日本沼虾微卫星引物筛选及群体遗传多样性分析  

Isolation and genetic diversity analysis of microsatellite DNA in Macrobrachium nipponense

作　　者：冯建彬[1,2] 马克异[1] 李家乐[1,3] 华雪铭[1] 丁怀宇[2] 

机构地区：[1]上海海洋大学省部共建水产种质资源发掘与利用教育郝重点实验室,上海201306 [2]淮阴师范学院江苏省环洪泽湖生态农业生物技术重点实验室,江苏淮安223300 [3]上海市高校水产养殖学E研究院,上海海洋大学水产与生命学院,上海201306

出　　处：《水产学报》2010年第5期688-695,共8页Journal of Fisheries of China

基　　金：江苏省苏北科技发展计划项目(BN2009036);江苏省环洪泽湖生态农业生物技术重点实验室开放课题(HZHL0803);上海海洋大学博士科研启动基金(B-8201-08-0279);上海市科委重点科技攻关计划项目(073205111);上海市重点学科建设项目(Y1101)

摘　　要：采用(CT)15、(CA)15两种生物素标记及磁珠富集法构建了太湖日本沼虾基因组微卫星富集文库。120个微卫星克隆中83个位点的核心序列重复在10个以上,长度介于191～580bp,平均为311bp;重复类型中,(CA/GT)n及(AG/TC)n分别占59.09%及20.45%,还检测到(GC)n、(AAG)n、(ACAA)n和(GGCAGA)n等17种类型,包括25.75%完美型、20.45%非完美型和53.80%复合性。用其中12条微卫星引物扩增吴江、滨湖、宜兴和吴兴4个群体240个个体的基因组DNA,各位点表现出较高的遗传多样性,除Mni86、Mni103和Mni114外,其它群体位点大都表现出显著的杂合子缺失,4个群体的遗传分化为低度分化(FST<0.05),4个群体中97.58%遗传变异存在于群体内,仅有2.42%的遗传变异来自于群体之间;吴江和吴兴群体亲缘关系最近,与宜兴和滨湖群体亲缘关系渐远。Microsatellite-enhanced genomic library of oriental river prawn Macrobrachium nipponense of Lake Taihu was constructed using repeat-enrichment method with biotin-labeled oligos [（CA）15 and （CT）15]and streptavidin magnetic beads. From a total of randomly selected 120 clones,83 clones with more than ten motif repeats were positive. And the sequence size ranges from 191 to 580 bp with an average of 311 bp. The （CA/ GT）n and （AG/TC）n were the abundant microsatellite motifs with a percentage of 59. 09% and 20. 45% in the isolated microsatellites. Other 17 types of repeat sequences were detected,including repeats consisting of （GC）n,（AAG）n,（ACAA）n and （GGCAGA）n. Characterization of these microsatellites showed that 25. 75% of sequences were classified as perfect type,20. 45% as imperfect type and 53. 80% as compound type. Genetic diversity of 4 wild stocks of M. nipponense in 4 sites of Lake Taihu,i. e. Wujiang （WJ）,Binhu （BH）,Yixing （YX） and Wuxing （WX）,were investigated using 12 microsatellite DNA loci from the isolated primers . The allele numbers,effective allele numbers,observed heterozygosity and expected heterozygosity under Hardy-Weinberg equilibrium were estimated to characterize genetic diversity. The result showed that all the 12 loci in the 4 stocks were highly polymorphic. Tests of departures from Hardy-Weinberg equilibrium indicated the loci except Mni86、Mni103 and Mni114 in each stock with significant heterozygosity deficiency. FST and AMOVA analysis across all stocks and loci indicated that there was low level of divergence among 4 stocks. The UPGMA clustering tree based on DA genetic distance demonstrated that Wujiang and Wuxing stocks have near relationship,and they have a distant relationship with Wuxi and Yixing stocks.

关 键 词：日本沼虾 微卫星引物 筛选 遗传多样性 

分 类 号：Q346.5[生物学—遗传学] S917[农业科学—水产科学]