Isolation and Characterization of Different Sensitive Mutants to UV-B Radiation from Activation Tagging Lines

Abstract

Activation tagging is another method for insertional mutagenesis. Because enhancers in T-DNA inserted into genome can activate transcription of nearby genes, it is expected that plant have dominant gain-of-function mutations and acquire resistant phenotype to environmental stress. We have created about 50,000 independent Arabidopsis lines transformed with a tetrameric enhancer T-DNA and performed functional analysis of genes of these lines in collaboration. We newly tried to isolate different sensitive mutants to UV-B radiation from this Arabidopsis activation tagging lines.
After 14 days plants of each line were irradiated with white light (6 W/m2) including UV-B (0.25 W/m2) for 35 hours, we searched plants that showed increased or diminished UV damage of appearance, especially bleaching of leafs, for 3 days under white fluorescent light in 16-h-light/8-h-dark cycles. As a result of selection of approximately 5,000 lines, we isolated some hypersensitive and resistant mutants to UV radiation.