基于微卫星标记的花鲈亲子鉴定技术

Paternity Test Technology of Lateolabrax maculatus Based on Microsatellite Markers

【目的】推进家系选育工作,构建准确、高效、经济的亲子鉴定技术,以准确获得花鲈(Lateolabrax maculatus)的系谱信息。【方法】选择8对有多态性的微卫星标记。应用微卫星荧光标记多重PCR和毛细管电泳检测技术,建立1组多重PCR反应体系。用软件GeneMarker和Cervus软件分析实验结果。【结果与结论】8个微卫星位点的平均等位基因数为10.125个,平均期望杂合度为0.657,平均观测杂合度为0.680。双亲未知时单亲累积排除概率(CE-1P)为97.21%,双亲之一已知时单亲累积排除概率(CE-2P)为99.74%。双亲未知时双亲对累积排除概率(CE-PP)为100.00%。174个子代和34个候选亲本的实际亲子鉴定结果显示,170个子代的似然率对数值均大于0。当置信度大于95%时,有143个子代均能匹配到父母本,实际鉴定率为82.18%。

【Objective】To get accurate genealogical information for the breeding of Lateolabrax maculatus and construct an accurate,efficient and low-cost paternity test technology based on microsatellite loci.【Method】8 pairs of polymorphic microsatellite markers were selected.A set of multiplex PCR reaction system was established by using fluorescent labeling multiplex PCR and capillary electrophoresis.The experimental results were analyzed with GeneMarker and Cervus software.【Result and Conclusion】The average number of alleles of 8 microsatellite loci was 10.125,the average polymorphism information content was 0.637,the average expected heterozygosity was 0.657,and the average observed heterozygosity was 0.680.Six microsatellite markers were highly polymorphic and two microsatellite markers were moderate.The cumulative exclusion probabilities of 8 microsatellite loci were 97.21%(CE-1P),99.74%(CE-2P)and 100.00%(CE-PP).The finnndings of the paternity tests among 34 candidate parents and 174 offsprings showed thatthe likelihood ratio logarithms of 170 offsprings were more than 0.When the confidence was greater than 95%,143 offsprings could match the parents,with successful identification rate of 82.18%.the paternity tests among 34 candidate parents and 174 offsprings showed thatthe likelihood ratio logarithms of 170 offsprings were more than 0.When the confidence was greater than 95%,143 offsprings could match the parents,with successful identification rate of 82.18%.