Inducible-targeted tagging system for localized insertional mutagenesis in arabidopsis thaliana

Abstract

The present study describes a system of inducible insertional mutagenesis based on the Ac-Ds family of transposons for targeted tagging in Arabidopsis thaliana. In this system, the Ac and Ds elements are carried within the same T-DNA and a heat shock inducible transposase fusion is utilized to control the levels of transposase gene expression, generating transpositions which can be subsequently stabilized without requiring crossing or segregation. I mapped 40 single copy lines by TAIL-PCR, which could be used as potential launch pads for heat shock mutagenesis. Using a starter line selected for detailed analysis, the efficiency of tagging over a 50-kilobase (kb) region in the genome was examined. Hits were obtained in the targeted genes with multiple alleles for most genes, with approximately equal numbers of hits detected in genes on either side of the T-DNA. These results establish the feasibility of our approach for localized saturation mutagenesis in Arabidopsis.