荧光标记微卫星技术用于凡纳滨对虾不同家系亲权关系鉴定

Application of Fluorescent Labeled Microsatellite Technology in Parentage Analysis of Litopenaeus vannamei

应用荧光标记微卫星技术对凡纳滨对虾(Litopenaeus vannamei)家系进行亲权鉴定。挑选14个多态信息含量较高的微卫星位点,以人工选育建立的凡纳滨对虾5个全同胞家系为试验材料,采用Cervus 3.0进行亲权分析,并根据家系内个体间的遗传距离进行UPGMA聚类分析。结果表明,14个微卫星位点的平均等位基因数为6,平均多态信息含量为0.6896,平均期望杂合度为0.7309,平均观测杂合度为0.7661,第一亲本、第二亲本和双亲的累计排除率分别为0.99721733、0.99996559和0.99999997;进一步模拟分析表明,要达到亲权鉴定的要求,在双亲性别已知时至少需要5个微卫星位点,双亲性别未知时至少需要6个微卫星位点;模拟分析及试验验证所选用的14个微卫星位点最多可以鉴定1 954个已知性别的亲本或1 203个未知性别的亲本。所选用的14个微卫星标记可在生产及科研试验中用于获取凡纳滨对虾系谱信息。

Fourteen microsatellite loci with high polymorphism were selected for parentage assignment in five full-sib families of selected white shrimp （Litopenaeus vannamei）by Cervus 3.0.The UPGMA dendrogram of the five families was constructed according to the genetic distance among 87 individuals.The results showed that the aver-age number of alleles per locus,mean polymorphism information content （PIC ）,mean observed heterozygosity, and mean expected heterozygosity were 6,0.6896,0.7309 and 0.7661,respectively.The combined exclusion probability of the first parent （CEP-1P）,the second parent （CEP-2P）and a parent pair （CEP-PP）were 0.99721733,0.99996559 and 0.99999997,respectively.Further simulation based on allele frequencies suggested that at least five microsatellite loci were needed to obtain accurate results of paternity test when the sex of both par-ents were known,and at least six microsatellite loci when the sex of both parents were unknown.The simulation analysis and experimental verification results showed that 1 954 known parents or 1 203 unknown parents could be detected using 14 microsatellite loci.Our study demonstrates that the 14 microsatellite loci could be used for pedi-gree analysis of Litopenaeus vannamei in breeding production and scientific experiments.