Gene trap strategy, an effective tool for identification of novel genes expressed in anther tissues in Arabidopsis thaliana

Abstract

Analysis of GUS expression patterns was previously carried out for 20,000 gene trap lines of Arabidopsis thaliana generated by the Kazusa DNA Research Institute. This analysis indicated that 51 lines showed GUS expression within stamens. In this paper, we further focused on 42 of these lines in order to examine the detailed expression patterns at different anther developmental stages. Seven lines showed GUS expression exclusively in the tapetum during the early stage of anther development, thirty lines in pollen grains at the tricellular pollen stage and five lines in filaments at the tricellular pollen stage. We identified four trapped-genes by sequencing the adjacent fragments to T-DNA insertions. RT-PCR analysis confirmed that the mRNA expression patterns in these four genes were consistent with the expression pattern observed by GUS activity. Our results demonstrated that the gene trap system is an efficient strategy for identifying stamen specific genes in Arabidopsis.