Constructing and phenotypic analyzing an activation-tagging Arabidopsis mutant pool.

Abstract

Activation tagging method is an effective tool of obtaining gain-of-function mutant and investigating the gene function,which plays an important role in plant functional genomics study. In this paper,we used Arabidopsis Columbia wild type as material to construct an activation tagging mutant pool by Agrobacterium tumefaciens-mediated transformation,the binary vector pCB260 contained two Ds elements,one GFP report gene and one basta resistance selection genes,which show more convenient and efficient to screen the transgenic plant. Until now,over ten thousand transformed plants were generated. Among them,about 50 dominant mutants with obvious phenotypes were isolated,including early or late flowering time,unmoral leaf shape and flower,sterility and thin seed capsule color. T-DNA flanking sequences of ten special mutants were validated by TAIL-PCR and sequencing,whose T-DNA insertion fragments distributed in all five chromosomes of Arabidopsis genome,respectively.