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基于 DNA 条形码分析和设计的 DNA 标记物鉴定和监测砂仁及其掺伪品。

Identification and Monitoring of Amomi Fructus and its Adulterants Based on DNA Barcoding Analysis and Designed DNA Markers.

机构信息

Department of Herbology, College of Korean Medicine, Wonkwang University, Iksan 54538, Korea.

Research Center of Traditional Korean Medicine, Wonkwang University, Iksan 54538, Korea.

出版信息

Molecules. 2019 Nov 19;24(22):4193. doi: 10.3390/molecules24224193.

Abstract

Amomi Fructus is one of the traditional medicines derived from the ripe fruits of the Zingiberaceae family of plants, which include , var. , and . Owing to their highly similar morphological traits, several kinds of adulterants of Amomi Fructus have been reported. Therefore, accurate and reliable methods of identification are necessary in order to ensure drug safety and quality. We performed DNA barcoding using five regions (ITS, , and intergenic spacer) of 23 Amomi Fructus samples and 22 adulterants. We designed specific DNA markers for Amomi Fructus based on the single nucleotide polymorphisms (SNPs) in the ITS. Amomi Fructus was well separated from the adulterants and was classified with the species of origin based on the detected SNPs from the DNA barcoding results. The AVF1/ISR DNA marker for produced a 270 bases amplified product, while the ALF1/ISF DNA marker produced a 350 bases product specific for . Using these DNA markers, the monitoring of commercially distributed Amomi Fructus was performed, and the monitoring results were confirmed by ITS analysis. This method identified samples that were from incorrect origins, and a new species of adulterant was also identified. These results confirmed the accuracy and efficiency of the designed DNA markers; this method may be used as an efficient tool for the identification and verification of Amomi Fructus.

摘要

砂仁是姜科植物的成熟果实之一,包括阳春砂、绿壳砂和海南砂。由于它们具有高度相似的形态特征,因此已经报道了几种砂仁的掺杂物。因此,为了确保药物的安全性和质量,需要准确可靠的鉴定方法。我们使用 23 个砂仁样品和 22 个掺杂物的五个区域(ITS、、和 内转录间隔区)进行了 DNA 条形码分析。我们根据 ITS 中的单核苷酸多态性(SNP)为砂仁设计了特定的 DNA 标记。根据 DNA 条形码结果中检测到的 SNP,砂仁与掺杂物很好地区分开来,并根据起源物种进行了分类。AVF1/ISR DNA 标记可产生 270 个碱基的扩增产物,而 ALF1/ISF DNA 标记可产生 350 个碱基的产物,特异性针对 。使用这些 DNA 标记物对商业分销的砂仁进行了监测,并通过 ITS 分析确认了监测结果。该方法鉴定了来源不正确的样品,并鉴定了一种新的掺杂物。这些结果证实了设计的 DNA 标记的准确性和效率;该方法可作为鉴定和验证砂仁的有效工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce8a/6891445/08e11d36961f/molecules-24-04193-g001.jpg

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