Aim of study: The diversity of ectomycorrhizal fungal communities (EM) above (EMFb) and below (EMMt) ground associated with Quercus cerris L., Q. pubescens Willd., and Pinus nigra J.F.Arnold was analyzed.Area of study: A 20 year-old orchard that produces Tuber aestivum truffles, located a few kilometers from Chiusi della Verna (latitude 43° 41’ 53’’; longitude 11° 56’ 9’’) in Tuscany (central Italy) was observed.Material and Methods: This investigation combined analyses of EMFb, EMMt, T. aestivum productivity, different host trees, and statistical data on community ecology.Main results: The EM communities showed high species richness and differed slightly in relation to both the host tree and their location above or below ground, providing frequent findings of Tricholoma and Tomentella, respectively. Positive correlations were found between the number of truffles and host trees, and between the weight and number of truffles and EMFb.Research highlights: Mycorrhizal fungi and truffle production are not in competition.Key words: Fungal communities; fruiting bodies; morphotypes; Tuber aestivum; competition; Italy.