药材天麻ITS2特异性引物的筛选  

Highly Specific DNA Barcoding Primers of ITS2 for Gastrodia elata

作　　者：马孝熙[1,2] 孙伟[1,2] 钱齐妮[3] 任伟超[1] 邬兰[1] 张雅琴[2] 宋明[2] 

机构地区：[1]中国中医科学院中药研究所,北京100700 [2]武汉理工大学化学工程学院,武汉430070 [3]重庆市药物种植研究所,重庆408435

出　　处：《世界科学技术-中医药现代化》2014年第2期325-328,共4页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：中国中医科学院中药研究所基本科研业务费自主选题项目(Z146):中草药DNA条形码的鉴定体系与中国药典标准;负责人:孙伟

摘　　要：目的:药材天麻由于隶属于单子叶植物兰科天麻属,天麻中含有大量多糖、蛋白质以及大量次生代谢物质,使其提取的DNA质量不高,影响后续的PCR扩增。且ITS2通用引物在单子叶植物部分物种通用性很差。本研究通过对DNA提取过程、PCR扩增过程进行优化,设计出一对用于鉴别天麻药材的ITS2引物,为中药材天麻的鉴定提供一种新方法。方法:以通用ITS2引物扩增出的两条序列为研究对象,经Primer Premier 5.0设计出3对特异性引物,通过对22份样品进行研究,筛选出一条高特异性的ITS2引物。结果:在54℃的退火温度下,TM2F-2R对天麻的鉴定效率高达90.9%。结论:TM2F-2R可作为天麻ITS2序列的特异性引物,为天麻的分子鉴定提供了一套准确、稳定的鉴定方法。This study aimed to design a pair of primers for amplifying internal transcribed spacer 2 region which was used to identify Gastrodia elata through optimizing of DNA extraction and PCR amplification process. Two sequences are used for research object by amplification of universal primers. Design three pairs of specific primers by Primer Premier 5.0 and select the highest specificity through the study of 22 samples. The results showed that identification efficiency of the primer named TM2F-2R is as high as 90.9% when Annealing Temperature is equal to 54 degrees Celsius. Therefore, TM2F-2R can be used as primers ITS2 sequences of G. elata, this article provides a set of accu-rate and stable identification methods for G. elata in the molecular identification.

关 键 词：天麻 ITS2 特异性引物 TM2F-2R 鉴定 

分 类 号：R284.1[医药卫生—中药学]