Project description:Transcriptome changes in 35S::KLU shoots at stage 1.03 compared to wild-type (WT) Col-0 shoots 35S::KLU and WT seedlings were grown on 1/2 MS medium, and 5 seedlings of each were harvested at stage 1.03. Samples were obtained from three independent experiments, and both genotypes were grown on the same plate. Samples were frozen in liquid nitrogen and ground with a Retsch machine and 3-mm metal balls. RNA was extracted with TriZol (Invitrogen) and further purified with the RNeasy Mini Kit (Qiagen). DNA digestion was done on columns with RNase-free DNase I (Roche). One µg of pure RNA samples were hybridized to Affymetrix ATH1 arrays at the VIB Microarray Facility (Leuven, Belgium).
Project description:Characterization of the impact of TT2 or MYB5 overexpression on gene expression. Col 0 plants were compared to 35S::TT2 and 35S::MYB5 overexpressors in control conditions
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:The Arabidopsis cytochrome P450 gene KLU (also known as CYP78A5) is predicted to produce a mobile molecule that has tremendous effects on the plant growth. To gain insight the mechanism of KLU affects plant growth, we use the RNA-seq to profile the transcriptome changes across wild-type,klu mutant, and overexpression of KLU.
Project description:Transcriptional profiling of Arabidopsis transgenic 35S:AtMYB102-SRDX plant under MS medium containing 200mM NaCl for 24h. Transgenic Arabidopsis (background : Col-0) overexpressing AtMYB102 fused to an active transcriptional repression domain (SRDX) showed strong salt tolerance.
Project description:Transcript profile of apices of 20 days-old Arabidopsis plants over expressing miR396b. We used ATH1 Affymetrics microarrays to obtain the transcription profile of plants overexpressing miR396b. We compared the transcriptome of 35S:miR396b plants vs wild-type plants (Col-0 accession), using ATH1 Affymetrics microarrays. For these experiments we collected vegetative apices together with the leaf primordia from plants of 20 days growth in short-day. For each genotype (Col-0, 35S:miR396b) we collected two samples.
Project description:Cd levels in the shoots, as well as in the roots were unexpectedly reduced in 35S:AtHMA4-expressing tobacco. Obtained results indicate that in the generation of the Cd-related phenotypes of transgenic plants substantial modifications of the host plant transcriptome was involved. Microarray based analysis was performed to compare expression profiles of the roots from tobacco expressing 35S-AtHMA4 with the wild-type (WT) plants, which were grown in the presence of 0.25 µM Cd. An effort was undertaken to understand which processes were modified in tobacco as a result of the expression of 35S:AtHMA4, which lead to decreased Cd uptake and lower accumulation in the shoots. Knowing underlying mechanisms is important for developing strategies to grow low cadmium tobacco.
Project description:The Arabidopsis cytochrome P450 KLUH (KLU)/CYP78A5 promotes organ growth in a non-cell autonomous manner. To identify genes regulated by KLU activity, homozygous klu-2 mutants carrying constructs for EtOH-inducible overexpression of wild-type KLU (35S::AlcR-AlcA::KLU) or of enzymatically inactive KLU protein (35S::AlcR-AlcA::KLUmut) were induced with EtOH and sampled at 90 min and 240 min after induction for gene expression changes. Keywords: Time course, genetic modification
Project description:A novel cold-inducible GSK3/Shaggy-like kinase cDNA (TaSK5) was isolated from winter wheat by a macroarray-based differential screening approach. Sequence analysis of TaSK5 revealed high similarity to Arabidopsis subgroup I GSK3/Shaggy-like kinases ASK-alpha, ASK-gamma and ASK-epsilon. Transgenic Arabidopsis plants overexpressing TaSK5 cDNA under the control of CaMV 35S promoter showed enhanced tolerance to salt and drought stresses. In contrast, the tolerance of the transgenic plants to freezing stress was not altered. To identify genes which are differentially regulated in the 35S:TaSK5 over-expressing Arabidopsis plants under non-stress conditions, we compared the genome-wide expression profiles of Col-0 and plants over-expressing TaSK5 using DNA microarrays. Sixty seven genes were found to be expressed at least 2-fold more strongly in 35S:TaSK5 plants than in Col-0, and 17 genes were found to be expressed at least 2-fold more strongly in Col-0 than in 35S:TaSK5 plants. Most of the TaSK5 up-regulated genes were also induced by abiotic stresses, including cold, salt and drought. These results support the involvement of TaSK5 in abiotic stress signal transduction. Keywords: transgenic vs wt Col.-0 comparison