Tuber melanosporum (Ascomycota, Pezizales) is an ectomycorrhizal fungus that produces highly appreciated hypogeous fruiting bodies called black truffles. The aim of this paper was to research the composition of ectomycorrhiza-associated fungal and bacterial communities in T. melanosporum oak plantations. Results of this paper showed the competitive effect of T. melanosporum on other fungal species, especially other mycorrhizal and pathogenic species. T. melanosporum was shown to be associated mainly with bacteria, some of them important for their properties as mycorrhizal helper bacteria. A dendrogram analysis of co-occurrence showed that T. melanosporum tended to co-occur with the following bacteria species: Singulisphaera limicola, Nannocistis excedens and Sporosarcina globispora. In addition, it was linked to fungal species such as Mortierella elongata, M. minutissima, Cryptococcus uzbekistanensis, C. chernovii and C. aerius. This study provides an exhaustive analysis of the diversity, structure and composition of fungal and bacterial communities associated with T. melanosporum to enhance understanding of the biology, composition and role of these communities in truffle plantations.

Summary: Cistus ladanifer scrublands, traditionally considered as unproductive, have nonetheless been observed to produce large quantities of king bolete ( Boletus edulis ) fruitbodies. These pyrophytic scrublands are prone to wildfires, which severely affect fungi, hence the need for fire prevention in producing C. ladanifer scrublands. In addition, B. edulis productions have severely decreased in the last years. A deeper understanding of the B. edulis life cycle and of biotic and abiotic factors influencing sporocarp formation is needed to implement management practices that facilitate B. edulis production. For example, some bacteria likely are involved in sporocarp production, representing a key part in the triple symbiosis (plant–fungus–bacteria). In this study, we used soil DNA metabarcoding in C. ladanifer scrublands to (i) assess the effect of site history and fire prevention treatment on bacterial richness and community composition; (ii) test if there was any correlation between various taxonomic groups of bacteria and mycelial biomass and sporocarp production of B. edulis ; and to (iii) identify indicator bacteria associated with the most productive B. edulis sites. Our results show that site history drives bacterial richness and community composition, while fire prevention treatments have a weaker, but still detectable effect, particularly in the senescent plots. Sporocarp production correlated positively with genera in Verrucomicrobia. Several genera, e.g. Azospirillum and Gemmatimonas , were identified as indicators of the most productive sites, suggesting a potential biological role in B. edulis fructification. This study provides a better understanding of the triple symbiosis (plant–fungus–bacteria) involved in C . ladanifer – B. edulis systems.