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一种基于简单实时聚合酶链反应(PCR)的检测方法,用于从当地人参种群中鉴定中国栽培人参品种大马牙。

A simple real-time polymerase chain reaction (PCR)-based assay for authentication of the Chinese Panax ginseng cultivar Damaya from a local ginseng population.

作者信息

Wang H, Wang J, Li G

机构信息

School of Life Science, Yantai University, Yantai, China.

School of Pharmacy, Yantai University, Yantai, China.

出版信息

Genet Mol Res. 2016 Jun 27;15(2):gmr8801. doi: 10.4238/gmr.15028801.

Abstract

Panax ginseng is one of the most important medicinal plants in the Orient. Owing to its increasing demand in the world market, cultivated ginseng has become the main source of medicinal material. Among the Chinese ginseng cultivars, Damaya commands higher prices and is grown in significant proportions among the local ginseng population. Due to the lack of rapid and accurate authentication methods, Damaya is distributed among different cultivars in the local ginseng population in China. Here, we identified a unique, Damaya-specific single nucleotide polymorphism (SNP) site present in the second intron of mitochondrial cytochrome c oxidase subunit 2 (cox2). Based on this SNP, a Damaya cultivar-specific primer was designed and an allele-specific polymerase chain reaction (PCR) was optimized for the effective molecular authentication of Damaya. We designed a method by combining a simple DNA isolation method with real-time allele-specific PCR using SYBR Green I fluorescent dye, and proved its efficacy in clearly discriminated Damaya cultivar from other Chinese ginseng cultivars according to the allelic discrimination analysis. Hence, this study provides a simple and rapid assay for the differentiation and conservation of Damaya from the local Chinese ginseng population.

摘要

人参是东方最重要的药用植物之一。由于其在世界市场上的需求不断增加,栽培人参已成为药材的主要来源。在中国参品种中,大马牙价格较高,且在当地人参群体中种植比例较大。由于缺乏快速准确的鉴定方法,大马牙在中国当地人参群体中与不同品种混在一起。在此,我们在细胞色素c氧化酶亚基2(cox2)线粒体的第二个内含子中鉴定出一个独特的、大马牙特有的单核苷酸多态性(SNP)位点。基于此SNP,设计了一个大马牙品种特异性引物,并优化了等位基因特异性聚合酶链反应(PCR)以实现大马牙的有效分子鉴定。我们设计了一种方法,将简单的DNA提取方法与使用SYBR Green I荧光染料的实时等位基因特异性PCR相结合,并通过等位基因鉴别分析证明了其在清晰区分大马牙品种与其他中国参品种方面的有效性。因此,本研究为从中国当地人参群体中鉴别和保存大马牙提供了一种简单快速的检测方法。

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