小麦穗部组织EST-SSR标记引物开发及其在小麦遗传多样性分析中的应用

Development of SSR Markers Derived from EST of Wheat Spikes and Their Use in Genetic Diversity Analysis of Wheat

为了筛选光温敏雄性不育系（PTGMS）小麦穗部的EST—SSR分子标记，探讨其EST—SSR标记用于小麦品种遗传差异研究的可行性，用SSRSCAN软件对PTGMS小麦穗部的3264条EST序列进行SSR位点查找，结果得到108条含有SSR标记的序列。设计64对引物并进行PCR扩增及非变性聚丙烯酰胺凝胶（PAGE）电泳。结果显示，64对引物中有41对能在42个小麦品种中扩出PCR条带，有多态性条带的为36对，具有较高的扩增效率。聚类分析结果显示，42个小麦品种的遗传相似系数在0．64～0．87之间，3个PT—GMS小麦品种具有较高的遗传相似系数。

The object of this research were to develop EST-SSR markers based on ESTs from the spikes of photo-thermo sensitive genic male sterile wheat lines, and test their application in wheat ge- netic diversity analysis. About 3264 EST sequences of PTGMS wheat spikes were screened using SSRSCAN software to search for SSR motifs, and 108 SSRs were obtained. 64 pairs of primers were designed with the Primer 5.0 Plus software and the PCR products of these primers were detected by PAGE. The results showed that among the designed 64 EST-SSR primers, 41 amplified distinct bands and expected products in 42 different wheat, and 36 were polymorphic. The dendrogram showed that the genetic similarity coefficient in 42 wheat cultivars was between 0. 64 and 0. 87, and 3 PTGMS wheat had a high genetic similarity coefficient. Due to high efficiency in developing SSR markers on ESTs of PTGMS wheat spikes, this varieties and wheat genetic analysis, EST-SSR markers might be a great h moting the hybrid wheat breeding. method would be of great importance in identification of wheat and as the specific ESTs from wheat spikes, the development of elp on study of wheat flower development, the yield traits, pro