基于微卫星多重PCR技术的黄喉拟水龟亲子鉴定
THE PARENTAGE ASSIGNMENT OF MAUREMYS MUTICA USING MULTIPLEX PCR OF MICROSATELLITES
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摘要: 利用黄喉拟水龟(Mauremys mutica)微卫星标记,筛选出16对微卫星引物,通过优化各引物比例、荧光接头浓度、退火温度和循环次数,建立了基于2组各含8个微卫星位点多重PCR体系的黄喉拟水龟亲子鉴定技术。应用2组微卫星多重PCR体系,通过ABI3130遗传分析仪以及cervus3.0软件对428只黄喉拟水龟进行了个体基因型检测和遗传多样性分析,结果显示,群体的平均等位基因数为14.190,平均多态信息含量为0.748,平均观测杂合度和期望杂合度分别为0.687、0.771。对89只候选母本及296只子代进行亲子鉴定分析,结果显示:在父本信息未知时,母本鉴定率为87%;母本获得的子代个数范围为1-12,个体间表现出巨大的差异,这为选择育种提供了物质基础。黄喉拟水龟多重PCR亲子鉴定技术的建立为群体遗传多样性分析、家系鉴定管理和选择育种提供了有效的技术手段。Abstract: In this study, we established a technique for the parentage assignment in Asian Yellow Pond Turtles(Mauremys mutica) based on two multiplex PCR panels. The primer ratio, the concentration of the fluorescent joint, the annealing temperature and the number of cycles were optimized in this technique. Each multiplex PCR panel had eight loci, 10 of which were developed in our lab and the other 6 were chosen from published literatures. With the multiplex PCR tool we performed the genotyping and the genetic diversity analysis on 428 individuals using ABl3I30 genetic an-alyzer and Cervus 3.0 software. The results showed that the average number of alleles was 14.190; the average poly-morphism information content(PIC) was 0.748; the average observed heterozygosity(Ho) and expected heterozygosity(He) were 0.687 and 0.771 respectively. The parentage assignment was performed on 89 candidate female parents and 296 offspring. Without information on the male parent, the identification rate was 87%. The numbers of offspring of different female parents were remarkably various, ranging from 1 to 12. This provided the material foundation for the selective breeding. This technique could be a highly efficient tool for the genetic diversity analysis, the marker-assisted family identification and management, and the selective breeding of Asian Yellow Pond Turtles.
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Keywords:
- Mauremys mutica /
- Microsatellite multiplex PCR /
- Parentage assignment
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