大黄鱼微卫星多重PCR体系的建立及其应用

利用本实验室开发的微卫星标记,通过优化退火温度、引物浓度、循环次数等条件,建立了3组大黄鱼微卫星多重PCR体系,每组包含3个微卫星位点,用3组微卫星多重PCR分析了一个大黄鱼选育群体JD-01的遗传多样性。结果显示,该群体的平均等位基因为12.111,平均有效等位基因为7.408,平均多态信息含量为0.846,平均观测杂合度和期望杂合度分别为0.825、0.868,香农多样性指数为2.165。运用Cervus 3.0软件,对已知系谱关系的9个大黄鱼家系及对应亲本进行了亲子鉴定分析,以验证3组微卫星多重PCR在亲子鉴定中的准确性。结果显示,使用该3组微卫星多重PCR体系进行亲子鉴定准确率为100%。大黄鱼微卫星多重PCR体系的建立为分析群体的遗传多样性、亲子鉴定和辅助家系管理提供了一种高效的技术手段。

Development and application of three multiplex PCR panels of microsatellites in large yellow croaker(Larimichthys crocea)

In this study, three multiplex PCR panels were established using microsatellite markers developed from our laboratory by optimizing annealing temperature, concentration of primers and times of cycle.A breeding group "JD-01" was analyzed using the three multiplex PCR panels.The results showed:the average number of alleles and effective number of alleles were 12.111 and 7.408 respectively.The average of polymorphism information content(PIC) were 0.846.The average of observed heterozygosity(H_o) and expected heterozygosity(H_e) was 0.825 and 0.868, respectively.The Shannon's diversity index was 2.165.By using Cervus 3.0 software, we verified parentage assignment that the pedigree relationship was known in 9 families of large yellow croaker.As a result, the accuracy rate of parentage assignment was 100% by using three multiple PCR panels.The development of microsatellite multiple PCR system provides a high efficiency technical method for genetic diversity analysis, parentage assignment and marker-assisted family management in large yellow croaker.