奶牛分子系谱构建或亲权鉴定的微卫星标记筛选  

Selection of Microsatellite Markers Used for Pedigree Tree Construction and/or Paternity Testing in Chinese Holstein Cows

作　　者：殷彬[1] 岳书俭 俞英[2] 曹巧巧[1] 王中华[1] 师科荣[1] 

机构地区：[1]山东农业大学动物科技学院山东省动物生物工程与疾病防治重点实验室,泰安271018 [2]中国农业大学动物科技学院农业部畜禽遗传育种重点实验室畜禽育种国家工程实验室,北京100193

出　　处：《畜牧兽医学报》2017年第4期595-604,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31402054);山东省自然科学基金(ZR2013CM013);山东省农业良种工程项目(2016LZGC030);国家现代奶业产业技术体系(CARS-37);山东省泰山学者种业创新人才团队计划培养项目(2014LZ07-04)

摘　　要：旨在利用较少的经过优化的遗传标记组合达到快速高效的亲缘鉴定,最终可有效应用于奶牛群体亲子鉴定与系谱构建。本研究从国际动物遗传学会(ISAG)数据库中初步选择了18个多态性较高的微卫星位点,经PCR优化验证获得可高效扩增的8个位点,并检测其在300头中国荷斯坦牛群体中的多态性分布。这些微卫星位点的亲子鉴定效力检测结果表明,8个标记平均等位基因数为14.63,平均多态性信息含量(PIC)为0.747,群体平均观察杂合度(Ho)为0.718,平均期望杂合度(He)为0.773。3种不同情形下进行亲子鉴定时8个位点的结合排除概率(CPE)均≥0.990。8对父子牛亲权鉴定结果表明,利用该8个标记可以识别并剔除系谱中的错误记录。结合排除概率的分析结果表明,当微卫星位点数增加至4个(TGLA227、TGLA122、BMC1207、BM103)时,3种不同情形下的CPE均≥0.949,可以满足个别案例牛亲子鉴定的要求。群体分子系谱构建的结果表明,当微卫星位点数大于或等于6个时,群体内近交系数和总群体近交系数均明显升高。因此,在生产实践中推荐使用TGLA227、TGLA122、BMC1207、BM103、INRA037、INRA134位点进行小规模群体(<200)的分子系谱构建或亲权鉴定。How to use less and optimized genetic markers to achieve convenient and efficient parentage determination for cows was our research purpose,so as to effectively apply them into paternity test and pedigree construction.Eighteen microsatellite markers with high polymorphisms in cattle recommended on the web of ISAG were selected in this study,8markers that could be efficiently amplified were genotyped in 300 Chinese Holstein cows from the Jiabao Farm,based on fluorescent primer and automatic sequencing techniques.The results of paternity testing efficiency showed that the average of allele number,polymorphism information content（PIC）,observed heterozygosity（Ho）,and expected heterozygosity（He）were 14.63,0.747,0.718and0.773,respectively.In 3different conditions of known and/or unknown parent-offspring,theircombined probabilities of exclusion（CPEs）were 0.990,0.999 and 0.999,respectively.The results of paternity testing from 8father-son showed that the 8markers could successively be used to identify correct parentage and eliminate error records in the pedigree.Combined probability of exclusion analysis showed that when the number of microsatellite markers increased to 4（TGLA227,TGLA122,BMC1207,BM103）,all CPEs under 3different situations were 0.949 or higher,which met the requirements of paternity tests in individual case.While,the results of molecular pedigree tree building at population level revealed that when the number of microsatellite sites increased to 6（TGLA227,TGLA122,BMC1207,BM103,INRA037 and INRA134）,inbreeding coefficient and total inbreeding coefficient within population were significantly increased,suggested that the 6 markers could efficiently be applied for pedigree tree construction and/or paternity testing for Chinese Holstein cows population（200 individuals）.

关 键 词：中国荷斯坦牛 微卫星位点 排除概率 亲子鉴定 

分 类 号：S823[农业科学—畜牧学] S813.1[农业科学—畜牧兽医]