Best practices in metabarcoding of fungi: From experimental design to results. Issue 10 (20th April 2022)

• Record Type: Journal Article
• Authors: Tedersoo, Leho; Bahram, Mohammad; Zinger, Lucie; Nilsson, R. Henrik; Kennedy, Peter G.; Yang, Teng; Anslan, Sten; Mikryukov, Vladimir
• Is Part Of: Molecular ecology. Volume 31:Issue 10(2022)
• Page Start: 2769
• Page End: 2795
• Publication Date: 2022-04-20
• Subjects: biodiversity -- bioinformatics -- community ecology -- experiment setup -- molecular identification -- statistical analyses; Molecular ecology -- Periodicals; Molecular population biology -- Periodicals; 576
• DOI: 10.1111/mec.16460
• Languages: English
• ISSNs: 0962-1083

Abstract

Abstract: The development of high‐throughput sequencing (HTS) technologies has greatly improved our capacity to identify fungi and unveil their ecological roles across a variety of ecosystems. Here we provide an overview of current best practices in metabarcoding analysis of fungal communities, from experimental design through molecular and computational analyses. By reanalysing published data sets, we demonstrate that operational taxonomic units (OTUs) outperform amplified sequence variants (ASVs) in recovering fungal diversity, a finding that is particularly evident for long markers. Additionally, analysis of the full‐length ITS region allows more accurate taxonomic placement of fungi and other eukaryotes compared to the ITS2 subregion. Finally, we show that specific methods for compositional data analyses provide more reliable estimates of shifts in community structure. We conclude that metabarcoding analyses of fungi are especially promising for integrating fungi into the full microbiome and broader ecosystem functioning context, recovery of novel fungal lineages and ancient organisms as well as barcoding of old specimens including type material.