基于ITS2条形码序列鉴定中药材两头尖及其混伪品
Identification of Anemone raddeana and Its Adulterants by ITS2 Sequence
任伟超 1马伟 2安超 3马孝熙 4邬兰 4孙伟4
作者信息
- 1. 黑龙江中医药大学药学院 哈尔滨 150040; 中国中医科学院中药研究所 北京 100700
- 2. 黑龙江中医药大学药学院 哈尔滨 150040
- 3. 住房城乡建设部城乡规划管理中心 北京 100835
- 4. 中国中医科学院中药研究所 北京 100700
- 折叠
摘要
目的：本研究利用ITS2序列对中药材两头尖及其混伪品进行DNA条形码鉴定，以确保该药材的质量及临床疗效。方法：采用试剂盒法对36份样品提取DNA，通过聚合酶链式反应（PCR）扩增其ITS2序列并双向测序，所得序列经过CodonCode Aligner拼接后，用软件MEGA5.1进行数据分析，包括变异位点分析、种内种间距离计算及系统发育树构建。结果：两头尖药材的ITS2序列长度为216 bp，种内最大K2P距离为0.014，种间的最小K2P距离为0.021；NJ树结果显示两头尖与其混伪品可明显区分，表现出良好的单系性。结论：ITS2序列作为DNA条形码能稳定、准确鉴别两头尖药材，为其鉴定提供新的技术手段。
Abstract
In this study, the internal transcribed spacer 2 of nuclear ribosomal DNA (ITS2) sequence was used for i-dentifying A nemone raddeana and its adulterants to ensure the quality of medicines and clinical efficacy. Genomic DNA was extracted from 36 samples using Genomic DNA kit and used as templates for Polymerase Chain Reaction (PCR). Sequence assembly and consensus sequence generation were performed by CodonCode Aligner. The intraspe-cific and interspecific genetic distances were computed and the neighbor-joining tree was constructed by MEGA 5.1 in accordance with the Kimura 2-Parameter (K-2P) model. Results: The length of ITS2 sequence of A nemone rad-deana was 216 bp. The Maximum intraspecific genetic distance was 0.014, the minimum interspecific genetic dis-tance was 0.021. The NJ tree showed that A . raddeana differ from its adulterants obviously. Conclusion: ITS2 se-quence was able to identify A . raddeana and its adulterants correctly stably and correctly, which provides a new tech-nique to its identification.
关键词
两头尖/ITS2/鉴定/混伪品Key words
A nemone raddeana/ITS2/identification/adulterants引用本文复制引用
基金项目
黑龙江中医药大学人才项目后备带头人支持计划(2012001)
出版年
2014
国家科技期刊平台
NSTL
维普
万方数据