Smita Kumar

Effect of arsenic on growth, oxidative stress, and antioxidant system in rice seedlings ☆

The physiological, biochemical, and proteomic changes in germinating rice seedlings were investigated under arsenic stress. A marked decrease in germination percentage, shoot, and root elongation as well as plant biomass was observed with arsenic treatments, as compared to control, whereas accumulation of arsenic and malondialdehyde (MDA) in seedlings were increased significantly with increasing arsenic concentration (both AsIII and AsV). The up-regulation of some antioxidant enzyme activities and the isozymes of superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), peroxidase (POD, EC 1.11.1.7), and glutathione reductase (GR, 1.6.4.2) substantiated that arsenic accumulation generated oxidative stress, which was more pronounced in As(III) treatment. We also studied the protective effect of reduced glutathione (GSH) and cysteine (Cys) to As(III)/As(V) stressed seedlings. Both GSH and Cys imparted enhanced tolerance to seedlings against arsenic stress. Seedlings growth improved while level of MDA declined significantly when GSH and Cys were supplemented to As(III)/As(V) treatments suggesting GSH and Cys-mediated protection against oxidative stress. The arsenic content was highest in roots of seedlings grown in As(III) in the presence of GSH/Cys. However, in case of As(V) plus GSH or Cys, the arsenic content in seedlings was highest in shoots. The results are suggestive of differential metabolism of As(III) and As(V) in rice.

Comparative transcriptome analysis of arsenate and arsenite stresses in rice seedlings

The effect of arsenic (As) exposure on genome-wide expression was examined in rice (Oryza sativa L., ssp. Indica). A group of defense and stress-responsive genes, transporters, heat-shock proteins, metallothioneins, sulfate-metabolizing proteins, and regulatory genes showed differential expression in rice seedlings challenged with arsenate (AsV) and arsenite (AsIII). AsV stress led to upregulation or downregulation of an additional set of genes in comparison to AsIII. Differential expression of several genes that showed the highest contrast in a microarray analysis was validated by following the quantitative changes in the levels of individual transcripts following challenge with AsV, AsIII, Cd, Cr, and Pb. Most of the selected genes responded to challenge by heavy metals such as arsenic. However, expression of one of the cytochrome P450 genes (Os01g43740) in rice root was induced by AsV but not by other heavy metals. Similarly, one glutaredoxin (Os01g26912) is expressed specifically in the AsIII-treated shoot.

Transcriptomic and metabolomic shifts in rice roots in response to Cr (VI) stress

BackgroundWidespread use of chromium (Cr) contaminated fields due to careless and inappropriate management practices of effluent discharge, mostly from industries related to metallurgy, electroplating, production of paints and pigments, tanning, and wood preservation elevates its concentration in surface soil and eventually into rice plants and grains. In spite of many previous studies having been conducted on the effects of chromium stress, the precise molecular mechanisms related to both the effects of chromium phytotoxicity, the defense reactions of plants against chromium exposure as well as translocation and accumulation in rice remain poorly understood.ResultsDetailed analysis of genome-wide transcriptome profiling in rice root is reported here, following Cr-plant interaction. Such studies are important for the identification of genes responsible for tolerance, accumulation and defense response in plants with respect to Cr stress. Rice root metabolome analysis was also carried out to relate differential transcriptome data to biological processes affected by Cr (VI) stress in rice. To check whether the Cr-specific motifs were indeed significantly over represented in the promoter regions of Cr-responsive genes, occurrence of these motifs in whole genome sequence was carried out. In the background of whole genome, the lift value for these 14 and 13 motifs was significantly high in the test dataset. Though no functional role has been assigned to any of the motifs, but all of these are present as promoter motifs in the Database of orthologus promoters.ConclusionThese findings clearly suggest that a complex network of regulatory pathways modulates Cr-response of rice. The integrated matrix of both transcriptome and metabolome data after suitable normalization and initial calculations provided us a visual picture of the correlations between components. Predominance of different motifs in the subsets of genes suggests the involvement of motif-specific transcription modulating proteins in Cr stress response of rice.

Arsenic tolerances in rice (Oryza sativa) have a predominant role in transcriptional regulation of a set of genes including sulphur assimilation pathway and antioxidant system.

World wide arsenic (As) contamination of rice has raised much concern as it is the staple crop for millions. Four most commonly cultivated rice cultivars, Triguna, IR-36, PNR-519 and IET-4786, of the West Bengal region were taken for a hydroponic study to examine the effect of arsenate (As(V)) and arsenite (As(III)) on growth response, expression of genes and antioxidants vis-à-vis As accumulation. The rice genotypes responded differentially under As(V) and As(III) stress in terms of gene expression and antioxidant defences. Some of the transporters were up-regulated in all rice cultivars at lower doses of As species, except IET-4786. Phytochelatin synthase, GST and γ-ECS showed considerable variation in their expression pattern in all genotypes, however in IET-4786 they were generally down-regulated in higher As(III) stress. Similarly, most of antioxidants such as superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) increased significantly in Triguna, IR-36 and PNR-519 and decreased in IET-4786. Our study suggests that Triguna, IR-36 and PNR-519 are tolerant rice cultivars accumulating higher arsenic; however IET-4786 is susceptible to As-stress and accumulates less arsenic than other cultivars.

Expression of a rice Lambda class of glutathione S-transferase, OsGSTL2, in Arabidopsis provides tolerance to heavy metal and other abiotic stresses.

Global industrial growth has contaminated the soil and water with many hazardous compounds, including heavy metals. These heavy metals are not only toxic to plants but also cause severe human health hazards when leach out into food chain. One of the approaches employed for the decontamination of environment includes identification and overexpression of genes involved in the detoxification mechanism of plants. Glutathione S-transferases (GSTs) are a superfamily of enzymes, principally known for their role in detoxification reactions. Different classes of GSTs have been used to develop plants with improved detoxification mechanism, but not much information is available for Lambda class of GSTs. Here, we studied expression of OsGSTLs in different rice genotypes under arsenic stress. The study suggests differential expression of these genes in arsenic sensitive and tolerant genotypes. Further, the role of one member of Lambda class OsGSTL2 was studied by expressing in heterologous system, Arabidopsis. Transgenic lines developed were analysed for their response to different abiotic stresses including heavy metals. Analysis suggests that OsGSTL2 provides tolerance for heavy metals and other abiotic stresses like cold, osmotic stress and salt. We conclude that OsGSTLs can be utilized for developing plant varieties tolerant to different abiotic stresses including heavy metals.

Omics and biotechnology of arsenic stress and detoxification in plants: current updates and prospective.

Arsenic (As), a naturally occurring metallic element, is a dreadful health hazard to millions of people across the globe. Arsenic is present in low amount in the environment and originates from anthropogenic impact and geogenic sources. The presence of As in groundwater used for irrigation is a worldwide problem as it affects crop productivity, accumulates to different tissues and contaminates food chain. The consumption of As contaminated water or food products leads to several diseases and even death. Recently, studies have been carried out to explore the biochemical and molecular mechanisms which contribute to As toxicity, accumulation, detoxification and tolerance acquisition in plants. This information has led to the development of the biotechnological tools for developing plants with modulated As tolerance and detoxification to safeguard cellular and genetic integrity as well as to minimize food chain contamination. This review aims to provide current updates about the biochemical and molecular networks involved in As uptake by plants and the recent developments in the area of functional genomics in terms of developing As tolerant and low As accumulating plants.

Differential expression and alternative splicing of rice sulphate transporter family members regulate sulphur status during plant growth, development and stress conditions

Sulphur, an essential nutrient required for plant growth and development, is mainly taken up by the plants as inorganic sulphate from the soil and assimilated into the sulphur reductive pathway. The uptake and transport of sulphate in plants is carried out by transporters encoded by the sulphate transporter gene family. Plant sulphate transporters have been classified with respect to their protein sequences, kinetic properties and tissue-specific localization in Arabidopsis. Though sulphate transporter genes from few other plants have also been characterized, no detailed study with respect to the structure and expression of this family from rice has been carried out. Here, we present genome-wide identification, structural and expression analyses of the rice sulphate transporter gene family. Our analysis using microarray data and MPSS database suggests that 14 rice sulphate transporters are differentially expressed during growth and development in various tissues and during biotic and abiotic stresses. Our analysis also suggests differential accumulation of splice variants of OsSultr1;1 and OsSultr4;1 transcripts during these processes. Apart from known spliced variants, we report an unusual alternative splicing of OsSultr1;1 transcript related to sulphur supply in growth medium and during stress response. Taken together, our study suggests that differential expression and alternative splicing of members of the sulphate transporter family plays an important role in regulating cellular sulphur status required for growth and development and during stress conditions. These findings significantly advance our understanding of the posttranscriptional regulatory mechanisms operating to regulate sulphur demand by the plant.

Sulfur mediated reduction of arsenic toxicity involves efficient thiol metabolism and the antioxidant defense system in rice

Arsenic (As) contamination is a global issue, with South Asia and South East Asia being worst affected. Rice is major crop in these regions and can potentially pose serious health risks due to its known As accumulation potential. Sulfur (S) is an essential macronutrient and a vital element to combat As toxicity. The aim of this study was to investigate the role of S with regards to As toxicity in rice under different S regimes. To achieve this aim, plants were stressed with AsIII and AsV under three different S conditions (low sulfur (0.5mM), normal sulfur (3.5mM) and high sulfur (5.0mM)). High S treatment resulted in increased root As accumulation, likely due to As complexation through enhanced synthesis of thiolic ligands, such as non-protein thiols and phytochelatins, which restricted As translocation to the shoots. Enzymes of S assimilatory pathways and downstream thiolic metabolites were up-regulated with increased S supplementation; however, to maintain optimum concentrations of S, transcript levels of sulfate transporters were down-regulated at high S concentration. Oxidative stress generated due to As was counterbalanced in the high S treatment by reducing hydrogen peroxide concentration and enhancing antioxidant enzyme activities. The high S concentration resulted in reduced transcript levels of Lsi2 (a known transporter of As). This reduction in Lsi2 expression level is a probable reason for low shoot As accumulation, which has potential implications in reducing the risk of As in the food chain.

Differential Expression of Rice Lambda Class GST Gene Family Members During Plant Growth, Development, and in Response to Stress Conditions

Glutathione S-transferases (GSTs; EC 2.5.1.18) are members of an isozyme family and catalyze the conjugation of the reduced tripeptide glutathione to a variety of hydrophobic and electrophilic substrates. Though members of different classes of the GST superfamily have been identified and characterized from many plant species including rice, no detailed information is available for the Lambda class gene family in rice. In this study, a genome-wide analysis was carried out to investigate expression patterns of three Lambda class GST members of rice including OsGSTL1, OsGSTL2 and OsGSTL3 in seedlings, at different growth and developmental stages as well as in response to various biotic and abiotic stresses. Expression analysis using microarray datasets and quantitative real-time reverse transcriptase polymerase chain reaction suggests that this gene family express differentially in various tissues, in response to hormones and during different biotic and abiotic stresses including heavy metals, cold, drought and salt stress. Massively Parallel Signature Sequencing (MPSS) analysis also showed differential expression of OsGSTLs during plant growth and development and under different stresses. Out of three members, maximum expression of OsGSTL2 was observed for the MPSS libraries in comparison to other members. We conclude that members of rice Lambda class GST family play an important role in plant growth and development and in combating different biotic and abiotic stresses.

Sulfur alleviates arsenic toxicity by reducing its accumulation and modulating proteome, amino acids and thiol metabolism in rice leaves.

Arsenic (As) contamination of water is a global concern and rice consumption is the biggest dietary exposure to human posing carcinogenic risks, predominantly in Asia. Sulfur (S) is involved in di-sulfide linkage in many proteins and plays crucial role in As detoxification. Present study explores role of variable S supply on rice leaf proteome, its inclination towards amino acids (AA) profile and non protein thiols under arsenite exposure. Analysis of 282 detected proteins on 2-DE gel revealed 113 differentially expressed proteins, out of which 80 were identified by MALDI-TOF-TOF. The identified proteins were mostly involved in glycolysis, TCA cycle, AA biosynthesis, photosynthesis, protein metabolism, stress and energy metabolism. Among these, glycolytic enzymes play a major role in AA biosynthesis that leads to change in AAs profiling. Proteins of glycolytic pathway, photosynthesis and energy metabolism were also validated by western blot analysis. Conclusively S supplementation reduced the As accumulation in shoot positively skewed thiol metabolism and glycolysis towards AA accumulation under AsIII stress.