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
方洁 1潘俊杰 1吕群丹 1周君美 1程科军1
作者信息
- 1. 丽水市农林科学研究院,浙江 丽水 323000
- 折叠
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.
Abstract
A total of 95 ITS2 sequences were obtained from 95 samples of Melastoma dodecandrum and confusable species like M.affine,M.can-didum,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 from 224 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Key words
Melastoma dodecandrum/confusable species/ITS2 sequence/DNA barcoding/identification引用本文复制引用
出版年
2024
NETL
NSTL
万方数据