赵新礼 舒泉湧 崔蕾 朱爱华

摘要 [目的]應用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。

关键词 倒卵叶五加;短柄五加;基原鉴定;DNA条形码;ITS2;psbA-trnH

中图分类号 R 282.5  文献标识码 A  文章编号 0517-6611（2021）15-0166-07

Abstract [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 onsite plants， using universal primers ITS2 and psbAtrnH， 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 psbAtrnH 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 interspecies genetic distance， and construct the NJ phylogenetic tree.[Result]The success rate of PCR amplification and sequencing of ITS2 and psbAtrnH was 100%.The plant morphology， ITS2 sequence and psbAtrnH 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 psbAtrnH：  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 psbAtrnH genetic markers， DNA barcode technology can accurately identify the 4 species of  Eleutherococcus  Maxim.;  E. brachypus  and  E. obovatus  should be the same plant.

Key words  Eleutherococcus obovatus;Eleutherococcus brachypus ;Base source identification;DNA barcoding;ITS2（internal transcribed spacer 2 sequence）;psbAtrnH（psbAtrnH intergenic spacer sequence）

基金項目 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）。

作者简介 赵新礼（1961—），男，陕西扶风人，主任药师，从事中药资源与栽培及分子生物学研究。

收稿日期 2021-01-27

倒卵叶五加与著名的中药材刺五加具有相似的化学成分和药理作用[1-2]，倒卵叶五加与短柄五加两者分布区域重叠，植物形态高度相似，在其产区内均作为倒卵叶五加药材使用，究竟两者为同一种植物还是2种植物长期以来存在争议[3-5]。……