Graduate School of Bioresources, Mie University, 1577 Kurima-machiya, Tsu, Mie, 514-8507, Japan.
Graduate School of Simulation Studies, University of Hyogo, 7-1-28 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan.
Sci Rep. 2020 Feb 20;10(1):3057. doi: 10.1038/s41598-020-59620-0.
Among terrestrial microorganisms, mushroom-forming fungi have been relatively well investigated, however the inconspicuous strains may be overlooked by conventional visual investigations causing underestimation of their phylogenetic diversity. Herein, we sought to obtain a comprehensive phylogenetic diversity profile for the early-diverging wood-decaying mushrooms Dacrymycetes, using an approach that combines fruiting-body collection, culture isolation, and environmental DNA (eDNA) metabarcoding of decaying branches. Among the 28 operational taxonomic units (OTUs) detected during a three-year investigation, 10 each were from fruiting bodies and cultured mycelia and 27 were detected as eDNA sequences. eDNA metabarcoding revealed various lineages across the Dacrymycetes phylogeny. Alternatively, fruiting-body and culture surveys uncovered only ~50% of the OTUs detected through eDNA metabarcoding, suggesting that several inconspicuous or difficult-to-isolate strains are latent in the environment. Further, eDNA and culture surveys revealed early-diverging clades that were not identified in the fruiting-body survey. Thus, eDNA and culture-based techniques can uncover inconspicuous yet phylogenetically important mushroom lineages that may otherwise be overlooked via typical visual investigations.
在陆地微生物中，蕈菌类真菌已经得到了相对较好的研究，然而，不显眼的菌株可能会被传统的视觉调查所忽视，导致其系统发育多样性被低估。在此，我们试图通过结合子实体收集、培养分离和腐朽树枝的环境 DNA(eDNA) 宏条形码的方法，获得早期木质腐朽蘑菇（齿菌目）的全面系统发育多样性特征。在为期三年的调查中，共检测到 28 个操作分类单位（OTU），其中 10 个来自子实体和培养菌丝，27 个被检测为 eDNA 序列。eDNA 宏条形码揭示了齿菌目中的各种谱系。相比之下，子实体和培养调查仅发现了 eDNA 宏条形码检测到的 OTU 的约 50%，表明环境中存在一些不显眼或难以分离的菌株。此外，eDNA 和培养调查揭示了早期分化的分支，这些分支在子实体调查中没有被识别。因此，eDNA 和基于培养的技术可以揭示不显眼但在系统发育上很重要的蘑菇谱系，否则这些谱系可能会被典型的视觉调查所忽视。
