Unknown,Transcriptomics,Genomics,Proteomics

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Differentiation of Arabidopsis guard cells: analysis of the networks incorporating the basic helix-loop helix transcription factor, FAMA

ABSTRACT: In plants, formation of functional stomatal guard cells is a highly regulated event so that this cell differentiation model constitutes an interesting genetic system to explore cell differentiation in response to cell cycle controllers or cell fate determinants such as transcription factors. The latest acting bHLH in the stomatal lineage circuit is FAMA that appears to be absolutely required to promote stomatal development by controlling the GMC to GC transition. In order for FAMA to trigger guard cell development, it probably initiates a gene activation cascade leading to cell differentiation and cessation of cell division. To identify and characterize FAMA inducible genes, transcriptome analysis using the Affymetrix ATH1 micro-array chip was carried out on inducible FAMA gain of function plants at 4hrs and 48 hrs post-induction. Transgenic plants with an estrogen-inducible FAMA expression cassette were used in a timecourse experiment to identify genes that were differentially regulated by FAMA. Two time points were analyzed, 4 hours and 48 hours post-induction. Seedlings were grown on vertically oriented plates in a 22 Â°C incubator under 24 hours light for 5 days. Plants were then transfered with forcepts to plates containing MS + Sucrose + 10 ÂµM estrogen (or new MS + sucrose plates for controls). After 4h or 48h on new plates, samples were collected, frozen in liquid nitrogen and stored at â??80 degree until enough material was obtained to do all RNA extractions in the same week. The Rneasy Plant Mini Kit (QIAGEN) for RNA isolation, and samples were labelled with standard kits, etc (get info) and hybridized to Affymetrix Ath1 array chips. 15 samples.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Dominique Bergmann

PROVIDER: E-GEOD-21786 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Drought and salt stress severely inhibit plant growth and development. However, the regulatory mechanisms of plants in response to these stresses are not fully understood. Here we find that the expression of a WRKY transcription factor WRKY46 is rapidly induced by drought, salt and oxidative stresses. Mutations of WRKY46 by T-DNA insertion lead to more sensitive to drought and salt stress, whereas, overexpression of WRKY46 exhibits hypersensitive in soil culture with higher water loss rate, but increased tolerance on the agar plates. ABA induced stomatal closing is impaired in the WRKY46 overexpressing line (OV46), which is potentially due to the lower ROS accumulation in the guard cells. Real-time qPCR and GUS activity assay further demonstrate that WRKY46 is expressed in guard cells, but its expression is not affected by dehydration treatment, suggesting different regulatory mechanisms for WRKY46 between guard cells and other WRKY46 expressed tissues. The stomatal movement and conductance assay indicate that WRKY46 is involved in light-dependent stomatal opening. Further microarray analysis reveals that WRKY46 regulates a set of genes involved in cellular osmoprotection and redox homeostasis under dehydration stress. Determinations of ROS and MDA content confirm its role in oxidative detoxification under stress. Furthermore, we find that WRKY46 modulates light-dependent starch metabolism in guard cells via regulating QQS gene expression. Taken together, we demonstrate that WRKY46 plays a role in modulating cellular osmoprotection and redox homeostasis under drought and salt stress, and functions independently in stomatal movement via regulating light-dependent starch metabolism and ROS levels in guard cells. We used microarrays to identify the certain downstream genes regulated by WRKY46 under normal and dehydration conditions. One-week-old Arabidopsis seedlings of wild-type Col-0 (WT) and WRKY46 overexpressing line (46T) with or without dehydration treatment for 1 h were harvested and used for RNA extraction and hybridization to Affymetrix Arabidopsis ATH1 microarrays. The experiment includes 3 biological replicates.

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Differentiation of Arabidopsis guard cells: analysis of the networks incorporating the basic helix-loop helix transcription factor, FAMA

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