Fine Mapping Of An Arabidopsis Thaliana Gene Controlling Microspore Development And Cloning Of Male Sterile Gene Promoters From Arabidopsis

An Arabidopsis partial male-sterile mutant was isolated from a library of mutants mutagenized by enthyl methane sulfonate (EMS) treatment. Genetic analysis indicated that the mutant was controlled by a single recessive nuclear gene named PMS15-16-2-3.Cytological observations of the anther development of pms15-16-2-3 mutant and the wild-type plant showed that the mutant middle layer degenerated later, both the tapetal cell morphology and the tetrads were abnormal, which resulted in few fertile pollen grains formed in the anther. The mutant PMS15-16-2-3 gene was mapped to a region of 28-kb in BAC T24C20 on chromosome 3 using a map-based cloning strategy.There are 8 genes in the region including At3g48120 , At3g48130, At3g48140, At3g48150, At3g48160, At3g48170, At3g48180and ATM. The function of genes including At3g48120,At3g48130,At3g48140 and At3g48180 is unknown. At3g48130 has hign expression in the anther comparing to other three genes which have a litter expression in the anther. At3g48150 is a gene that is related to ubiquitin protein ligase that regulates mitotic progression and exit by controlling the stability of cell cycle regulatory proteins. At3g48160 encods E2F-like protein, an inhibitor of the endocycle.At3g48170 is a member of the ALDH gene superfamily of Arabidopsis. AtATM encodes a homolog of the human ATM gene, which is mutated in ataxia telangiectasia, a chromosome instability disorder. No genes involved in microspore formation were reported in this region besides AtATM, so we believe that PMS15-16-2-3 gene could be a new gene controlling microspore development in Arabidopsis. The promoter of male sterile gene ms1 was amplified by PCR from genome of Arabidopsis thaliana Columbia, which had the same sequence as published previously, and then cloned into T-vector. The expression vector was constructed by inserting the promoter into pCAMIBA-1301.The promotor-1301 was directly introduced into Agrobacterium tumetaciens LBA4404 which was used to mediate the transformation of promoter fragment into Arabidopsis thaliana. These two vectors was detected by PCR and digested with restriction endonucleases and were proved right. We had trans-gene plants of MS1,DEX1,FAD7and MYB32.This will provide further insight into the promotor especial expression in Arabidopsis thaliana and reciprocity with other male sterile gene.