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当前位置: 首页 > 文章 > 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 核农学报 2020 (8) 1655-1665
Position: Home > Articles > Application of DNA Barcoding and Multiple Real-time Fluorescence Quantitative PCR for Authenticating Dendrobium officinale Journal of Nuclear Agricultural Sciences 2020 (8) 1655-1665

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

作  者：
陈雪燕;刘艳艳;谭晴晴;任金瑞;姜欣欣;姜秀梅;刘中华;张全芳
单  位：
关键词：
铁皮石斛;多重荧光PCR;探针;鉴定
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。
译  名：
Application of DNA Barcoding and Multiple Real-time Fluorescence Quantitative PCR for Authenticating Dendrobium officinale
作  者：
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;Shandong Jinan MeiYuan Dendrobium Officinale Cultivation Co.LTD;Shandong Wosen Agricultural Technology Co.LTD;
关键词：
Dendrobium officinale;;multiple real-time fluorescence quantitative PCR;;probe;;identification
摘  要：
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.

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