Puneet Singh Chauhan

Induction of plant defense enzymes and phenolics by treatment with plant growth-promoting rhizobacteria Serratia marcescens NBRI1213

In greenhouse experiments, plant growth–promoting rhizobacteria (PGPR) Serratia marcescens NBRI1213 was evaluated for plant growth promotion and biologic control of foot and root rot of betelvine caused by Phytophthora nicotianae. Bacterization of betelvine (Piper betle L.) cuttings with S. marcescens NBRI1213 induced phenylalanine ammonia-lyase, peroxidase, and polyphenoloxidase activities in leaf and root. Qualitative and quantitative estimation of phenolic compounds was done through high-performance liquid chromatography (HPLC) in leaf and root of betelvine after treatment with S. marcescens NBRI1213 and infection by P. nicotianae. Major phenolics detected were gallic, protocatechuic, chlorogenic, caffeic, ferulic, and ellagic acids by comparison of their retention time with standards through HPLC. In all of the treated plants, synthesis of phenolic compounds was enhanced compared with control. Maximum accumulation of phenolics was increased in S. marcescens NBRI1213–treated plants infected with P. nicotianae. In a greenhouse test, bacterization using S. marcescens NBRI1213 decreased the number of diseased plants compared with nonbacterized controls. There were significant growth increases in shoot length, shoot dry weight, root length, and root dry weight, averaging 81%, 68%, 152%, and 290%, respectively, greater than untreated controls. This is the first report of PGPR–mediated induction of phenolics for biologic control and their probable role in protecting betelvine against P. nicotianae, an important soil-borne phytopathogenic fungus.

Plant growth-promoting bacteria Bacillus amyloliquefaciens NBRISN13 modulates gene expression profile of leaf and rhizosphere community in rice during salt stress.

Growth and productivity of rice and soil inhabiting microbial population is negatively affected by soil salinity. However, some salt resistant, rhizosphere competent bacteria improve plant health in saline stress. Present study evaluated the effect of salt tolerant Bacillus amyloliquefaciens NBRISN13 (SN13) inoculation on rice plants in hydroponic and soil conditions exposed to salinity. SN13 increased plant growth and salt tolerance (NaCl 200 mM) and expression of at least 14 genes under hydroponic and soil conditions in rice. Among these 14 genes 4 (NADP-Me2, EREBP, SOSI, BADH and SERK1) were up-regulated and 2 (GIG and SAPK4) repressed under salt stress in hydroponic condition. In greenhouse experiment, salt stress resulted in accumulation of MAPK5 and down-regulation of the remaining 13 transcripts was observed. SN13 treatment, with or without salt gave similar expression for all tested genes as compared to control. Salt stress caused changes in the microbial diversity of the rice rhizosphere and stimulated population of betaine-, sucrose-, trehalose-, and glutamine-utilizing bacteria in salt-treated rice rhizosphere (SN13 + salt). The observations imply that SN13 confers salt tolerance in rice by modulating differential transcription in a set of at least 14 genes. Stimulation of osmoprotectant utilizing microbial population as a mechanism of inducing salt tolerance in rice is reported for the first time in this study to the best of our knowledge.

Trichoderma: a potential bioremediator for environmental clean up

Environmental awareness has resulted in development of regulatory measures that aim to straighten past mistakes and protect the environment from future contamination and exploitation. However, much consideration and research needs to go into the decision-making process for an effective clean up of a particular contaminated site. Each technology developed has its advantages and limitations for the treatment of specific contaminants. Bioremediation and phytoremediation in association with microbes are innovative technologies having a potential to alleviate numerous environmental pollution problems. Owing to its dominant presence in contaminated sites, the application of the fungi in bioremediation is well documented. The genus Trichoderma is genetically very diverse with a number of capabilities among different strains with agricultural and industrial significance. It is also tolerant to a range of recalcitrant pollutants including heavy metals, pesticides, and polyaromatic hydrocarbons. This review presents an updated overview of application of Trichoderma for biological or phytobial remediation of environmental contaminants.

Gene expression profiling through microarray analysis in Arabidopsis thaliana colonized by Pseudomonas putida MTCC5279, a plant growth promoting rhizobacterium

Plant growth promotion is a multigenic process under the influence of many factors; therefore an understanding of these processes and the functions regulated may have profound implications. Present study reports microarray analysis of Arabidopsis thaliana plants inoculated with Pseudomonas putida MTCC5279 (MTCC5279) which resulted in significant increase in growth traits as compared with non-inoculated control. The gene expression changes, represented by oligonucleotide array (24652 genes) have been studied to gain insight into MTCC5279 assisted plant growth promotion in Arabidopsis thaliana. MTCC5279 induced upregulated Arabidopsis thaliana genes were found to be involved in maintenance of genome integrity (At5g20850), growth hormone (At3g23890 and At4g36110), amino acid synthesis (At5g63890), abcissic acid (ABA) signaling and ethylene suppression (At2g29090, At5g17850), Ca+2 dependent signaling (At3g57530) and induction of induced systemic resistance (At2g46370, At2g44840). The genes At3g32920 and At2g15890 which are suggested to act early in petal, stamen and embryonic development are among the downregulated genes. We report for the first time MTCC5279 assisted repression of At3g32920, a putative DNA repair protein involved in recombination and DNA strand transfer in a process of rapid meiotic and mitotic division.

Uncultured bacterial diversity in tropical maize (Zea mays L.) rhizosphere

Structure of maize (Zea mays L.) rhizosphere bacteria was evaluated to explore the feasibility of identifying novel rhizosphere bacteria using culture‐independent method based on direct amplification and analysis of 16S rRNA gene (rRNA) sequences and especially to obtain a better understanding of bacterial community structure and diversity from maize. A total of 274 sequences were analyzed and assigned 48.00% Proteobacteria, 10.30% Actinobacteria, 9.90% Bacteroidetes, 6.60% Verrucomicrobia, 4.80% Acidobacteria, 1.80% Firmicutes, 1.50% Chloroflexi, 1.50% TM7, 1.10% Deinococcus‐Thermus, 0.70% Planctomycetes, 0.70% Gemmatimonadetes and 0.40% Cyanobacteria. Economically important phyla Actinobacteria was second most dominant group after Proteobacteria, in our clone library. It would be interesting to hypothesize that root exudates from maize rhizosphere favors growth of Actinobacteria like microbes to eliminate pathogenic bacteria and decompose plant matter, for enhanced plant and soil health. An additional 12.8% of clone library (35 operational taxonomical units (OTUs) from 43 clones) with less than 94% similarity to any GenBank sequence could not be assigned to any known phylum and may represent unidentified bacterial lineages and suggests that a large amount of the rhizobacterial diversity remains to be characterized by culturing. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Developmental stage-dependent differential gene expression of superoxide dismutase isoenzymes and their localization and physical interaction network in rice ( Oryza sativa L.)

Superoxide dismutase (SOD) isoenzymes are essential for scavenging excess reactive oxygen species in living organisms. So far, expression pattern of SOD isoenzymes genes along leaf development plus their sub-cellular localization and physical interaction network have not yet been clearly elucidated. Using multiple bioinformatics tools, we predicted the sub-cellular localizations of SOD isoforms and described their physical interactions in rice. Using in silico approaches, we obtained several evidences for existence of seven SOD genes and a SOD copper chaperone gene. Their transcripts were differentially expressed along with different developmental stage of rice leaf. Finally, we performed quantitative real time-polymerase chain reaction (qRT-PCR) to validate in silico differential expression pattern of SOD genes experimentally. Expression of two cytosolic cCuZn-SODs was high during the whole vegetative stage. Two plastidic Fe-SODs were found and their expression levels were very low and started to increase from the late vegetative stage. Their expression patterns were very similar to each other, indicating the formation of heterodimer. However, their expression patterns are different from those for ArabidopsisFe-SODs. The expression of pCuZn-SOD was very high in the early developmental stage, but qRT-PCR results were different, which remains for further study. From the results on the differential expression of SOD genes, we can understand the role of each SOD gene and even predict their role under certain circumstances based on in silico analysis.

Synergistic effect of Pseudomonas putida and Bacillus amyloliquefaciens ameliorates drought stress in chickpea (Cicer arietinum L.)

Two plant growth promoting rhizobacteria (PGPR) Pseudomonas putida NBRIRA and Bacillus amyloliquefaciens NBRISN13 with ability to tolerate abiotic stress along with multiple PGP traits like ACC deaminase activity, minerals solubilisation, hormones production, biofilm formation, siderophore activity were evaluated for their synergistic effect to ameliorate drought stress in chickpea. Earlier we have reported both the strains individually for their PGP attributes and stress amelioration in host plants. The present study explains in detail the possibilities and benefits of utilizing these 2 PGPR in consortium for improving the chickpea growth under control and drought stressed condition. In vitro results clearly demonstrate that both the PGPR strains are compatible to each other and their synergistic growth enhances the PGP attributes. Greenhouse experiments were conducted to evaluate the effect of inoculation of both strains individually and consortia in drought tolerant and sensitive cultivars (BG362 and P1003). The growth parameters were observed significantly higher in consortium as compared to individual PGPR. Colonization of both PGPR in chickpea rhizosphere has been visualized by using gfp labeling. Apart from growth parameters, defense enzymes, soil enzymes and microbial diversity were significantly modulated in individually PGPR and in consortia inoculated plants. Negative effects of drought stress has been ameliorated and apparently seen by higher biomass and reversal of stress indicators in chickpea cultivars treated with PGPR individually or in consortia. Findings from the present study demonstrate that synergistic application has better potential to improve plant growth promotion under drought stress conditions.

Pseudomonas putida NBRIC19 dihydrolipoamide succinyltransferase (SucB) gene controls degradation of toxic allelochemicals produced by Parthenium hysterophorus

Aims:  The aim of this study was to identify the gene responsible for degradation of toxic allelochemicals of Parthenium by generating Tn5‐induced mutant of Pseudomonas putida NBRIC19. Furthermore, the study characterizes the mutant at physiological, biochemical and molecular level that helped in understanding the mechanisms of reducing the allelopathic inhibition of Parthenium by Ps. putida NBRIC19.

Impact of salinity-tolerant MCM6 transgenic tobacco on soil enzymatic activities and the functional diversity of rhizosphere microbial communities

The development of genetically modified plants for agriculture has provided numerous economic benefits, but has also raised concern over the potential impact of transgenic plants upon the environment. The rhizosphere is the soil compartment that is directly under the influence of living roots; it constitutes a complex niche likely to be exploited by a wide variety of bacteria potentially influenced by the introduction of transgenes in genetically modified plants. In the present study, the impact of overexpression of the salinity stress-tolerant minichromosome maintenance complex subunit 6 (MCM6) gene upon functional diversity and soil enzymatic activity in the rhizosphere of transgenic tobacco in the presence and absence of salt stress was examined. The diversity of culturable bacterial communities and soil enzymes, namely, dehydrogenases and acid phosphatases, was assessed and revealed no significant (or only minor) alterations due to transgenes in the rhizosphere soil of tobacco plants. Patterns in principal components analysis showed clustering of transgenic and non-transgenic tobacco plants according to the fingerprint of their associated bacterial communities. However, the presence of MCM6 tobacco did not cause changes in microbial populations, soil enzymatic activities or the functional diversity of the rhizosphere soil microbial community.

Genome-wide analysis of rice dehydrin gene family: Its evolutionary conservedness and expression pattern in response to PEG induced dehydration stress

Abiotic stresses adversely affect cellular homeostasis, impairing overall growth and development of plants. These initial stress signals activate downstream signalling processes, which, subsequently, activate stress-responsive mechanisms to re-establish homeostasis. Dehydrins (DHNs) play an important role in combating dehydration stress. Rice (Oryza sativa L.), which is a paddy crop, is susceptible to drought stress. As drought survival in rice might be viewed as a trait with strong evolutionary selection pressure, we observed DHNs in the light of domestication during the course of evolution. Overall, 65 DHNs were identified by a genome-wide survey of 11 rice species, and 3 DHNs were found to be highly conserved. The correlation of a conserved pattern of DHNs with domestication and diversification of wild to cultivated rice was validated by synonymous substitution rates, indicating that Oryza rufipogon and Oryza sativa ssp. japonica follow an adaptive evolutionary pattern; whereas Oryza nivara and Oryza sativa ssp. indica demonstrate a conserved evolutionary pattern. A comprehensive analysis of tissue-specific expression of DHN genes in japonica and their expression profiles in normal and PEG (poly ethylene glycol)-induced dehydration stress exhibited a spatiotemporal expression pattern. Their interaction network reflects the cross-talk between gene expression and the physiological processes mediating adaptation to dehydration stress. The results obtained strongly indicated the importance of DHNs, as they are conserved during the course of domestication.