基于“传统+分子生物学”联用技术鉴定云南华宁产四数九里香药材  

Identification of the National Medicine Murraya Tetramera Huang Based on the Combination Technology of“Traditional+Molecular Biology”

作　　者：周永福[1,2,3] 姚如杰 黎学明 Zhou Yongfu;Yao Rujie;Li Xueming(Application Technology Promotion Center of Chongqing Industry Polytechnic College,Chongqing 401120,China;Technology Center,Chongqing Zi-Guang Chemical Co.,Ltd.,Chongqing 401121,China;College of Chemistry and Chemical Engineering,Chongqing University,Chongqing 400044,China)

机构地区：[1]重庆工业职业技术学院应用技术推广中心,重庆401120 [2]重庆紫光化工股份有限公司技术中心,重庆401121 [3]重庆大学化学化工学院,重庆400044

出　　处：《亚太传统医药》2023年第11期60-65,共6页Asia-Pacific Traditional Medicine

基　　金：重庆紫光化工股份有限公司博士后项目(KQY20220530)。

摘　　要：目的:鉴定民族药四数九里香药材。方法:采用“传统+分子生物学”联用技术:即在传统鉴定评价的基础之上,再采用DNA试剂盒提取样本DNA,结合九里香属植物ITS2基因序列设计引物,对样本进行PCR扩增,对所得产物进行双向测序,测序结果与GenBank中的九里香属植物ITS2基因序列进行比对,采用Mega 6.0软件计算序列间的遗传距离K2P,采用邻接法构建系统进化树,进一步对样本进行鉴别。结果:检测样本与九里香属植物K2P遗传距离介于0.017~0.048之间,其中与四数九里香(Murraya tetramera)K2P遗传距离最小为0.017,采用邻接法构建的系统树显示九里香属植物具有较好的单系性,均分别独立聚为1个分支,可以显著区分九里香属植物,检测样本与四数九里香(Murraya tetramera)聚为一支。结论:ITS2基因序列分析结果与传统评价结果相互佐证鉴定四数九里香,将为云南华宁产四数九里香的分类学研究提供科学线索。Objective:In order to identify the ethnic medicine Murraya tetramera Huang;Methods:In this paper,the“traditional+molecular biology”combined technology was used to identify the Murraya tetramera Huang:DNA samples were extracted by DNA extraction kit.Primers were designed to amplify ITS2 gene sequence of Murraya tetramera Huang.Two-way sequencing of ITS2 gene sequence was performed.The results were obtained with ITS2 gene sequence downloaded by GenBank.The genetic distance of K2P(Kimura-2-parameter)between sequences was calculated by Mega 6.0 software.Neighbor-joining method was used.The system tree was constructed to further identify the experimental samples.Results:The genetic distance of K2P between the tested samples and Jiulixiang plants ranged from 0.017~0.048.The genetic distance between the tested samples and Murraya tetramera K2P was the smallest,which was 0.017.The phylogenetic tree constructed by adjacency method showed that the plants had good monophyletic character and clustered independently into one branch.Conclusion:The results of ITS2 gene sequence identification and appearance evaluation mutually support each other,which will provide scientific clues for the taxonomic research of the Murraya tetramera Huang produced in Huaning,Yunnan.

关 键 词：“传统+分子生物学”技术 四数九里香 DNA分子鉴定 ITS2 

分 类 号：R284.1[医药卫生—中药学]