Entoloma hirtipes in Türkiye: First molecular confirmation based on ITS from Çanakkale

Entoloma hirtipes (Schumach.) M.M. Moser is a member of the Entolomataceae characterized by considerable morphological variability and taxonomic complexity, making accurate species delimitation challenging. In this study, a macrofungal specimen collected from Çanakkale Province in northwestern Türkiye was identified using an integrative taxonomic approach combining detailed macro- and micromorphological observations with ITS rDNA-based phylogenetic analyses. Macroscopic and microscopic examinations included the assessment of basidiospores, basidia, cheilocystidia, and pileipellis structures, which were compared with published descriptions. The examined material exhibited diagnostic features consistent with E. hirtipes, including a hygrophanous pileus, a fibrillose stipe surface, and heterodiametric angular basidiospores. The ITS region was amplified using the ITS1/ITS4 primer pair and analysed together with reference sequences retrieved from GenBank. Bayesian phylogenetic analysis recovered the newly generated sequence within a strongly supported E. hirtipes clade (PP = 1.00), fully corroborating the morphological identification. The combined morphological and molecular evidence enabled the reliable identification of the examined material as E. hirtipes. This study provides the first ITS rDNA-based molecular confirmation of E. hirtipes from Türkiye, expands the documented distribution of the species within the country, and demonstrates the importance of integrative taxonomy for the accurate delimitation of morphologically similar species in Entolomataceae.