摘要
目的:应用DNA条形码技术对青海冬虫夏草和新疆虫草进行鉴别,考查ITS序列和18S序列作为DNA条形码的可行性。方法:利用ITS序列和18S序列对冬虫夏草和新疆虫草的DNA提取物进行PCR扩增并测序,并与Gen Bank数据库中下载的共23条近缘虫草的ITS序列和18S序列进行比对;利用MEGA 6.0分析软件计算的K-2-P距离,构建虫草样品和参比序列的18S、ITS序列的NJ树。结果:通过遗传距离和NJ树分析可知,本研究所用20份青海冬虫夏草和新疆虫草样本确定为虫草类,与其近缘虫草各自聚为一支;ITS序列构建的NJ树能够从种属水平上将青海冬虫夏草和新疆虫草成功鉴别开来。结论:ITS序列较18S序列更适合作为冬虫夏草和新疆虫草的条形码进行鉴定研究。
This study aimed to identify Cordyceps sinensis（Berk.） Sacc from Qinghai Province and Cordyceps grsacilis（Grev.） Dur et Mont from Xinjiang Uygur Autonomous Region by DNA barcoding method. The feasibility of ITS sequence and 18 S sequence was examined as a DNA barcoding. The DNA samples were extracted and purified. The PCR amplification and bidirectional sequencing of the materials were carried out to obtain ITS and 18 S sequences. Then ITS and 18 S sequences achieved together with 23 ITS and 18 S sequences from Gen Bank database were aligned. MEGA program（version 6.0） was employed to calculate K2 P and built NJ trees of ITS and 18 S sequences. From the analysis of the experimental data,the 20 samples in this study were almost clustered together as its closely related genera formed a monophyletic group. The NJ tree constructed by ITS sequencescan identify the two species of Cordyceps in the species’ level. It was concluded that the ITS sequence was more suitable for the identification of Qinghai Cordyceps sinensis and Xinjiang Cordyceps grsacilis compared with the 18 S sequence.
出处
《世界科学技术-中医药现代化》
2016年第5期826-832,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然基金委地区科学基金项目(81460637):利用"组分中药"模式探讨哈萨克药材准噶尔乌头在体内的毒性标记物
负责人:张帆
新疆医科大学校级科研创新基金项目(XJC201312):基于DNA条形码技术的新疆虫草
冬虫夏草的鉴定
负责人:马红梅
