三疣梭子蟹多态性微卫星DNA标记的筛选及评价

Screening and evaluation of Portunus trituberculatus polymorphic microsatellite DNA markers

根据前人FIASCO方法构建的三疣梭子蟹微卫星富集文库,对含微卫星的DNA序列设计了71对引物并对其进行了多态性筛选,筛选出21对多态的微卫星引物,对三疣梭子蟹1个野生群体的30个个体进行遗传多样性检测,同时对引物进行评价。21个位点共获得了188个等位基因,平均每个位点扩增得到8.9个等位基因。不同引物获得的等位基因数差异较大,从3～13个不等,其中Pot8、Pot37、Pot48、Pot53、Pot54、Pot66六个位点分别获得了11、12、12、11、13、11个等位基因,而Pot46仅获得了3个等位基因。等位基因的大小分布在131～312bp,基本符合引物设计时理论产物长度。21个微卫星位点的期望杂合度的范围为0.659～0.889,PIC值均高于0.5,表明它们都有很高的杂合度,均可用于三疣梭子蟹种群遗传结构分析,为三疣梭子蟹品种选育、种系评估提供更多的微卫星DNA信息。

A microsatellite enriched library of P. trituberculatu was constructed following the FIASCO method. Seventy-one pairs of primers were designed according to the sequences containing microsatellite motifs. Twenty-one loci were polymorphic after the evaluation of 71 loci based on genetic diversity analysis of a wild population comprising of 30 individuals for. A to- tal of 188 alleles were obtained with an average of 8.9 alleles per locus. The number of alleles per locus amplified was significantly different, ranging from 3 to 13. The number of alleles obtained for the loci Pot8, Pot37, Pot48, Pot53, Pot54, Pot66 was 11,12,12,11,13 and 11, respectively, and Pot46 got only three alleles. Allele size distribution was in the range of 131 312bp, which is basically in line with the design theory of primer product length. Twenty-onemicrosatellite loci of expected heterozygosity were in the range of 0. 659 ~ 0. 889, and PIC val- ues were higher than 0.5. The results indicated that they have a high heterozygosity, which can be used for P. trituberculatus population genetic structure, species breeding, and germqine as- sessment of microsatellite DNA.