DNA条形码与实时荧光定量PCR技术在铁皮石斛鉴定中的应用  

Application of DNA Barcoding and Multiple Real-time Fluorescence Quantitative PCR for Authenticating Dendrobium officinale

作　　者：陈雪燕[1] 刘艳艳[1] 谭晴晴[1] 任金瑞 姜欣欣 姜秀梅 刘中华 张全芳[1] CHEN Xueyan;LIU Yanyan;TAN Qingqing;REN Jinrui;JIANG Xinxin;JIANG Xiumei;LIU Zhonghua;ZHANG Quanfang(Biotechnology Research Center,Shandong Academy of Agricultural Sciences/Laboratory of Applied Life Science,Shandong Academy of Agricultural Sciences,Jinan,Shandong 250100;Shandong Jinan MeiYuan Dendrobium Officinale Cultivation Co.LTD,Jinan,Shandong 250100;Shandong Wosen Agricultural Technology Co.LTD,Dezhou,Shandong 255039)

机构地区：[1]山东农业科学院生物技术研究中心/山东省农业科学院应用生命科学实验室,山东济南250100 [2]山东济南梅园铁皮石斛种植有限公司,山东济南250100 [3]山东沃森农业科技有限公司,山东德州255039

出　　处：《核农学报》2020年第8期1655-1665,共11页Journal of Nuclear Agricultural Sciences

基　　金：山东省创新公共服务平台专项计划(2018JGX111),山东省重点研发计划(2018GSF119002),济南市科技局项目(201711203)。

摘　　要：为建立铁皮石斛准确、高效的鉴定体系,本研究以101份石斛属和蝴蝶兰属植物样品为试验材料,通过对比ITS、psbA-trnH、matK及rbcL基因在石斛属植物中的鉴定能力,筛选出ITS作为本研究最理想的DNA条形码。以ITS序列作为靶基因,设计铁皮石斛特异引物和特异探针,以及石斛属通用引物和探针,利用实时荧光定量PCR(TaqMan)技术,建立铁皮石斛多重实时荧光PCR检测新体系,通过特异性、灵敏度和实际样本验证,发现参试样品中的25份铁皮石斛均可被有效鉴定,与其他鉴定方法相比,该方法具有特异性强、灵敏度高(高出普通PCR 100倍)、重复性好且高效经济的优势。本研究结果对铁皮石斛的资源保护与利用起到了积极作用。In order to establish an accurate and efficient identification system for Dendrobium officinale,101 Dendrobium and Phalaenopsis plants were used as experimental materials.ITS was screened as the most suitable DNA barcoding sequence for this study by comparing the identification ability of ITS,psbA-trnH,matK and rbcL genes in Dendrobium plants.The specific primers and probes were designed for Dendrobium officinale and Dendrobium plants.A new multiplex real-time fluorescent PCR detecting system of Dendrobium officinale was established by real-time fluorescence quantitative TaqMan technique.The results showed that 25 samples of Dendrobium officinale could be accurately identified by verification of specificity,sensitivity and actual samples.Compared with other identification methods,the method has the advantages of strong specificity,high sensitivity(100 times than ordinary PCR),good repeatability,high efficiency and economy.It plays a positive role in the protection and utilization of Dendrobium officinale resources.

关 键 词：铁皮石斛 多重荧光PCR 探针 鉴定 

分 类 号：S567.239[农业科学—中草药栽培]