倒卵叶五加药材的基原鉴定研究  

Study on the Base Source Identification of Eleutherococcus obovatus Radix et Rhizoma seu Caulis

作　　者：赵新礼 舒泉湧 崔蕾 朱爱华 ZHAO Xin-li;SHU Quan-yong;CUI Lei(Shaanxi Institute of Traditional Chinese Medicine(Shaanxi Medical Information Center),Xianyang,Shaanxi 712000;Shaanxi Key Laboratory of Traditional Chinese Medicine and Natural Medicine Research and Development,Xi’an,Shaanxi 710075)

机构地区：[1]陕西中药研究所(陕西医药信息中心),陕西咸阳712000 [2]陕西省中药与天然药物研发重点实验室,陕西西安710075

出　　处：《安徽农业科学》2021年第15期166-172,180,共8页Journal of Anhui Agricultural Sciences

基　　金：陕西省科技统筹创新工程项目(2016KTTSSF 01-01-02);陕西省咸阳市重大科技专项(2014K 01-17)。

摘　　要：[目的]应用DNA条形码鉴定技术对短柄五加等五加属植物进行研究,以弄清倒卵叶五加和短柄五加的物种关系。[方法]通过对馆藏标本与现地植物形态进行观测,采用通用引物ITS2和psbA-trnH对采自陕甘宁地区的短柄五加等50个样本进行扩增后双向测序,并从GenBank数据库下载了五加属等相关植物的序列,分别通过ITS2 database或其序列注释,获得ITS2和psbA-trnH序列,随后进行BLAST比对鉴定分析;用MEGA 6.06软件对所得序列进行比对,分析变异位点,计算GC含量、种内和种间遗传距离,构建NJ系统发育树。[结果]ITS2和psbA-trnH序列的PCR扩增和测序成功率均为100%;倒卵叶五加和短柄五加的植物形态、ITS2序列和psbA-trnH序列完全相同。ITS2序列:短柄五加和糙叶五加仅有一个碱基的差异,蜀五加与藤五加序列相同;psbA-trnH序列:短柄五加与糙叶五加的差异显著,短柄五加与蜀五加序列相同。[结论]同时使用ITS2、psbA-trnH这2个标记的DNA条形码可以准确鉴别短柄五加等五加属植物;短柄五加与倒卵叶五加为同一种植物。[Objective]The molecular identification technique of DNA barcode was used to identify the species of several plants of Eleutherococcus Maxim.in order to understand the relationship between Eleutherococcus brachypus and Eleutherococcus obovatus.[Method]By observing the morphology of collection specimens and on-site plants,using universal primers ITS2 and psbA-trnH,50 samples including Eleutherococcus brachypus collected from Shaanxi,Gansu and Ningxia were amplified and sequenced in both directions,and downloaded the sequence of Araliacea and other related plants from GenBank database,obtained ITS2 and psbA-trnH sequence through ITS2 database or its sequence annotation,and then performed BLAST comparison and identification analysis.MEGA 6.06 software was used to compare the obtained sequences,analyze the mutation sites,calculate the GC content,intra-and inter-species genetic distance,and construct the NJ phylogenetic tree.[Result]The success rate of PCR amplification and sequencing of ITS2 and psbA-trnH was 100%.The plant morphology,ITS2 sequence and psbA-trnH sequence of Eleutherococcus brachypus and Eleutherococcus obovatus were identical.ITS2 sequence:Eleutherococcus brachypus and Eleutherococcus henryi had only one base difference,and the sequence of Eleutherococcus leucorrhizus var.setchuenensis was the same as that of Eleutherococcus leucorrhizus.The sequence of psbA-trnH:Eleutherococcus brachypus and Eleutherococcus henryi sequence were significantly different,and the sequence of Eleutherococcus brachypus and Eleutherococcus leucorrhizus var.setchuenensis was the same.[Conclusion]At the same time,using ITS2 and psbA-trnH genetic markers,DNA barcode technology can accurately identify the 4 species of Eleutherococcus Maxim.;E.brachypus and E.obovatus should be the same plant.

关 键 词：倒卵叶五加 短柄五加 基原鉴定 DNA条形码 ITS2 PSBA-TRNH 

分 类 号：R282.5[医药卫生—中药学]