Tuber P. Micheli ex F.H. Wigg. species have always attracted people's attention, with their high diversity, culinary and economic interest, strong aromas as well as the importance of plant ecosystem and animal nutrition. Interest in truffle species has been increasing in recent years in Turkey. Although some truffle species have been known previously in Turkey, many species are yet to be identified. Tuber macrosporum Vittad. samples were collected from Edirne and Tekirdağ regions in 2017 in a field study conducted to find new truffle species. Tuber macrosporum samples were firstly identified by macro and microscopic features, and this result was supported as 99% by DNA analyses when compared to GenBank. A short description of the newly reported species is given along with its macro and microphotographs, and spore images taken by a scanning electron microscope (SEM). Additionally, ITS based evolutionary history of the species is provided with phylogenetic trees.