铁线莲属蒙药材的rDNA-ITS序列分析

rDNA-ITS SEQUENCE ANALYSIS OF THE TRADITIONAL MONGOLIAN MEDICINAL PLANTS IN CLEMATIS L.

目的:比较研究铁线莲属入蒙药的6种药用植物(芹叶铁线莲、长瓣铁线莲、黄花铁线莲、棉团铁线莲、辣蓼铁线莲、短尾铁线莲)的rDNA-ITS序列差异性及其规律,寻找6种植物药之间的分子鉴别方法。方法:从铁线莲属蒙药材中提取总DNA,用PCR法扩增ITS,运用Clustal X、MEGA等软件比较和分析ITS的序列特征。结果:铁线莲属入蒙药的6种药用植物ITS序列长度为548～552bp,具变异位点57个,其中信息位点23个,特异性鉴别位点33个。序列已提交至NCBI数据库,登录号为JN809679-JN809684。结论:ITS序列对铁线莲属入蒙药的6种药用植物具有较好的分辨性,为铁线莲属蒙药材分子鉴别提供了参考。

Objective ： To investigate the patterns and differences of the rDNA ITS sequences of six traditional Mongolian medicinal plants in Clematis L. （C. aethusifolia, C. macropetala, C. intricata, C. hexapetala C. mandshurica and C. brevicaudata）and to establish a molecular biological method in iden- tification of these plants. Methods ： DNA was extracted from the dry leaves of the plants, ITS gene frag- ments were amplified by PCR and sequenced. The sequences were compared and analyzed using Clustal X and MEGA softwares. Results： The length of ITS region varied from 548 - 552 bp with 57 variable sites ,23 parsimony information sites and 33 singleton sites in the 6 plants in Clematis L. The sequences were submitted to NCBI database with the registry numbers of JN809679 - JN809684. Conclusion：ITS sequences could be used to identify the six species in Clematis L. and also could provide a reference for molecular identification.