《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (7): 2003-2011.doi: 10.16431/j.cnki.1671-7236.2019.07.015

• 遗传繁育 • 上一篇    下一篇

利用微卫星标记鉴定德州驴亲子关系

王敏1,2, 张新浩3, 崔冉1,2, 刘宇1,2, 杨莉3, 李海静3, 赵春江1,2   

  1. 1. 中国农业大学动物科技学院, 北京 100193;
    2. 中国农业大学马研究中心, 北京 100193;
    3. 东阿阿胶股份有限公司, 国家胶类中药工程技术研究中心, 东阿 252201
  • 修回日期:2018-12-17 出版日期:2019-07-20 发布日期:2019-07-22
  • 通讯作者: 李海静, 赵春江 E-mail:lihaijing@dongeejiao.com;cjzhao@cau.edu.cn
  • 作者简介:王敏(1993-),女,山东临沂人,博士,研究方向:动物遗传育种,E-mail:1105002815@qq.com;张新浩(1988-),男,山东聊城人,硕士,研究方向:动物遗传育种,E-mail:704589857@qq.com
  • 基金资助:

    山东省农业良种工程:地方良种皮、肉专门化新品系选育(2017LZGC020);校企重点研发项目:东阿黑驴育种综合配套技术研究(性状综合选育部分)(201605410411094)

Parentage Testing of Dezhou Donkey with Microsatellite Marker

WANG Min1,2, ZHANG Xinhao3, CUI Ran1,2, LIU Yu1,2, YANG Li3, LI Haijing3, ZHAO Chunjiang1,2   

  1. 1. College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;
    2. Equine Center of China Agricultural University, Beijing 100193, China;
    3. National Engineering Research Center for Gelatin-based Traditional Chinese Medicine, Dong E E Jiao Co., Ltd., Donge 252201, China
  • Revised:2018-12-17 Online:2019-07-20 Published:2019-07-22

摘要:

试验旨在建立一套适用于德州驴亲子关系的鉴定体系。选取13个微卫星基因座作为标记,采集了53头德州驴血液样本,其中子代驴驹16头,候选父本13头,候选母本24头,用酚-仿法抽提血液基因组进行PCR扩增和基因扫描,并利用Peak Scanner Software v1.0软件读取基因型分型结果。对微卫星基因座的遗传多样性进行分析,利用似然法(Cervus 3.0软件)和排除法对个体间的亲子关系进行了鉴定。结果显示,13个微卫星基因座的平均等位基因数、平均观测杂合度(Ho)、平均期望杂合度(He)和平均多态信息含量(PIC)分别为6.846、0.689、0.671和0.625。期望杂合度与观测杂合度之差在0.002~0.088之间,差值较小。13个微卫星基因座的累计排除概率(EP)达到0.990以上。微卫星基因座具有高度多态性和较高的排除概率,适用于遗传分析和个体鉴定。利用Cervus 3.0软件基于似然法分析得到了16头子代驴驹的最似亲本,结合排除法对这16头驴驹及其最似亲本进行基因型比对,最终在53头德州驴中确定了11个亲子对。本试验建立了以13个微卫星位点作为核心标记,将似然法和排除法相结合作为主要分析方法的德州驴亲子关系鉴定体系,为育种工作提供参考资料。

关键词: 德州驴; 微卫星; 亲子鉴定

Abstract:

This study was aimed to establish the parentage testing system of Dezhou donkey.13 microsatellite loci were selected as markers.A total of 53 blood samples of Dezhou donkey were collected,including 16 offsping,13 candidate fathers and 24 candidate mothers samples.The genomic DNA was obtained with phenol and chloroform method.The microsatellite loci were amplified by PCR and conducted gene scanning,the genotyping result was read using Peak Scanner Software v1.0.The genetic diversity of microsatellite loci were analyzed,the parentage relationship between individuals were identified using the likelihood-based (Cervus 3.0 software)and exclusion-based methods.The results showed that the average of alleles number,observed heterozygosity (Ho),expected heterozygosity (He) and polymorphism information content (PIC) were 6.846,0.689,0.671 and 0.625.The difference between expected heterozygosity and observed heterozygosity varied from 0.002 to 0.088.Their combined exclusion probability reached 0.990.The microsatellite loci showed high polymorphic and high exclusion probabilities,which made them suitable for genetic analysis and parentage testing.Most similar parents of 16 offspring donkeys were identified using Cervus 3.0 software.The genotypes of the 16 donkeys and their most likely parents were compared to exclude unrelated individuals.The parents of 11 donkeys were identified.In this experiment,13 microsatellite loci were used as the core markers,and the likelihood-based and exclusion-based methods were used as the main analytical method for the parentage testing system of Dezhou donkey,which provided the useful information for breeding work.

Key words: Dezhou donkey; microsatellite; parentage testing

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