Project description:rs09-06_silencing-muntants - silencing mutants - Find endogenous targets of SDE3, SDE5 and IR71. - Lines IR71 KO n°3, smd8#25, sde3 (Col-0), sde5 (Col-0), Suc-SUL(Col-0) and Col-0 were grown for 11 days on MS solid medium, seedlings were then transferred in MS liquid medium and harvested 2.5 days after. Keywords: genotype comparaison 8 dye-swap - CATMA arrays
Project description:rs09-10_fwa - oct07. Impact of loss of DNA methylation and small interference RNAs (siRNAs) on gene expression. Mutants impaired for the DNA methylation maintenance pathway and for the RNA interference machinery were compared to mutants simultaneously impaired for both machineries to estimate their impact on genome activity. 8 dye-swaps. Gene knockout.
Project description:ra09-01_qtlleafgrowth - sg1-sg3 - What are the transcriptional consequences of the allelic contrast at SG3, SG1 and EGO3 loci mapped for its effect on shoot growth and identified at the gene level - Plants were grown in the in vitro conditions where we have mapped and cloned the concerned QTL and which allows us to see the phenotype linked to the segregation of the natural alleles. We are comparing 2 genotypes descending from a RIL from the Bur x Col RIL set, homozygous and identical everywhere in the genome except for the 23.4 kb at SG1 loci where one genotype is Bur (97.4 and 175.4) and the other is Col (97.1 and 175.1). 4 dye-swap - genotype comparison
Project description:gnp_blan06_torpten - rnai tor time course - The impact of the TOR pathway on growth and stress responses. Modification of translational and transcriptional profiles in Tor-inducible RNAi mutants and identification of TOR targets. 28 dye-swaps - normal vs. rnai mutant comparison.
Project description:au09-03_gamma-irradiation - 2 doses of ionising radiations (x-rays) - Metabolic pathways involved in the response of plants to ionising radiation treatment. - Arabidopsis thaliana (Col-0) seeds were grown in Petri dishes under sterile conditions until they had 2-rosette-leaves (on average 10 days). Then they received a dose of 10 or 40 gray of X-ray (Faxitron HP). Plant were harvested 2 and 26 hours after irradiation, immediatly frozen and stored at -80°C until RNA extraction (Rneasy plant mini Kit, Qiagen) . Experiment was carried in duplicates and a non irradiated control was done for each time. 8 dye-swap - dose response,time course
Project description:rs10-05_tcv - gene profiling of turnip crinkle virus (tcv) sirna - 1. What are the genes (including miRNA precursors) that are differentially regulated in a set of viral siRNA in A.thaliana? 2. There are also differentially regulated during an evolution and a fitness process? - This is a plant evolution project on TCV in which, the biological questions are: 1. What are the genes (including miRNA precursors) that are differentially regulated in Col0 and dcl234 mutant in wt conditions? 2. What are the genes (including miRNA precursors) that are differentially regulated in a set of viral siRNA in Col0 and dcl234 mutant? 3. There are also differentially regulated between the plant generations 1 (G1) and 11 (G11)? 4. What are the genes (including miRNA precursors) that are differentially regulated in a set of viral siRNA after fitness experiment? 24 dye-swap - gene knock out,treated vs untreated comparison
Project description:adt05-04_drn - drn/drn-l expressing cells vs non-expressing cells - DRN targets - In primordia timing (pt) mutant background DRN::erGFP transgenic cell culture (somatic embryos suitable to regenerate plants) has been protoplasted and protoplasts have been sorted for GFP+ and GFP- cells. Total RNA was extracted and sent to Evry for comparison (samples DRN a,b,c). DRN-L::erGFP transgenic cell culture has been protoplasted and protoplasts have been sorted for GFP+ and GFP- cells (samples DRN-L-2-1 and DRN-L-3-1). Keywords: null 5 dye-swap - CATMA arrays
Project description:ra10-04_roots - ws vs clca2/35a2/352.7 - What are the transcriptional adaptations induced by genetically modifying vacuolar nitrate transport - Plants were grown for 12 days after germination in vertical Petri dishes onto a 2mM NO3 medium (derived from Armengaud et al 2004) in long days (16h light at 65 µE). Samples consisted of pooled roots from at least 30 to 50 plants per sample. The experiment has been repeated 4 times (exp IFX1-IFX3-CPLA2-CPLA3)and samples were harvested around 11h in the morning (ie 3h after illumation). 10 dye-swap - gene knock out, genotype comparison, normal vs transgenic comparison
Project description:One of the key innovations of the flowering plants is their female reproductive organ, the carpel. Here we show that a mechanism controlling carpel margin development in the model flowering plant Arabidopsis thaliana was recruited from light-regulated processes. This recruitment followed the loss from the basic Helix-Loop-Helix transcription factor SPATULA (SPT) of a domain previously responsible for its negative regulation by phytochrome. We propose that the loss of this domain was a prerequisite for the light-independent expression in female reproductive tissues of a genetic module that also promotes shade avoidance responses in vegetative organs. Striking evidence for this proposition is provided by the restoration of wild type carpel development to spt mutants by low red/far-red light ratios, simulating vegetation shade, which we show to occur via PHYB, PIF4 and PIF5. Our data illustrate the potential of modular evolutionary events to generate rapid morphological change, and thereby provide a molecular basis for neo-Darwinian theories that describe this non-gradualist phenomenon. Furthermore, the effects shown here of light quality perception on carpel development lead us to speculate on the potential role of light-regulated mechanisms in plant organs that, like the carpel, form within the shade of surrounding tissues. The SPATULA (SPT) coding sequence was fused to the 5’-extremity of a sequence encoding the 75 C-terminal residues of the viral VP16 transcriptional activator and inserted into the pG0229-35S:GR plant transformation vector between the CaMV-35S promoter and sequences encoding the rat glucocorticoid receptor (GR), so as to conserve the entire reading frame. The resulting plasmid was transferred to Agrobacterium tumefaciens C58/pMP90 cells and used to transform A. thaliana Col-0 plants by standard methods. A homozygous, single-copy transformant was identified, from which two populations of ten T3 descendents were grown and treated by dipping of inflorescences for 2 min in cycloheximide (CHX; 10 µg/ml) containing Silwet L-77 surfactant (0.01% v/v). This treatment reduced translation to approximately 5% of its native level in inflorescence tissues, as measured by the in vivo incorporation of [35S]-methionine into proteins. One hour later, inflorescence tissues of the two plant populations were dipped for 2 min in CHX solution, as described above, with and without dexamethasone (DEX; 10 µM). After a further 2h, treated inflorescences, excluding open flowers, were harvested and pooled prior to RNA extraction for global gene expression analyses. Microarray analyses were performed using CATMA microarrays each containing 31776 gene-specific tags corresponding to 22089 A. thaliana genes. Three biological replicates were performed, based on three entirely separate induction experiments involving T3 plants from the same T2 parent. A transformed line containing a construction in which SPT had been replaced by an initiation codon and nuclear localization signal (NLS) was used to verify that the targets identified did not respond to the nuclear translocation of VP16-GR protein alone (35S:NLS-VP16-GR plants). In this NLS control experiment, two biological replicates were performed. One technical replicate with fluorochrome reversal was performed for each biological replicate. The labeling of cRNAs with Cy3-dUTP or Cy5-dUTP (Perkin-Elmer-NEN Life Science Products), the hybridization of these cRNAs to microarrays, and the subsequent scanning of microarrays, was performed as described by Lurin et al. (2004) Plant Cell 16, 2089-2103.
Project description:Transcriptomic study of lpp2 KO mutant in response to abcsisic acid - Fully developed rosette leaves were immersed for 1 min in a solution of 10-5 M ABA with 0.1% DMSO and 0.05%silwet. RNA are extracted after a 3h incubation. This treatment is compared to control (1 min immersion in 0.1% DMSO and 0.05%silwet) then 3h incubation. Keywords: treated vs untreated comparison 4 dye-swap - CATMA arrays