川乌与草乌的ITS序列分析

Study on ITS Sequences of Herbal Medicine of Chuanwu and Caowu

目的比较川乌、草乌及市场流通的混淆种类的内转录间隔区(ITS)序列,分析该片段在乌头类药材的DNA分子鉴别中的意义。方法利用PCR扩增和ABI377自动测序。结果川乌、草乌及其混淆种类之间在内转录间隔区(ITS)序列特征、碱基频率和遗传距离等方面都存在一定差异。结论ITS序列可准确鉴定川乌和草乌,以及草乌与其混淆种类,该方法更具简单性和实用性。

OBJECTIVE To analyze the ITS sequences of Chinese Aconitum species,including A, carmichaeli,A, kusnezoffii and those which are medicinally possibly used as their substitutes ,and to evaluate the utility of ITS sequences in the molecular authentication of the herbal medicinal aconites. METHODS ITS of 20 taxa of Aconitum were amplified using PCR systems and directly sequenced by an ABI 377 automated sequencer. RESULTS The sequencer analysis showed that the ITS 1 ranged from 248 to 252 bp, and the ITS 2 from 214 -217 bp,and the size of the 5.8S coding region was 164 bp for all the species studied. Chuanwu,Caowu and their adulterant species were shown to be different from each other in their characters of ITS sequences ,base frequencies and the pairwise genetic distances. CONCLUSION ITS region could be adopted as a molecular marker for differentiating medicinal Aconitum species.