摘要
DNA条形码技术的快速发展将促进我国中药材的质量控制和标准化建设,是国际生物分类和鉴定研究中引人注目的新方向,但目前为止该技术相关报道多采用中药材基原植物叶片进行鉴定研究,直接应用于中药材鉴定较少,业内广泛关注的采用DNA条形码技术鉴定中药材过程涉及的①如何从根茎类中药材中提取DNA;②不同产地样本种内变异情况;③采用DNA条形码鉴定中药材时的稳定性和准确性等研究报道不足。在此背景下,本文选取多基原中药材秦艽作为研究对象,广泛收集其主产区不同基原和其常见混伪品及其近缘种共86个样品,改良传统DNA试剂盒提取方法,对比植物DNA条形码热点候选序列ITS(internal transcribedspacer),psbA-trnH,matK,rbcL和ITS2优劣,结果显示改良后的试剂盒法可使秦艽药材DNA全部提取成功,基于ITS2序列100%成功鉴定秦艽药材及其混伪品,同时基于本研究建立的鉴定流程成功将药店购买的秦艽药材样品鉴定到基原物种。
DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials： （1） the extraction methods for total DNA of the rhizomes of the medicinal materials; （2） intra-specific variation among samples from different places of origin; （3） accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions （ITS2, psbA-trnH, matK, rbcL, and ITS） were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode＇s utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.
出处
《药学学报》
CAS
CSCD
北大核心
2012年第12期1710-1717,共8页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(81130069,81073001)
863计划资助项目(2012AA021602)
教育部长江学者和创新团队发展计划资助项目(IRT1150)
关键词
秦艽药材
ITS2
物种鉴定
稳定性
准确性
Gentianae Macrophyllae Radix
ITS2
species identification
stability
accuracy
