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Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom fungi and reveals thresholds for fungal species and higher taxon delimitation
Studies in Mycology ( IF 16.5 ) Pub Date : 2018-05-30 , DOI: 10.1016/j.simyco.2018.05.001
D. Vu , M. Groenewald , M. de Vries , T. Gehrmann , B. Stielow , U. Eberhardt , A. Al-Hatmi , J.Z. Groenewald , G. Cardinali , J. Houbraken , T. Boekhout , P.W. Crous , V. Robert , G.J.M. Verkley

Species identification lies at the heart of biodiversity studies that has in recent years favoured DNA-based approaches. Microbial Biological Resource Centres are a rich source for diverse and high-quality reference materials in microbiology, and yet the strains preserved in these biobanks have been exploited only on a limited scale to generate DNA barcodes. As part of a project funded in the Netherlands to barcode specimens of major national biobanks, sequences of two nuclear ribosomal genetic markers, the Internal Transcribed Spaces and 5.8S gene (ITS) and the D1/D2 domain of the 26S Large Subunit (LSU), were generated as DNA barcode data for ca. 100 000 fungal strains originally assigned to ca. 17 000 species in the CBS fungal biobank maintained at the Westerdijk Fungal Biodiversity Institute, Utrecht. Using more than 24 000 DNA barcode sequences of 12 000 ex-type and manually validated filamentous fungal strains of 7 300 accepted species, the optimal identity thresholds to discriminate filamentous fungal species were predicted as 99.6 % for ITS and 99.8 % for LSU. We showed that 17 % and 18 % of the species could not be discriminated by the ITS and LSU genetic markers, respectively. Among them, ∼8 % were indistinguishable using both genetic markers. ITS has been shown to outperform LSU in filamentous fungal species discrimination with a probability of correct identification of 82 % vs. 77.6 %, and a clustering quality value of 84 % vs. 77.7 %. At higher taxonomic classifications, LSU has been shown to have a better discriminatory power than ITS. With a clustering quality value of 80 %, LSU outperformed ITS in identifying filamentous fungi at the ordinal level. At the generic level, the clustering quality values produced by both genetic markers were low, indicating the necessity for taxonomic revisions at genus level and, likely, for applying more conserved genetic markers or even whole genomes. The taxonomic thresholds predicted for filamentous fungal identification at the genus, family, order and class levels were 94.3 %, 88.5 %, 81.2 % and 80.9 % based on ITS barcodes, and 98.2 %, 96.2 %, 94.7 % and 92.7 % based on LSU barcodes. The DNA barcodes used in this study have been deposited to GenBank and will also be publicly available at the Westerdijk Institute's website as reference sequences for fungal identification, marking an unprecedented data release event in global fungal barcoding efforts to date.



中文翻译:

大规模产生和分析丝状真菌DNA条形码可增强对王国真菌的覆盖率,并揭示真菌物种的阈值和较高的分类群定界

物种鉴定是生物多样性研究的核心,近年来,生物多样性研究已青睐基于DNA的方法。微生物资源中心是微生物学中各种高质量参考材料的丰富来源,然而,这些生物库中保存的菌株仅在有限的范围内被利用才能生成DNA条形码。作为荷兰资助的对主要国家生物库标本,两个核糖体遗传标记,内部转录空间和5.8S基因(ITS)以及26S大亚基(LSU)的D1 / D2域的序列进行条形码处理的项目的一部分被生成为大约的DNA条码数据。最初分配给ca的100000真菌菌株。在乌得勒支的韦斯特代克真菌生物多样性研究所保存的CBS真菌生物库中有17,000种。使用超过12,000种ex-type的24,000条DNA条形码序列和7300种被接受物种的手动验证的丝状真菌菌株,用于区分丝状真菌物种的最佳同一性阈值预计为ITS的99.6%和LSU的99.8%。我们表明,不能通过ITS和LSU遗传标记分别区分17%和18%的物种。其中,使用这两种遗传标记无法区分约8%。ITS已被证明在丝状真菌种类鉴别方面优于LSU,正确识别的机率为82%对77.6%,聚类质量值为84%对77.7%。在较高的分类学分类中,已证明LSU具有比ITS更好的区分能力。群集质量值为80%,在顺序水平上,LSU在识别丝状真菌方面优于ITS。在一般水平上,两个遗传标记产生的聚类质量值均较低,表明在属水平上进行分类学修订的必要性,并且有可能应用更保守的遗传标记甚至整个基因组。根据ITS条码,预测在属,科,纲和类水平上的丝状真菌鉴定的分类学阈值为94.3%,88.5%,81.2%和80.9%,基于LSU为98.2%,96.2%,94.7%和92.7%条码。这项研究中使用的DNA条码已保存到GenBank中,也将在Westerdijk Institute的网站上公开提供,作为真菌鉴定的参考序列,这标志着迄今为止全球真菌条形码工作中前所未有的数据发布事件。在一般水平上,两个遗传标记产生的聚类质量值均较低,表明在属水平上进行分类学修订的必要性,并且有可能应用更保守的遗传标记甚至整个基因组。根据ITS条码,预测在属,科,纲和类水平上的丝状真菌鉴定的分类学阈值为94.3%,88.5%,81.2%和80.9%,基于LSU为98.2%,96.2%,94.7%和92.7%条码。这项研究中使用的DNA条码已保存到GenBank中,也将在Westerdijk Institute的网站上公开提供,作为真菌鉴定的参考序列,这标志着迄今为止全球真菌条形码工作中前所未有的数据发布事件。在一般水平上,两个遗传标记产生的聚类质量值均较低,表明在属水平上进行分类学修订的必要性,并且有可能应用更保守的遗传标记甚至整个基因组。根据ITS条码,在属,科,纲和类水平上预测的丝状真菌鉴定的分类阈值为94.3%,88.5%,81.2%和80.9%,而基于LSU的分类阈值为98.2%,96.2%,94.7%和92.7%条码。这项研究中使用的DNA条码已保存到GenBank中,也将在Westerdijk Institute的网站上公开提供,作为真菌鉴定的参考序列,这标志着迄今为止全球真菌条形码工作中前所未有的数据发布事件。表明在属水平上进行分类学修订的必要性,以及可能应用更保守的遗传标记甚至整个基因组的必要性。根据ITS条码,预测在属,科,纲和类水平上的丝状真菌鉴定的分类学阈值为94.3%,88.5%,81.2%和80.9%,基于LSU为98.2%,96.2%,94.7%和92.7%条码。这项研究中使用的DNA条码已保存到GenBank中,也将在Westerdijk Institute的网站上公开提供,作为真菌鉴定的参考序列,这标志着迄今为止全球真菌条形码工作中前所未有的数据发布事件。表明在属水平上进行分类学修订的必要性,以及可能应用更保守的遗传标记甚至整个基因组的必要性。根据ITS条码,预测在属,科,纲和类水平上的丝状真菌鉴定的分类学阈值为94.3%,88.5%,81.2%和80.9%,基于LSU为98.2%,96.2%,94.7%和92.7%条码。这项研究中使用的DNA条码已保存到GenBank中,也将在Westerdijk Institute的网站上公开提供,作为真菌鉴定的参考序列,这标志着迄今为止全球真菌条形码工作中前所未有的数据发布事件。根据ITS条码,在属,科,纲和类水平上预测的丝状真菌鉴定的分类阈值为94.3%,88.5%,81.2%和80.9%,而基于LSU的分类阈值为98.2%,96.2%,94.7%和92.7%条码。这项研究中使用的DNA条码已保存到GenBank中,也将在Westerdijk Institute的网站上公开提供,作为真菌鉴定的参考序列,这标志着迄今为止全球真菌条形码工作中前所未有的数据发布事件。根据ITS条码,预测在属,科,纲和类水平上的丝状真菌鉴定的分类学阈值为94.3%,88.5%,81.2%和80.9%,基于LSU为98.2%,96.2%,94.7%和92.7%条码。这项研究中使用的DNA条码已保存到GenBank中,也将在Westerdijk Institute的网站上公开提供,作为真菌鉴定的参考序列,这标志着迄今为止全球真菌条形码工作中前所未有的数据发布事件。

更新日期:2018-05-30
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