利用EST-SSR通用性和EST数据库筛选三叶草EST-SSR分子标记

Screening of EST-SSR Marker using the Transferability of EST-SSR and the EST Database in Clover(Trifolium L.)

为加速分子标记在三叶草中的应用,利用三叶草EST数据库（GenBank/dbEST）和EST-SSR的通用性,开展了三叶草EST-SSR分子标记筛选及利用研究。从五种作物和三叶草的EST数据中共筛选出445对EST-SSR引物,挑选107对引物用于5个三叶草品种的遗传多样性分析,其中表现多态性的引物为103对（96.3%）,检测到的等位变异为293个,每对引物的等位变异数目为1-6（平均2.8）,PIC值最高、最低、平均分别为0.79、0.28和0.55。聚类分析表明,在相似系数0.618水平可分为三大类,类Ⅰ由海发、瑞文德和碧盛组成,类Ⅱ为杂三叶,类Ⅲ为红三叶。

To accelerate the application of molecular markers in clover,screening and utilization of clover EST-SSR markers were carried out using two ways of transferring from other crops and designing from clover EST itself.445EST-SSRs were developed by screening from 5crops and clover EST database itself.107EST-SSR primers were selected randomly to investigate the genetic relationship of the 5clover cultivars.Of which,103 pairs of primers（96.3%）showed polymorphism.293 alleles with an average of 2.8alleles per primer were found.The PIC（Polymorphism information contents）for each polymorphic primer varied from 0.28 to 0.79,with an average of 0.55.Clustering analysis indicated that the 5clover cultivars were clustered into 3groups based on 0.618 of GS.GroupⅠincluded white clover haifa,rivendel and zapican;Group Ⅱ and Group Ⅲ were alsike clover and red clover respectively.