Proteomics reveals differential expression of proteins related to a variety of metabolic pathways by genistein-induced Bradyrhizobium japonicum strains

Batista, Jesiane Stefânia da Silva; Hungría, Mariangela

doi:10.1016/j.jprot.2011.10.032

Cited by 30 publications

(24 citation statements)

References 37 publications

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“…Notably, all the flagellar assembly proteins except NGR_c28210 (the second copy of MotB) were down-regulated in bacteroids. This is in contrast to the induction of flagellar proteins in Bradyrhizobium japonicum by genistein application and the requirement of flagella in biofilm formation and competitive nodulation of S. meliloti [42], [43], [44], [45]. However, it was also reported that nonflagellated mutants of S. meliloti formed normal nitrogen-fixing nodules on alfalfa [46], suggesting that flagella are dispensable for function of bacteroids.…”

Section: Resultsmentioning

confidence: 89%

High-Resolution Transcriptomic Analyses of Sinorhizobium sp. NGR234 Bacteroids in Determinate Nodules of Vigna unguiculata and Indeterminate Nodules of Leucaena leucocephala

Tian

Chen

et al. 2013

PLoS ONE

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The rhizobium-legume symbiosis is a model system for studying mutualistic interactions between bacteria and eukaryotes. Sinorhizobium sp. NGR234 is distinguished by its ability to form either indeterminate nodules or determinate nodules with diverse legumes. Here, we presented a high-resolution RNA-seq transcriptomic analysis of NGR234 bacteroids in indeterminate nodules of Leucaena leucocephala and determinate nodules of Vigna unguiculata. In contrast to exponentially growing free-living bacteria, non-growing bacteroids from both legumes recruited several common cellular functions such as cbb3 oxidase, thiamine biosynthesis, nitrate reduction pathway (NO-producing), succinate metabolism, PHB (poly-3-hydroxybutyrate) biosynthesis and phosphate/phosphonate transporters. However, different transcription profiles between bacteroids from two legumes were also uncovered for genes involved in the biosynthesis of exopolysaccharides, lipopolysaccharides, T3SS (type three secretion system) and effector proteins, cytochrome bd ubiquinol oxidase, PQQ (pyrroloquinoline quinone), cytochrome c550, pseudoazurin, biotin, phasins and glycolate oxidase, and in the metabolism of glutamate and phenylalanine. Noteworthy were the distinct expression patterns of genes encoding phasins, which are thought to be involved in regulating the surface/volume ratio of PHB granules. These patterns are in good agreement with the observed granule size difference between bacteroids from L. leucocephala and V. unguiculata.

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Section: Resultsmentioning

confidence: 89%

High-Resolution Transcriptomic Analyses of Sinorhizobium sp. NGR234 Bacteroids in Determinate Nodules of Vigna unguiculata and Indeterminate Nodules of Leucaena leucocephala

Tian

Chen

et al. 2013

PLoS ONE

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“…In this case, thioredoxin transfers reducing power to damaged proteins, restoring their reduced state [71]. Finally, thioredoxin was synthesized under high-temperature conditions, confirming its induction as a general response to stress [72]; it is also induced in the early stages of symbiotic interaction in B. japonicum [73] and in the plant interaction with G. diazotrophicus [74]. …”

Section: Resultsmentioning

confidence: 98%

Proteomic profiling of Rhizobium tropiciPRF 81: identification of conserved and specific responses to heat stress

et al. 2012

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BackgroundRhizobium tropici strain PRF 81 (= SEMIA 4080) has been used in commercial inoculants for application to common-bean crops in Brazil since 1998, due to its high efficiency in fixing nitrogen, competitiveness against indigenous rhizobial populations and capacity to adapt to stressful tropical conditions, representing a key alternative to application of N-fertilizers. The objective of our study was to obtain an overview of adaptive responses to heat stress of strain PRF 81, by analyzing differentially expressed proteins when the bacterium is grown at 28°C and 35°C.ResultsTwo-dimensional gel electrophoresis (2DE) revealed up-regulation of fifty-nine spots that were identified by MALDI-TOF/TOF-TOF. Differentially expressed proteins were associated with the functional COG categories of metabolism, cellular processes and signaling, information storage and processing. Among the up-regulated proteins, we found some related to conserved heat responses, such as molecular chaperones DnaK and GroEL, and other related proteins, such as translation factors EF-Tu, EF-G, EF-Ts and IF2. Interestingly, several oxidative stress-responsive proteins were also up-regulated, and these results reveal the diversity of adaptation mechanisms presented by this thermotolerant strain, suggesting a cross-talk between heat and oxidative stresses.ConclusionsOur data provide valuable protein-expression information relevant to the ongoing genome sequencing of strain PRF 81, and contributes to our still-poor knowledge of the molecular determinants of the thermotolerance exhibited by R. tropici species.

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“…IPG strips (pH 4–7, 13 cm, GE Healthcare, Uppsala, Sweden) were rehydrated overnight with aliquots of 350 μg of solubilized proteins. Next, the strips were submitted to isoelectric focalization and SDS-PAGE as described by Batista and Hungria [72]. Gels were stained overnight with Comassie Brilliant Blue (CBB) R-350 (GE Healthcare), destained in a solution of 40% ethanol and 10% acetic acid and scanned (ImageScanner LabScan v. 5.0).…”

Section: Methodsmentioning

confidence: 99%

“…Spots which showed a significantly higher volume in the inoculated condition were excised and processed as described before [72], with trypsin (Gold, mass spectrometry grade, Promega, Madison, WI) at 37°C overnight. Tryptic peptides (0.5 μL) were mixed with saturated solution of HCCA (α-cyano-4-hydroxy-cinamic acid) in 50% acetonitrile, 0.1% TFA (trifluoroacetic acid).…”

Section: Methodsmentioning

confidence: 99%

Transcriptional analysis of genes involved in nodulation in soybean roots inoculated with Bradyrhizobium japonicumstrain CPAC 15

et al. 2013

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BackgroundBiological nitrogen fixation in root nodules is a process of great importance to crops of soybean [Glycine max (L.) Merr.], as it may provide the bulk of the plant’s needs for nitrogen. Legume nodulation involves several complex steps and, although studied for many decades, much remains to be understood.ResultsThis research aimed at analyzing the global expression of genes in soybean roots of a Brazilian cultivar (Conquista) inoculated with Bradyrhizobium japonicum CPAC 15, a strain broadly used in commercial inoculants in Brazil. To achieve this, we used the suppressive subtractive hybridization (SSH) technique combined with Illumina sequencing. The subtractive library (non-inoculated x inoculated) of soybean roots resulted in 3,210 differentially expressed transcripts at 10 days after inoculation were studied. The data were grouped according to the ontologies of the molecular functions and biological processes. Several classes of genes were confirmed as related to N2 fixation and others were reported for the first time.ConclusionsDuring nodule formation, a higher percentage of genes were related to primary metabolism, cell-wall modifications and the antioxidant defense system. Putative symbiotic functions were attributed to some of these genes for the first time.

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Proteomics reveals differential expression of proteins related to a variety of metabolic pathways by genistein-induced Bradyrhizobium japonicum strains

Cited by 30 publications

References 37 publications

High-Resolution Transcriptomic Analyses of Sinorhizobium sp. NGR234 Bacteroids in Determinate Nodules of Vigna unguiculata and Indeterminate Nodules of Leucaena leucocephala

High-Resolution Transcriptomic Analyses of Sinorhizobium sp. NGR234 Bacteroids in Determinate Nodules of Vigna unguiculata and Indeterminate Nodules of Leucaena leucocephala

Proteomic profiling of Rhizobium tropiciPRF 81: identification of conserved and specific responses to heat stress

Transcriptional analysis of genes involved in nodulation in soybean roots inoculated with Bradyrhizobium japonicumstrain CPAC 15

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