摘要
目的:本研究应用ITS2序列作为DNA条形码,建立余甘子及其混淆品分子鉴定方法。方法:收集8种共17份余甘子及其混淆品植物样本,提取基因组DNA,扩增ITS2基因并双向测序。所得序列采用Codon Code Aligner进行拼接,同时,从Gen Bank数据库中获3条相关ITS2序列用于比较分析。利用MEGA 6.06软件分析其种内及种间遗传距离,并构建系统进化树。结果:余甘子ITS2序列长度为208 bp,与其他各物种种间变异位点较多,遗传距离较大,为0.174-0.823。聚类树结果显示,余甘子基原植物独聚一支,与其他混淆品能够容易区分。结论:本研究采用ITS2序列能够有效鉴别余甘子及其混淆品基原植物,可为保证临床用药真实安全提供技术支撑。
In order to guarantee the species correction of Phyllanthus emblica, a molecular identification method taken ITS2 as DNA barcode has been verified. Samples from P. emblica and its adulterants were collected, and their ITS2 regions were amplified and sequenced. Sequences were then assembled by CodonCode Aligner and analyzed with MEGA 6.06. The Kimura 2-Parameter （K2P） distances were computed and the phylogenetic tree was built using the neighbor-joining （NJ） and maximum likelihood （ML） trees. It was found that the length of ITS2 sequence of P. emblica was 208 bp. The K2P distance of the inter-specific samples between P. emblica and its adulterants ranged from 0.174 to 0.823. The phylogenetic tree indicated that P. emblica and its adulterants could be distinguished clearly. In conclusion, it was demonstrated that the ITS2 sequence was convevient for uncovering the molecular evidence behind the identification of P. emblica and its adulterants, and provided a fundation for identifying the original plants, which secured the safety and effectiveness of clinical medication.
出处
《世界科学技术-中医药现代化》
2016年第8期1419-1423,共5页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
广东省中医院院内专项(2015KT1817):岭南中草药DNA条形码分子鉴定和生态适宜性研究
负责人:黄志海
关键词
余甘子
ITS2序列
分子鉴定
DNA条形码
Phyllanthus emblica, ITS2, molecular identification, adulterants, DNA barcode
