铁皮石斛的PCR-RFLP鉴别方法  

Identification of Dendrobium officinale Kimura et Migo by PCR-RFLP

作　　者：王孟颖 万林春 赵雯 饶毅[1] 曾黎明 WANG Meng-ying;WAN Lin-chun;ZHAO Wen;RAO Yi;ZENG Li-ming(Jiangxi University of Traditional Chinese Medicine,Nanchang Jiangxi 330004,China;Jiangxi Provincial Institute for Drug Control,NMPA key Laboratory of Quality Evaluation of Traditional Chinese Patent Medicine,Jiangxi Provincial Engineering Research Center for Drug and Medical Device Quality,Nanchang 330029,China;College of biological science and engineering,Jiangxi Agricultural University,Nanchang,Jiangxi 330045,China)

机构地区：[1]江西中医药大学,南昌330004 [2]江西省药品检验检测研究院,国家药品监督管理局中成药质量评价重点实验室,江西省药品与医疗器械质量工程技术研究中心,南昌330029 [3]江西农业大学生物科学与工程学院,南昌330045

出　　处：《药物分析杂志》2023年第3期509-515,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：江西省重大科技研发专项项目-中药材大品种铁皮石斛产业链关键技术研究(20203KBC28W019)。

摘　　要：目的:建立铁皮石斛专有的分子鉴别方法。方法:通过对铁皮石斛及其近缘种石斛的叶绿体matk基因序列进行对比分析,根据鉴别位点设计特异的铁皮石斛鉴别引物,通过聚合酶链反应-限制性片段长度多态性方法(PCR-RFLP)技术进行鉴别。结果:本试验中鉴别引物可成功扩增铁皮石斛及其近缘种石斛,PCR产物为331 bp,铁皮石斛的PCR产物可被Tru9 Ⅰ限制性内切酶切开,而近缘种石斛不能被切开,可通过琼脂糖凝胶电泳成功鉴别。结论:本试验建立的PCR-RFLP鉴别方法可以鉴别铁皮石斛及其近缘种石斛。Objective:To establish a specific molecular identification method for Dendrobium officinale Kimura et Migo.Methods:Through the comparative analysis of chloroplast matk gene sequences of Dendrobium officinale Kimura et Migo and its closely related species,the specific primers were designed according to the distinguish sites,and the PCR-RFLP technique was used to identify Dendrobium officinale Kimura et Migo.Results:In this experiment,Dendrobium officinale Kimura et Migo and its closely related species were successfully amplified by the identification primers.The PCR product was 331 bp.The PCR product of Dendrobium officinale Kimura et Mi-go could be cut by Tru9 I restriction enzyme,but the PCR product of closely related species could not be cut.It could be successfully identified by agarose gel electrophoresis.Conclusion:The PCR-RFLP method established in this experiment can identify Dendrobium officinale Kimura et Migo and its closely related species.

关 键 词：铁皮石斛 PCR-RFLP法 分子鉴别 MATK基因 近缘物种 

分 类 号：R917[医药卫生—药物分析学]