摘要
目的:利用DNA条形码鉴定技术快速、准确地鉴别中药材薄荷及其密切相关种。方法:提取薄荷药材及其易混品的DNA、对ITS2序列进行PCR扩增和测序,应用CodonCode Aligner V3.0对测序峰图进行校对拼接,去除低质量序列及引物区,获得ITS2序列。利用MEGA5.0软件计算物种种内种间Kimura 2-parameter(K2P)遗传距离。采用相似性搜索法、最近距离法、构建NJ系统聚类树等方法进行鉴定分析。结果:薄荷药材与其他密切相关种之间的K2P遗传距离分布于0.071～0.231,大于薄荷种内K2P遗传距离(0～0.006);3种鉴定方法也表明ITS2序列适合鉴别薄荷药材与其密切相关种。结论:ITS2条形码可有效鉴别中药材薄荷及其易混品,为快速、准确地鉴定薄荷提供了科学依据。
Objective： To identify Menthae Haplocalycis Herba and its closely related species using DNA barcoding technique.Method： Total genomic DNA was isolated from Mentha canadensis and its closely related species.Nuclear DNA ITS2 sequences were amplified,and purified PCR products were sequenced.Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner V3.0.The Kimura 2-Parameter（K2P） distances were calculated using software MEGA 5.0.Identification analyses were performed using BLAST1,Nearest Distance and neighbor-joining（NJ） methods.Result： The intra-specific genetic distances of M.canadensis were ranged from 0 to 0.006,which were lower than inter-specific genetic distances between M.canadensis and its closely related species（0.071-0.231）.All the three methods showed that ITS2 could discriminate M.canadensis from its closely related species correctly.Conclusion： The ITS2 region is an efficient barcode for identification of Menthae Haplocalycis Herba,which provides a scientific basis for fast and accurate identification of the herb.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2012年第8期1114-1117,共4页
China Journal of Chinese Materia Medica
基金
工信部中药材生产扶持项目[工信部消费(2011)340]
关键词
DNA条形码
ITS2
薄荷
DNA barcoding
ITS2
Menthae Haplocalycis Herba
