生物多样性 ›› 1997, Vol. 05 ›› Issue (3): 173-178.  DOI: 10.17520/biods.1997028

• 论文 • 上一篇    下一篇

用RFLP和PCR-RFLP技术研究东北虎和华南虎线粒体DNA多态性*

吴平,周开亚,王亚明,黄恭情,徐麟木   

  1. (南京师范大学生物系,  南京 210097)
    (苏州市动物园,  苏州 215001)
    (南京玄武湖动物园,  南京 210009)
  • 收稿日期:1996-05-23 修回日期:1996-10-20 出版日期:1997-08-20 发布日期:1997-08-20

Mitochondrial DNA polymorphism between Siberian tiger and South China tiger revealed by mtDNA RFLP and PCR-RFLP

Wu Ping, Zhou Kaiya, Wang Yaming, Huang Gongqing, Xu linmu   

  1. 1) Department of Biology , Nanjing Normal University , Nanjing  210097
    2) Suzhou Zoo , Suzhou 215001
    3) Xuanwuhu Zoo , Nanjing 210009
  • Received:1996-05-23 Revised:1996-10-20 Online:1997-08-20 Published:1997-08-20

摘要:

采用mtDNA RFLP和PCR-RFLP技术研究了东北虎和华南虎的mtDNA的多态性。在mtDNA RFLP研究中,分离纯化了东北虎和华南虎肝、肾和心脏组织的mtDNA,用20种识别6碱基对的限制性内切酶消化, 结果只有1种限制性内切酶(XbaⅠ)检测到多态性片段,其余19种限制性内切酶消化产生的限制性格局在东北虎和华南虎完全一致。在PCR-RFLP研究中,用PCR技术分别扩增了东北虎和华南虎mtDNA的控制区(control region),用8种识别4碱基对的限制性内切酶分别对扩增产物进行消化,结果只有1种限制性内切酶(RsaⅠ) 检测到多态性片段。mtDNA RFLP及PCR-RFLP的结果均提示东北虎和华南虎之间的遗传距离极小。这可能与下列因素有关:两者分布区间无天然隔离屏障;具有强扩散能力;近几百年才被相互隔离。

Abstract

MtDNA RFLP and PCR-RFLP techniques were used to reveal polymorphism between Siberian tiger (Panthera tigris altaica) and  South  China  tiger (Panthera tigris amoyensis).  In mtDNA RFLP study,  mitochondrial DNA was extracted from liver, kidney and heart tissues  of Siberian tiger  and South China tiger  and  purified. Restriction  endonuclease  fragment patterns of mtDNA were digested with 20 enzymes recognized 6-bp. Only 1 (XbaⅠ) out  of  20  restriction  enzymes  revealed  polymorphic fragments.  The restriction pattern generated by 19 of the enzymes in  Siberian  tiger were identical with that  in  South  China  tiger. In PCR-RFLP study, D-loop was amplified and 8 endonucleases recognized 4-bp were used to digest PCR product, 1 out of 8 endonucleases (RsaⅠ)  revealed polymorphic fragments.  Genetic  distance between Siberian tiger and South China tiger  calculated based on the two results was very small.  This  is  probably  due  to  the  following  factors: there was no natural barrier between the two  subspecies; they have strong dispersal ability and were separated only  within  several  hundred years.