Expression profiling of soybean genes in response to drought
ABSTRACT: This SuperSeries is composed of the following subset Series: GSE29663: Expression profiling of soybean genes in response to drought (vegetative stage) GSE40604: Expression profiling of soybean genes in response to drought (reproductive stage) Refer to individual Series
Project description:Drought-responsive genes in soybean leaves were successfully identified using Affymetrix Soybean Gene 1.0 ST arrays on leaves samples of reproductive-stage soybean plants. R1 soybean plants planted in pots were imposed drought by withholding water for 5 days until the soil moisture content dropped to 5%, and 3rd trifoliates (now at the R2 stage) were collected for expression profiling. Soybean plants were grown in pots. When the plants reached the R1 stage (started flowering), drought treatment was imposed by withholding water. The soil moisture content was monitored during the process until the 5th day of water withholding, when soil moisture content reached 5%. The 3rd trifoliate (counting from shoots), now at the R2 stage, was collected for total RNA extraction, while other 3rd trifoliates of similar chlorophyl index were collected for leaves water content determination to identify the severity of the stress. Total RNA from 3rd trifoliates were used for expression profiling using Affymetrix Soybean Gene 1.0 ST arrays. Four biological repeats per treatment were performed, three biological repeats were chosen for expression profiling.
Project description:Drought-responsive genes in soybean leaves were successfully obtained using soybean gene 1.0 ST array. Leaf samples from the vegetative stage of soybean plants were used. V6 soybean plants planted in the pots were imposed drought by withholding water for 6 days until the soil moisture content drop to 5% and trifolium 4th were collected for expression profiling
Project description:Soybean (Glycine max) is susceptible to root rot when subjected to continuous cropping, and this disease can seriously diminish the crop yield. Herein, isobaric tag for relative and absolute quantitation (iTRAQ) labeling and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were employed for proteomic analysis of continuously cropped soybean inoculated with the arbuscular mycorrhizal (AM) fungus Funneliformis mosseae. Differential expression of proteins in soybean roots was determined following 1 year of continuous cropping. A total of 131 differentially expressed proteins (DEPs) were identified in F. mosseae-treated samples, of which 49 and 82 were up- and down-regulated, respectively. The DEPs were annotated with 117 Gene Ontology (GO) terms, with 48 involved in biological processes, 31 linked to molecular functions, and 39 associated with cell components. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis mapped the DEPs to 113 mainly metabolic pathways including oxidative phosphorylation, glycolysis and amino acid metabolism. Expression of glucan 1,3-beta-glucosidase, chalcone isomerase, calcium-dependent phospholipid binding and other defense-related proteins was up-regulated by F. mosseae, suggesting inoculation promotes the growth and development of soybean and increases disease resistance. The findings provide an experimental basis for further research on the molecular mechanisms of AM fungi in resolving problems associated with continuous soybean cropping.
Project description:To understand the responses of plants to environmental stresses will help mitigate the problems via creating stress-tolerant crop cultivars. We have carried out comparative expression analysis of roots of two soybean varieties Williams 82 and DT2008 that have constrasting drought-responsive phenotype under dehydration and well-watered (control) conditions. Affymetrix’s whole Soybean Gene Expression Microarray (66K) was used. The Williams 82 and DT2008 soybean plants were grown for 14 days in the vermiculite soil under greenhouse conditions. The whole plants of 14-d-old plants were detached and exposed to dehydration on KimTowel papers for 0 (well-watered, control), 2 and 10 h. All roots of independent 14-d-old plants were collected. Total RNA was prepared and used for the microarray hybridization. Three independent biological replicates were used for each plant sample.
Project description:Environmental stresses such as drought, salinity and both high and low temperature are frequently faced by crops all over the world and can be considered major limiting factors for plant geographical distribution and productivity. Breeding has allowed creation of crops more adapted to some of the adverse environmental conditions and to overcome the geographical limitation without major consequences to productivity. However, due to the climate changes observed in the last few decades, some agricultural areas have experienced the “green seed problem” characterized by chlorophyll retention in mature seeds. This is problematic in oil seed crops such as soybean and canola since it is related to lower seed and oil quality, resulting in serious financial losses. Besides the environmental factors, there are also genetic components controlling the susceptibility of different cultivars to green seed production. Understanding the molecular mechanisms controlling chlorophyll retention in seeds is crucial to allow advanced molecular breeding techniques and genetic engineering as a way to increase tolerance to this growing problem. We have used maturing soybean seeds of the cultivar MG/BR 46, harvested in R6, R7 and R8, produced under non-stressed and stressed environmental conditions, to understand he molecular basis of chlorophyll degradation and, consequently, its retention during soybean seed maturation. Soybean seeds were produced under stressed and non-stressed conditions and harvested in 3 different stages of maturation: R6 non-stressed, R7 non-stressed, R8 non-stressed, R6 stressed, R7 stressed, R8 stressed.
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:Genome-wide Transcriptional Analysis of Genes Associated with Drought Stress in Gossypium herbaceum root This experiment was designed to investigate the molecular mechanism associated with drought tolerance in root tissue of Gossypium herbaceum. The gene expression profiles of the root tissue using Affymetrix Cotton Genome Array were compared with drought tolerant and drought sensitive genotype of G.herbaceum under drought stress and watered condition. Many genes in various molecular function or biological processes were over- or under-represented between drought tolerant and sensitive genotype, suggesting various molecular mechanism and biochemical pathways are interlinked and tolerant genotype have developed multiple mechanisms as an adaptory behavior against drought stress. The transcriptional responses of root tissue in drought tolerant and sensitive genotype of Gossypium herbaceum under drought stress have been investigated. Physiological responses to drought stress, such as stomatal conductance, water use efficiency, root bending assay on different mannitiol concentration were also measured as indicators of imposed drought stress. Total RNA was isolated from root tissue from both genotype under drought stress and normal irrigated condition with three biological replicates
Project description:BiP overexpression confers resistance to drought, through an as yet unknown mechanism that is related to ER functioning, has been described that the delay in leaf senescence by BiP overexpression might relate to the absence of the response to drought. We used microarrays to detail the global programme of gene expression underlying in soybean leaves overexpressing BiP and with different water potentials to elucidate the mechanisms by which BiP protects plant cells against cell death induced by dehydration. BiP overexpressing plants and wild type as a control were subjected to dry slowly for 25 days. Third or fourth trifoliate leaves were collected from plants in water potential next 1.0 MPa, 1.5 MPa and 1.7 MPa for test weak and strong stress and subject RNA extraction and hybridization on Affymetrix microarrays maintaining the hybridization of two chips per treatment.
Project description:Water availability is the biggest single limitation on plant productivity worldwide. In Arabidopsis, adjustments to drought stress, involving changes in metabolism and gene expression drive increased drought tolerance and initiate diverse drought avoidance and escape responses. To address regulatory processes that integrate these complex responses we hypothesised that we needed to identify genes that govern early responses to drought. To this end, we produced a high-resolution time series transcriptomics dataset, coupled with detailed physiological and metabolic analyses of plants subjected to a slow transition from well-watered to the onset of drought conditions. 1825 differentially expressed genes (DEGs) were identified which showed no significant enrichment in gene ontology terms associated with dehydration responses and abscisic acid (ABA) regulation, confirming that the gene expression time series had targeted events prior to severe drought stress. Initial changes in gene expression coincided with a drop in carbon assimilation, not the later increase in foliar ABA content. Thus the early physiological and gene expression responses to drought were not driven by changes in leaf ABA content. In order to identify gene regulatory networks (GRNs) linked to early events, we used Bayesian network modelling of differentially expressed transcription factor (TF) genes. This approach identified AGAMOUS-LIKE 22 as key hub gene in a TF GRN. AGL22 is involved in the transition from vegetative state to flowering. Loss of AGL22 expression affected flowering time and drying rate providing a link between early changes in metabolism and the subsequent initiation of developmental responses to stress that govern plant productivity. A novel experimental design strategy (A Mead et al, in preparation), based on the principle of the “loop design”, was developed to enable efficient extraction of information about key sample comparisons using a two-color hybridization experimental system. With 108 distinct samples (four biological replicates, at each of 13 time points in control and droughted conditions, plus a shared time zero set) to be compared, the experimental design included 216 two-color microarray slides, allowing four technical replicates of each sample to be observed. One third of the slides were devoted to assessment of changes in gene expression between time points, using a simple loop design to link samples from time points across the sampled times, directly comparing samples collected on adjacent sampling times (i.e. day 1 with day 2, day 2 with day 3, etc.). With the remaining slides, four separate loops were constructed comparing treatments, biological replicates and sampling times. All of the samples in the microarray experiment were Col-0.
Project description:Systems responses of mature leaves from 4 reference cultivars of a larger collection of European potato cultivars (Solanum tuberosum L.) are investigated by metabolome profiling and RNA-Sequencing. The chosen reference cultivars, Milva, Alegria, Desiree, and Saturna, vary in ascending order in regard to drought tolerance. Systems analyses are based on 3 independent field trials and 3 paralleled greenhouse trials. Robust responses across all cultivars and conditions to natural seasonal drought stress comprise proline, raffinose, galactinol, arabitol, arabinonic acid, chlorogenic acid, and 102 transcripts which consist to a high proportion of heat shock proteins and genes with signaling or regulatory functions, such as a homolog of abscisic acid receptor PYL4. Constitutive differences of the tolerant cultivars, Desiree and Saturna, compared to the sensitive cultivars include arbutin (hydroquinone-beta-D-glucopyranoside), octopamine (p-hydroxyphenylethanolamine), ribitol and 248 differential transcripts. Many of these transcripts are disease related, receptor kinases, or regulatory genes, for example a homolog of the Arabidopsis FOUR LIPS MYB-regulator of stomatal cell proliferation. Functional enrichment analyses imply that heat stress is a major acclimation component of potato leaves to agronomical relevant drought stress. Enhanced leaf heat stress is a result of drought caused by loss of transpiration cooling. This effect and CO2-limitation are the main dilemmas of drought- or ABA-induced stomatal closure. Constitutive differences between tolerant and sensitive cultivars indicate partially synergistic interactions of drought and biotic stress responses. We suggest that drought tolerance of the potato reference cultivars may be caused by general resistance mechanisms which are part of previously selected pathogen tolerance. Transcriptome profiling by RNA-sequencing of 48 leaf samples from 4 potato cultivars grown under control or drought stress conditions in 6 independent experiments