Discriminatory Power Evaluation of Nuclear Ribosomal RNA Barcoding Sequences Through Ophiocordyceps sinensis Related Samples.
Discriminatory Power Evaluation of Nuclear Ribosomal RNA Barcoding Sequences Through Ophiocordyceps sinensis Related Samples.
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DOI：
10.3389/fmicb.2018.02498
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发表时间：
2018
影响因子：
5.2
通讯作者：
Liu B
中科院分区：
文献类型：
--
作者：
Zhang P;Cui S;Ren X;Kang S;Wei F;Ma S;Liu B
由于冬虫夏草的成本大幅上升，假冒伪劣产品可能对健康产生不利影响，快速准确的物种级DNA条码识别系统可能是一种有效的方法，并显著提高监管能力。以43个野生中华稻曲霉子实体、纯培养物、商品菌丝发酵粉和伪品为材料，用Simpson判别指数测定了核糖体RNA基因簇中3个亚基序列的识别力。根据Simpson‘s判别指数(D=0.972)，43个样本中内转录间隔区(ITS)序列的变异度和识别力最高，其次是大亚基(LSU；D=0.963)和小亚基(SSU；D=0.921)。在ITS-1、ITS-2和ITS-5.8S序列中，ITS-2序列对43个样本的识别率最高。所有中华按蚊样本在95%的相似性下被聚为一个唯一的ITS序列簇，2个中华按蚊样本和6个非中华按蚊样本被聚为一类。中华绒毛虫样本显示了虚假声明。结果表明，ITS区可为中华稻曲病菌样品提供准确的物种鉴定，特别是当宏观和微观方法不能应用于深加工的商品时。由于中华按蚊相关产品的鉴定是保证其安全性和有效性的关键，因此在下一次修订《中国药典》时，应考虑通过ITS序列比对或物种特异性靶标的独特的聚合酶链式反应扩增来鉴定中华按蚊。
Since the cost of Ophiocordyceps sinensis has increased dramatically and the counterfeits may have adverse effect to health, a rapid and precise species-level DNA barcoding identification system could be a potent approach and significantly enhance the regulatory capacity. The discrimination power of three subunits sequences from nuclear ribosomal RNA gene cluster were determined by Simpson’s index of discrimination using 43 wild O. sinensis fruiting bodies, pure cultures, commercial mycelium fermented powder and counterfeits. The internal transcribed spacer (ITS) sequences showed the highest variance and discrimination power among 43 samples, as determined by Simpson’s index of discrimination (D = 0.972), followed by large subunit (LSU; D = 0.963) and small subunit (SSU; D = 0.921). ITS-2 sequences showed the highest discrimination power for 43 samples among ITS-1, ITS-2, and 5.8S region of ITS sequences. All O. sinensis samples were grouped into a unique ITS sequence cluster under 95% similarity and two O. sinensis samples and six non-O. sinensis samples showed false claims. Our data showed that the ITS region could provide accurate species identification for O. sinensis samples, especially when macroscopic and microscopic method could not be applied in the highly processed commercial products. Since the authentication of O. sinensis related products is essential to ensure its safety and efficacy, identification of O. sinensis through ITS sequence comparison or unique PCR amplification of the species specific target, such as the ITS region, should be considered in the next revision of Chinese pharmacopeia.
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