摘要
以32份马铃薯或番茄晚疫病菌基因组DNA为模板,分析了琼脂糖凝胶电泳法和聚丙烯酰胺凝胶电泳法对10对SSR引物多态性检测的影响,同时比较了聚丙烯酰胺凝胶电泳两种银染法(常规银染法和快速银染法)的差异。结果显示,聚丙烯酰胺凝胶电泳较琼脂糖凝胶电泳分离效果好,具有更高的多态性检出率,其中快速银染法与常规银染法的检出结果相同,但常规银染步骤繁琐,整个过程要用1-2h,而快速银染只需约15min,且染色背景较浅,条带清晰。聚类分析显示,聚丙烯酰胺凝胶电泳快速银染法可将供试晚疫病菌株分布于更多的聚类组,显示出更高的遗传变异度。可见,聚丙烯酰胺凝胶电泳快速银染法结合SSR分子标记技术,可有效地用于马铃薯或番茄晚疫病菌群体遗传多样性分析。
Two gel electrophoresis systems（3% agarose gel electrophoresis and 12% polyacrylamide gel electrophoresis）and two coloration methods of polyacrylamide gel electrophoresis systems were used to comparatively analyze polymorphic level based on 10 SSR primers and 32 isolates of Phytophthora infestans.The results showed that the number of allele and gene polymorphism of 12% polyacrylamide gel electrophoresis（PAGE）were higher than that of 3% agarose gel electrophoresis.For the improved method of PAGE sliver staining,its background color was lighter,more strap clear,and it had same ratio of polymorphism and took only about 15 min,much faster than the conventional method of PAGE sliver staining which has complicated steps and will take 1-2 h.The results of cluster analysis revealed that 12% polyacrylamide gel electrophoresis with the improved method of sliver staining have a higher genetic diversity than that of 3% agarose gel electrophoresis with EB staining.With adoption of SSR marker techniques,the improved method of PAGE sliver staining could be effectively applied to assess the genetic diversity among isolates of Phytophthora infestans on potato or tomato.
出处
《生物技术通报》
CAS
CSCD
北大核心
2010年第11期171-176,共6页
Biotechnology Bulletin
基金
国家自然科学基金项目(30600401
30971892)
福建省自然科学基金项目(2009J01098
2010J01109)
福建省科技厅项目(2009R10028-3)
福建省财政专项-福建省农业科学院科技创新团队建设基金项目(STIF-Y07)
关键词
晚疫病菌
SSR
琼脂糖凝胶电泳
聚丙烯酰胺凝胶电泳
Phytophthora infestans Simple sequence repeat Agarose gel electrophoresis Polyacrylamide gel electrophoresis
