北苍术种子DNA条形码研究及序列特征分析  

DNA Barcoding for Atractylodes chinensis(DC.)Koidz.Seeds and its Sequence Characteristics Analysis

作　　者：谢红波 赵晴 穆威杉 史萌萌 石林春[2] 刘金欣[1] 赵春颖[1] 李云峰[1] XIE Hong-bo;ZHAO Qing;MU Wei-shan;SHI Meng-meng;SHI Lin-chun;LIU Jin-xin;ZHAO Chun-ying;LI Yun-feng(Chengde Medical University,Hebei Key Laboratory of Study and Exploitation of Chinese Medicine,Hebei Chengde,067000,China;Chinese Academy of Medical Sciences&Peking Union Medical College,Institute of Medicinal Plant Development,Beijing,100193,China)

机构地区：[1]承德医学院河北省中药研究与开发重点实验室,河北承德067000 [2]中国医学科学院&北京协和医学院药用植物研究所

出　　处：《承德医学院学报》2022年第1期13-18,共6页Journal of Chengde Medical University

基　　金：承德医学院国家自然科学基金项目培育基金(202011);承德医学院重大项目科研专项(KY2020003);“河北省中药研究与开发重点实验室”开放性课题(ZYKF202002);河北省中医药管理局2017年度中医药类科研计划课题(2017079)。

摘　　要：目的基于DNA条形码技术对北苍术种子进行鉴定研究及序列特征分析,探讨北苍术种子DNA条形码技术鉴定的可行性。方法收集北苍术种子12份,利用体视显微镜观察北苍术的外观形态特征;通过DNA提取、聚合酶链式反应(PCR)和双向测序获得其ITS2和psbA-trnH序列;利用CodonCode Aligner V9.0.1对其峰图质量进行评价,使用MEGA-X进行多序列比对和变异位点分析。结果北苍术种子的DNA提取、PCR扩增和测序成功率均为100%,且取得良好的测序结果;分别各获得12条北苍术种子的ITS2序列和psbA-trnH序列,其中获得的ITS2序列仅存在2个插入/缺失位点,psbA-trnH序列包含3个变异位点和4个单倍型,表明不同产地的北苍术种子种内变异相对较小。结论基于ITS2序列和psbA-trnH序列的DNA条形码研究和序列分析,为北苍术种子的物种鉴定和遗传多样性研究提供了参考。Objective Identification and sequence characteristics analysis of Atractylodes chinensis(DC.)Koidz.seeds were executed based on DNA barcoding technology,to explore the feasibility of the DNA barcoding technology for the authentication of Atractylodes chinensis(DC.)Koidz.seeds.Methods A total of 12 seeds of Atractylodes chinensis(DC.)Koidz.were collected,and the morphological features of seeds were described by stereomicroscope.The ITS2 and psbA-trnH sequences were obtained after DNA extraction,polymerase chain reaction(PCR)and bi-directional sequencing.The quality of chromatographic traces was evaluated using Codoncode Aligner V9.0.1,and multi-sequence alignment and mutation site analysis were performed by MEGA-X.Results The results of DNA extraction,PCR amplification and sequencing were successfully obtained with a rate of 100%,and the sequencing output files were high quality.Moreover,12 ITS2 sequences and 12 psbA-trnH sequences of Atractylodes chinensis(DC.)Koidz.seeds were obtained,respectively.Among which,two insertion/deletion sites were found in ITS2 sequences,and 12 psbA-trnH sequences could be divided into four haplotypes,including three variable sites.It indicated that the intraspecific variation of Atractylodes chinensis(DC.)Koidz.seeds from different habitats was relatively conservative.Conclusion The research of DNA barcoding and sequence characteristics analysis based on ITS2 and psbA-trnH sequences can provide support for species identification and genetic diversity of Atractylodes chinensis(DC.)Koidz.seeds.

关 键 词：北苍术 种子 DNA条形码 ITS2 PSBA-TRNH 序列特征 

分 类 号：R28[医药卫生—中药学]