天然产物研究与开发 ›› 2018, Vol. 30 ›› Issue (12): 2193-2202.doi: 10.16333/j.1001-6880.2018.12.025

• 开发研究 • 上一篇    下一篇

重楼法定基原与其同属近缘种的多重PCR鉴别体系研究

张开元1,饶文霞1,尹显梅1,陈蓉2,3,崔欣2,唐卓2*,尹鸿翔3*   

  1. 1成都中医药大学药学院,成都 611137;2中国科学院成都生物研究所天然产物中心,成都 610041;3成都中医药大学民族医药学院,成都 611137
  • 出版日期:2019-01-02 发布日期:2019-01-02
  • 基金资助:

    国家自然科学基金(81573545、81001606);霍英东教育基金 (121045);四川省科技厅应用基础研究项目(2017JY0136);四川省崇州市现代农业创新驱动试点区(2015-CX00-00004-ZF)

Multiplex PCR System for the Identification of Paridis Rhizoma’s Legal Origins and Its Closely Related Species from Genus Paris

ZHANG Kai-yuan1, RAO Wen-xia1, YIN Xian-mei1, CHEN Rong2,3, CUI Xin2, TANG Zhuo2*, YIN Hong-xiang3*   

  1. 1College of pharmacy,Chengdu University of TCM,Chengdu 611137,China; 2Natural Products Research Center,Chengdu Institute of Biology,Chinese Academy of Sciences,Chengdu 610041,China; 3College of ethnic medicine,Chengdu University of TCM,Chengdu 611137,China
  • Online:2019-01-02 Published:2019-01-02

摘要: 本研究通过分析重楼法定基原(七叶一枝花和云南重楼)与其同属近缘种ITS序列的差异设计了3对特异性引物。利用多重聚合酶链反应(PCR)原理建立了一套能快速、准确鉴别重楼法定基原与其同属近缘种的方法。最终,七叶一枝花样本可扩增出片段大小为210 bp的特异性片段,云南重楼种下的两类基因型(YN-I 、YN-II)样本可分别扩增出248 bp和149 bp的特异性片段。在法定基原得到阳性鉴定的同时,其他同属近缘种无任何片段,避免了干扰。该方法首次实现了,在一套多重PCR反应体系中同时鉴别重楼药典法定基原与其同属近缘种,对于提升中药重楼的质量控制技术水平具有重要意义。

关键词: 重楼, 法定基原, 同属近缘种, 多重PCR, 鉴定, 七叶一枝花, 云南重楼

Abstract: In this study,three specific primers pairs were designed bases on comparison of the difference of the ITS gene sequences of Rhizoma Paridis’ legal origins (Paris polyphylla var. chinensis and P.polyphylla var.yunnanensis) from their closely related species.A fast and accurate method was established to identify Rhizoma Paridis’ legal origins and their closely related species according to the principles of multiplex polymerase chain reaction(PCR).Eventually,the size of 210 bp specific bands were amplified from the P.polyphylla var. chinensis;the sizes of 248 bp and 149 bp specific bands were amplified from P.polyphylla var. yunnanensis samples of two kinds of genotypes (YN-I and YN-II) respectively.The positive identification of legal origins were obtained while their closely related species weren’t amplified any bands to interfere with the result.For the first time,this method realized authentification of Rhizoma Paridis’ legal origins in one multiple PCR identification system simultaneously.This method is of great significance for improving the technical level of quality control of Rhizoma Paridis.

Key words: Rhizoma Paridis, legal origins, closely related species, multiplex polymerase chain reaction, identification, Paris polyphylla var. chinensis, Paris polyphylla var.yunnanensis

中图分类号: 

R932