使用SPE软件筛选绿尾虹雉微卫星引物及其在亲缘关系鉴定上的应用  

Screening of Polymorphic Microsatellite Loci in Lophophorus lhuysii by SPE and Its Application on Kinship Testing

作　　者：王立 李严 岳碧松[2] 陈黎 沈富军 寇洁 王也 侯蓉 张亮 WANG Li;LI Yan;YUE Bisong;CHEN Li;SHEN Fujun;KOU Jie;WANG Ye;HOU Rong;ZHANG Liang(Sichuan Key Laboratory of Conservation Biology for Endangered Wildlife,Chengdu Research Base of Giant Panda Breeding,Chengdu 610081,China;Key Laboratory of Bio-Resources and Eco-Environment,Ministry of Education,College of Life Sciences,Sichuan University,Chengdu 610065,China;Sichuan Fengtongzhai National Nature Reserve Administration,Yaan,Sichuan Province 625700,China)

机构地区：[1]四川省濒危野生动物保护生物学重点实验室,成都大熊猫繁育研究基地,成都610081 [2]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610065 [3]四川蜂桶寨国家级自然保护区管护中心,四川雅安625700

出　　处：《四川动物》2022年第4期371-378,共8页Sichuan Journal of Zoology

摘　　要：圈养绿尾虹雉Lophophorus lhuysii多为一雄配多雌,但由于雄鸟更换,雌鸟产蛋期交错,且无特定产蛋位,导致亲子关系难以确认。本研究从绿尾虹雉基因组中广泛筛选微卫星位点,得到13对引物。采用微卫星PCR引物特异性鉴定程序(SPE)对引物序列进行评估,排除1例PCR产物长度相近的特殊多位点扩增。利用获得的12个微卫星标记在18份绿尾虹雉蛋壳/羽毛DNA样品中进行遗传多样性分析,同时以其中15份蛋壳样品(2018-2019年搜集)为检材,进行了亲缘(同胞)关系鉴定。结果表明,这12个微卫星标记可用于绿尾虹雉亲缘关系的鉴定,对提高小种群的维持能力,保持种群遗传多样性具有重要意义。Chinese monals(Lophophorus lhuysii)are often socially housed in captivity with 2 females and 1 male in the unit. The exchange of males during the breeding season and the unfixed nest site for laying eggs have led to the vague recording of parentage information of L. lhuysii in the absence of genetic testing. To solve this problem,13 microsatellite loci from the genome of L. lhuysii were isolated by routine method. In addition,the multiple amplification conditions of the primers were verified by SPE software. Specifically,the length of two multiple amplification PCR products differed by a single base was identified. The remaining 12 microsatellite loci were genotyped in 18 L. lhuysii eggshell/feather DNA samples. The sibling relationship between 15 individuals born between 2018-2019 was successfully completed. These markers can be used in the systematic control of kinship determination for L. lhuysii,and can also be used as powerful tools for the genetic management of L. lhuysii.

关 键 词：绿尾虹雉 微卫星 SPE软件 多位点扩增 亲缘关系鉴定 

分 类 号：Q959.7[生物学—动物学] Q953