基于4种序列的辽产苍术属植物DNA条形码鉴定

DNA Barcoding Identification of Atractylodes from Liaoning Province Based on Four Sequences

目的:采用4种序列对辽宁产苍术属植物进行鉴定,进而选择适合苍术属植物DNA条形码鉴定的序列。方法:使用试剂盒提取辽宁产4种苍术属植物关苍术Atractylodes japonica Koidz.ex Kitam、朝鲜苍术A.coreana(Nakai)Kitam、茅苍术A.lancea(Thunb.)DC.、北苍术A.chinensis(Bunge)Koidz.共43份样品的总DNA,进行聚合酶链式反应(PCR)扩增及产物测序;使用DNAMAN 8.0软件对测得的双向序列进行拼接;使用MEGA 6.0软件进行序列比对;使用邻接法构建系统聚类树。结果:仅内转录间隔区2(ITS2)序列将4种苍术属植物各分为一支,聚类效果较好,psb A-trn H、mat K、rbc L聚类混乱,物种分离度低。结论:ITS2序列可作为苍术属植物DNA条形码鉴定的有效序列片段,苍术野生品和栽培品、叶裂与叶不裂均聚在一起,ITS2序列片段不能区分。

Objective:Four sequences were used to identify the plants of Atractylodes from Liaoning Province,and the suitable sequences were selected.Methods:Genomic DNAs were extracted from 43 samples of A.japonica Koidz.ex Kitam,A.coreana(Nakai)Kitam,A.lancea(Thunb.)DC.and A.chinensis(Bunge)Koidz.,followed by sequences amplification through polymerase chain reaction(PCR).The sequencing results were splicing and comparison of the measured bidirectional sequences using DNAMAN 8.0.MEGA 6.0 software was used for sequence comparison,and adjacency method was used to construct the system cluster tree.Results:Only the internal transcribed spacer 2(ITS2)sequence divided the four species of Atractylodes into one branch,and the clustering effect was good.The clustering of psb A-trn H,mat K and rbc L was chaotic,and the species separation degree was low.Conclusion:ITS2 sequence can be used as an effective sequence fragment for the identification of Atractylodes plants by DNA barcoding.Wild and cultivated Atractylodes,leaf cleft and leaf uncleft are all clustered together,and ITS2 sequence fragments cannot be distinguished.