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党参药材及其混伪品的ITS/ITS2条形码鉴定研究
引用本文:赵莎,辛天怡,侯典云,庞晓慧,陈睿旸,高建平.党参药材及其混伪品的ITS/ITS2条形码鉴定研究[J].世界科学技术-中医药现代化,2013,15(3):421-428.
作者姓名:赵莎  辛天怡  侯典云  庞晓慧  陈睿旸  高建平
作者单位:北京中医药大学中药学院 北京 100102;中国医学科学院/北京协和医学院/药用植物研究所 北京 100193;中国医学科学院/北京协和医学院/药用植物研究所 北京 100194;中国医学科学院/北京协和医学院/药用植物研究所 北京 100195;北京中医药大学东方医院 北京 100078;山西医科大学药学院 太原 030001
基金项目:科学技术部国家科技支撑计划(2011BAI07B07):华北区党参、锁阳、金银花等质量标准提高及其综合利用研究,负责人:高建平。
摘    要:为简便有效地鉴定党参药材及其混伪品并验证ITS/ITS2 序列作为DNA 条形码鉴定药材的稳定性和准确性,本研究选用党参药材及其混伪品作为研究对象,对33 份药材样本提取基因组DNA,通过PCR 扩增ITS 序列,采用比对法、最小距离法对序列鉴定能力进行评估。通过序列比对,分析变异位点信息确定不同的单倍型并计算种内和种间K2P 距离。ITS2 序列采用基于隐马尔可夫模型的HMMer(Hidden Markov Model)注释方法获得。结果表明,党参药材3 个基原物种ITS 序列长度为654-655 bp,ITS2 序列长度均为239 bp,ITS/ITS2 序列种内平均K2P 遗传距离均远小于其与混伪品的种间平均K2P 遗传距离,因此ITS/ITS2 序列作为DNA 条形码能稳定、准确鉴别党参药材及其混伪品,为其鉴定提供新的技术手段。

关 键 词:党参  ITS/ITS2  分子鉴定  DNA条形码
收稿时间:2013/5/24 0:00:00
修稿时间:6/4/2013 12:00:00 AM

Identification of Codonopsis Radix and Its Adulterants Using the ITS/ITS2 Barcodes
Zhao Sh,Xin Tianyi,Hou Dianyun,Pang Xiaohui,Chen Ruiyang and Gao Jianping.Identification of Codonopsis Radix and Its Adulterants Using the ITS/ITS2 Barcodes[J].World Science and Technology-Modernization of Traditional Chinese Medicine,2013,15(3):421-428.
Authors:Zhao Sh  Xin Tianyi  Hou Dianyun  Pang Xiaohui  Chen Ruiyang and Gao Jianping
Affiliation:School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100102, China;Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China;Dongfang Hospital of Beijing University of Chinese Medicine, Beijing 100078, China;College of Pharmacy, Shanxi Medical University, Taiyuan 030001, China
Abstract:The ITS/ITS2 barcodes were used to simply and effectively identify Codonopsis Radix and its adulterants. In this study, ITS (internal transcribed spacer of unclear ribosomal DNA) regions were amplified using PCR(polymerase chain reaction) from thirty-three samples of Codonopsis Radix and ITS2 regions were obtained from the ITS sequences using the hidden Markov model (HMMer)-based annotation methods. The sequences of ITS/ITS2 regions were aligned and the genetic distances were computed by MEGA5.0. Species identification efficiency of ITS/ITS2 sequences were evaluated using BLAST1 and nearest distance methods. The results indicated that The sequences lengths of ITS regions of Codonopsis Radix were 654-655 bp, and the lengths of ITS2 regions were 239 bp. The intraspecific genetic distances among Codonopsis Radix were obviously lower than the interspecific genetic distance between Codonopsis Radix and its adulterants. Therefore, ITS/ITS2 regions can stably and accurately distinguish Codonopsis Radix and its adulterants.
Keywords:Codonopsis Radix  ITS/ITS2  Molecular Identification  DNA barcoding
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