Co-operative Research on Genes Associated with Self-incompatibility

目的：为了解高等植物自交不亲和的分子机制，分析与自交不亲和有关的蛋白质和dna及其功能。结果：油菜属孢子体自交不亲和植物，克隆并测序了s -座糖蛋白（SLG）和s受体蛋白激酶（SRK）的多个基因。此外，还分离到了它们的相关基因SRA和SRB。遗传分析表明，前者与s位点相关，后者与s位点无关。基因组DNA分析表明，存在许多与SRK和SLG相关的基因痕迹。其中至少有一个基因与s位点有关。为了分析这些植物的s基因型，采用PCR方法设计了RFLP系统。使用该系统可以确定大多数s基因型。针对芸苔属植物的转化，对几个品种进行了转化和再生能力的试验。一些特殊的品种被发现对这一目的有用。在lpomea自交不亲和系统中，分离到了与芸苔SRK和烟草S-RNase相似的基因，但这些基因都没有与s位点连锁。从蛋白质的2d电泳分析中发现了与s基因型相关的特异性位点，并将进一步分析。对日本梨配子体系统进行了研究，分离出若干s - rnase，并确定了其氨基酸序列。我们还分离并分析了一个与s位点无关的类似RNase。分析了不同rna酶对日本梨花粉管伸长活性的影响。在百合的情况下，一个与cAMP相关的系统被认为可以抑制自花花粉在花柱中的伸长。

Aims : To understand the molecular mechanisms fo self-incompatibility systems of higher plants, proteins and the DNAs associated with the self-incompatibility were analyzed and their functions were studied.Results : As for Brassica plants, which is belonging to the sporophytic self-incompatibility, several genes of SLG (S-locus glycoprotein) and SRK (S-receptor protein kinase) were cloned and sequenced. Furthermore their related genes, SRA and SRB were also isolated. Analysis of the inheritance of these genes revealed that the former groups are linked to S-locus and the latters are not. Analysis of the genomic DNA indicated that so many traces of the related genes to SRK and SLG are present. At least one gene among them was linked to S-locus. To analyze the S-genotypes of the plants, an RFLP system using a PCR method was devised. Using this system most of the S-genotypes can be determined. For the transformation of Brassica plants, several varieties were tested for the ability of transformation and regeneration. Some specific varieties were revealed useful for this purpose. In the self-incompatibility system of lpomea, similar genes with SRK in Brassica and S-RNase in tobacco were isolated, but these genes were not linked to S-locus. From protein analysis with 2D-electrophoresis specific spots linked to S-genotypes were found and will be analyzed further. For a gametophytic system, Japanese pear was studied and several S-RNases were isolated and their amino acid sequences were established. A similar RNase but not linked to the S-locus was also isolated and analyzed. The effects of various RNases were analyzed for their activity of pollen tubes elongation of Japanese pear. In the case of lilium, a system associated with cAMP was suggested to inhibit elongation of the self pollens throught the style.