We assessed culturable soil microfungal diversity in various habitats around Hornsund, Spitsbergen in the High Arctic, using potato dextrose agar (PDA) medium. Thermal growth classification of the fungi obtained was determined by incubating them in 4°C and 25°C, permitting separation of those with psychrophilic, psychrotolerant and mesophilic char− acteristics. In total, 68 fungal isolates were obtained from 12 soil samples, and grouped into 38 mycelial morphotypes. Intergenic spacer regions of these morphotypes were sequenced, and they represented 25 distinct taxonomic units, of which 21 showed sufficient similarity with available sequence data in NCBI to be identified to species level. Soil under ornithogenic influence showed the highest species diversity, including sequences assigned to Mortierella macrocystis, M. elongata, Mortierella sp., Cudoniella sp., Varicosporium elodeae, Beauveria bassiana, Geomyces pannorum, Penicillium sp. and Atradidymella muscivora. Fourteen taxa were classified as psychrophilic, seven mesophilic, and four psychrotolerant.