Cellular and moleculr biological study of self-incompatibility in plants

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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.