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基于 ITS 序列的多重 SCAR 标记鉴定白芷药材。

Authentication of the Herbal Medicine Angelicae Dahuricae Radix Using an ITS Sequence-Based Multiplex SCAR Assay.

机构信息

Herbal Medicine Research Division, Korea Institute of Oriental Medicine, Daejeon 34054, Korea.

出版信息

Molecules. 2018 Aug 24;23(9):2134. doi: 10.3390/molecules23092134.

Abstract

The accurate identification of plant species is of great concern for the quality control of herbal medicines. The Korean Pharmacopoeia and the Pharmacopoeia of the People's Republic of China define Angelicae Dahuricae Radix ( in Korean and in Chinese) as the dried roots of or var. belonging to the family Apiaceae. Discrimination among species on the basis of morphological characteristics is difficult due to their extremely polymorphic traits and controversial taxonomic history. Furthermore, dried roots processed for medicinal applications are indistinguishable using conventional methods. DNA barcoding is a useful and reliable method for the identification of species. In this study, we sequenced the internal transcribed spacer (ITS) region of nuclear ribosomal RNA genes in , var. , and the related species and . Using these sequences, we designed species-specific primers, and developed and optimized a multiplex sequence-characterized amplified region (SCAR) assay that can simply and rapidly identify respective species, and verify the contamination of adulterant depending on the polymerase chain reaction (PCR) amplification without sequencing analysis in a single PCR reaction. This assay successfully identified commercial samples of Angelicae Dahuricae Radix collected from Korean and Chinese herbal markets, and distinguished them from adulterants. This multiplex SCAR assay shows a great potential in reducing the time and cost involved in the identification of genuine Angelicae Dahuricae Radix and adulterant contamination.

摘要

准确识别植物物种是控制草药质量的关键。《韩国药典》和《中华人民共和国药典》将白芷(在韩语中为 ,在中文中为 )定义为伞形科 属或 的干燥根。由于其形态特征具有极高的多态性,且分类历史存在争议,因此基于形态特征对 物种进行区分具有一定难度。此外,经过加工用于药用的干根也无法通过常规方法进行鉴别。DNA 条形码是一种用于物种鉴定的有效且可靠的方法。本研究中,我们对核核糖体 RNA 基因的内部转录间隔区(ITS)进行了测序,涉及 、 和相关物种 。利用这些序列,我们设计了物种特异性引物,并开发和优化了多重序列特征扩增区域(SCAR)分析方法,该方法可以简单快速地鉴定出各自的物种,并通过聚合酶链反应(PCR)扩增来验证是否存在掺杂物,而无需进行测序分析,从而在单个 PCR 反应中完成。该方法成功地鉴定了从韩国和中国草药市场采集的白芷商业样本,并将其与掺杂物区分开来。该多重 SCAR 分析方法在减少鉴定真正白芷及其掺杂物所需的时间和成本方面具有很大的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/daf3/6225120/3490a3594d34/molecules-23-02134-g001.jpg

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