Gene expression profiling of drought-sensitive (IR64) and tolerant (N22) rice cultivars
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ABSTRACT:
ORGANISM(S): Oryza sativa
PROVIDER: GSE183894 | GEO | 2023/12/31
REPOSITORIES: GEO
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Project description:Traditional rice varieties found in India have many desirable characteristics. Amongst them, their differential responses to abiotic and biotic stresses are of great agricultural importance. Drought or osmotic stress is one of the major abiotic stresses afflicting crop plants in India. Indigenous varieties like Dagad deshi have been found to be drought resistant and, thereby, are being studied in great detail by plant breeders and biotechnologists alike. In this study, we have analyzed the transcriptomes of two contrasting cultivars, i.e. Dagad deshi (tolerant) and IR20 (susceptible), under control and stress conditions to elucidate the differences in their responses to drought stress using Affymetrix microarray platform. Hydroponically grown seven-day-old seedlings of Dagad deshi and IR20 were subjected to drought stress by placing them on 3 mm Whatmann sheets under light for 3 h and 6 h at 28±1oC. For control samples, seedlings were kept in RGM (root growth medium) for 6 h at 28±1oC. RNA extracted from each sample was hybridized on rice Affymetrix microarrays. Three biological replicates of each sample were used for microarray analysis. Overall, eighteen samples were analyzed representing control, 3 h and 6 h of dehydration stress for each cultivar (Dagad deshi and IR20).
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Project description:Traditional rice varieties found in India have many desirable characteristics. Amongst them, their differential responses to abiotic and biotic stresses are of great agricultural importance. Drought or osmotic stress is one of the major abiotic stresses afflicting crop plants in India. Indigenous varieties like Dagad deshi have been found to be drought resistant and, thereby, are being studied in great detail by plant breeders and biotechnologists alike. In this study, we have analyzed the transciptomes of two contrasting cultivars, i.e. Dagad deshi (tolerant) and IR20 (susceptible), under control and stress conditions to elucidate the differences in their responses to drought stress using Affymetrix microarray platform.
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Project description:Drought tolerance is a key trait for increasing and stabilizing barley productivity in dry areas worldwide. Identification of the genes responsible for drought tolerance in barley (Hordeum vulgare L.) will facilitate understanding of the molecular mechanisms of drought tolerance, and also genetic improvement of barley through marker-assisted selection or gene transformation. To monitor the changes in gene expression at transcription levels in barley leaves during the reproductive stage under drought conditions, the 22K Affymetrix Barley 1 microarray was used to screen two drought-tolerant barley genotypes, Martin and Hordeum spontaneum 41-1 (HS41-1), and one drought-sensitive genotype Moroc9-75. Seventeen genes were expressed exclusively in the two drought-tolerant genotypes under drought stress, and their encoded proteins may play significant roles in enhancing drought tolerance through controlling stomatal closure via carbon metabolism (NADP malic enzyme (NADP-ME) and pyruvate dehydrogenase (PDH), synthesizing the osmoprotectant glycine-betaine (C-4 sterol methyl oxidase (CSMO), generating protectants against reactive-oxygen-species scavenging (aldehyde dehydrogenase (ALDH), ascorbate-dependant oxidoreductase (ADOR), and stabilizing membranes and proteins (heat-shock protein 17.8 (HSP17.8) and dehydrin 3 (DHN3). Moreover, 17 genes were abundantly expressed in Martin and HS41-1 compared with Moroc9-75 under both drought and control conditions. These genes were likely constitutively expressed in drought-tolerant genotypes. Among them, 7 known annotated genes might enhance drought tolerance through signaling (such as calcium-dependent protein kinase (CDPK) and membrane steroid binding protein (MSBP), anti-senescence (G2 pea dark accumulated protein GDA2) and detoxification (glutathione S-transferase (GST) pathways. In addition, 18 genes, including those encoding Δl-pyrroline-5-carboxylate synthetase (P5CS), protein phosphatase 2C-like protein (PP2C) and several chaperones, were differentially expressed in all genotypes under drought; thus, they were more likely general drought-responsive genes in barley. These results could provide new insights into further understanding of drought-tolerance mechanisms in barley.
2009-05-13 | GSE15970 | GEO
Project description:Drought tolerance is a key trait for increasing and stabilizing barley productivity in dry areas worldwide. Identification of the genes responsible for drought tolerance in barley (Hordeum vulgare L.) will facilitate understanding of the molecular mechanisms of drought tolerance, and also genetic improvement of barley through marker-assisted selection or gene transformation. To monitor the changes in gene expression at transcription levels in barley leaves during the reproductive stage under drought conditions, the 22K Affymetrix Barley 1 microarray was used to screen two drought-tolerant barley genotypes, Martin and Hordeum spontaneum 41-1 (HS41-1), and one drought-sensitive genotype Moroc9-75. Seventeen genes were expressed exclusively in the two drought-tolerant genotypes under drought stress, and their encoded proteins may play significant roles in enhancing drought tolerance through controlling stomatal closure via carbon metabolism (NADP malic enzyme (NADP-ME) and pyruvate dehydrogenase (PDH), synthesizing the osmoprotectant glycine-betaine (C-4 sterol methyl oxidase (CSMO), generating protectants against reactive-oxygen-species scavenging (aldehyde dehydrogenase (ALDH), ascorbate-dependant oxidoreductase (ADOR), and stabilizing membranes and proteins (heat-shock protein 17.8 (HSP17.8) and dehydrin 3 (DHN3). Moreover, 17 genes were abundantly expressed in Martin and HS41-1 compared with Moroc9-75 under both drought and control conditions. These genes were likely constitutively expressed in drought-tolerant genotypes. Among them, 7 known annotated genes might enhance drought tolerance through signaling (such as calcium-dependent protein kinase (CDPK) and membrane steroid binding protein (MSBP), anti-senescence (G2 pea dark accumulated protein GDA2) and detoxification (glutathione S-transferase (GST) pathways. In addition, 18 genes, including those encoding Δl-pyrroline-5-carboxylate synthetase (P5CS), protein phosphatase 2C-like protein (PP2C) and several chaperones, were differentially expressed in all genotypes under drought; thus, they were more likely general drought-responsive genes in barley. These results could provide new insights into further understanding of drought-tolerance mechanisms in barley. Seven flag leaves of a replication for each genotype were harvested at 0 d, 1 d, 3 d and 5 d after reach 10% of AWC in the soil to constitute a single biological replicate. These flag leaves were employed for RNA isolation by using Trizol reagent following the manufacturer’s protocol (Invitrogen, Karlsruhe, Germany). The RNA was further purified using RNeasy Kit (Qiagen, Hilden, Germany). RNA yield and quality were determined by using an Agilent 2100 Bioanalyzer (Agilent Techologies, Boblingen, Germany). A table of the average, log2 RMA signal intensity values of three biological replicates for each Sample is linked below as a supplementary file.
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Project description:Gene expression profiling of drought-sensitive (IR64) and tolerant (N22) rice cultivars
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Project description:We investigated the physiological and gene expression response of drought-tolerant (IR57311 and LC-93-4; subgroup indica) and drought-sensitive (Nipponbare and Taipai 309; subgroup japonica) rice (Oryza sativa) cultivars to 18 days of drought stress in climate chamber experiments. Rice plants were grown under water sufficient and water limiting conditions in three independent experiments in a controlled climate chamber with 12 h day length at 600 µE m-2 s-1; temperature was 26°C in the light and 22°C at night, with a relative humidity of 75% in the light and 70% at night. Leaf material for expression profiling analysis was harvested five hours after the beginning of the light period after 18 days of stress or control treatment. Normalization and statistical testing was performed using the R package limma (R 2.3.1, limma version 2.7.3; (Smyth, 2005)). The methods Robustspline for within array normalization and Quantile for between array normalization were applied. A linear model with the effects genotype, drought treatment, genotype x drought, dye was fitted to normalized data in limma that models the systemic variation in the data. Afterwards, for the comparisons of interest, moderated t-statistics that use an empirical Bayes method were calculated. Differentially expressed genes were identified using the decideTests function (method global, Benjamini & Hochberg fdr corrected p-value <0.05 in limma) Keywords: stress reponse Growth design. Three independent experiments were performed. The design was a split-plot design with five blocks per drought or control treatment in each experiment. Each treatment and cultivar was represented by five replicate pots with one plant per pot. Pots were randomized within the blocks. Block position was rotated daily. Hybridisation design: A total of 28 samples, each representing mRNA from four parallel plants, were hybridized to 14 arrays. The hybridization design was optimized for the estimation of the effects of condition and the condition x cultivar interaction taking a variance minimization approach. In a two-step procedure, a smaller design for one sensitive and one tolerant cultivar was enlarged to encompass all four cultivars by integrating eight additional arrays. The optimization was programmed using R [R Development Core Team 2006] and carried out on the 16-node Beowulf Linux-Cluster at the University of Potsdam. R-scripts are available upon request and from the Bioinformatics Team at the MPI of Molecular Plant Physiology (see the website for more information). Allocation of the three biological replicates of each of the combinations of treatment and cultivar was completed such that a balanced distribution with respect to the labeling was achieved. Each combination of condition and cultivar from all experiments has been used at least once.
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