基于通用DNA条形码序列的黄精属药用植物分子鉴定  

Molecular authentication of medicinal Polygonatum Species utilizing the universal DNA barcode sequences

作　　者：张明英 李依民 程文萍 高静 颜永刚 杨琳 胡锦航 张岗 ZHANG Ming-ying;LI Yi-min;CHENG Wen-ping;GAO Jing;YAN Yong-gang;YANG Lin;HUJin-hang;ZHANG Gang(Shaanxi Qinling Application Development and Engineering Center of Chinese Herbal Medicine,College of Pharmacy,Shaanxi University of Chinese Medicine,Xi’an 712046,China;Shaanxi Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Shaanxi University of Chinese Medicine,Xianyang 712083,China;Key Laboratory for Research of"Qin Medicine"of Shaanxi Administration of Traditional Chinese Medicine,Shaanxi University of Chinese Medicine,Xi’an 712046,China)

机构地区：[1]陕西中医药大学药学院陕西省秦岭中草药应用开发工程技术研究中心,陕西西安712046 [2]陕西中医药大学陕西省中药资源产业化协同创新中心,陕西咸阳712083 [3]陕西中医药大学陕西省中医药管理局“秦药”研发重点实验室,陕西西安712046

出　　处：《中草药》2023年第1期235-244,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(82003898);陕西省自然科学基础研究计划项目(2022JM-458);陕西中医药大学校级科研课题(2020GP34);陕西中医药大学博士科研启动经费(104080001);陕西中医药大学“秦药”品质评价及资源开发学科创新团队项目(2019-QN01)。

摘　　要：目的 分析4个通用植物DNA条形码序列(trn H-psb A、mat K、rbc L和ITS2)及其组合对黄精属药用植物的物种鉴定分辨率,挖掘适用于黄精属种间鉴定的高分辨率分子标记。方法 以《中国药典》2020年版中收录的黄精属药用植物黄精Polygonatum sibiricum、滇黄精P. kingianum、多花黄精P. cyrtonema、玉竹P. odorati及其地方常见同属替代品、混伪品共12种79个野生个体为对象,将4个通用DNA条形码序列独立、联合分析,评估其种间、种内变异情况,并分别基于建树法(tree-based method)和PWG距离法(PWG-distance method)评估不同条形码及其组合的物种鉴定分辨率。结果 ITS2序列扩增成功率低,trn H-psb A、mat K、rbc L序列的引物在黄精属植物中通用性较好;3组叶绿体序列的种间变异依次为mat K>trn H-psb A>rbc L,种内变异差异不显著,种间、种内遗传距离无明显的Barcoding gap;各条形码独立及联合分析的物种鉴定分辨率普遍偏低,其中,组合条形码trn H-psb A+mat K+rbc L在建树法分析中的分辨率最高,为25%,trn H-psb A+rbc L在距离法分析中的分辨率最高,为50%。结论 4个通用DNA条形码序列及其组合都并非黄精属药用植物不同种间有效区分鉴定的理想分子标记,但多序列联合分析能在一定程度上提高物种鉴定成功率。Objective To investigate the species discrimination power of the four universal plant DNA barcodes(trn H-psb A, mat K,rbc L and ITS2) and corresponding multi-barcode combinations in Polygonatum, and to explore high-resolution molecular markers suitable for Polygonatum. Methods Seventy-nine wild individuals from 12 species, representing all the four medicinal species of Polygonatum(P. sibiricum, P. kingianum, P. cyrtonema, P. odoratum) included in the Chinese Pharmacopoeia(2020 Edition) and their local commonly used substitutions and inauthentic adulterants, were sampled. The interspecific and intraspecific genetic variation were estimated, tree-based and PWG-distance methods were applied to evaluate the species discrimination efficiency of each barcode sequence and their combinations. Results The primers of trn H-psb A, mat K and rbc L all showed good universality while most of the individuals failed to obtain ITS2 sequence in PCR amplification. The interspecific genetic variation of the three chloroplast sequences was mat K>trn H-psb A>rbc L, while their intraspecific genetic difference was not significant, and no obvious Barcoding gap was detected. All these barcode sequences including their combinations only get limited species resolution. Among which, the combination of trn H-psb A+mat K+rbc L possessed the best species-resolving power of 25% in tree-based method,trn H-psb A+rbc L showed the highest resolution degree of 50% in PWG-distance method. Conclusion None of the four barcode sequences nor their combinations were ideal molecular markers to address the problems of medicinal Polygonatum species authentication. Nonetheless, as the number of sequence increases, the degree of species resolution improves.

关 键 词：黄精属 DNA条形码 药用植物 物种鉴定 黄精 滇黄精 多花黄精 玉竹 

分 类 号：R286.12[医药卫生—中药学]