DNA meta-barcoding using rbcL based mini-barcode revealed presence of unspecified plant species in Ayurvedic polyherbal formulations
DNA meta-barcoding using rbcL based mini-barcode revealed presence of unspecified plant species in Ayurvedic polyherbal formulations
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DOI：
10.1002/pca.3026
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发表时间：
2021-02-01
影响因子：
3.3
通讯作者：
Joshi, Madhvi
中科院分区：
文献类型：
--
作者：
Pandit, Ramesh;Travadi, Tasnim;Joshi, Madhvi
导言阿育吠陀利用药用植物的有益特性。高要求加上植物药供应不足，以及缺乏对其准确鉴定的知识，导致草药产品中掺假。在复杂的草药配方中，鉴定变得更加困难。本文分析了四种不同的中草药配方。目的利用高通量下一代测序技术，检测基于rbcL基因的植物DNA微型条形码，以鉴别中药制剂中的靶向和非靶向植物。30种正宗药用植物的混合物被用来检查迷你条形码的物种分辨能力。从草药制剂中提取DNA，制备扩增子文库，并在IonS5系统上进行测序。利用各种生物信息学工具对数据进行分析。结果对照样本的分析揭示了DNA微型条码的最佳分辨率。对商业样品的数据分析显示，只有一种草药配方含有所有植物，并与所列内容相匹配。在两种配方中，分别只检测到21株植物中的10株和20株植物中的11株。此外，在这些配方中还检测到了几种未列出的植物。两种配方含有分配给非目标植物的20%的读数。总体而言，21.98%的读数分配给非目标植物。结论本研究清楚地展示了元条码在复杂草药基质质量控制中的成功应用和潜力。结果表明，该方法可用于中药炮制品的药物警戒。
Introduction Ayurveda takes advantage of the beneficial properties of medicinal plants. High demands in combination with inadequate availability of botanicals and a lack of knowledge with respect to their precise identification lead to adulterations in herbal products. Identification becomes more difficult in complex herbal formulations. Four different polyherbal formulations have been analyzed for the present paper. The targeted plants have different pharmacological properties for various ailments.Objective We aimed to examine the rbcL gene based plant DNA mini-barcode to identify target and non-target plants in polyherbal formulations by using high-throughput next generation sequencing.Methods Degenerate primers of the selected mini-barcode region have been identified from the literature. A blend of 30 authentic medicinal plant species was used to examine the species resolution capacity of the mini-barcode. DNA was isolated from herbal formulations, an amplicon library was prepared, and sequencing was performed on an IonS5 system. Data were analyzed using various bioinformatics tools.Results Analysis of control pooled samples revealed the optimum resolving power of the DNA mini-barcode. Data analysis of the commercial samples revealed that only one herbal formulation contained all plants and matched with listed contents. In two formulations, only 10 out of 21 and 11 out of 20 plants were detected, respectively. Additionally, several non-listed plants were also detected in these formulations. Two formulations contained >20% reads assigned to non-target plants. Overall, 21.98% of the reads were assigned to non-target plants.Conclusion The present study clearly demonstrated the successful application and potential of meta-barcoding in the quality control of complex herbal matrices. The results strongly suggest that this approach can be used in pharmacovigilance of processed herbal products.