A truffle is the fruiting body of a subterranean ascomycete fungus, predominantly one of the many species of the genus Tuber. In addition to Tuber, over one hundred other genera of fungi are classified as truffles including Geopora, Peziza, Choiromyces, and Leucangium. These genera belong to the class Pezizomycetes and the Pezizales order. Several truffle-like basidiomycetes are excluded from Pezizales, including Rhizopogon and Glomus. Truffles are ectomycorrhizal fungi, so they are usually found in close association with tree roots. Spore dispersal is accomplished through fungivores, animals that eat fungi. These fungi have significant ecological roles in nutrient cycling and drought tolerance.

An ectomycorrhiza is a form of symbiotic relationship that occurs between a fungal symbiont, or mycobiont, and the roots of various plant species. The mycobiont is often from the phyla Basidiomycota and Ascomycota, and more rarely from the Zygomycota. Ectomycorrhizas form on the roots of around 2% of plant species, usually woody plants, including species from the birch, dipterocarp, myrtle, beech, willow, pine and rose families. Research on ectomycorrhizas is increasingly important in areas such as ecosystem management and restoration, forestry and agriculture.