摘要
目的应用ITS2条形码序列鉴别贯叶金丝桃及其混伪品。方法提取贯叶金丝桃及其混伪品DNA,使用ITS2通用引物进行PCR扩增测序,去除低质量碱基和拼接获得ITS2条形码目的序列。基于MEGA 6.0中的K2P模型,计算贯叶金丝桃及其混伪品的种内和种间遗传距离并构建系统发育树(NJ树),同时预测ITS2二级结构。结果贯叶金丝桃ITS2序列长度为236 bp,种内最大K2P遗传距离为0.004,与混伪品的种间K2P遗传距离分布为0.107~0.490;由NJ树可知,所有贯叶金丝桃均聚为一大支,呈现出明显的单系性,能很好地与其混伪品分开,同时贯叶金丝桃与其混伪品在ITS2的二级结构上差异明显。结论应用ITS2条形码序列可准确鉴别贯叶金丝桃及其混伪品,可用于贯叶金丝桃鉴别。
Objective To identify Hypericum perforatum and its adulterants by ITS2 bar code sequence. Methods The DNA of Hypericum perforatum and its adulterants were extracted and sequenced after ITS2 universal primers were amplified by PCR. Low-quality bases were removed and spliced to obtain ITS2 bar code sequence. Based on the K2P model in MEGA 6. 0,intraspecific and interspecific genetic distances were calculated and the phylogenetic tree（ NJ tree） was constructed,and the secondary structure of ITS2 was predicted. Results ITS2 sequence length of Hypericum perforatum was 236 bp,the maximum K2P genetic distance within the species was0. 004, and the interspecific K2 P genetic distance between Hypericum perforatum and its adulterants was 0. 107-0. 490. From the NJ tree, all the Hypericum perforatum clustered into a large branch, showing a clear monophyletic, which could be well separated from its adulterants. Meanwhile, there were significant differences in ITS2 secondary structure between Hypericum perforatum and its adulterants.Conclusion The ITS2 bar code sequence can accurately identify Hypericum perforatum and its adulterants, which can be used for identification of Hypericum perforatum.
作者
杨宋琪
唐小龙
陈海燕
柯潇
Yang Songqi;Tang Xiaolong;Chen Haiyan;Ke Xiao(Sichuan Jishengtang Pharmaceutical Co., Ltd., Chengdu, Sichuan, China 610041)
出处
《中国药业》
CAS
2018年第11期1-4,共4页
China Pharmaceuticals
基金
四川省重大产业技术创新专项[2012CD00082]
关键词
贯叶金丝桃
ITS2
鉴别
混伪品
Hypericum perforatum
TS2
identification
adulterants
