葡萄EST-SSR引物的开发及部分种质聚类分析

Development of EST-SSR Primers and Cluster Analysis of Some Cultivars in Grape

借助SSRs软件查找葡萄属含有SSR的EST序列,从葡萄属EST-SSR数据库中开发新型的EST-SSR标记,进行可利用性筛选,并设计引物。共开发出28对引物,从中筛选到16对多态性引物对62份葡萄材料进行PCR扩增。计算相似系数并构建聚类树状图,通过聚类分析研究葡萄属不同品种之间的亲缘关系,进一步验证EST-SSR在葡萄种质分类鉴定中的可行性。结果表明:绝大多数材料间的相似系数大于0.74,62份供试材料在相似系数0.658处可分为4个类群,大多数亲缘关系较近的种质资源都聚集在一起,相似系数种内较种间大,与传统分类情况基本一致。

To develop the new EST-SSR marks from the EST-SSR database of grape germplasm,the statistic software of SSRs were utilized to search the sequences of EST containing SSRs,then the available were selected and the corresponding primers were designed and filtered. All the obtained 16 pairs of pleomorphic primers were used to carry out PCR amplification for 62 grape materials. Using UPGMA method to construct dendrogram based on the similarity coefficient calculated by the software of NTSYS. To invstigate the genetic relationships of grape germplasm resources and farther validate the feasibility of EST-SSR on classifying grape germplasm resources,the clustered result was analyzed. The result indicated that the similarity coefficient of majority germplasm was more than 0. 74,and 62 grape materials were classified into 4 groups at 0. 658. Most grape germplasm resources having close kinship were clustered together,and the similarity coefficient of intraspecies was bigger than interspecies. The result was consistent with that acquired from traditional taxonomy.