文章摘要
基于DNA条形码技术鉴定蒙药材刺柏叶及其混淆品
Identification of Juniperus Folium the Mongolian medicine and its adulterants based on DNA barcoding technique
投稿时间：2023-10-23  修订日期：2024-04-03
DOI：
中文关键词: 蒙药材  刺柏叶  混淆品  DNA条形码  鉴别
英文关键词: Mongolian medicine  Juniperus Folium  DNA barcoding  adulterants  identification
基金项目:
作者单位邮编
娜仁图雅 内蒙古药品检验研究院 010000
刘杰 中国食品药品检定研究院 
籍学伟 内蒙古药品检验研究院 
徐继民 中国食品药品检定研究院 
郑健* 中国食品药品检定研究院 102629
韩塔娜 内蒙古药品检验研究院 
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中文摘要:
      目的：基于DNA条形码技术鉴别蒙药材刺柏叶及其混淆品。方法：采用国际通用的条形码序列ITS2、psbA-trnH、matK和rbcL,对刺柏叶及其混淆品圆柏叶共计11份样品进行DNA提取、扩增,采用CodonCode Aligner进行序列拼接,并用MEGA软件对拼接序列进行变异位点分析,NJ聚类分析,并计算其平均种内、种间遗传距离。结果: 4对引物的PCR扩增产物测序成功率分别为ITS2 (100%)、psbA-trnH (100%)、rbcL (100%)、matK (0%)；ITS2序列和psbA-trnH序列均可通过变异位点比较区分刺柏叶及其混淆品；NJ聚类分析的结果显示, psbA-trnH序列的NJ聚类树中刺柏叶与圆柏叶均能分别聚为一支,且psbA-trnH序列的平均种间遗传距离明显大于平均种内遗传距离。结论： psbA-trnH序列能有效区分圆柏叶与刺柏叶,可作为鉴别刺柏叶及其混淆品圆柏叶的条形码序列,为蒙药材刺柏叶及其混淆品的鉴别提供支持。
英文摘要:
      Objective: To identify the original plant of Juniperus Folium from its adulterants by DNA barcoding. Method: The DNA extraction and amplification of 11 samples of Juniperus Folium and the adulterants Sabina Folium were carried out by using the internationally recognized barcode sequences ITS2, psbA-trnH, matK and rbcL. CodonCode Aligner was adopted to align the sequence, and MEGA software was used to analyze the mutation sites and Neighbor-Joining cluster analysis of the aligner sequence, and to calculate its average intra-specific and inter-specific genetic distance. Results: The success rates of PCR amplification products were ITS2 (100%), psbA-trnH (100%), rbcL (100%) and matK (0%), respectively. The ITS2 sequences and psbA-trnH sequences could distinguish the Juniperus Folium and its adulterants by comparison of variation sites. The results of Neighbor-Joining cluster analysis showed that the psbA-trnH sequences of Juniperus Folium and its adulterants could be clustered into one branch respectively, and the average interspecific genetic distance of psbA-trnH sequences were significantly greater than the average intraspecific genetic distance. Conclusion: The psbA-trnH sequences can effectively distinguish Juniperus Folium from Sabina Folium, and can be used as barcode sequence to identify Juniperus Folium and the adulterants Sabina Folium, which would provide support for the identification of Mongolian medicinal materials Juniperus Folium and the adulterants Sabina Folium.
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