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提取梨 (PyrusserotinaRehd .)自交不亲和品种“二十世纪”(基因型为S2 S4 )、自交亲和的突变品种“奥嗄二十世纪”(S2 SSM4 ,SM =Stylar_partmutant;花柱部分突变 )及其亲和后代花柱的可溶性蛋白。经等电聚焦电泳 (IEF_PAGE)分析表明 ,“奥嗄二十世纪”及其后代花柱仍存在SSM4 蛋白 ,但其含量逐代减少 ,同时发现“奥嗄二十世纪”的SSM4 基因仅在柱头表达 ,而“二十世纪”的S4 基因表达的部位除了柱头外 ,还包括花柱上部及花柱下部 ,且表达量呈现从柱头到花柱下部下降的趋势。S蛋白经等电聚焦电泳的凝胶板进行RNase活性染色处理 ,也得到相同的结果。从花柱 (包括柱头 )中纯化出的S蛋白经SDS_PAGE电泳后进行RNase活性染色的结果表明 ,S4 与SSM4 蛋白的分子量相近 (约 30kD) ,并且均具有RNase活性。进一步以酵母RNA为基质测定的比活性也基本相等 ,约为 2 75U·min-1·mg-1蛋白。在离体条件下 ,上述两种S蛋白 (S_RNase)也以相同的程度抑制S4 或SSM4 花粉发芽及花粉管伸长。研究证明 ,自交亲和突变品种“奥嗄二十世纪”的SSM4 基因也具有原始自交不亲和品种“二十世纪”S4 基因的功能。因此 ,其自交亲和的原因可归结为SSM4 基因的表达量较少及SSM4 基因仅在柱头中表达的缘故。
The self-incompatibility mutant “Pythium triticina” (S2 SSM4, SM = Stylar_partmutant), a self-compatible mutant “Twentieth Century” (genotype S2 S4) ) And its affinity progeny style soluble protein. Analysis by isoelectric focusing electrophoresis (IEF_PAGE) showed that there was still SSM4 protein in the “嗄 嗄 twentieth century” and its descendants, but the content of SSM4 decreased gradually. At the same time, it was found that the SSM4 gene of 嗄 嗄 twentieth century was only expressed in stigma While in the twentieth century, the expression of S4 gene also included the upper part of the style and the lower part of the style in addition to the stigma. The expression of S4 gene decreased from the stigma to the lower part of the style. The same result was obtained when S protein was stained with RNase by isoelectric focusing electrophoresis gel plate. The results of RNase activity staining of S protein purified from style (including stigma) by SDS-PAGE showed that the molecular weight of S4 and SSM4 protein were similar (about 30 kD) and both had RNase activity. The specific activity of yeast RNA as a substrate was also about the same, about 2 75 U · min -1 · mg -1 protein. In ex vivo, the two S proteins (S_RNase) also inhibited pollen germination and pollen tube elongation of S4 or SSM4 to the same extent. Studies have shown that the SSM4 gene of the self-compatible mutant “Austrian Twentieth Century” also has the function of the S4 gene of the original self-incompatible variety “Twentieth Century”. Thus, the reason for their self-compatibility can be attributed to the low expression of the SSM4 gene and the expression of the SSM4 gene only in the stigma.