Project description:Cabernet Sauvignon shoot tip total protein. Treatments were control (well-watered) versus no watering. Samples were taken at 4, 8, 12, and 16 days following start of drought stress in treatment vines. Total protein was phenol extracted and Lys-C/trypsin digested using trifluoroethanol FASP method. Total peptides analyzed by gas phase fractionation using four fractions on an LTQ XL mass spectrometer (Thermo). A protein database was compiled from three sources 10 September 2011: 1) all reviewed V. vinifera protein entries in UniProt (151 sequences); 2) V. vinifera proteins predicted by the International Grape Genome Program (29749 sequences identified by UniProt search “Taxonomy:29760 AND author:vitulo AND reviewed:no”); 3) mitochondrial proteins associated in UniProt (81 non-redundant sequences). Spectrum-peptide matching was performed with X!Tandem and the GPM Cyclone (www.thegpm.org) in automated mode using MudPit merging. The GPM Cyclone XE and X!Tandem Cyclone version 2011.12.01.1. were used. Default ion trap parameters were used with the exceptions of MS error (+3, -1 Da), the inclusion of point mutations, the inclusion of reversed sequences, and a protein expect value of -1. Approximately 27,000 spectra per sample were assigned to peptides. Normalized spectral abundance factors (NSAF) were calculated according to (7). Protein identifications were filtered and protein and peptide FDRs calculated according to (8). Protein identifications were excluded if they did not matched at least one spectrum in all three biological replicates and at least a total of six spectra among the replicates.
Project description:Potted Cabernet Sauvignon vines in the greenhouse were exposed to irrigated controls, non-irrigated water-deficits, and saline treatments for 16 days. Plant shoot tips were harvested every 4 days (0,4,8,12, and 16 days) to measure the progression of changes of global gene expression due to the stress. PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Grant Cramer. The equivalent experiment is VV2 at PLEXdb. time: Day 0 - stress: Control(3-replications); time: Day 4 - stress: Control(3-replications); time: Day 4 - stress: Water-Deficit(3-replications); time: Day 4 - stress: Salinity(3-replications); time: Day 8 - stress: Control(3-replications); time: Day 8 - stress: Water-Deficit(3-replications); time: Day 8 - stress: Salinity(3-replications); time: Day 12 - stress: Control(3-replications); time: Day 12 - stress: Water-Deficit(3-replications); time: Day 12 - stress: Salinity(3-replications); time: Day 16 - stress: Control(3-replications); time: Day 16 - stress: Water-Deficit(3-replications); time: Day 16 - stress: Salinity(3-replications)
Project description:The drought-tolerant soybean variant HeiNong 44 was analyzed via proteome analysis. Soybean was exposed to drought stress for 0, 8, and 24 h, and protein samples were extracted for detection of differentially expressed proteins. Protein sequencing of leaf tissues under drought stress yielded a total of 549 differentially expressed proteins: 75 and 320 upregulated proteins as well as 70 and 84 downregulated proteins were obtained after 8 and 24 h of drought stress, respectively.
Project description:Constitutive active form of DREB2A, a transcriptional regulator involving to plant drought and high-salinity stress response, was overexpressed in Arabidopsis, and gene expression pattern was compared between transgenic plants and wild type plants.
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
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.
Project description:Three rice major tissues, namely flag leaf, shoot and panicle, were involved in this study. Each tissue had two kinds stress treatment, drought and high salinity, in 3 different time courses. For drought treated samples, an additional water recovery was applied. Each experiment had three replicates. Keywords: Comparison of gene expression in three tissues with stress treatment and without treatment To globally elucidate potential genes involved in drought and high-salinity stresses responses in rice, an oligomer microarray covering 37,132 genes including cDNA or EST supported and putative genes was applied to study the expression profiling of shoot, flag leaf, and panicle under drought or high-salinity treatment. Three rice major tissues, namely flag leaf, shoot and panicle, were involved in this study. Each tissue had two kinds stress treatment, drought and high salinity, in 3 different time courses. For drought treated samples, an additional water recovery was applied. Each experiment had three replicates.