Publications of NIPGR Scientists
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Item ath-miR164c influences plant responses to the combined stress of drought and bacterial infection by regulating proline metabolism(Elsevier B.V., 2020) Gupta, Aarti; Patil, Mahesh; Qamar, Aarzoo; Senthil-Kumar, MuthappaPlants under combined stresses exhibit a prominent shift in molecular responses compared with plants exposed to the same stresses independently. Profiling responses to individual and combined stressors at the gene expression level have identified several genes with intersecting responses to these stressors. However, the upstream regulators at the intersection of plant responses to individual and combined stresses are not known. Here, using the transcriptome of Arabidopsis thaliana under individual and combined drought and Pseudomonas syringae infection, we identified several genes whose expression overlaps between individual and combined stresses. To study the key regulator of such an overlapping gene, we predicted that the expression of 1-Pyrroline-5-carboxylate synthase 1 (AtP5CS1) is regulated by ath-miR164c at post-transcriptional level. Our results from the stem-loop RT-PCR based expression analysis revealed significant downregulation of ath-miR164c in response to P. syringae infection under both well-irrigated (pathogen only) and drought stress (combined stress) conditions. Furthermore, an Arabidopsis loss-of-function mutant of the miRNA ath-miR164c exhibited resistance to pathogen infection under combined stress, unlike the wild-type plants, implicating the role of ath-miR164c in regulating plant immunity. AtP5CS1 gene expression and proline accumulation were enhanced in the ath-miR164c mutant plants relative to the wild-type plants, demonstrating that ath-miR164c regulates AtP5CS1 of the proline biosynthesis pathway, which was also validated by 5’RLM-RACE results. This miRNA-mediated modulation of AtP5CS1 gene expression under combined stress fills crucial gaps in identifying the key convergent players in the current understanding of plant stress responses.Item AtGBF3 confers tolerance to Arabidopsis thaliana against combined drought and Pseudomonas syringae stress(Elsevier B.V., 2019) Dixit, Sandeep Kumar; Gupta, Aarti; Fatima, Urooj; Senthil-Kumar, MuthappaIn field conditions, plants are often exposed to a combination of abiotic and biotic stresses, for instance, drought and pathogen infection. Transcriptome studies on Arabidopsis thaliana and other plants under individual and combined drought and pathogen stresses have unveiled the activation of shared molecular defense mechanisms. These shared plant responses are characterized by commonly regulated genes under both individual as well as combined stresses. Therefore, the identification of commonly regulated genes during individual and combined stress conditions can reveal plant responses towards combined stress. Available transcriptome studies on combined-stressed plants have hinted at G-Box Binding Factor 3 (GBF3) as one of the regulatory components of the shared response. However, the mechanistic understanding of the role of AtGBF3 under combined drought and pathogen stress is not yet decoded. In the current study, we used genetic approaches to identify the role of AtGBF3 in conferring tolerance to individual and combined drought and pathogen stress. Atgbf3 mutant plants showed increased susceptibility, while AtGBF3-overexpressing plants were tolerant under individual and combined drought and Pseudomonas syringae pv. tomato infection stresses as compared to wild-type plants. We further analyzed the global transcriptome of Atgbf3 mutant plants under combined stress to identify its downstream targets. We also established a high-throughput method to apply combined polyethylene glycol and pathogen stress on Murashige and Skoog medium-grown plants to further validate the role of AtGBF3 in combined stress.Item Current understanding of regulation of GBF3 under abiotic and biotic stresses and its potential role in combined stress tolerance(Taylor & Francis Group, 2019) Dixit, Sandeep Kumar; Gupta, Aarti; Senthil-Kumar, MuthappaG-box binding factors (GBFs) belong to basic leucine zipper (bZIP) super family of transcription factors. Among six reported members of Arabidopsis thaliana GBFs, AtGBF3 transcripts has been shown to accumulate to high levels in dry seed, stamens, mature pollen and siliques. Amino acid sequence analysis of AtGBF3 reveals presence of proline rich region at N-terminus and basic leucine zipper domain at C-terminus. Earlier, it has been demonstrated that GBFs bind to the G-box element found within the promoter of stress responsive genes including alcohol dehydrogenase (AtAdh) and activate its transcription. Expression profile of AtGBF3 depicts increase in mRNA levels in plants under drought, osmotic, heat, salt and cold stress and biotic stresses. In silico analysis reveals GBF3 localization in nucleus and cytoplasm. Cues from localization studies on GBF3 homologs suggest that upon stress signal perception, GBF3 is phosphorylated and localized to nucleus where it can regulate expression of stress related genes. Till now target genes of GBF3 and mechanism of their regulation during individual and combined abiotic - biotic stresses is not comprehensively reviewed. Through our analysis in this review, we propose GBF3 as major player in integrating abscisic acid and salicylic acid cross talk during combined stress.Item GBF3 transcription factor imparts drought tolerance in Arabidopsis thaliana(Nature Publishing Group, 2017) Ramegowda, Venkategowda; Gill, Upinder Singh; Sivalingam, Palaiyur Nanjappan; Gupta, Aarti; Gupta, Chirag; Govind, Geetha; Nataraja, Karaba N.; Pereira, Andy; Udayakumar, Makarla; Mysore, Kirankumar S.; Senthil-Kumar, MuthappaDrought transcriptome analysis of finger millet (Eleusine coracana) by cDNA subtraction identified drought responsive genes that have a potential role in drought tolerance. Through virus-induced gene silencing (VIGS) in a related crop species, maize (Zea mays), several genes, including a G-BOX BINDING FACTOR 3 (GBF3) were identified as candidate drought stress response genes and the role of GBF3 in drought tolerance was studied in Arabidopsis thaliana. Overexpression of both EcGBF3 and AtGBF3 in A. thaliana resulted in improved tolerance to osmotic stress, salinity and drought stress in addition to conferring insensitivity to ABA. Conversely, loss of function of this gene increased the sensitivity of A. thaliana plants to drought stress. EcGBF3 transgenic A. thaliana results also suggest that drought tolerance of sensitive plants can be improved by transferring genes from far related crops like finger millet. Our results demonstrate the role of GBF3 in imparting drought tolerance in A. thaliana and indicate the conserved role of this gene in drought and other abiotic stress tolerance in several plant species.Item Transcriptome changes in Arabidopsis thaliana infected with Pseudomonas syringae during drought recovery(Nature Publishing Group, 2017) Gupta, Aarti; Senthil-Kumar, MuthappaField-grown plants experience cycles of drought stress and recovery due to variation in soil moisture status. Physiological, biochemical and transcriptome responses instigated by recovery are expected to be different from drought stress and non-stressed state. Such responses can further aid or antagonize the plant’s interaction with the pathogen. However, at molecular level, not much is known about plantpathogen interaction during drought recovery. In the present study, we performed a microarray-based global transcriptome profiling and demonstrated the existence of unique transcriptional changes in Arabidopsis thaliana inoculated with Pseudomonas syringae pv. tomato DC3000 at the time of drought recovery (drought recovery pathogen, DRP) when compared to the individual drought (D) or pathogen (P) or drought recovery (DR). Furthermore, the comparison of DRP with D or DR and P transcriptome revealed the presence of a few common genes among three treatments. Notably, a gene encoding proline dehydrogenase (AtProDH1) was found to be commonly up-regulated under drought recovery (DR), DRP and P stresses. We also report an up-regulation of pyrroline-5-carboxylate biosynthesis pathway during recovery. We propose that AtProDH1 influences the defense pathways during DRP. Altogether, this study provides insight into the understanding of defense responses that operate in pathogen-infected plants during drought recovery.Item Global profiling of phytohormone dynamics during combined drought and pathogen stress in Arabidopsis thaliana reveals ABA and JA as major regulators(Nature Publishing Group, 2017) Gupta, Aarti; Hisano, Hiroshi; Hojo, Yuko; Matsuura, Takakazu; Ikeda, Yoko; Mori, Izumi C.; Senthil-Kumar, MuthappaGlobal transcriptome studies demonstrated the existence of unique plant responses under combined stress which are otherwise not seen during individual stresses. In order to combat combined stress plants use signaling pathways and ‘cross talk’ mediated by hormones involved in stress and growth related processes. However, interactions among hormones’ pathways in combined stressed plants are not yet known. Here we studied dynamics of different hormones under individual and combined drought and pathogen infection in Arabidopsis thaliana by liquid chromatography-mass spectrometry (LC-MS) based profiling. Our results revealed abscisic acid (ABA) and salicylic acid (SA) as key regulators under individual drought and pathogen stress respectively. Under combined drought and host pathogen stress (DH) we observed non-induced levels of ABA with an upsurge in SA and jasmonic acid (JA) concentrations, underscoring their role in basal tolerance against host pathogen. Under a non-host pathogen interaction with drought (DNH) stressed plants, ABA, SA and JA profiles were similar to those under DH or non-host pathogen alone. We propose that plants use SA/JA dependent signaling during DH stress which antagonize ABA biosynthesis and signaling pathways during early stage of stress. The study provides insights into hormone modulation at different time points during combined stress.Item Transcriptomic changes under combined drought and nonhost bacteria reveal novel and robust defenses in Arabidopsis thaliana(Elsevier B.V., 2017) Choudhary, Aanchal; Gupta, Aarti; Ramegowda, Venkategowda; Senthil-Kumar, MuthappaPlants in the natural conditions are often challenged by a combination of two or more stressors. A combination of drought and pathogen is one of the most pressing threats to the plant’s growth and survival in the field, and thus warrants a mechanistic understanding. Susceptible plant-pathogen interaction, owing to effector-mediated suppression of plant defense responses, limits its scope for combined stress studies. In the present study, we have investigated the morpho-physiological responses of Arabidopsis thaliana to simultaneous drought and nonhost bacterial pathogen Pseudomonas syringae pv. tabaci. Combined stress treatment provoked an early and more pronounced hypersensitive response in the plant as compared to the non-host pathogen treatment. We have further deciphered the molecular basis for the robust defense response observed under combined stress by transcriptomic profiling carried out using whole-genome microarray. We found that the enhanced resistance to the combined stress is accompanied by a massive transcriptional reprogramming involving several transcripts specifically responding to the stress combination. A prominent over-representation of genes involved in basal defense-related machinery was observed under the combined stress. Genes involved in various defense signaling cascades, accumulation of secondary metabolites and those encoding for receptor-like kinases were highly up-regulated under the combined stress. Up-regulated genes related to redox homeostasis and hypersensitive response (HR)-mediated cell death were also found to be markedly enriched under combined stress. We also compared the global gene expression profile of A. thaliana subjected to combined drought-nonhost bacteria to those treated with a combination of drought-host bacteria Pseudomonas syringae pv. tomato DC3000. A significant induction of genes responding to drought as well as bacteria was observed during both the interactions. However, the amplitude of induction was more pronounced under the combination of drought and nonhost bacteria. Our results also indicate that plant activates multiple defense pathways upon exposure to combined stress which strengthens the overall basal immunity of the plant, characterized by a stronger HR response.Item Concurrent stresses are perceived as new state of stress by the plants: overview of impact of abiotic and biotic stress combinations(Springer, 2017) Gupta, Aarti; Senthil-Kumar, MuthappaCrop plants under natural conditions often encounter abiotic and biotic stresses either individually or in combination, single or multiple times in their life cycle. During their concurrence, different stressors interact with each other over the plant interface leading to altered plant responses. Initial stressor can modulate plant physiology and thereby influences plant response towards another stressor. Consequent to the stress interaction, plants encountering concurrent stress show different responses in comparison to the plants exposed to the individual stresses. Additionally, plant defence responses are somewhat skewed towards one stressor during concurrent occurrence of stresses. Such different responses are the cognate ‘net effect’ of combined stress felt by the plant. The net effect exhibited by plants under combined stress is unique to each stress combination. Thus, in lieu of the combined stress responses, which are different from the individual stress responses, the combined stress has been proposed as a new state of stress. Plant responses towards this new state are not just dictated by either of the individual stresses alone but by more complex interaction. In this chapter, we present an overview of the combined stresses with emphasis on drought and bacterial stressors and discuss the stress interaction effect and net effect.Item Concurrent drought stress and vascular pathogen infection induce common and distinct transcriptomic responses in chickpea(Frontiers Media S.A., 2017) Sinha, Ranjita; Gupta, Aarti; Senthil-Kumar, MuthappaChickpea (Cicer arietinum); the second largest legume grown worldwide is prone to drought and various pathogen infections. These drought and pathogen stresses often occur concurrently in the field conditions. However, the molecular events in response to that are largely unknown. The present study examines the transcriptome dynamics in chickpea plants exposed to a combination of water-deficit stress and Ralstonia solanacearum infection. R. solanacearum is a potential wilt disease causing pathogen in chickpea. Drought stressed chickpea plants were infected with this pathogen and the plants were allowed to experience progressive drought with 2 and 4 days of R. solanacearum infection called short duration stress (SD stresses) and long duration stress (LD stresses), respectively. Our study showed that R. solanacearum multiplication decreased under SD-combined stress compared to SD-pathogen but there was no significant change in LD-combined stress compared to LD-pathogen. The microarray analysis during these conditions showed that 821 and 1039 differentially expressed genes (DEGs) were unique to SD- and LD-combined stresses, respectively, when compared with individual stress conditions. Three and fifteen genes were common among all the SD-stress treatments and LD-stress treatments, respectively. Genes involved in secondary cell wall biosynthesis, alkaloid biosynthesis, defense related proteins, and osmo-protectants were up-regulated during combined stress. The expression of genes involved in lignin and cellulose biosynthesis were specifically up-regulated in SD-combined, LD-combined, and LD-pathogen stress. A close transcriptomic association of LD-pathogen stress with SD-combined stress was observed in this study which indicates that R. solanacearum infection also exerts drought stress along with pathogen stress thus mimics combined stress effect. Furthermore the expression profiling of candidate genes using real-time quantitative PCR validated the microarray data. The study showed that down-regulation of defense-related genes during LD-combined stress resulted in an increased bacterial multiplication as compared to SD-combined stress. Overall, our study highlights a sub-set of DEGs uniquely expressed in response to combined stress, which serve as potential candidates for further functional characterization to delineate the molecular response of the plant to concurrent drought-pathogen stress.Item Understanding the impact of drought on foliar and xylem invading bacterial pathogen stress in chickpea(Frontiers Media S.A., 2016) Sinha, Ranjita; Gupta, Aarti; Senthil-Kumar, MuthappaIn field conditions, plants are concurrently exposed to multiple stresses, where one stressor impacts the plants’ response to another stressor, and the resultant net effect of these stresses differs from individual stress response. The present study investigated the effect of drought stress on interaction of chickpea with Pseudomonas syringae pv. phaseolicola (Psp; foliar pathogen) and Ralstonia solanacearum (Rs; xylem inhabiting wilt causing pathogen), respectively, and the net-effect of combined stress on chlorophyll content and cell death. Two type of stress treatments were used to study the influence of each stress factor during combined stress, viz., imposition of drought stress followed by pathogen challenge (DP), and pathogen inoculated plants imposed with drought in course of pathogen infection (PD). Drought stress was imposed at different levels with pathogen inoculum to understand the influence of different stress intensities on stress interaction and their net impact. Drought stressed chickpea plants challenged with Psp infection (DPsp) showed reduced in planta bacterial number compared to Psp infection alone. Similarly, Rs infection of chickpea plants showed reduced in planta bacterial number under severe drought stress. Combined drought and Psp (DPsp) infected plants showed decreased cell death compared to plants infected only with Psp but the extent of cell death was similar to drought stressed plants. Similarly, chlorophyll content in plants under combined stress was similar to the individual drought stressed plants; however, the chlorophyll content was more compared to pathogen only infected plants. Under combined drought and Rs infection (DRs), cell death was similar to individual drought stress but significantly less compared to only Rs infected plants. Altogether, the study proposes that both stress interaction and net effect of combined stress could be majorly influenced by first occurring stress, for example, drought stress in DP treatment. In addition, our results indicate that the outcome of the two stress interaction in plant depends on timing of stress occurrence and nature of infecting pathogen.
