Project description:We examined global expression profiles of 7-days old 35S-TrAP transgenic plants compared to Col-0 wild-type using an Affymetrix ATH1 GeneChip and identified 586 genes that are differentially expressed in the 35S-TrAP transgenic plants (q<0.005). Of these, 295 transcripts were elevated whereas 291 were reduced (Figure 2E). We performed real-time PCR and RNA blot assays to validate the microarray results for the differentially expressed genes (DEGs).
Project description:Keeping imbibed seeds at low temperatures for a certain period, so called seed vernalization (SV) treatment, promotes seed germination and subsequent flowering in various plants. Vernalization-promoting flowering requires GSH. However, the expression patterns analyzed by GeneChip arrays showed that increased GSH biosynthesis partially mimics SV treatment in Arabidopsis thaliana. SV treatment (keeping imbibed seeds at 4°C for 24 h) induced a specific pattern of gene expression and promoted subsequent flowering in wild-type plants. A similar pattern was observed at 22°C in transgenic plants (35S-GSH1 plants) overexpressing the γ-glutamylcysteine synthetase gene GSH1, coding an enzyme limiting GSH biosynthesis, under the control of the cauliflower mosaic virus 35S promoter. This pattern was strengthened at 4°C but flowering was less responsive to SV treatment. There was a difference in the transcript behaviour of the flowering repressor FLC between wild-type and 35S-GSH1 plants. Unlike other genes responsive to SV treatment, SV-dependent decrease in FLC in wild-type plants was reversed in 35S-GSH1 plants. SV treatment increased GSSG level in wild-type seeds, whereas GSSG level was high in 35S-GSH1 plants, even at a non-vernalizing temperature. Taking into consideration that low temperatures stimulate GSH biosynthesis and bring about oxidative stress, GSSG is considered to trigger low temperature response, but enhanced GSH synthesis was not enough for mimicking SV treatment. To complete it, it essentially required the cellular redox retransition from the oxidized to the reduced state that is observed after the seed vernalization treatment.
Project description:SLIM1 has a well established role in regulating transcriptional responses to sulfur deficiency in Arabidopsis thaliana. In order to investigate the impact of SLIM1 expression under sufficient nutrient conditions, we generated 35S::SLIM1 over-expression lines. SLIM1OX plants were found to have larger rosette area, bolt earlier, and enter developmental senescence earlier than Col-0 and slim1KO (slim1-cr) plants. RNA-seq followed by differential expression analysis was performed on rosette tissue at three timepoints.
Project description:We performed RNA sequencing of Pseudomonas syringae pv. tomato (Pst) DC3000-infected A. thaliana Col-0 and 35S::CBP60g plants at normal (23C) and elevated (28C) temperatures. 4-week-old plants were pre-incubated at 23C and 28C for 48h and then leaves were syringe-infiltrated with DC3000 bacterial inoculum. Plants were incubated at their respective temperatures (23C or 28C) for another 24h post-inoculation before tissue collection for RNA extraction. RNA samples for submitted for RNA sequencing and we found different clusters of DC3000-regulated genes that were similarly or differentially regulated between Col-0 and 35S::CBP60g at elevated temperature. Temperature-downregulated DC3000-induced genes in Col-0 plants that were restored in 35S::CBP60g plants were enriched for immunity/defense-related genes, including those essential for host salicylic acid (defense hormone) biosynthesis and accumulation.
Project description:Expression data from 7-day-old Arabidopsis emf2 mutant, rescued emf2 mutant harboring 35S::BoEMF2.1, WT Columbia ecotype and WT harboring 35S::BoEMF2.1 named transWT