sinorhizobium , Agrobacterium , Phyllobacterium and Brady rhizobium . The

  6. 臭氧-接种生物滤池组合工艺去除饮用水中典型致嗅物质%Ozonation-Inoculated Biofiltration for Removal of the Typical Taste and Odor Compounds in Drinking Water

    Institute of Scientific and Technical Information of China (English)

    袁蓉芳; 周北海; 施春红; 顾军农; 李玉仙

    2013-01-01

    通过在生物滤池表面接种MIB(2-甲基异茨醇)及geosmin(土臭素)降解菌,增强生物滤池的作用,并探讨臭氧-生物滤池组合工艺对MIB和geosmin的处理效果.结果表明:单独接种生物滤池可使ρ(MIB)和ρ(geosmin)从初始的500 ng/L分别降至125和112 ng/L,MIB和geosmin的去除效果先随EBCT(空床停留时间)的延长而显著增加,但当EBCT大于20 min后无明显变化;随着滤料深度的增加,滤池生物量逐渐降低,对污染物的去除率增加缓慢.在接种生物滤池前增加臭氧单元,当EBCT为20min、臭氧投加量为2 mg/L时,臭氧-接种生物滤池组合工艺可去除84%的MIB和94%的geosmin,其中接种生物滤池单元中生物量随滤池深度的增加呈先增后减的趋势,滤料深度为100~200 mm时,单位高度滤料的去除率最高.采用臭氧-接种生物滤池组合工艺可有效去除水中的MIB和geosmin.%2-methylisoborneol (MIB) and geosmin are two of the most common taste and odor compounds which can not be readily removed by conventional drinking water treatment processes.The objective of this study was to enhance the biofiltration of MIB and geosmin by inoculating the sand filter with MIB and geosmin degraders,and to examine the removal efficiencies of MIB and geosmin using inoculated filter and non-inoculated filter in the presence and absence of ozonation.The degraders were Micrococcus sp.,Flavobacterium sp.,Brevibacterium sp.and Pseudomonas sp.as MIB degraders,and Chryseobacterium sp.,Sinorhizobium sp.,and Stenotrophomonas sp.as geosmin degraders.These bacteria were isolated from granular activated carbon in a commercial water plant.For the filter inoculated with a consortium of MIB and geosmin biodegraders,MIB and geosmin contents were decreased to 125 and 112 ng/L from 500 ng/L,respectively.Initially,removal efficiencies of MIB and geosmin rapidly increased with the extension of empty bed contact time (EBCT),and resulted in no obvious change when EBCT

  7. Resposta do Feijoeiro Comum à Inoculação com Rizóbio e Suplementação com Nitrogênio Mineral em Dois Biomas Brasileiros

    Directory of Open Access Journals (Sweden)

    Luciana Fernandes de Brito

    2015-08-01

    Full Text Available O feijoeiro comum (Phaseolus vulgaris L. pode se beneficiar da fixação biológica de N2, mas respostas inconsistentes da cultura à inoculação com rizóbio indicam a necessidade de aplicação de N mineral complementar. Este trabalho teve por objetivo avaliar a resposta do feijoeiro à inoculação com rizóbio, associada à suplementação com N mineral, nos biomas Cerrado e Mata Atlântica. Foram conduzidos quatro experimentos de campo, dois em Santo Antônio de Goiás, GO, um em Valença, RJ e um em Macaé, RJ, onde a inoculação com estirpes comerciais de rizóbio foi comparada à inoculação com a estirpe BR 923 de Sinorhizobium sp., à adubação com N mineral e à suplementação com N na semeadura e em cobertura. A avaliação da população nativa de rizóbio indicou 105 células g-1 no solo na área experimental em Goiás, anteriormente cultivada com feijão, e 102células g-1 em Valença, anteriormente mantida com pastagem. Nos dois experimentos em Goiás, o rendimento de grãos, da ordem de 2.100 kg ha-1, não diferiu entre os tratamentos testemunha absoluta, inoculação com rizóbio ou aplicação de 120 kg ha-1 de N. Em Valença, a inoculação com estirpes comerciais forneceu rendimentos da cultivar Ouro Negro superiores à testemunha absoluta, na ausência de adubação de cobertura; na presença de 40 kg ha-1 de N em cobertura, a inoculação com rizóbio proporcionou rendimento de 3.420 kg ha-1, superior aos demais tratamentos. Na média das diferentes fontes de N na semeadura, a adubação de cobertura aumentou a produção de grãos de 2.367 para 2.542 kg ha-1. Em Macaé, em solo com alto teor de matéria orgânica, os maiores rendimentos foram obtidos com inoculação das estirpes comerciais associada a 40 kg ha-1 de N em cobertura, com efeito deletério da adubação de 80 kg ha-1 de N no plantio. Concluiu-se que em áreas sem cultivo prévio de feijão, a inoculação com estirpes comerciais de rizóbio aumentou o