Identification of transcription factors regulating plant secondary metabolism through the integration of functional genomics and metabolomics

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The aim of this project (a collaboration between IFR and JIC) is to identify new genes encoding transcription factors regulating secondary metabolism in plants. Because of the exceptional resources associated with the model plant, Arabidopsis thaliana, this work will focus on identification of the function of Arabidopsis transcription factors. Transcription factors offer exceptionally powerful tools to manipulate plant metabolism, either through genetic engineering or through marker assisted breeding. Specifically the project aims to: 1) Identify those transcription factors regulating secondary metabolism from a pre-selected pool of 38 candidate genes. Genes with duplicate functions in controlling secondary metabolism will be identified by systematic over-expression and metabolic fingerprinting primarily using LC/MS and NMR. 2) Following the identification of unique regulatory functions through over- expression and metabolic fingerprinting, the precise regulatory role of one representative of each unique activity will be determined using metabolic profiling (GC/MS, LC/MS, LC/MS/MS, LC/UV) of knock-out mutants compared to controls. Profiling will be guided by the fingerprinting data from the prior over-expression studies. 3) Integrate the metabolic consequences of regulatory gene over-expression and knock out to identify the regulatory roles of each sub-family of transcriptional regulators. 4) Identify the target genes for two distinct transcriptional regulators using whole genome transcript profiling of tissues either inhibited in activity of either transcription factor or over-expressing them, compared to controls. It is possible that if regulators of relatively uncharacterised pathways are chosen, new biosynthetic genes may be discovered during this part of the project.

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