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Quantitative proteomics analysis reveals the tolerance of wheat to salt stress in response to Enterobacter cloacae SBP-8.

ABSTRACT: Salinity stress adversely affects the plant growth and is a major constraint to agriculture. In the present study, we studied the role of plant growth promoting rhizobacterium (PGPR) Enterobacter cloacae SBP-8 possessing ACC deaminase activity on proteome profile of wheat (Triticum aestivum L.) under high salinity (200 mM NaCl) stress. The aim of study was to investigate the differential expressed protein in selected three (T-1, T-2, T-3) treatments and absolute quantification (MS/MS analysis) was used to detect statistically significant expressed proteins. In this study, we investigated the adaptation mechanisms of wheat seedlings exposed to high concentration of NaCl treatment (200 mM) for 15 days in response to bacterial inoculation based on proteomic data. The identified proteins were distributed in different cellular, biological and molecular functions. Under salt stress, proteins related to ion-transport, metabolic pathway, protein synthesis and defense responsive were increased to a certain extent. A broader comparison of the proteome of wheat plant under different treatments revealed that changes in some of the metabolic pathway may be involved in stress adaption in response to PGPR inoculation. Hierarchical cluster analysis identified the various up-regulated/down-regulated proteins into tested three treatments. Our results suggest that bacterial inoculation enhanced the ability of wheat plant to combat salt stress via regulation of transcription factors, promoting antioxidative activity, induction of defense enzymes, lignin biosynthesis, and acceleration of protein synthesis.

SUBMITTER: Singh RP

PROVIDER: S-EPMC5587313 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Quantitative proteomics analysis reveals the tolerance of wheat to salt stress in response to Enterobacter cloacae SBP-8.

Singh Rajnish Prakash RP Runthala Ashish A Khan Shahid S Jha Prabhat Nath PN

PloS one 20170906 9

Salinity stress adversely affects the plant growth and is a major constraint to agriculture. In the present study, we studied the role of plant growth promoting rhizobacterium (PGPR) Enterobacter cloacae SBP-8 possessing ACC deaminase activity on proteome profile of wheat (Triticum aestivum L.) under high salinity (200 mM NaCl) stress. The aim of study was to investigate the differential expressed protein in selected three (T-1, T-2, T-3) treatments and absolute quantification (MS/MS analysis) w ...[more]

PMID: 28877183

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Project description:The present study investigated the physiological and biochemical characteristics of Enterobacter cloacae HG-1 isolated from saline-alkali soil. We further studied the effect of this strain on the salt tolerance of wheat and on the community structure of nitrogen-fixing bacteria in rhizosphere soil. We determined that the investigated strain had high nitrogen fixation activity and produced iron carriers, 1-aminocyclopropane-1-carboxylic acid deaminase, and plant hormones. The metabolites of this strain contained 2,3-butanediol, [R-(R*, R*)], 2-heptanone, and other growth-promoting and antibacterial substances. The strain was also highly salt-tolerant (10% NaCl). After the inoculation of wheat with the HG-1 strain, we recorded increases in root length, plant height, fresh weight, and dry weight of 19.15%, 18.83%, 16.67%, and 17.96%, respectively, compared with uninoculated plants (P < 0.05). Compared with the leaves of uninoculated plants, the proline concentration in the leaves of inoculated plants increased by 12.43% (P < 0.05), the malondialdehyde level decreased by 27.26% (P < 0.05), K+ increased by 20.69%, Ca2+ increased by 57.53% and Na+ decreased by 31.43% (all P<0.05). Furthermore, we detected that inoculation with the HG-1 strain did not affect the species composition of nitrogen-fixing bacteria in wheat rhizosphere soil at the phylum level. However, the average relative abundance of Proteobacteria was significantly increased, whereas the abundance of Verrucomiorobia was significantly decreased compared with uninoculated plants. At the genus level, we detected 32 genera in control samples and 27 genera in inoculated samples, and the species diversity and relative abundance of samples inoculated with the HG-1 strain decreased compared with uninoculated plants. Inoculated samples had lower abundances of Azospirillum, Rhodomicrobium, and Anabaena. Our study demonstrated that the inoculation of wheat with E. cloacae HG-1 could promote the growth of wheat under salt stress and increase salt stress tolerance. The results of this study investigating the interaction among soil, plants, and microorganisms supplement agricultural microbial databases and could provide a reference for the development of microbial-based saline soil improvement programs.

Project description:BackgroundEnterobacter cloacae (EC) is a Gram-negative bacterium that has been utilized extensively in biotechnological and environmental science applications, possibly because of its high capability for adapting itself and surviving in hazardous conditions. A search for the EC from agricultural and industrial areas that possesses high capability to tolerate and/or accumulate cadmium ions has been conducted in this study. Plausible mechanisms of cellular adaptations in the presence of toxic cadmium have also been proposed.MethodsNine strains of EC were isolated and subsequently identified by biochemical characterization and MALDI-Biotyper. Minimum inhibitory concentrations (MICs) against cadmium, zinc and copper ions were determined by agar dilution method. Growth tolerance against cadmium ions was spectrophotometrically monitored at 600 nm. Cadmium accumulation at both cellular and protein levels was investigated using atomic absorption spectrophotometer. Proteomics analysis by 2D-DIGE in conjunction with protein identification by QTOF-LC-MS/MS was used to study differentially expressed proteins between the tolerant and intolerant strains as consequences of cadmium exposure. Expression of such proteins was confirmed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Bioinformatics tools were applied to propose the functional roles of cadmium-binding protein and its association in cadmium tolerance mechanisms.ResultsThe cadmium-tolerant strain (EC01) and intolerant strain (EC07) with the MICs of 1.6 and 0.4 mM, respectively, were isolated. The whole cell lysate of EC01 exhibited approximately two-fold higher in cadmium binding capability than those of the EC07 and ATCC 13047, possibly by the expression of Cd-binding proteins. Our proteomics analysis revealed the higher expression of DUF326-like domain (a high cysteine-rich protein) of up to 220 fold in the EC01 than that of the EC07. Confirmation of the transcription level of this gene by qRT-PCR revealed a 14-fold induction in the EC01. Regulation of the DUF326-like domain in EC01 was more pronounced to mediate rapid cadmium accumulation (in 6 h) and tolerance than the other resistance mechanisms found in the ATCC 13047 and the EC07 strains. The only one major responsive protein against toxic cadmium found in these three strains belonged to an antioxidative enzyme, namely catalase. The unique proteins found in the ATCC 13047 and EC07 were identified as two groups: (i) ATP synthase subunit alpha, putative hydrolase and superoxide dismutase and (ii) OmpX, protein YciF, OmpC porin, DNA protection during starvation protein, and TrpR binding protein WrbA, respectively.ConclusionAll these findings gain insights not only into the molecular mechanisms of cadmium tolerance in EC but also open up a high feasibility to apply the newly discovered DUF326-like domain as cadmium biosorbents for environmental remediation in the future.

Dataset Information

Quantitative proteomics analysis reveals the tolerance of wheat to salt stress in response to Enterobacter cloacae SBP-8.

Publications

Quantitative proteomics analysis reveals the tolerance of wheat to salt stress in response to Enterobacter cloacae SBP-8.

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