基于DNA条形码序列的辽宁省龙胆药材鉴定研究  

Study on identification of medicinal plant Gentiana scabra Bge.in Liaoning province based on DNA barcode sequences

作　　者：薛嘉宁 赵容[1] 于莹 高铭泽 尹海波[1] Xue Jianing;Zhao Rong;Yu Ying;Gao Mingze;Yin Haibo(School of Pharmacy,Liaoning University of Traditional Chinese Medicine,Dalian 116600,China)

机构地区：[1]辽宁中医药大学药学院,大连116600

出　　处：《国际中医中药杂志》2024年第1期103-107,共5页International Journal of Traditional Chinese Medicine

基　　金：辽宁省道地DNA指纹图谱建立项目(2022003)。

摘　　要：目的基于内转录间隔区(ITS)碱基序列,运用DNA条形码技术对辽宁省9个地区道地药用植物龙胆Gentiana scabra Bge.药材进行鉴别分析。方法利用DNA试剂盒提取法提取26批龙胆样本药用部位DNA,对ITS序列部分进行PCR扩增并双向测序,从Genbank下载药用植物龙胆的其他来源和外群序列,运用SeqMan 7.1.0软件对测序结果进行拼接,使用MEGA 7.0软件分析数据,计算K2P遗传距离,并利用邻接(NJ)法建立系统发育树进行分析。结果根据NJ聚类树结果可知,不同来源的龙胆样品可聚为一大支,其中坚龙胆和三花龙胆分别聚为一支,区别较明显;龙胆与条叶龙胆聚为一支,亲缘关系较近,且结合变异位点与遗传距离可知,龙胆与条叶龙胆的碱基序列相似,种间差异较小。26批样本中仅1个样品出现种内变异,其余样本碱基序列均相同,"清原龙胆"与辽宁省其他地区龙胆样品无明显差异。结论使用ITS序列的DNA条形码技术可用于龙胆药材及其不同来源的基原植物的种内和种间鉴别,且成功率较高。Objective To identify and analyze the genuine medicinal plant Gentiana scabra Bge.from 9 regions in Liaoning Province using DNA barcode technology based on the base sequence of internal transcribed spacer.Methods DNA was extracted from the medicinal parts of 26 Gentiana scabra Bge.samples by using DNA kit extraction method.The ITS sequence was amplified through polymerase chain reaction(PCR),and then two-way sequencing was carried out.Other sources and outgroup sequences of the medicinal plant Gentiana scabra Bge.were downloaded from Genbank.After the sequencing results were spliced by using SeqMan 7.1.0 software,MEGA 7.0 software was used to analyze and compare the data,and calculate the genetic distance of K2P(Kimura 2-parameter).The phylogenetic tree was established by Neighbor-Joining(NJ)method for analysis.Results According to the results of NJ cluster tree,all Gentiana scabra Bge.samples from different sources were clustered into one large branch,and Gentiana scabra Franch.and Gentiana triflora Pall.were clustered into one branch respectively,with obvious differences;Gentiana scabra and Gentiana manshurica Kitag.were clustered into one branch,and the genetic relationship was relatively close.In combination with the variation site and genetic distance,the base sequences of Gentiana scabra and Gentiana manshurica were very similar,and the interspecific differences were very small.Except for the intraspecific variation of only one sample collected in Liaoning Province,the base sequences of the other samples were the same,and there was no difference between"Gentiana scabra Bge.in Qingyuan"and Gentiana scabra Bge.samples from other regions in Liaoning Province.Conclusion The DNA barcode technology of ITS sequence can be used to differentiate and identify medicinal plant Gentiana scabra Bge.and its original plants from different sources with a high success rate.

关 键 词：龙胆 碱基序列 DNA条形码 分类学 辽宁省 

分 类 号：R282.5[医药卫生—中药学]