两种电泳方法在分析新陆早系列品种SSR标记结果中的比较与应用

Comparation and Application of the Two Electrophoresis on SSR of Earliness Cultivars in Upland Cotton in Xinjiang

为了研究两种电泳方法在新陆早系列材料SSR分析不同阶段使用效果的可行性,本研究利用2.5%琼脂糖凝胶和6%聚丙烯酰胺凝胶对新陆早系列材料SSR扩增结果进行检测。结果显示,两种电泳方式都能够得到较为清晰的扩增条带,但在获得的条带总数及多态性比率上有所差异。琼脂糖凝胶电泳共检测出30条条带,其中有18条为多态性条带,多态性比率为60%;而在聚丙烯酰胺电泳方法共检测出50条条带,多态性条带为33条,多态性比率为66%。基于两种电泳方法所获得的SSR标记位点,利用SPSS和PopGen32软件计算遗传相似系数,并构建遗传关系聚类图。分析结果显示,两种电泳方法所获得的聚类结果大致相同,在个别品种的分析结果上存在着一定程度上的差异。其中,聚丙烯酰胺凝胶获得的分析结果与实际遗传背景拟合性较好,所以,在最终的SSR标记分析过程中采用6%的聚丙烯酰胺凝胶进行分析效果更好,而琼脂糖凝胶电泳更加适合早期SSR标记引物的大量筛选。为新陆早系列材料SSR标记快速、准确分析提供了研究方法与技术支持。

In order to study the feasibility of two kinds of electrophoresis for analyszing, the result of SSR, the amplification production were detected by agarose gel electrophoresis and polyacrylamide gel electrophoresis respectively. The result showed that two electrophoresis methods were able to get clear amplified bands, but the number and polymorphic were different he band number that checked by agarose gel electrophoresis was 30, and the rate ofpolymorphism was 60%; The band number checked by polyacrylamide gel electrophoresis was 50, and the rate of polymorphism was 66%; Dendrograms demonstrating were constructed by SPSS16.0 and Popgen32 with the marker data, it was showed that the trend of dendrograms demonstrating analysis by two electrophoresis was same, but the result analyzed by polyacrylamide gel electrophoresis was nearly to the actual genetic background. So, the polyacrylamide gel electrophoresis was fitting to final SSR analysis, and agarose gel electrophoresis was suitable for early screening of primers. The result would provide a rapid, accurate analysis research method and technical support for SSR of Earliness Cultivars in Upland Cotton in Xinjiang.