四川农业大学学报 ›› 2011, Vol. 29 ›› Issue (02): 218-224.

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中药当归及其混伪品的rDNA ITS序列分析与鉴别

王晓丽   

  1. 泸州医学院
  • 收稿日期:2011-03-27 修回日期:2011-04-13 出版日期:2011-06-30 发布日期:2011-06-30
  • 通讯作者: 王晓丽
  • 作者简介:

      张春,女,27岁,博士,讲师,主要从事药用植物分子鉴定及分子生药学研究,

    Tel: 15196088137, Fax: (0830)3162291, E-mail: zc83good@126.com

  • 基金资助:
    泸州医学院自然科学基金项目

Authentication of Angelica sinensis and adulterants by analysis of rDNA ITS sequences

Zhang 0chun   

  1. Luzhou Medical College
  • Received:2011-03-27 Revised:2011-04-13 Online:2011-06-30 Published:2011-06-30
  • Contact: Zhang 0chun

摘要: 摘要:目的:通过研究比较当归及其混伪品中药材rDNA ITS序列的差异和规律,建立当归与其混伪品药材之间的分子鉴别方法。方法:对rDNA ITS序列进行PCR扩增、测序,结合GenBank中相关材料的序列,利用ClustalW、MEGA4、PAUP 4.0b10等软件进行序列分析,并利用最大简约法构建系统树。结果:所有材料的ITS序列长度为598-601bp。ITS1区总位点数为216,38个为简约信息位点(占17.6%);ITS2区总位点数为225,31个为简约信息位点(占13.8%)。当归各个样品间遗传距离在0.000-0.003之间,当归与混伪品间遗传距离在0.074-0.135之间。当归与混伪品药材间的碱基差异显著,共分离出13个当归的特异鉴别位点。在系统发育树中,当归与混伪品材料分别聚在不同的分支中。结论:rDNA ITS序列特征可作为当归及其混伪品药材鉴别的有效分子标记;当归的归属问题有待商榷。

关键词: 关键词:当归, 混伪品;rDNA ITS序列;DNA分子鉴别

Abstract: Abstract: Objective:The rDNA ITS sequences were analyzed for studying the difference between Angelica sinensis and adulterants and establishing the molecular biological method for the identification of A. sinensis and the others. Methods: rDNA ITS regions (ITS1, 5.8S, ITS2) were amplified and sequenced. Sequences of 6 A. sinensis and its related species from Genbank were analyzed by using software of ClustalW, MEGA4 and PAUP 4.0b10 etc.. The phylogenetic tree was constructed by Maximum Parsimony (MP) method. Results: The lengths of ITS sequences varied from 598 to 601 bp. Of 216 total sites in ITS1, 38 were parsimoniously informative (17.6%); Of 225 total sites in ITS2, 31 were parsimoniously informative (13.8%). The distance among 6 A. sinensis was 0.000 to 0.003. The distance between A. sinensis and the adulterants was 0.074 to 0.135. The differences of rDNA ITS regions among A. sinensis and their adulterants are obvious. 13 distinct variable sites can be used as specific authenticable sites. In the MP tree, A. sinensis and their adulterants were grouped in the different clades. Conclusion:The rDNA ITS sequence can be used as good marker for authenticating A. sinensis from their adulterants. The taxonomic position of A. sinensis should be reconsidered.

Key words: Keywords:Angelica sinensis, adulterants; rDNA ITS region; DNA molecular authentication