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Identifying the ‘unidentified’ fungi: a global-scale long-read third-generation sequencing approach

Fungal Diversity 2020年第4期103卷 273-293页

作者： Leho Tedersoo Sten Anslan Mohammad Bahram Urmas Kõljalg Kessy Abarenkov Institute of Ecology and Earth Sciences Tartu University14a Ravila50411 TartuEstonia Zoological Institute Braunschweig University of TechnologyMendelssohnstr.438106 BrunswickGermany Department of Ecology Swedish University of Agricultural SciencesUlls väg 1675651 UppsalaSweden Natural History Museum of Tartu University 14a Ravila50411 TartuEstonia

Molecular identification methods,in particular high-throughput sequencing tools,have greatly improved our knowledge about fungal diversity and biogeography,but many of the recovered taxa from natural environments cannot be identified to species or even higher taxonomic *** study addresses the phylogenetic placement of previously unrecognized fungal groups by using two complementary approaches:(i)third-generation amplicon sequencing analysis of DNA from global soil samples,screening out ITS reads oftop50 sequences).For the global soil samples,we chose to amplify the full rRNA gene operon using four partly overlapping amplicons and multiple newly developed primers or primer combinations that cover nearly all fungi and a vast majority of non-fungal *** extracted the rRNA 18S(SSU)and 28S(LSU)genes and performed phylogenetic analyses against carefully selected reference *** SSU and LSU analyses placed most soil sequences and top50 sequences to known orders and classes,but tens of monophyletic groups and single sequences remained outside described ***,the LSU analyses recovered a few small groups of sequences that may potentially represent novel *** conclude that rRNA genes-based phylogenetic analyses are efficient tools for determining phylogenetic relationships of fungal taxa that cannot be placed to any order or class using ITS sequences ***,in many instances,longer rRNA gene sequences and availability of both SSU and LSU reads are needed to improve taxonomic *** leveraging third-generation sequencing from global soil samples,we successfully provided phylogenetic placement for many previously unidentified sequences and broadened our view on the fungal tree of life,with 10-20%new order-level taxa.I

关键词： Soil fungi top 50 most wanted fungi Phylogenetic diversity PacBio sequencing 18S rRNA gene(small subunit) 28S rRNA gene(large subunit)

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