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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,psb A-trn H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psb A-trn H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psb A-trn H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。
Abstract：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 psb A-trn H 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 lindleyanum were also comprehensively analyzed to deduced the origin of Pleurospermum Lindleyanum samples. Results The total DNA was extracted successfully. The ITS2 primers and psb A-trn H 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 psb A-trn H 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 morphological characteristics and modern DNA barcoding.
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基本信息:
中图分类号:R282.5
引用信息:
[1]朱金芳,张平,胡江兰,等.天山棱子芹的DNA条形码分子鉴定[J].新疆医科大学学报,2021,44(08):948-952.
基金信息:
新疆维吾尔自治区十三五重大专项(2016A03005)
2021-02-28
2021
2021-07-27
2021
1
2021-08-15
2021-08-15