摘要
为了探索鉴定桑白皮及其混伪品的新方法,本实验提取桑白皮药材及其混伪品的DNA,对ITS2序列进行PCR扩增和双向测序,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MEGA4.0软件计算物种种内种间Kimura 2-pa-rameter（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。结果表明桑白皮药材与其混伪品之间的K2P遗传距离分布于0.003~0.343,大于桑种内K2P遗传距离0,NJ树也显示桑白皮可与其混伪品明显分开。因此,ITS2条形码可有效鉴别中药材桑白皮及其混伪品,为快速准确地鉴定桑白皮提供了科学依据和新的方法。
To find a new method to identify Mori Cortex and its adulterants by analysis their ITS2 sequence of barcode,total genomic DNA was isolated from Mori Cortex and its adulterants.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 us ing software MEGA 4.0.Identification analyses were performed using BLAST1,Nearest Distance and neighbor joining（NJ） methods.The results showed the intra-specific genetic distance of Mori Cortex was 0,which were lower than inter-specific genetic distances between Mori Cortex and its closely related species（0.003-0.343）.The ITS2 region is an efficient barcode for identification of Mori Cortex and its closely related species,which provides a scientific basis for fast and accurate identification and new method of Mori Cortex.
出处
《世界科学技术-中医药现代化》
北大核心
2013年第3期393-396,共4页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
科学技术部国家科技支撑计划项目(2011BAI07B08):中药材质量评价体系建立及其质量标准示范研究
负责人:杨秀伟
