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微卫星分型法应用于豢养繁殖长江江豚的父权鉴定

ISSN号：1673-6273
期刊名称：《现代生物医学进展》
时间：0
分类：Q959.8[生物学—动物学]
作者机构：[1]南昌大学生命科学学院,江西南昌330031, [2]中国科学院水生生物研究所,湖北武汉430072, [3]中国科学院研究生院,北京100039
相关基金：科技部973项目（2007CB411600）,国际合作项目（2004DFB03000）,国家自然科学基金项目（30730018,30570252）

作者：冯俊伟[1], 郑劲松[2], 周钊[2,3], 林刚[1], 王丁[2]

关键词：长江江豚, 微卫星, 基因分型, 父权鉴定, Yangtze finless porpoise, Microsatellite, Genotyping, Paternity determination

中文摘要：

目的：鉴定武汉白鱀豚馆及铜陵淡水豚国家级自然保护区两个豢养长江江豚繁殖群体中出生的6头幼豚的生物学父亲。方法：选择8对江豚物种特异性微卫星引物对两个待鉴定群体的14个DNA样品进行了荧光标记PCR扩增,将纯化后的扩增产物在ABI3130遗传分析仪上进行基因分型,并根据GeneScanRox500内标确定不同等位基因的大小,随后对待鉴定对象进行等位基因分析,并计算主要多态性参数。结果：所采用的8个微卫星座位在待鉴定的两个江豚群体中均表现出多态性。在母本已知的条件下,利用其中6个微卫星座位的等位基因数据,通过排除法成功地鉴定出两个繁殖群体中出生的6头幼豚的生物学父亲。结论：本研究首次成功地将6个物种特异性微卫星标记应用于豢养繁殖长江江豚的父权鉴定,从而为该物种微卫星亲子鉴定技术体系的建立及迁地保护繁殖群体遗传谱系的构建奠定了技术基础。

英文摘要：

Objective：This study was conducted to identify the biological fathers of six young Yangtze finless porpoises born in the two captive breeding colonies of Wuhan Baiji Dolphinarium and Tongling Freshwater Cetacean National Natural Reserve. Methods：Eight polymorphic microsatellite primer pairs,which had been specially developed for Yangtze finless porpoise or finless porpoise,were fluorescently labeled and applied to amplify the 14 DNA samples from the two breeding colonies. PCR products were purified,and then genotyped on an ABI3130 Genetic Analyzer with the GeneScan Rox 500 internal ladder to accurately decide the size of each allele. Subsequently,analysis was conducted on the genotypes of all individuals. Results：All 8 microsatellite loci showed polymorphism in the two captive breeding colonies. Besides,on the condition that all the mother-offspring relationships were known definitely,the true fathers for the six offspring were easily and successfully identified by excluding the impossible fathers via analyzing these alleles generated by 6 out of the 8 microsatellite loci. Conclusions：In this study,for the first time,six specific microsatellite markers were successfully applied in paternity determination of six captivity-bred Yangtze finless porpoises. It provides a solid technical base for the establishment of an effective parentage identification system,and also for the construction of accurate studbooks which are essentially important for the genetic management of any ex situ conservation Yangtze finless porpoise populations.

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