基于psbA-trnH序列的辽宁省黄精药材DNA条形码研究  

DNA Barcoding of Polygonatum in Liaoning Province Based on psbA-trnH Sequences

作　　者：高铭泽 于莹 薛嘉宁 尹海波[1] 赵容[1,2] GAO Ming-ze;YU Ying;XUE Jia-ning;YIN Hai-bo;ZHAO Rong(School of Pharmacy,Liaoning University of Traditional Chinese Medicine,Dalian 116600,China;State Key Laboratory of Dao-di Herbs,Beijing 100700,China)

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600 [2]道地药材国家重点实验室,北京100700

出　　处：《中国现代中药》2023年第3期493-498,共6页Modern Chinese Medicine

基　　金：中央本级重大增减支项目(2060302);辽宁省道地DNA指纹图谱建立项目(2022003)。

摘　　要：目的:基于psbA-trnH序列,运用DNA条形码技术对辽宁省12个地区及省外2个地区黄精药材样本进行鉴别分析,为辽宁省黄精药材的鉴别和遗传多样性分析提供参考。方法:利用DNA试剂盒提取法,从38个黄精样本的药用部位中提取DNA,对psbA-trnH部分进行聚合酶链式反应(PCR)扩增及双向测序,并从GenBank下载药用植物黄精的不同来源和外群序列,运用SeqMan软件对测序结果进行拼接,使用MEGA软件对数据进行分析比对,计算K2P(Kimura 2-parameter)遗传距离,并采用邻接法(NJ)建立系统发育树进行分析。结果:根据NJ系统发育树结果可知,不同来源的所有黄精样品聚为一大支,外群猪牙花聚为一支,区别较明显;辽宁省、湖北省及贵州省黄精与美国国家生物技术信息中心(NCBI)所下载的黄精聚为一支,并且结合变异位点与遗传距离可知,辽宁省、湖北省及贵州省的黄精物种为黄精Polygonatum sibiricum Red.,种间差异较小,辽宁省所收集的黄精有4个样品出现了变异位点,其余样本碱基序列均相同。结论:使用psbA-trnH序列DNA条形码技术能够对黄精药材不同的基原植物进行区分鉴别且成功率较高,可用于对黄精属物种进行种内和种间鉴别,为保障辽宁省黄精药材来源的准确鉴定提供参考。Objective:To distinguish the Polygonatum samples from 12 producing regions in Liaoning Province and 2 regions outside Liaoning by DNA barcoding based on psbA-trnH sequences,so as to provide a theoretical basis for the identification and genetic diversity analysis of Polygonatum in Liaoning.Methods:DNA was extracted from the medicinal parts of 38 Polygonatum samples by DNA extraction kits.Bidirectional sequencing was performed after polymerase chain reaction(PCR)amplification of psbA-trnH sequences.The sequences of different Polygonatum sources and the outgroup were downloaded from GenBank.A neighbor-joining(NJ)phylogenetic tree was established based on Kimura 2-parameter(K2P)after the sequence assembly by SeqMan and sequence alignment in MEGA 11.0.Results:According to the phylogenetic tree,all the Polygonatum samples from different sources were clustered into one clade,and the outgroup(Erythronium japonicum)was in a separate clade.The sequences of the Polygonatum samples from Liaoning,Hubei,and Guizhou and those downloaded from GenBank shared the same clade.This result,together with the mutation sites and genetic distance,indicated that the Polygonatum species in Liaoning,Hubei,and Guizhou was Polygonatum sibiricum Red.,with small interspecific differences.Four samples collected in Liaoning had mutation sites,and the sequences of the rest samples were the same.Conclusion:The DNA barcoding based on psbA-trnH sequences can distinguish Polygonatum plants of different origins with a high success rate,which can be used for the intraspecific and interspecific identification of Polygonatum and provide a reference for the accurate identification of Polygonatum resources in Liaoning.

关 键 词：黄精 PSBA-TRNH DNA条形码 

分 类 号：R282.[医药卫生—中药学]