Bioorganic Chemical Research on Self-Incompatibility of Plants.

1.抗S-和NS-糖蛋白抗血清的制备：用纯化的S_8或NS-糖蛋白免疫小鼠，获得抗血清。这些抗血清对S-和NS-糖蛋白具有相对特异性。并获得了抗S-糖蛋白的单克隆抗体。这些抗血清和抗体可用于研究S-和NS-糖蛋白在芸苔属植物中的分布以及这些糖蛋白在遗传杂交中的行为.用抗S-糖蛋白的抗血清，用免疫胶体金法分析了S-糖蛋白在油菜乳头细胞中的定位。金颗粒在成熟花的乳突细胞的细胞壁中积累。这一结果支持了S-糖蛋白参与花粉识别系统的假设.从B的柱头上。以油菜S_8菌株为材料，制备cDNA文库，用合成的DNA探针筛选S_8和NS糖蛋白的cDNA。获得了S_8-糖蛋白和NS-糖蛋白各一个克隆，并测定了NS-糖蛋白的全序列。NS-糖蛋白的位点独立于S-基因的位点，因此与自交不亲和性的表达无关。NS糖蛋白与B的S位点相关糖蛋白具有很高的同源性。在物种中高度保守的甘蓝。这些数据表明，NS-糖蛋白应该在物种间的识别中发挥一定的作用。

1. Preparation of antisera against S- and NS-glycoproteins : Mice were immunized with purified S_8 or NS-glycoproteins and antisera were obtained. These antisera were relatively specific to S- and NS-glycoproteins. Furthermore, a monoclonal antibody against S-glycoprotein was obtained. These antisera and antibody were useful in studying the distribution of S- and NS-glycoproteins in Brassica species and the behavior of these glycoproteins in genetic crossing.2. Using an antiserum for S-glycoprotein, the localization of S-glycoprotein in the papillar cells of Brassica campestris was analyzed by immunogold method. The gold particles were accumulated in the cell wall of the papillar cells of mature flowers. This result supports the hypothesis that S-glycoproteins should be involved in the recognition system of the self pollens.3. From the stigmas of B. campestris S_8 strain, the cDNA library was prepared and cDNAs of S_8- and NS-glycoproteins were screened by synthetic DNA probes. Each one clone for S_8- and NS-glycoprotein was obtained and the full sequence of NS-glycoprotein was determined. The locus of the NS-glycoproteins was independent from that of S-genes and therefore unrelated with the expression of self-incompatibility. The NS-glycoprotein was proved to be quite homologous with S-locus-related glycoproteins of B. oleracea which are highly conserved among the species. These data suggested that NS-glycoproteins should play some roles in the recognition among the species.