Distinct compartmentalization of microbial community and potential metabolic function in the fruiting body of Tricholoma matsutake

he uniquely compartmentalized fruiting body structure of the ectomycorrhizal fungus (EMF) Tricholoma matsutake, is a hotspot of microbial habitation and interaction. However, microbial diversity within this microniche structure of the EMF is rarely investigated. Furthermore, there is limited information concerning microbiomes associated with sporomes belonging to the ubiquitous fungal phylum Basidiomycota, particularly with respect to fungus-EMF interactions. In this study, we conducted high throughput sequencing, using ITS (fungal) and 16S rRNA (bacterial) marker genes to characterize and compare fruiting body microbiomes in the outer (pileipellis and stipitipellis) and inner layers (pileum context, stipe context, and lamellae) of the fruiting body of T. matsutake. Our results show the number of unique bacterial operational taxonomic units (OTUs) among the different compartments ranged from 410 to 499 and was more than double that of the shared/common OTUs (235). Micrococcales, Bacillales, Caulobacter, and Sphingomonas were the primary significant bacterial taxa within the different compartments of the dissected T. matsutake fruiting body. Non-parametric multivariate analysis of variance showed significant compartmental differences for both the bacterial and the fungal community structure within the T. matsutake fruiting body. The metabolic profiling revealed putative metabolisms (of amino acids, carbohydrates, and nucleotides) and the biosynthesis of secondary metabolites to be highly enriched in outer layers; in the inner parts, the metabolisms of energy, cofactors, vitamins, and lipids were significantly higher. This study demonstrates for the first time the distinct compartmentalization of microbial communities and potential metabolic function profiles in the fruiting body of an economically important EMF T. matsutake.Funding text 1: This work was funded by the Guizhou Science & Technology Pillar Program (No. 2019-2336), Special fund for innovation capacity construction of Guizhou research institutions (No. 2018-4002), Guizhou High-level Talents Program, CAS “Light of West China” Program, and Basic Research— General Program of Yunnan Province, China. Funding text 2 This work was funded by the Guizhou Science & Technology Pillar Program (No. 2019-2336), Special fund for innovation capacity construction of Guizhou research institutions (No. 2018-4002), Guizhou High-level Talents Program, CAS ?Light of West China? Program, and Basic Research?General Program of Yunnan Province, China