摘要
目的应用DNA条形码技术对采集的天山棱子芹样本基原进行分子鉴定。方法提取样品总DNA并以ITS2扩增序列为主,psbA-trnH扩增序列为参考,对天山棱子芹样本DNA进行扩增,通过琼脂糖凝胶电泳检测总DNA及引物扩增结果,并进行测序。将两引物扩增成功并测序后的序列导入美国国立生物技术信息中心(national center of biotechnology information,NCBI)数据库比对,并对样本ITS2序列及其同科属植物序列进行NJ树聚类分析,再结合形态学特征推断天山棱子芹样本的基原。结果总DNA提取成功,ITS2引物及psbA-trnH引物均扩增成功。NCBI数据库比对结果显示,样本ITS2序列与伞形科棱子芹属天山棱子芹(Hymenidium nanum)的相似度(98.2%)高于ps⁃bA-trnH序列与瘤果芹属裂苞瘤果芹(Trachydium involucellatum)的相似度(93.92%);NJ树聚类分析结果显示,样本与天山棱子芹聚为一支,支持率为92%;形态学特征比对结果表明,样本与天山棱子芹形态特征吻合。结论综合传统的形态学与现代DNA条形码鉴定结果,最终确定样本的基原为天山棱子芹。
Objective To identify origin of Pleurospermum lindleyanum by DNA barcoding technique.Methods The total DNA of the samples was extracted and the DNA of Pleurospermum lindleyanum samples were amplified with ITS2 main sequences and psbA-trnH reference sequences.Total DNA and primer amplification results were detected by agarose gel electrophoresis and sequenced.The amplified and sequenced sequences of the two primers were imported into NCBI database for comparison.Meanwhile,the ITS2 sequences of the samples and the downloaded sequences of the same family plants were clustered by NJ tree.Then,the morphological characteristics of Pleurospermum lindleyan⁃um were also comprehensively analyzed to deduced the origin of Pleurospermum Lindleyanum samples.Results The total DNA was extracted successfully.The ITS2 primers and psbA-trnH primers were amplified successfully.The comparison results of NCBI database showed that the similarity between ITS2 amplified sequence and Hymenidium nanum(98.2%)was higher than that between psbA-trnH amplified sequence and Trachydium involucellatum(93.92%).The results of NJ tree cluster analysis showed that the samples and Hymenidium nanum were grouped together,and the support rate was 92%.The comparison results of morphological characteristics showed that morphological characteristics of samples were consistent with those of Pleurospermum lindleyanum.Conclusion It could be eventually deduced that the origin of samples is Pleurospermum lindleyanum via comprehensively analyzing the results of traditional morpholog‐ical characteristics and modern DNA barcoding.
作者
朱金芳
张平
胡江兰
阿吉艾克拜尔·艾萨
ZHU Jinfang;ZHANG Ping;HU Jianglan;Ajiakber Aisa(Key Laboratory of Utilization of Unique Medical Resource in Xinjiang Technical Institute of Physics and Chemistry,Chinese Academy of Sciences,Urumqi 830011;School of Food Science and Pharmacy,Xinjiang Agricultural University,Urumqi 830052,China)
出处
《新疆医科大学学报》
CAS
2021年第8期948-952,共5页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区十三五重大专项(2016A03005)。
关键词
天山棱子芹
DNA条形码
分子鉴定
ITS2
Pleurospermum lindleyanum
DNA barcoding
molecular identification
ITS2
