| Description: | BACKGR: In mp leaves the vasculature is reduced, while other tissues seem unaffected. Transcript levels of vascular specific genes should therefore be specifically and dramatically reduced. Further, we have observed that a number of auxin inducible procambial genes are hyper auxin-inducible in 35S:MP plants. Differential profiling of mRNA pools from mp mutant versus 35S:MP cotyledon/young rosette leaves should provide a unique opportunity for the identification of vascular specific genes and auxin inducible genes. This approach is not covered by earlier experiments in mp mutant background, because it will not be restricted to rapid auxin responses and involves a 35S:MP background. RATION: The MP gene encodes an .auxin response. factor and a large number of auxin inducible genes are expressed at low levels in mp mutants and at excessive levels in 35S:MP plants.Northern analysis shows that this is also the case for procambial genes, reflecting the auxin inducibility of vascular differentiation. The experiment aims at identifying genes not rapidly auxin-induced in vascular differentiation. Because of the combined effects of the genotype specific anatomical differences and the genotype enhanced auxin inducibility, we expect extreme differences in transcript abundance. This could help overcoming limitations in assessing minor differences for a large number of genes, whose precise profiles can subsequently been sorted out on smaller arrays. TREATM/GENOT: We suggest assessment at 4 hour after application of 20uM IAA to 35S:MP cotyledons/young rosettes versus 0uM IAA mp mutant cotyledons/young rosettes. PRIOR/LINKS: single experiment/high priority. Open for interaction. TISSUE/RNA: RNA prepared by a senior post-doc, from immersed plants. We have >20 genes with known profiles to test each RNA sample. These internal controls may also help in further microarray optimization. SIGN: Identification of vascular genes is a worldwide enterprise, predominantly done in woody species. This approach in Arabidopsis is cost efficient and the further analysis of identified genes can take advantage of the enormous genomic resources available in Arabidopsis PERSP: Our long-term goal is understanding signaling in vascular development using mutant based profiles. DOC: We can provide quantified Northern data to demonstrate the feasibility of the approach. RELEASE: pref.3 months LIT: EMBO J 17, 1405-1411 |
NASCArrays Information at The BAR
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