scholarly journals Expression and Functional Roles of Bradyrhizobium japonicum Genes Involved in the Utilization of Inorganic and Organic Sulfur Compounds in Free-Living and Symbiotic Conditions

2011 ◽  
Vol 24 (4) ◽  
pp. 451-457 ◽  
Author(s):  
Masayuki Sugawara ◽  
Gopit R. Shah ◽  
Michael J. Sadowsky ◽  
Oleg Paliy ◽  
Justin Speck ◽  
...  
Keyword(s):  
Bradyrhizobium Japonicum ◽  
Gene Clusters ◽  
Sulfur Source ◽  
Organosulfur Compounds ◽  
In Planta ◽  
Growth And Survival ◽  
Functional Roles ◽  
Organic Sulfur Compounds ◽  
Sulfatase Activity ◽  
Bradyrhizobium Spp

Strains of Bradyrhizobium spp. form nitrogen-fixing symbioses with many legumes, including soybean. Although inorganic sulfur is preferred by bacteria in laboratory conditions, sulfur in agricultural soil is mainly present as sulfonates and sulfur esters. Here, we show that Bradyrhizobium japonicum and B. elkanii strains were able to utilize sulfate, cysteine, sulfonates, and sulfur-ester compounds as sole sulfur sources for growth. Expression and functional analysis revealed that two sets of gene clusters (bll6449 to bll6455 or bll7007 to bll7011) are important for utilization of sulfonates sulfur source. The bll6451 or bll7010 genes are also expressed in the symbiotic nodules. However, B. japonicum mutants defective in either of the sulfonate utilization operons were not affected for symbiosis with soybean, indicating the functional redundancy or availability of other sulfur sources in planta. In accordance, B. japonicum bacteroids possessed significant sulfatase activity. These results indicate that strains of Bradyrhizobium spp. likely use organosulfur compounds for growth and survival in soils, as well as for legume nodulation and nitrogen fixation.

Strain distribution and in planta production of an extracellular polysaccharide depolymerase from Bradyrhizobium japonicum

1992 ◽  
Vol 38 (8) ◽  
pp. 857-861 ◽  
Author(s):  
Michael F. Dunn ◽  
Arthur L. Karr
Keyword(s):  
Bradyrhizobium Japonicum ◽  
Root Nodules ◽  
Extracellular Polysaccharide ◽  
Extracellular Polysaccharides ◽  
Neutral Sugar ◽  
In Planta ◽  
In Vitro Production ◽  
Polysaccharide Composition ◽  
Vitro Production

Thirty-four strains of Bradyrhizobium japonicum were screened for the in vitro production of an extracellular polysaccharide depolymerase active against the B. japonicum acidic extracellular polysaccharide that contains mannose, glucose, galactose, and 4-O-methylgalactose as neutral sugar components. Over 90% of tested strains producing this type of extracellular polysaccharide also produced the extracellular polysaccharide depolymerase, whereas strains producing a compositionally different extracellular polysaccharide did not. In addition, representatives of species related to B. japonicum by extracellular polysaccharide composition or host range were also phenotypically depolymerase negative. Depolymerase was also present in soybean root nodules formed by B. japonicum strain 2143. In contrast to the cell-associated depolymerase activity found in free-living cells of this strain, most of the depolymerase activity present in nodules is free of the bacteroids. The widespread occurrence of the depolymerase among B. japonicum strains and the spatiotemporal distribution of its activity in planta are consistent with the enzyme playing a role in the removal of surface extracellular polysaccharide from the microorganism during the infection of nodulation process. Key words: Bradyrhizobium japonicum, soybean, extracellular polysaccharides, extracellular polysaccharide depolymerase, bacteroids.

The complete denitrification pathway of the symbiotic, nitrogen-fixing bacterium Bradyrhizobium japonicum

2005 ◽  
Vol 33 (1) ◽  
pp. 141-144 ◽  
Author(s):  
E.J. Bedmar ◽  
E.F. Robles ◽  
M.J. Delgado
Keyword(s):  
Nitric Oxide ◽  
Nitrous Oxide ◽  
Bradyrhizobium Japonicum ◽  
Mutational Analysis ◽  
Control Level ◽  
Gene Clusters ◽  
Alternative Form ◽  
Nitrate Respiration ◽  
Regulatory Cascade ◽  
Denitrification Genes

Denitrification is an alternative form of respiration in which bacteria sequentially reduce nitrate or nitrite to nitrogen gas by the intermediates nitric oxide and nitrous oxide when oxygen concentrations are limiting. In Bradyrhizobium japonicum, the N2-fixing microsymbiont of soya beans, denitrification depends on the napEDABC, nirK, norCBQD, and nosRZDFYLX gene clusters encoding nitrate-, nitrite-, nitric oxide- and nitrous oxide-reductase respectively. Mutational analysis of the B. japonicum nap genes has demonstrated that the periplasmic nitrate reductase is the only enzyme responsible for nitrate respiration in this bacterium. Regulatory studies using transcriptional lacZ fusions to the nirK, norCBQD and nosRZDFYLX promoter region indicated that microaerobic induction of these promoters is dependent on the fixLJ and fixK2 genes whose products form the FixLJ–FixK2 regulatory cascade. Besides FixK2, another protein, nitrite and nitric oxide respiratory regulator, has been shown to be required for N-oxide regulation of the B. japonicum nirK and norCBQD genes. Thus nitrite and nitric oxide respiratory regulator adds to the FixLJ–FixK2 cascade an additional control level which integrates the N-oxide signal that is critical for maximal induction of the B. japonicum denitrification genes. However, the identity of the signalling molecule and the sensing mechanism remains unknown.

scholarly journals Methanosarcina acetivorans contains a functional ISC system for iron-sulfur cluster biogenesis

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Thomas M. Deere ◽  
Divya Prakash ◽  
Faith H. Lessner ◽  
Evert C. Duin ◽  
Daniel J. Lessner
Keyword(s):  
Parent Strain ◽  
Biochemical Characterization ◽  
Sulfur Metabolism ◽  
Gene Clusters ◽  
Sulfur Source ◽  
Methanosarcina Acetivorans ◽  
Cysteine Desulfurase ◽  
The Core ◽  
Iron Sulfur ◽  
Core Components

Abstract Background The production of methane by methanogens is dependent on numerous iron-sulfur (Fe-S) cluster proteins; yet, the machinery involved in Fe-S cluster biogenesis in methanogens remains largely unknown. Methanogen genomes encode uncharacterized homologs of the core components of the ISC (IscS and IscU) and SUF (SufBC) Fe-S cluster biogenesis systems found in bacteria and eukaryotes. Methanosarcina acetivorans contains three iscSU and two sufCB gene clusters. Here, we report genetic and biochemical characterization of M. acetivorans iscSU2. Results Purified IscS2 exhibited pyridoxal 5′- phosphate-dependent release of sulfur from L-cysteine. Incubation of purified IscU2 with IscS2, cysteine, and iron (Fe2+) resulted in the formation of [4Fe-4S] clusters in IscU2. IscU2 transferred a [4Fe-4S] cluster to purified M. acetivorans apo-aconitase. IscU2 also restored the aconitase activity in air-exposed M. acetivorans cell lysate. These biochemical results demonstrate that IscS2 is a cysteine desulfurase and that IscU2 is a Fe-S cluster scaffold. M. acetivorans strain DJL60 deleted of iscSU2 was generated to ascertain the in vivo importance of IscSU2. Strain DJL60 had Fe-S cluster content and growth similar to the parent strain but lower cysteine desulfurase activity. Strain DJL60 also had lower intracellular persulfide content compared to the parent strain when cysteine was an exogenous sulfur source, linking IscSU2 to sulfur metabolism. Conclusions This study establishes that M. acetivorans contains functional IscS and IscU, the core components of the ISC Fe-S cluster biogenesis system and provides the first evidence that ISC operates in methanogens.

The 73-kb pIAA plasmid increases competitive fitness of Pseudomonas syringae subspecies savastanoi in oleander

1993 ◽  
Vol 39 (7) ◽  
pp. 659-664 ◽  
Author(s):  
Sara E. Silverstone ◽  
David G. Gilchrist ◽  
Richard M. Bostock ◽  
Tsune Kosuge
Keyword(s):  
Population Density ◽  
Pseudomonas Syringae ◽  
Indoleacetic Acid ◽  
Leaf Tissue ◽  
Insertion Mutant ◽  
Wild Type ◽  
In Planta ◽  
Bacterial Strains ◽  
Growth And Survival ◽  
Iaa Production

Pseudomonas syringae subsp. savastanoi causes tumors on olive and oleander by producing the plant growth regulators indoleacetic acid (IAA) and cytokinins following infection of the plant. The contribution of IAA production to the ability of P. syringae subsp. savastanoi to grow and survive in oleander leaf tissue was studied. Bacterial strains differing only with respect to IAA production were characterized. Growth and survival of wild-type and two mutant strains of P. syringae subsp. savastanoi in oleander leaf tissue were monitored by weekly colony counts and IAA plate assays. Growth rate of the three strains in culture and in planta did not differ significantly. However, the wild-type strain reached a higher population density and maintained its maximum density at least 9 weeks longer than either mutant population. An insertion mutant containing the IAA plasmid (pIAA), but incapable of IAA production, did not maintain a higher population density than a strain cured of the IAA plasmid. The pIAA-cured strain maintained a higher population density when coinoculated with an IAA-producing strain than when inoculated alone. These results suggest that IAA production may contribute to the fitness of P. syringae subsp. savastanoi in oleander tissue and that the iaa operon alone may be responsible for the competitive advantage of cells harboring pIAA.Key words: indoleacetic acid, bacterial ecology.

scholarly journals A Role for Bradyrhizobium japonicum ECF16 Sigma Factor EcfS in the Formation of a Functional Symbiosis with Soybean

2012 ◽  
Vol 25 (1) ◽  
pp. 119-128 ◽  
Author(s):  
S. B. Stockwell ◽  
L. Reutimann ◽  
M. L. Guerinot
Keyword(s):  
Stress Responses ◽  
Sigma Factor ◽  
Bradyrhizobium Japonicum ◽  
Critical Role ◽  
Negative Regulator ◽  
In Planta ◽  
Cellular Processes ◽  
Core Enzyme ◽  
Σ Factor ◽  
Target Promoters

Alternative sigma (σ) factors, proteins that recruit RNA polymerase core enzyme to target promoters, are one mechanism by which bacteria transcriptionally regulate groups of genes in response to environmental stimuli. A class of σ70 proteins, termed extracytoplasmic function (ECF) σ factors, are involved in cellular processes such as bacterial stress responses and virulence. Here, we describe an ECF16 σ factor, EcfS (Blr4928) from the gram-negative soil bacterium Bradyrhizobium japonicum USDA110, that plays a critical role in the establishment of a functional symbiosis with soybean. Nonpolar insertional mutants of ecfS form immature nodules that do not fix nitrogen, a defect that can be successfully complemented by expression of ecfS. Overexpression of the cocistronic gene, tmrS (blr4929), phenocopies the ecfS mutant in planta and, therefore, we propose that TmrS is a negative regulator of EcfS, a determination consistent with the prediction that it encodes an anti-σ factor. Microarray analysis of the ecfS mutant and tmrS overexpressor was used to identify 40 transcripts misregulated in both strains. These transcripts primarily encode proteins of unknown and transport-related functions and may provide insights into the symbiotic defect in these strains.

Plant natural products: a primerThe present review is one in the special series of reviews on animal–plant interactions.

2010 ◽  
Vol 88 (7) ◽  
pp. 601-614 ◽  
Author(s):  
M. A. Bernards
Keyword(s):  
Natural Products ◽  
Structural Diversity ◽  
Structural Type ◽  
Biological Functions ◽  
Plant Interactions ◽  
In Planta ◽  
Functional Roles ◽  
Chemical Mechanisms ◽  
Know How ◽  
Plant Natural Products

Over the course of evolution, plants have adapted various structural and chemical mechanisms to protect themselves and interact with their environment. The chemical mechanisms are largely based on the secondary metabolites or natural products. Although plant natural products are generally divided into three main categories (terpenoids, alkaloids, and phenylpropanoids) that are based on structural type and biosynthetic origin, there are many other smaller categories of unique compounds. Many important in planta biological functions can be attributed to plant natural products, in large part, owing to their tremendous structural diversity. To understand the functional roles of plant natural products, both as protective compounds and interorganismal signals, it is important to know how they are formed in plants. This minireview provides a general background about the three main categories of plant natural products, their biosynthetic origins, and their structural diversity.

scholarly journals Loss of the nodule-specific cysteine rich peptide, NCR169, abolishes symbiotic nitrogen fixation in the Medicago truncatula dnf7 mutant

2015 ◽  
Vol 112 (49) ◽  
pp. 15232-15237 ◽  
Author(s):  
Beatrix Horváth ◽  
Ágota Domonkos ◽  
Attila Kereszt ◽  
Attila Szűcs ◽  
Edit Ábrahám ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Medicago Truncatula ◽  
Symbiotic Nitrogen Fixation ◽  
Root Nodules ◽  
Nitrogen Fixing ◽  
In Planta ◽  
Functional Roles ◽  
Absolute Requirement ◽  
Cell Layers

Host compatible rhizobia induce the formation of legume root nodules, symbiotic organs within which intracellular bacteria are present in plant-derived membrane compartments termed symbiosomes. In Medicago truncatula nodules, the Sinorhizobium microsymbionts undergo an irreversible differentiation process leading to the development of elongated polyploid noncultivable nitrogen fixing bacteroids that convert atmospheric dinitrogen into ammonia. This terminal differentiation is directed by the host plant and involves hundreds of nodule specific cysteine-rich peptides (NCRs). Except for certain in vitro activities of cationic peptides, the functional roles of individual NCR peptides in planta are not known. In this study, we demonstrate that the inability of M. truncatula dnf7 mutants to fix nitrogen is due to inactivation of a single NCR peptide, NCR169. In the absence of NCR169, bacterial differentiation was impaired and was associated with early senescence of the symbiotic cells. Introduction of the NCR169 gene into the dnf7-2/NCR169 deletion mutant restored symbiotic nitrogen fixation. Replacement of any of the cysteine residues in the NCR169 peptide with serine rendered it incapable of complementation, demonstrating an absolute requirement for all cysteines in planta. NCR169 was induced in the cell layers in which bacteroid elongation was most pronounced, and high expression persisted throughout the nitrogen-fixing nodule zone. Our results provide evidence for an essential role of NCR169 in the differentiation and persistence of nitrogen fixing bacteroids in M. truncatula.

scholarly journals Rhodococcus opacus expresses the xsc gene to utilize taurine as a carbon source or as a nitrogen source but not as a sulfur source

Microbiology ◽  
2004 ◽  
Vol 150 (6) ◽  
pp. 1859-1867 ◽  
Author(s):  
Karin Denger ◽  
Jürgen Ruff ◽  
David Schleheck ◽  
Alasdair M. Cook
Keyword(s):  
Gene Cluster ◽  
Gene Clusters ◽  
Pcr Primers ◽  
Sole Source ◽  
Amino Acid Sequences ◽  
Rhodococcus Opacus ◽  
Sulfur Source ◽  
Gram Positive Bacteria ◽  
Sulfoacetaldehyde Acetyltransferase ◽  
Different Levels

The Gram-positive bacteria Rhodococcus opacus ISO-5 and Rhodococcus sp. RHA1 utilized taurine (2-aminoethanesulfonate) as the sole source of carbon or of nitrogen or of sulfur for growth. Different gene clusters and enzymes were active under these different metabolic situations. Under carbon- or nitrogen-limited conditions three enzymes were induced, though to different levels: taurine-pyruvate aminotransferase (Tpa), alanine dehydrogenase (Ald) and sulfoacetaldehyde acetyltransferase (Xsc). The specific activities of these enzymes in R. opacus ISO-5 were sufficient to explain the growth rates under the different conditions. These three enzymes were purified and characterized, and the nature of each reaction was confirmed. Analyses of the genome of Rhodococcus sp. RHA1 revealed a gene cluster, tauR-ald-tpa, putatively encoding regulation and oxidation of taurine, located 20 kbp from the xsc gene and separate from two candidate phosphotransacetylase (pta) genes, as well as many candidate ABC transporters (tauBC). PCR primers allowed the amplification and sequencing of the tauR-ald-tpa gene cluster and the xsc gene in R. opacus ISO-5. The N-terminal sequences of the three tested proteins matched the derived amino acid sequences of the corresponding genes. The sequences of the four genes found in each Rhodococcus strain shared high degrees of identity (>95 % identical positions). RT-PCR studies proved transcription of the xsc gene when taurine was the source of carbon or of nitrogen. Under sulfur-limited conditions no xsc mRNA was generated and no Xsc was detected. Taurine dioxygenase (TauD), the enzyme catalysing the anticipated desulfonative reaction when taurine sulfur is assimilated, was presumed to be present because oxygen-dependent taurine disappearance was demonstrated with taurine-grown cells only. A putative tauD gene (with three other candidates) was detected in strain ISO-5. Regulation of the different forms of metabolism of taurine remains to be elucidated.

scholarly journals Characterisation of a novel endophyte NRPS gene and its role in endophyte-grass symbioses

2007 ◽  
Vol 13 ◽  
pp. 491-493
Author(s):  
H. Harzer ◽  
R.D. Johnson ◽  
S. Rasmussen ◽  
C.R. Voisey ◽  
L.J. Johnson
Keyword(s):  
Fungal Endophytes ◽  
Gene Clusters ◽  
Gene Replacement ◽  
In Planta ◽  
Dna Library ◽  
Peptide Synthetase ◽  
Genomic Dna Library ◽  
Targeted Gene Replacement ◽  
Fungal Secondary Metabolite

Symbiotic grass associations with fungal endophytes (genera Neotyphodium and Epichloë) display enhanced fitness as well as prolonged field persistence over their endophyte free equivalents. Perennial ryegrass, an important agronomic grass, is typically associated with the N. lolii endophyte. The endophyte lives within the intercellular spaces without inducing any symptoms in the plant. The aim of this study is to elucidate the biosynthetic function of fungal secondary metabolite gene clusters. Non-ribosomal peptide synthetase genes (NRPSs) of unknown function were targeted, as these genes are commonly associated with the production of bioactive peptides some of which are ecologically important. Some novel endophyte NRPS genes have been identified using a degenerate PCR screen; one of these, NRPS5 will be discussed here. Clones were obtained by screening a fosmid Epichloë festucae genomic DNA library and we are currently determining gene function by using targeted gene replacement followed by an assessment in vitro and in planta using metabolomics and appropriate bioassay screens. Keywords: endophyte, NRPS, secondary metabolism

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