三角帆蚌微卫星位点筛选及多态性分析

采用磁珠富集法,以生物素标记的(CA)10寡核苷酸为探针,构建了三角帆蚌(Hyriopsis cumingii)基因组微卫星富集文库。根据微卫星位点的侧翼序列设计引物,随机挑选扩增出与预期大小相符的32对引物,引物荧光标记后对24个三角帆蚌个体分别进行PCR扩增,共筛选出20对多态性较好的引物。结果表明,20个微卫星位点的等位基因数为5～20,有效等位基因数2.321 3～13.260 3。观察杂合度和期望杂合度分别为0.318 2～1.000 0和0.582 5～0.946 1;PIC值0.547 2～0.919 9;其中14个位点符合Hardy-Weinberg平衡(P>0.05)。本研究筛选的20个微卫星标记可作为三角帆蚌遗传多样性、种群遗传结构等研究的理想分子标记。

Screening and Genetic Diversity Analysis of Microsatellite in Hyriopsis cumingii

Microsatellite-enhanced genomic library of the Hyriopsis cumingii was constructed using repeat-enrichment method with biotin-labeled oligo(CA)10and streptavidin coated magnetic beads.32 pairs of primers which could amplify expected PCR products were designed from the microsatellite flanking sequences and labeled by fluorophore,20 microsatellite loci showed high levels of genetic polymorphism testing on 24 individuals.The number of alleles per locus ranged from 5 to 20,the number of effective alleles per locus ranged from 2.3213 to 13.2603.The ranges of observed and expected heterozygosity were 0.3182 to 1.000 0 and 0.582 5 to 0.946 1,respectively;polymorphism information content(PIC) ranged from 0.5472 to 0.9199.14 of 20 markers accorded with Hardy-Weinberg equilibrium(P>0.05).These 20 microsatellite loci would be used as the ideal molecular markers in the studies of genetic diversity,population genetic structure and other relevant research in H.cumingii.