民族药用植物地菍及其常见易混种的DNA条形码鉴定  

Identification of Melastoma dodecandrum and Confusable Species by DNA Barcoding

作　　者：方洁 潘俊杰 吕群丹 周君美 程科军 FANG Jie;PAN Junjie;LU Qundan;ZHOU Junmei;CHENG Kejun(Lishui Institute of Agriculture and Forestry Sciences of Zhejiang,Lishui 323000,China)

机构地区：[1]丽水市农林科学研究院,浙江丽水323000

出　　处：《浙江林业科技》2024年第5期73-79,共7页Journal of Zhejiang Forestry Science and Technology

基　　金：丽水市科技项目(2020zdhz01)。

摘　　要：目的:基于ITS2条形码序列对民族药用植物地菍Melastoma dodecandrum及其常见易混种进行分子鉴定,以保证药材使用的正确性和临床药效。方法:分别在浙江、江西、福建和广西等地采集地菍及其7种常见易混种多花野牡丹M.affine、野牡丹M.candidum、展毛野牡丹M.normale、毛菍M.sanguineum、少花柏拉木Blastus pauciflorus、锦香草Phyllagathis cavaleriei和秀丽野海棠Bredia amoena样品95份,提取DNA并进行PCR扩增,测序并获取ITS2序列,同时下载GenBank相应物种的ITS2序列30条。使用Gene Tool软件分析ITS2序列长度、GC含量和变异位点等情况,利用Clustal X和MEGA 7.0软件计算遗传距离和构建邻接(NJ)系统发育聚类树。结果:地菍及其常见易混种ITS2序列长度为224~236 bp,GC含量为60.6%~69.2%;地菍只有1种单倍型,种内K2P遗传距离显著小于其他7个易混种;邻接(NJ)进化树中地菍可与其他物种成功区分。结论:基于ITS2序列的DNA条形码技术能够成功鉴定地菍及其常见易混种,可为地菍的生产、使用和监管提供有力的安全保障。A total of 95 ITS2 sequences were obtained from 95 samples of Melastoma dodecandrum and confusable species like M.affine,M.candidum,M.normale,M.sanguineum,Blastus pauciflorus,Phyllagathis cavaleriei,Bredua amoena collected from Zhejiang,Jiangxi,Fujian and Guangxi provinces.And 30 ITS2 sequences of corresponding species were downloaded from GenBank.Sequence characteristics such as length,GC contents,and variable sites were analyzed by Gene Tool software while genetic distance calculation and neighbor-joining(NJ) phylogenetic tree was built by Clustal X and MEGA 7.0.The result demonstrated that the length of ITS2 sequences of M.dodecandrum and confusable species ranged from224 to 236 bp with GC content of 60.6%-69.2%.The intra-specific K2P genetic distance of M.dodecandrum was significantly lower than that among the other seven confusable species.It was obvious that the ITS2 sequences of M.dodecandrum formed into one separate clade in the phylogenetic tree,while the other seven confusable species clustered into other clades.

关 键 词：民族药用植物 地菍 ITS2序列 DNA条形码 鉴定 

分 类 号：S567[农业科学—中草药栽培]