Whole-Genome Analysis of SSB Larvae Infestation Regulated Rice Genes
ABSTRACT: Rice, the world’s 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: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:Infestation with white-backed planthopper (WBPH) to rice caused induced resistance to rice pathogens but brown planthopper (BPH) infestation induce weaker resistance to rice pathogens. We compared changes in gene expression in rice plants infested with WBPH and BPH to gain some insight into the WBPH-induced resistance to rice pathogens. An analysis, using microarrays, of gene expression in rice plants infested with these planthoppers revealed that WBPH infestation caused high induction of many defense-related genes including pathogenesis-related (PR) genes than BPH infestation. Furthermore, hydroperoxide lyase 2 (OsHPL2) which is an enzyme to produce C6 volatiles was induced by WBPH infestation, but not by BPH infestation. Experiment Overall Design: Agilent rice oligo microarray was used to investigate the gene expression profiling in rice plants infested with WBPH or BPH. Total RNA was extracted from pooled leaf blades infested with WBPH or BPH for 24 h and from mock-treated pooled leaf blades. Total RNA (200 ng) was labeled with Cy-3 or Cy-5 using an Agilent low RNA input linear amplification kit. Fluorescently labeled targets were hybridized to Agilent rice oligo microarrays. Hybridization and wash processes were performed according to the manufacturer’s instructions, and hybridized microarrays were scanned using an Agilent DNA microarray scanner. Agilent Feature Extraction software was employed for the image analysis and data extraction processes. Fold changes in expression level in each treatment were compared with those of the respective mock-treated controls. In each treatment, the experiment was performed independently three times.
Project description:High temperature markedly reduces the yields and quality of rice grains. To identify the mechanisms underlying heat stress-induced responses in rice grains, proteomic technique was used. Khao Dawk Mali 105 rice grains at the milky, doughy, and mature stages of development after flowering were treated at 40 °C for 3 days. Aromatic compounds were decreased in rice grains under heat stress. The protein abundance involved in glycolysis and tricarboxylic acid cycle, including glyceraldehyde 3-phosphate dehydrogenase and citrate synthase, was changed in milky and doughy grains after heat treatment; however, no changes in mature grains. The abundance involved in amino acid metabolism was increased in doughy grains, but decreased in milky grains. In addition, the abundance involved in starch and sucrose metabolism, such as starch synthase, ADP-glucose pyrophosphorylase, granule-bound starch synthase, and alpha amylase, was decreased in milky grains, but increased in doughy grains. A number of redox homeostasis-related proteins, such as ascorbate peroxidase and peroxiredoxin, were increased in developing rice grains treated with heat stress. These results suggest that in response to heat stress, the abundance of numerous proteins involved in redox homeostasis and carbohydrate biosynthetic pathways may play a major role in the development of KDML105 rice grains.
Project description:Plants require a distinctive cohort of enzymes to coordinate division and cell expansion. Proteomic analysis now enables interrogation of immature leaf bases where these processes occur. Hence we investigated proteins in tissues sampled from leaves of a drought-tolerant rice (IAC1131) to provide insights into the effect of soil drying on gene expression when compared with the drought-sensitive Nipponbare. Shoot growth zones were dissected to count dividing cells and extract protein for subsequent Tandem Mass Tags (TMT) quantitative proteomic analysis. Gene Ontology (GO) annotations of differentially expressed proteins provided insights into responses of Nipponbare and IAC1131 to drought. Soil drying did not affect the proportion of mitotic cells in IAC1131. More than 800 proteins across most functional categories were up-regulated in drought (and down-regulated on re-watering) in IAC1131, including those involved in organization of the meristem and subsequent cell formation. On the other hand, the proportion of dividing cells in Nipponbare was severely impaired during drought and fewer than 200 proteins responded in abundance when the growing zones underwent a drying cycle. However, those proteins involved in oxidation state and response to external stimuli were more likely to be upregulated by drought, even in Nipponbare.
Project description:Lipopolysaccharide is a Microbe Associated Molecular Pattern (MAMP) that is known to induce defense responses in plants. We have shown that treatment of rice leaves with Xoo LPS induces callose deposition, reactive oxygen production and enhances resistance against subsequent infection by the pathogen. We have performed transcriptional profiling of rice leaves that are treated with Xoo LPS to identify differentially expressed genes. Xoo LPS was injected into mid-veins of rice leaves and RNA was isolated 15 hours later.
Project description:An ERF transcription factor, Submergence-1A (Sub1A), dramatically enhances the tolerance to prolonged submergence in rice. For instance, rice accessions which lack Sub1A (e.g. M202) die within 7-10 d of complete submergence. By contrast, genotypes which posses Sub1A (e.g. M202(Sub1)) can endure submergence stress for 14 d. In this study, the two near isogenic lines with and without Sub1A were subjected to microarray analysis using Affymetrix Gene Chip technology. This analysis provided beneficial information to elucidate general response to submergence stress and to estimate Sub1A-dependent defense response to the stress at mRNA accumulation level. Aerial tissue of 14-d-old plants which were submerged for 24 h were subjected to RNA extraction and hybridization on Affymetrix microarrays. Non-submerged plants were used as control. Two independent biological replicates were analyzed for each treatment/genotype.
Project description:Comparative transcriptome analysis was performed to study differentially expressed genes (DEGs) between CK and Cocytodes caerulea Guenée challenged (CH) leaves of ramie.We obtained 40.2 and 62.8 million reads for CH and CK libraries respectively.De novo assembling of these reads generated 26,759 and 29,988 unigenes respectively. After a integrate assembly for all data of these two libraries, a total of 46,533 unigenes with an average length of 845 bp were obtained.A total of 1179 genes were identified as DEGs, 657 and 1230 of them were up- and down- regulated respectively, in response to C.caerulea infestation. Leaf samples of Cocytodes caerulea infested (T2) and un-infested (T1) ramie were RNA-Seq sequenced to compare differented expressed genes.
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. 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:Lipopolysaccharide is a Microbe Associated Molecular Pattern (MAMP) that is known to induce defense responses in plants. In rice we have shown that Xoo LPS induce callose deposition, reactive oxygen production and induced resistance response. The exopolysaccaride (EPS) secreted by Xoo might be involved in supressing these defense responses. We have performed transcriptional profiling of rice leaf gene expression changes after treatment with Xoo strains BXO1003 (LPS-, EPS-), BXO1002 (LPS+ EPS-) and BXO43 (wild type) along with milliQ treated leaves to identify the genes that are differentially expressed. RNA was isolated from mid veins of rice leaves 15 hours after injecting them with Xoo strains BXO1003 (LPS-, EPS-), BXO1002 (EPS-), BXO43 (wild type) or milli-Q water. The rice gene expression in each of the treatment was normalized based on the gene expression in the milli-Q treatment.
Project description:Two sets of wheat lines near-isogenic to Lr34 were used to compare gene expression profiles of wheat: 1. with and without Lr34 gene; 2. rust and mock inoculation; 3. distal and basal portion of the flag leaves. The two sets of wheat near-isogenic lines were used to subtract genetic background variations and to enrich Lr34-regulated gene expression profiles. The study is aimed to better understand the mechanisms of the well-known durable leaf rust resistance gene, Lr34, mediated resistance at the transcriptome level. Experiment Overall Design: Wheat near-isogenic lines, Jupateco with Lr34 (JUR), Jupateco without Lr34 (JUS), Thatcher with Lr34 (THR), and Thatcher without Lr34 (THS) were used. Thatcher lines were rust (I) or mock (M) inoculated. Jupateco lines were mock inoculated. Distal (T) and basal (B) half of the leaves were harvested and processed separately. Three biological replications were applied to each treatment.