Cellular and moleculr biological study of self-incompatibility in plants

为了探讨甘蓝型油菜自交不亲和系统的分子机制，本课题旨在研究与S基因座相关的基因的结构和功能。1.甘蓝型油菜自交不亲和性相关基因的分析。从甘蓝型油菜S8和S12品系柱头的cDNA文库中，克隆了S糖蛋白(SLG)和S受体蛋白(SK)基因，并建立了它们的序列。此外，还克隆了SLG12、SLG9和SRK9的基因组DNA，并对其进行了序列分析。2.SLG8反义基因的转化利用农杆菌介导法将SLG8的反义表达载体转化到白菜中。在获得的转化子中，自交不亲和性在反义基因的作用下被打破。3.SRK蛋白激酶结构域的表达。SRK蛋白的胞内结构域与GST融合表达。在体外系统中，表达的蛋白具有自我磷酸化活性。

To calrify the molecular mechanism of self-incompatibility system of Brassica, this project aimed the analysis of the structures and functions of genes presumed to be associated with the S-locus.1.Analysis of genes associated with self-incompatibility of Brassica.From the cDNA libraries of stigma of B.campestris S8 and S12 strains, clones encoding SLG (S-locus glycorotein) and SRK (S-receptor kinase) were isolated and their siquences were established. Furthermore, the genomic DNA of SLG12, SLG9 and SRK9 were also cloned ANd their sequences were analyzed.2.Transformation of the antisense gene of SLG.A antisense constract for SLG8 was transformed to a strain of B.rapa with Agrobacterium-mediated system. In a transoformant obtained, the self-incompatibility was broken down with the action of the antisense gene. The gene was actively inherited to hte next generation through the crossing.3.Expression of the kinase domain of SRK.The inner cellular domain of SRK,kinase domain, was expresses as a fusion protein with GST.The expressed protein rvealted the activity of auto-phosphorylation in an in-vitro system.