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ITS 区为鉴定草药补充剂中的灵芝/灵芝(Ganoderma)提供了可靠的 DNA 条码。

The ITS region provides a reliable DNA barcode for identifying reishi/lingzhi (Ganoderma) from herbal supplements.

机构信息

Department of Biology, Santa Clara University, Santa Clara, California, United States of America.

Oregon's Wild Harvest, Redmond, Oregon, United States of America.

出版信息

PLoS One. 2020 Nov 12;15(11):e0236774. doi: 10.1371/journal.pone.0236774. eCollection 2020.

Abstract

The dietary supplement industry is rapidly growing yet, a recent study revealed that up to 60% of supplements may have substituted ingredients, some of which can be harmful contaminants or additives. When ingredients cannot be verified morphologically or biochemically, DNA barcoding complemented with a molecular phylogenetic analysis can be a powerful method for species authentication. We employed a molecular phylogenetic analysis for species authentication of the commonly used fungal supplement, reishi (Ganoderma lingzhi), by amplifying and sequencing the nuclear ribosomal internal transcribed spacer regions (ITS) with genus-specific primers. PCR of six powdered samples and one dried sample all sold as G. lucidum representing independent suppliers produced single, strong amplification products in the expected size-range for Ganoderma. Both best-hit BLAST and molecular phylogenetic analyses clearly identified the presence of G. lingzhi DNA in all seven herbal supplements. We detected variation in the ITS sequences among our samples, but all herbal supplement samples fall within a large clade of G. lingzhi ITS sequences. ITS-based phylogenetic analysis is a successful and cost-effective method for DNA-based species authentication that could be used in the herbal supplement industry for this and other fungal and plant species that are otherwise difficult to identify.

摘要

膳食补充剂行业发展迅速,但最近的一项研究表明,高达 60%的补充剂可能含有替代成分,其中一些可能是有害的污染物或添加剂。当成分无法通过形态学或生物化学进行验证时,DNA 条形码与分子系统发育分析相结合可以成为物种鉴定的有力方法。我们采用分子系统发育分析,使用属特异性引物扩增和测序核核糖体内部转录间隔区(ITS),对常用真菌补充剂灵芝(Ganoderma lingzhi)进行物种鉴定。对六种粉末样品和一种干燥样品进行 PCR,这些样品均代表独立供应商销售的赤芝(G. lucidum),均在 Ganoderma 的预期大小范围内产生单一、强扩增产物。最佳 BLAST 比对和分子系统发育分析均清楚地表明,所有七种草药补充剂中均存在灵芝 DNA。我们在样品中检测到 ITS 序列的变异,但所有草药补充剂样品均落在灵芝 ITS 序列的一个大亚群内。基于 ITS 的系统发育分析是一种成功且具有成本效益的基于 DNA 的物种鉴定方法,可用于草药补充剂行业,以及其他难以识别的真菌和植物物种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4351/7660467/33808ca2f544/pone.0236774.g001.jpg

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