Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of Arabidopsis cell cultures treated with ABA or lipids to identify diacylglycerol pyrophosphate regulated genes in ABA signaling

ABSTRACT: Identification of diacylglycerol pyrophosphate regulated genes in ABA signaling.

The specific plant phosphorylated form of phosphatidic acid (PA), diacylglycerol pyrophosphate (DGPP) was recently shown to be a second messenger in abscisic acid (ABA) signaling. The aim of the project is to identify among the set of ABA-regulated genes the ones also regulated by DGPP and/or PA.
Five ml of 3 days-old suspension cells was incubated with ABA or lipids for 3 h under the conditions of culture. Lipids were emulsified by sonication for 1 mi, four times, at 4°C, in one ml of culture medium then added to 4 ml suspension cells. Cells were filtrated under vacuum, frozen in liquid nitrogen and RNA extracted. Dioleoyl PA and dioctanoyl PA are from Sigma, dioleoyl DGPP and dioctanoyl DGPP are from Avanti Polar Lipids.

INSTRUMENT(S): Axon GenePix 4000A scanning hardware

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: ludivine taconnat

PROVIDER: E-MEXP-473 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Title : Characterization of genes differentially expressed in roots of transgenic arabidopsis lines expressing the p25 protein of beet necrotic yellow vein virus. Biological question : Rhizomania ("crazy root") is a severe disease of sugar beet caused by beet necrotic yellow vein virus (BNYVV), which is transmitted by the soil-inhabiting fungus Polymyxa betae. Symptoms of virus infection are characterized by a constricted tap root and a massive proliferation of fine rootlets that often undergo necrosis. BNYVV RNA-3 encodes a 25 kDa (p25) which is an important determinant of leaf symptom phenotype. It also governs BNYVV invasion of the plant root system and induction of rootlet proliferation in sugar beet. In order to obtain a better understanding of molecular aspects of disease development in roots and to characterize specific host genes involved in response to viral infection, transgenic Arabidopsis overexpressors of p25 viral protein was obtained and better characterized. It was shown that transgenic plants that efficiently expressed p25 protein produced more lateral roots. Comparative analysis (microarray) was performed between wild type Arabidopsis roots and transgenic Arabidopsis roots expressing p25 protein, in order to identify Arabidopsis genes differentially expressed in response to p25 viral protein. Experiment description: Seeds were surface sterilized, chilled at 4C for 4 days, and then germinated and grown on square Petri plates containing sterilized Murashige and Skoog (MS) medium with 1% sucrose. Such stock plates were arranged vertically in plastic racks and placed into growth chamber. After 5 days, plants were transferred carefully onto fresh MS medium big round plates. On each plate, 60 Wild Type (WT) plantlets were transferred on the half right of the plate, and 60 transgenic plantlets (B, E or T lines) were transferred on the half left of the plate. Plates were arranged horizontally and placed into growth chamber. Experiment 1 : 5 plates containing WT0A control plants and B0A transgenic plants. Experiment 2 : 5 plates containing WT1 control plants and B transgenic plants. 5 plates containing WT2 control plants and E transgenic plants. 5 plates containing WT3 control plants and T transgenic plants. Plants were harvested after 7 days (experiment 1) or 12 days (experiment 2), and WT roots or transgenic roots were pooled and conserved at -80C.

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Transcription profiling of Arabidopsis cell cultures treated with ABA or lipids to identify diacylglycerol pyrophosphate regulated genes in ABA signaling

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