黄口荔枝螺转录组数据的微卫星标记开发与分析

Characterization and analysis of microsatellite markers in Thais luteostoma using next generation sequencing

以黄口荔枝螺转录组测序所得到的拼接序列为基础,利用MISA软件进行微卫星分析,利用Primer5.0设计引物160对。结果显示,83对引物能够成功扩增,引物在黄口荔枝螺渔山列岛群体多态性检测中发现,37个SSR位点能够表现出多态性,一共获得了129个等位基因,各位点等位基因数为2~5个,平均每个位点等位基因数为3.49个,观测杂合度H_o,期望杂合度H_e,多态信息含量PIC范围分别介于0.000~0.800、0.146~0.645、0.139~0.573之间,有9个表现为高度多态,20个表现为中度多态,8个表现为低度多态。实验结果进行Hardy-Weinberg平衡检测后,利用Bonferroni correction法进行校正,有29个位点显著偏离,结果表明渔山列岛黄口荔枝螺群体总体遗传多样性水平较低。

Thais luteostoma is one of the most important marine shellfish species, which widely inhabits throughout the southern and northern coastal areas in China. In this paper, we developed a series of microsatellite markers us- ing a transcriptome-based sequencing to provide a foundation for genetic research on Thais luteostoma. We se- quenced a large number of ESTs and screened 160 potential microsatellites of nucleotide repeats using MISA soft- ware. We successfully obtained clear reproducible bands for 83 microsatellite loci. These were amplified in 30 wild individuals sampled from Yushan Island, Ningbo, Zhejiang province. 37 SSR loci were polymorphic （number of alleles per locus ranged from 2 to 5, with an average of 3.49）. The observed heterozygosity ranged from 0.000 to 0.800, while the expected heterozygosity varied from 0.146 to 0.645. Polymorphic information content ranged from 0.139 to 0.573, and 29 SSR loci deviated from Hardy-Weinberg equilibrium. Among these, 9 loci had a PIC of 〈0.5, so were classified as highly polymorphic, additional 20 loci were moderately polymorphic with PICs ranging from 0.25 to 0.5. The PIC of 8 loci was 〈0.25, indicating that these were classified as low polymorphic loci. The results show that the genetic diversity of Thais luteostoma is low in Yushan Island.