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利用随机扩增多态性DNA技术鉴定五种药用八角莲

傅荣昭, 邵鹏柱, 王骏, 毕培曦, 施大文, 孙勇如*   

  1. 1.中国科学院遗传研究所, 北京 100101;
    2.香港中文大学生化系, 香港、新界、沙田;
    3.香港中文大学中药研究中心, 香港、新界、沙田;
    4.上海医科大学药学院, 上海 200032
  • 收稿日期:1999-06-14 修回日期:1999-10-20 出版日期:2000-06-15 发布日期:2000-06-15
  • 通讯作者: 孙勇如*

RAPD Differentiation of Five Medicinal Dysosma Species

Fu Rongchao, Shaw Pangchui, Wang Jun, But Paulhay, Shi Dawen, Sun Yongru*   

  1. 1. Institute of Genetics, Chinese Academy of Sciences, Beijing 100101;
    2. Department of Biochemitry Chinese University of Hong Kong Shatin, N T, Hong Kong, China;
    3. Chinese Medicinal Material Research Centre, Chinese University of Hong Kong, Shatin, N. T, Hong Kong, China;
    4. School of Pharmacy Shanghai Medical University Shanghai 200032
  • Received:1999-06-14 Revised:1999-10-20 Online:2000-06-15 Published:2000-06-15
  • Contact: Sun Yongru*

摘要: 现有的传统中药材鉴定方法有时很难可靠地用于药用植物八角莲的鉴定研究。本文在利用作者新建立的从干燥材料中分离高质量DNA的技术和比较不同聚合酶链反应(PCR)模板浓度的基础上, 利用随机扩增多态性DNA(RAPD)技术对五种药用八角莲的7个样品进行了分子鉴定。在筛选的42个引物中, 12个引物产生的多态性条带, 可靠地区分了这7个八角莲样品, 包括3个不同地区的D.versipellis, 1D. majorense, 1D pleiantha, 1D.furfuracea 1D, veitchii。每个样品都找到了1~3个稳定可靠的特异分子标记。此外, 本文发现DfurfuraceaD, versipellis的亲缘关系最远, Dveitchii较近, 而与D.majorense最近。D.veitchiiD.majorense的亲缘关系比与D,versipellis, D.furfuraceaD.pleiantha更近。结果表明RAPD技术能够有效地用于这7个药用八角莲样品的分子鉴定。

关键词: DNA, RAPD指纹, 干燥药材, 八角莲

Abstract: Traditional methods of authentication are not always reliable in differentiation of Dysosma species. We explored the potentiality of random amplified polymorphic DNA (RAPD) technique for the differentiation of the medicinal plants. Among the forty-two primers screened, twelve were able to generate reproducible bands that distinguished the seven samples of five Dysosma species, D. versipellis (Hance) M. Cheng from 3 localities, D.majorense (Gagnep.) Ying, D. pleiantha (Hance) Woodson, D. furfuracea S.Y Bao, and D. veitchii (Hemsl. et Wils.) Fu. Characteristic RAPD marker bands unique to each of the seven Dysosma samples were identified. RAPD polymorphism was lower among the intra-specific samples of D. versipellis, but higher in inter-specific samples. Our results demonstrate that RAPD marker technique can effectively authenticate these seven Dysosma samples.

Key words: DNA, DNA, RAPD fingerprints, RAPD fingerprints, Dried medicinal materials, Dried medicinal materials, Dysosma, Dysosma

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*To whom correspondence should be addressed. Tel: (86)10-64859071; Fax: (86)10-64914896; E-mail: liwenbin@bj.col.com.cn
The present address of the first author: 317 Biotech Bldg, Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, 14853.