Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Control, low water potential (drought) stress and stress release transcriptome analysis of mutants impaired in proline (Delta1-pyrroline-5-carboxylate synthase1-4; p5cs1-4) or abscisic acid (aba2-1) synthesis.

ABSTRACT: mRNA sequencing was used to identify genes differentially expressed in Delta1-pyrroline-5-carboxylate synthase1-4 (p5cs1-4) mutants deficient in low water potential-induced proline accumulation and the Abscisic Acid (ABA) deficient mutant aba2-1 compared to Col-0 wild type. Three treatments were analyzed for each genotype: unstressed control, a 96 hour low water potential (-1.2 MPa) treatment and a stress-release treatment where seedlings were removed from the stress treatment and transferred back to control media for 10 hours. Approximately 100 genes were found to be significantly up-regulated in p5cs1-4 compared to wild type in both the control and stress treatments with an equal number down regulated. There was also a substantial transcription response to the stress release treatment which varied between wild type, p5cs1-4 and aba2-1. Wild type (Col-0), p5cs1-4 and aba2-1 Arabidopsis thaliana seedlings were germinated on agar plates and after seven days of growth, seedlings were transferred either to fresh plates of control media or to low water potential (-1.2 MPa) stress plates. Samples were collected 96 h after transfer (Control and stress treatments). At 96 h, additional seedlings were transferred from the stress treatment back to control agar plates and samples collected 10 h after this stress release. Three independent biological experiments and mRNA extractions were performed for each genotype/treatment (27 mRNA extractions. After quality check and quantitation, the mRNA samples were pooled to produce a set of 9 samples for sequencing.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Verslues Paul

PROVIDER: E-GEOD-75933 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:To better undersand the effects of drought stress on wheat developing seeds, the transcription profile of early developing wheat seeds under control and drought stress conditions were comparatively analyzed by using the Affymetrix wheat geneChip. Drought stress is a major yield-limiting factor for wheat. Wheat yields are particularly sensitive to drought stress during reproductive development. Early seed development stage is an important determinant of seed size, one of the yield components. We specifically examined the impact of drought stress imposed during postzygotic early seed development in wheat. We imposed a short-term drought stress on plants with day-old seeds and observed that even a short-duration drought stress significantly reduced the size of developing seeds as well as mature seeds. Drought stress delayed the developmental transition from syncytial to cellularized stage of endosperm. Coincident with reduced seed size and delayed endosperm development, a subset of genes associated with cytoskeleton organization was misregulated in developing seeds under drought-stressed. Several genes linked to hormone pathways were also differentially regulated in response to drought stress in early seeds. Notably, drought stress strongly repressed the expression of wheat storage protein genes such as gliadins, glutenins and avenins as early as 3 days after pollination. Our results provide new insights on how some of the early seed developmental events are impacted by water stress, and the underlying molecular pathways that can possibly impact both grain size and quality in wheat. Winter wheat cultivar Redland, PI 502907 (Triticum aestivum L.) was used for this study. Seedlings were vernalized at 4°C for 6 weeks and then transplanted to a one gallon pot of soil-sand mixture (3:1, v/v) and grown in a growth chamber under the following conditions: relative humidity, 50–70%; 16-h light/8-h dark photoperiod; 21°C daytime temperature and 18°C nights. Plants were watered regularly twice daily at the rate of 100ml/ per pot. Because, wheat has an asynchronous fertilization pattern for ovlues in the inflorescence, each floret needs to be specifically marked for timing the fertilization and stress induction. After spikes developed, unfertilized ovules were monitored and observed for the fertilization process. Closed wheat spikes with anthers outside were marked as fertilized. Drought stress was imposed 24h after the fertilization (HAF). Drought stress treatment was initiated by discontinuing watering on the drought treatment plants while control plants were regularly watered twice daily. Stress treatment was applied at 48 HAF and relieved at 96 HAF. The microarray study focuses on 24 HAF to 72 HAF in control and drought stress conditions. We started to impose drought stress 24HAF.

Dataset Information

Control, low water potential (drought) stress and stress release transcriptome analysis of mutants impaired in proline (Delta1-pyrroline-5-carboxylate synthase1-4; p5cs1-4) or abscisic acid (aba2-1) synthesis.

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