摘要
目的:中国药典收载的鸡血藤、大血藤、滇鸡血藤及相关物种均有"血藤"之称,其各自的来源不同,功效主治不同,不能混用。为确保其临床用药的安全、有效,本研究应用DNA条形码序列psbA-trnH对鸡血藤、大血藤、滇鸡血藤及其混伪品进行鉴定研究。方法:通过对样本进行基因组DNA提取,PCR扩增psbA-trnH序列。所得序列经拼接后,用MEGA5.0和DNAMAN进行种内种间序列分析,构建NJ树,对psbA-trnH序列在血藤类药材中的鉴定能力进行评估。结果:鸡血藤、大血藤和滇鸡血藤序列长度和GC含量各不相同,psbA-rrnH种内序列相似度高,均高于它们与混伪品间的序列相似性。鸡血藤、大血藤、滇鸡血藤与其他非药典收载的血藤类药材的种间最小K2P距离分别为0.137、0.426和0.003,均大于其种内最大遗传距离0.014、0.004和0。在NJ树中,鸡血藤、大血藤均单独聚为一支,能明显的与其他混伪品区分开,滇鸡血藤与异形南五味子聚为一支。结论:psbA-trnH序列能准确鉴定鸡血藤、滇鸡血藤、大血藤药材及其混伪品,该条形码分子鉴定方法可作为传统鉴定方法的有效补充。
Spatholobi Caulis,Sargentodoxae Caulis and Kadsurae Caulis recorded in the Chinese Pharmacopoeia were all called ＂Xueteng＂.Because of their different sources and efficacies,they cannot be misused.In order to ensure their clinical safety and efficacy,the psbA-trnH region was used to identify Spatholobi Caulis,Sargentodoxae Caulis,Kadsurae Caulis and their adulterants.Total genomic DNA was extracted,and the psbAtrnH region was amplified and sequenced.The intra- and inter-specific genetic distances were computed using MEGA5.0 and DNAMAN.The neighbor-joining（NJ） tree was constructed.The identification competence of psbA-trnH in ＂Xueteng＂ herbs was assessed.The results showed that these quence lengths and GC contents of Spatholobi Caulis,Sargentodoxae Caulis,Kadsurae Caulis were different.The intraspecific sequences had high similarities,which was higher than its interspecific sequence similarity compared to its adulterants.The minimum interspecific distances of Spatholobi Caulis,Sargentodoxae Caulis,Kadsurae Caulis were 0.137,0.426 and 0.003,respectively.All of them were more than their maximum intraspecific distances,which were 0.014,0.004 and 0.In NJ tree,Spatholobi Caulis and Sargentodoxae Cauliswere formed into one single branch,which can be clearly distinguished from other adulterants.Kadsurae Caulisand Kadsura heteroclite were formed into one single branch.It was concluded that the psbA-trnH region can accurately distinguish Spatholobi Caulis,Kadsurae Caulis,Sargentodoxae Caulisfrom their adulterants.This DNA barcoding molecular identification method can be used as an effective complement to the traditional identification methods.
出处
《世界科学技术-中医药现代化》
2016年第1期40-45,共6页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
科学技术部"863"计划(2012AA021602):基于DNA条形码的珍稀药用
野生等资源快速检测技术及产品研发
负责人:陈士林
