Establishment of reliable pedigree information of different lineages is crucial for selective breeding and stock enhancement programs. However, owing to the absence of a reliable assay for parentage assignment, such information is lacking in orange‐spotted grouper (Epinephelus coioides), which is an economically important coral‐reef fish. In this study, we developed two multiplex microsatellite polymerase chain reaction systems that consisted of 12 microsatellite markers (seven in system A and five in system B) for parentage assignment of orange‐spotted groupers. The theoretical exclusion probability per locus ranged from 0.079 to 0.436 when both parents were unknown (E‐1P) and from 0.191 to 0.614 when one parent was known (E‐2P). The combined exclusion probability values of 12 loci were 98.8% (E‐1P) and 100% (E‐2P), respectively. The accuracy of the parentage test for a mixed family (15 dams and 12 sires, 226 offspring) was 100%. In addition, two wild populations (79 individuals) were added as interferential offspring, only four of which were misassigned to the stock. The success rate of assignment was 98.69%. We provide a reliable microsatellite‐based assay for parentage assignment in orange‐spotted groupers, which will be useful for selective breeding and stock enhancement programs for this species and maybe other groupers.

中文翻译：

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橙斑石斑鱼（石斑鱼）亲子分配的多重微卫星测定法的发展

建立不同谱系的可靠谱系信息对于选择育种和种群改良计划至关重要。但是，由于缺乏可靠的亲子鉴定方法，橙斑石斑鱼（Epinephelus coioides）缺乏此类信息。），这是一种具有重要经济意义的珊瑚礁鱼。在这项研究中，我们开发了两个多重微卫星聚合酶链反应系统，该系统由12个微卫星标记组成（系统A中为七个，系统B中为五个），用于桔斑石斑鱼的亲本分配。当两个父母都不认识时（E-1P），每个基因座的理论排除概率在0.079至0.436之间；当一个父母已知（E-2P）时，每个基因座的理论排除概率在0.191至0.614之间。12个基因座的组合排除概率值分别为98.8％（E-1P）和100％（E-2P）。混合家庭（15个水坝和12个父亲，226个后代）的亲子鉴定的准确性为100％。此外，添加了两个野生种群（79个个体）作为有干扰的后代，其中只有四个被误分配到了种群中。作业成功率为98.69％。