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T-DNA based gene tagging is an efficient method to isolate plant genes of interest. To facilitate the isolation of particular types of genes the tagging vectors may be provided with promoterless reporter gene constructs. Such promoter trap vectors allow screening for genes with particular type of expression pattern by assessing the expression of the reporter gene. We generated two promoter trap vectors during this project, one with the GUS reporter and one with a combined GUS-GFP reporter. Both of these were used to generate transgenic lines of Arabidopsis. The project resulted in generation of a large number of transgenic promoter trap lines (over 1000).
These lines have been tested for stress-induced reporter gene expression stress using mainly low temperature and oxidative stress treatment for the screens. Several transgenic lines with cold responsive GUS expression were generated as well as two lines were the GUS expression was responsive to paraquat (a herbicide generating oxidative stress).
In summary we have identified stress-induced genes of Arabidopsis using the promoter trap approach. The tagging of the genes will facilitate their subsequent cloning and, future use in studying the process of stress adaptation in plants.
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