Tuber macrosporum, a large spore species is first time confirmed from Poland. It grows naturally in southern part of Poland, in calcareous soils. The fruit bodies of the species were found in mixed fresh forest with dominant tree species such as:Pinus sylvestris, Carpinus betulus and Quercus robur. The soil chemistry and texture of the site was analysed. Fruit body of the truffle is conserved in the collection of biological material in Ecology Department of Forest Research Institute herbarium.

Key words: Tuber macrosporum, habitat, phylogeny.