Expression analysis of CaMsrB2-transformed rice (Oryza sativa L. cv. ILMI) without/with Bar selection marker under drought stress conditions
Ontology highlight
ABSTRACT: Methionine sulfoxide reductases catalyze the reduction of MetSO back to the correct Met residue. Previously, the gene of Capsicum annuum methionine sulfoxide reductase B2 was isolated and CaMSRB2-overexpressing tomato shows enhanced growth, which may trigger increased resistance to the pathogens. To assess the role of this enzyme in rice, we generated transgenic lines under the control of the rice Rab21 (responsive to ABA protein 21) promoter with/without Bar marker gene. Several physiological tests such as MV and Fv/Fm, indicators of an oxidative stress-inducing agent and a potential maximal PSII quantum yield, respectively strongly suggested CaMSRB2 confers drought tolerance to rice. Using 3M-bM-^@M-2-tiling microarray covering the whole rice genes, we carried out genome-wide expression analyses with CaMsrB2-transformed rice (Oryza sativa L. cv. ILMI). Rice was grown in port for six weeks and treated with drought by water withholding for two days. A total of 15 chips were used for the microarray experiment. RNA was extracted from plants just before and 2 days after the duration of water withdrawal for the control and the comparison, respectively. Experiments were performed with three or two biological replicates.
Project description:Using 3′-tiling microarray covering the rice genes, we carried out genome-wide expression analyses of total and polysome-fractionated samples treated with drought, salt and cold at 2-week-old rice leaves. compared to non-treated negative control (NC) samples. Our study is the first report which elucidated the global alternation of polysome association in response to abiotic stress. A total of 24 chips were used for microarray. At 2h, total and polysomal RNAs were extracted from drought, salt and cold at 2-week-old rice leaves with three biological replicates.
Project description:Rice tungro bacilliform virus (RTBV) is determinant of the rice tungro disease symptom. We compare the gene responses by RTBV infection between RTBV susceptive (TN1) and resistant cultivar (TW16). TN1 infected with RTBV shows the mild stunting and leaf yellowing, although TW16 shows only mild stunting at only early infection stage, and recovers the symptoms. Keywords: virus infection, disease response Comparison between RTBV and mock infected rice. Biological replicates: 3 control, 3 infected, independently grown and harvested. 1 samples derived from 5 plants grown under same conditons
Project description:Detailed information: Rice (*Oryza sativa* L. cv. Nipponbare) is a drought-susceptible species which is well suited for studies of abiotic stress response because of the comprehensive bioinformatics resource available. By withholding water from the entire root system of young rice plants, or half the root system only, it was possible to infer the relative impact of signals arriving from roots growing in wet and dry soil on the shoot proteome. The global proteome of shoots had 685 proteins in common to all three drought treatments but there were major shifts in abundance of individual proteins within 16 functional categories. The dominant changes were analyzed more deeply. First, we investigated transport and cell component organization, where some proteins were up-regulated by drought but many more down-regulated. Proteins involved in protein metabolism were up-regulated in general by drought when they were responsible for protein degradation but those involved in protein synthesis were down-regulated when water was withheld. Stress-related proteins behaved very consistently by increasing in droughted plants but notably some proteins were most abundant when roots of the same plant were growing in both wet and dry soil. This suggests that drought signals are complex interactions and not simply the additive effect of water supply to the roots. Changes in carbohydrate-processing proteins were consistent with the passive accumulation of soluble sugars in shoots under drought, with hydrolysis of sucrose and starch synthesis both enhanced. Data analysis information: The result raw files were converted to mzXML format and processed through the global proteome machine (GPM) software (version 2.1.1) of the X!Tandem algorithm (freely available at http://www.thegpm.org). The 16 gel fractions were processed serially for each experiment and the output files were generated as non-redundant, merged files with protein identifications with log (e) values less than -1, for each individual gel fraction. A protein database compiled from NCBI *O*. *sativa* with 26938 protein sequences (August 2011) was used in GPM to search the tandem mass spectra; the database also included common trypsin and human peptide contaminants. False discovery rates (FDR) were evaluated by searching against a reversed sequence database. Search parameters included MS and MS/MS tolerances of +2 Da and +0.2 Da, carbamidomethylation of cysteine as fixed modifications, oxidation of methionine as variable modifications and tolerance of two missed tryptic cleavages and K/R-P cleavages.
Project description:Jasmonates is inductively produced as a major plant hormone responsible for defense reactions in plants against both biotic and abiotic stresses, such as pathogen infection and mechanical wounding. We identified JA-inducible genes in the wild-type rice leaves 0 - 4 h after JA treatment using 44k microarray. Expression profiling in the wild-type rice leaves treated with jasmonic acid for 0.5, 1, 2, and 4 h was compared with that in the untreated wild-type rice leaves using two-color method with three biological replicates.
Project description:Methionine sulfoxide reductases catalyze the reduction of MetSO back to the correct Met residue. Previously, the gene of Capsicum annuum methionine sulfoxide reductase B2 was isolated and CaMSRB2-overexpressing tomato shows enhanced growth, which may trigger increased resistance to the pathogens. To assess the role of this enzyme in rice, we generated transgenic lines under the control of the rice Rab21 (responsive to ABA protein 21) promoter with/without Bar marker gene. Several physiological tests such as MV and Fv/Fm, indicators of an oxidative stress-inducing agent and a potential maximal PSII quantum yield, respectively strongly suggested CaMSRB2 confers drought tolerance to rice. Using 3′-tiling microarray covering the whole rice genes, we carried out genome-wide expression analyses with CaMsrB2-transformed rice (Oryza sativa L. cv. ILMI). Rice was grown in port for six weeks and treated with drought by water withholding for two days.
Project description:Methionine sulfoxide reductases catalyze the reduction of MetSO back to the correct Met residue. Previously, the gene of Capsicum annuum methionine sulfoxide reductase B2 was isolated and CaMSRB2-overexpressing tomato shows enhanced growth, which may trigger increased resistance to the pathogens. To assess the role of this enzyme in rice, we generated transgenic lines under the control of the rice Rab21 (responsive to ABA protein 21) promoter with/without Bar marker gene. Several physiological tests such as MV and Fv/Fm, indicators of an oxidative stress-inducing agent and a potential maximal PSII quantum yield, respectively strongly suggested CaMSRB2 confers drought tolerance to rice. Using 3′-tiling microarray covering the whole rice genes, we carried out genome-wide expression analyses with CaMsrB2-transformed rice (Oryza sativa L. cv. ILMI). Rice was grown in port for six weeks and treated with drought by water withholding for two days.
Project description:Plasma membrane NADPH oxidases (NOXs) are major producers of reactive oxygen species (ROS) in plant cells under normal growth and stress conditions. Rice NOXs have multiple homologs but their functional mechanisms are largely unknown. We used microarrays to detail the global gene expression profiles in rice wild-type (WT, Dongjin) and a mutant osnox2 which loss the functions of OsNOX2 protein under drought and identified distinct classes of genes between the two type rice plants under both normal growth and drought stressed conditions. The youngest fully expanded leaves from 2.5-month-old WT and osnox2 plants (three replicates each), grown under normal growth (soil moisture, 47.3%) and drought conditions (soil moisture, 8.5%), were used for RNA extraction and hybridization on Affymetrix microarrays. Control: normal growth condition; Drought: drought stress condition.
Project description:Rice, the worldM-bM-^@M-^Ys most important food crop, is attacked by multiple herbivores and pathogens.the rice striped stem borer (SSB) Chilo suppressalis is one of another most important rice insect pests. Here, we use Affymetrix Whole-Genome rice arrays to detect SSB infestation responsive genes. RNA samples from five damaged stems of SSB-challenged 24 h rice plants or five stems from unchallenged plants (for control samples) were used to array analysis. Two replicate biological experiments of SSB treatment rice arrays and one control samples array were peformed.
Project description:Jasmonates is inductively produced as a major plant hormone responsible for defense reactions in plants against both biotic and abiotic stresses, such as pathogen infection and mechanical wounding. Jasmonoyl isoleucine is known to be a bioactive compound of jasmonate and plays a pivotal role for plant defenses. We identified OsJAR1M-bM-^HM-^Rrelated JA-inducible genes in osjar1 tos17 mutant (osjar1-2) rice leaves 0 - 2 h after JA treatment using 44k microarray. Expression profiling in the wild-type rice leaves treated with jasmonic acid for 0, 0.5, 1, and 2 h was compared with that in the osjar1 mutant leaves treated with jasmonic acid for 0, 0.5, 1, and 2 h using two-color method with three biological replicates.
Project description:Rice is a critically important food source but yields worldwide are vulnerable to periods of drought. We exposed eight genotypes of upland and lowland rice (Oryza sativa L. ssp. japonica and indica) to drought stress at the late vegetative stage and harvested leaves for protein extraction and subsequent label-free shotgun proteomics. Gene ontology analysis revealed some differentially expressed proteins were induced by drought in all eight genotypes; we speculate that these play a universal role in drought tolerance. However, some highly genotype-specific patterns of response to drought suggest that some mechanisms of metabolic reprogramming are not universal. Such proteins had largely uncharacterized functions, making them biomarker candidates for drought tolerance screens.