Fluorescence Visual Identification of Four Main Varieties of Tibetan Medicinal Berberis cortex Based on Site-Specific PCR Technology.

IF 3 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Phytochemical Analysis Pub Date : 2025-06-18 DOI:10.1002/pca.70002

Yan Chen, Hongying Li, TingTing Zhao, Jiaqi Cui, Wenjun Song, Huilin Li, Gang Fan, Rong Chen

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引用次数: 0

Abstract

Background: Berberis cortex is a typical multi-origin species in Tibetan medicine, with varying medicinal effects among different species. Establishing a rapid and accurate identification method for the major species of Tibetan medicinal B. cortex, including Berberis vernae Schneid. (SY), Berberis diaphana Maxim. (XH), Berberis kansuensis Schneid. (GS), and Berberis dictyophylla Franch. (CHZ), is conducive to the quality control of B. cortex.

Methods: Site-specific PCR visualization using SYBR Green I was developed based on rbcL SNP sites for SY, GS, and CHZ. For XH, which cannot be differentiated through rbcL and other barcoding SNP sites, RAPD-PCR methodology was employed to screen for species-specific sequences. A site-specific amplification visualization system was subsequently developed based on the identified sequence.

Results: The developed site-specific PCR visualization systems demonstrated excellent sensitivity, with all systems capable of detecting genomic DNA at concentrations as low as 100 fg. These systems were employed to analyze 16 batches of actual B. cortex samples. The analysis revealed that four samples were identified as SY, six samples as GS, two samples as CHZ, and three samples as XH. All results were concordant with sequencing data. One remaining negative sample was confirmed through sequencing as adulterated with Phellodendri Chinensis Cortex. Further authentication of 10 batches of Tibetan patent medicines containing B. cortex revealed that 2 batches contained both SY and GS, one batch contained both SY and CHZ, three batches contained exclusively GS, and three batches contained XH.

Conclusion: A site-specific PCR fluorescence visual identification system has been developed for the authentication of four major B. cortex species, enabling accurate identification of botanical origins in both B. cortex crude materials and Berberis-containing Tibetan patent medicines.

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基于位点特异性PCR技术的4个主要藏药小檗皮品种的荧光视觉鉴定

背景：小檗皮是藏医药中典型的多源种，不同种间药效差异较大。建立了包括小檗在内的主要藏药种皮的快速、准确鉴别方法。（SY）， Berberis diaphana Maxim。（XH）， kansuensis Berberis Schneid。（GS）和小檗（beris dictyophylla Franch）。（CHZ），有利于白芷的质量控制。方法：基于SY， GS和CHZ的rbcL SNP位点，使用SYBR Green I开发了位点特异性PCR可视化。对于无法通过rbcL和其他条形码SNP位点进行区分的XH，采用RAPD-PCR方法筛选物种特异性序列。随后，基于已确定的序列开发了位点特异性扩增可视化系统。结果：所开发的位点特异性PCR可视化系统表现出优异的灵敏度，所有系统都能够检测低至100 fg浓度的基因组DNA。这些系统被用于分析16批实际的B. cortex样品。分析发现4个样品为SY， 6个样品为GS， 2个样品为CHZ， 3个样品为XH。所有结果与测序数据一致。剩余1份阴性样品经测序确认为掺假黄柏皮。对10批含黄皮的藏成药进行进一步鉴定，发现2批同时含有SY和GS， 1批同时含有SY和CHZ， 3批只含GS， 3批含XH。结论：建立了4种主要黄皮药材的位点特异性PCR荧光视觉鉴定系统，可准确鉴定黄皮药材和含小檗碱的藏成药的植物来源。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Phytochemical Analysis 生物-分析化学

CiteScore

6.00

自引率

6.10%

发文量

审稿时长

1.7 months

期刊介绍： Phytochemical Analysis is devoted to the publication of original articles concerning the development, improvement, validation and/or extension of application of analytical methodology in the plant sciences. The spectrum of coverage is broad, encompassing methods and techniques relevant to the detection (including bio-screening), extraction, separation, purification, identification and quantification of compounds in plant biochemistry, plant cellular and molecular biology, plant biotechnology, the food sciences, agriculture and horticulture. The Journal publishes papers describing significant novelty in the analysis of whole plants (including algae), plant cells, tissues and organs, plant-derived extracts and plant products (including those which have been partially or completely refined for use in the food, agrochemical, pharmaceutical and related industries). All forms of physical, chemical, biochemical, spectroscopic, radiometric, electrometric, chromatographic, metabolomic and chemometric investigations of plant products (monomeric species as well as polymeric molecules such as nucleic acids, proteins, lipids and carbohydrates) are included within the remit of the Journal. Papers dealing with novel methods relating to areas such as data handling/ data mining in plant sciences will also be welcomed.