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利用叶绿体基因组的matK、rbcL和ITS区域的通用DNA条形码,通过扩增子长度多态性对药用植物进行分子鉴定。

Molecular identification of medicinal plants with amplicon length polymorphism using universal DNA barcodes of the -, and - regions.

作者信息

Thakur Vishwa Vijay, Tiwari Sharad, Tripathi Niraj, Tiwari Gyanendra

机构信息

Biotechnology Centre, Jawaharlal Nehru Agriculture University, Jabalpur, 482004 India.

Department of Plant Breeding and Genetics, Jawaharlal Nehru Agriculture University, Jabalpur, 482004 India.

出版信息

3 Biotech. 2019 May;9(5):188. doi: 10.1007/s13205-019-1724-6. Epub 2019 Apr 25.

Abstract

Amplification success and species discrimination efficiency of universal DNA barcode primers (-, , , , and ) was evaluated in 46 representative medicinal plant species of 28 families on agarose gel. The results showed that amplicons length polymorphism revealed by the primers -, - and can simultaneously discriminate all the 46 species under study precisely. Some of the plant species included in this study are used as potential adulterants to other plant species. We were able to successfully discriminate the plant species from their potential substitute. is an adulterant of which has been successfully discriminated by all these three markers. Another example of adulteration between and was successfully discriminated by , on the basis of variability in amplicon length of these two medicinal herbs. Further, and are also adulterants for each other and variability in amplicon length between these two species was revealed by - markers. A colour code distance matrix based on amplicon length polymorphism was designed to select primers which can effectively discriminate plants species on the basis of their amplicon length. Discrimination of plant species with the universal markers on agarose gel is a noble and inexpensive approach as it does not require sequencing of amplicons. This procedure will provide a way for the development of diagnostic markers to identify adulteration not only in herbal drug formulations but also in food material.

摘要

在琼脂糖凝胶上,对28个科的46种代表性药用植物物种评估了通用DNA条形码引物(-、-、-、-、-和-)的扩增成功率和物种鉴别效率。结果表明,引物-、-和-所揭示的扩增子长度多态性能够同时精确鉴别所有46种被研究的物种。本研究中包含的一些植物物种被用作其他植物物种的潜在掺假物。我们能够成功地将这些植物物种与其潜在替代品区分开来。-是-的掺假物,这三种标记物均已成功鉴别出二者。-和-之间掺假的另一个例子,基于这两种药用植物扩增子长度的变异性,被-、-成功鉴别。此外,-和-也互为掺假物,-标记物揭示了这两个物种之间扩增子长度的变异性。设计了一个基于扩增子长度多态性的颜色编码距离矩阵,以选择能够根据扩增子长度有效鉴别植物物种的引物。在琼脂糖凝胶上使用通用标记物鉴别植物物种是一种既高尚又经济的方法,因为它不需要对扩增子进行测序。这一程序将为开发诊断标记物提供一条途径,不仅用于识别草药制剂中的掺假,也用于识别食品原料中的掺假。

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