The aim of this paper was to provide an overview of the recent research concerning truffle biology, ecology and cultivation in Serbia and worldwide. Truffles are ectomycorrhizal fungi, primarily from to the genus Tuber (Ascomycota: Pezizales) which form underground fruiting bodies. Unless they form a symbiotic relationship with plant roots, truffles do not produce fruiting bodies. Survival and reproductive success of truffles is affected by numerous ecological factors (climate, soil characteristics and vegetation). They emit aromatic volatile organic compounds to attract free living organisms such as insects and mice that serve as vectors for truffle dispersal. Truffles use volatile signaling molecules throughout their life cycle to regulate interactions with microorganisms and plant roots. The most cultivated species of truffles in the world is the species T. melanosporum, followed by T. aestuvum. Inoculation with spores is the most common method for production of plants colonized with truffle mycorrhizas. It is recommended to plant seedlings with a high degree of truffle colonization in order to increase the production of fruiting bodies in plantation. Truffle plantations should be established using local genetic resources. In order to preserve natural truffle populations in Europe, the focus should be shifted from searching for truffles in nature to the establishment of plantations with mycorrhized seedlings.