The article provides results of our revision of three herbarium specimens of the genus Tulostoma collected in Ukraine and deposited in the Mycological Herbarium of V.N. Karazin Kharkiv National University, CWU (Myc). Previously, they have been identified as Tulostoma kotlabae, T. niveum, and T. subsquamosum and reported as newly recorded species in Ukraine. For the examined specimens, we analyzed the nucleotide sequences of the ITS region of ribosomal DNA. For one specimen, the large subunit (LSU) of rDNA was also obtained. Based on the nucleotide sequencing results, one specimen (GB00121) was re-identified as Tulostoma melanocyclum and the other two specimens (GB00125 and GB00129) – as T. simulans. The latter species is reported in Ukraine for the first time. Given that genus Tulostoma has been subject to a number of recent taxonomic changes, it would be reasonable to make a revision of other herbarium specimens from Ukraine using molecular methods.