Ophiostomatales (Ascomycota) contains many species, most of which are associated with bark beetles. Some members of this order are plant or animal pathogens, while others colonize soil, different plant tissues, or even carpophores of some Basidiomycota. However, little is known about soil-inhabiting Ophiostomatales fungi. A survey of these fungi associated with soil under beech, oak, pine, and spruce stands in Poland yielded 623 isolates, representing 10 species: Heinzbutiniagrandicarpa, Leptographiumprocerum, L.radiaticola, Ophiostomapiliferum, O.quercus, Sporothrixbrunneoviolacea, S.dentifunda, S.eucastaneae, and two newly described taxa, namely Sporothrixroztoczensissp. nov. and S.silvicolasp. nov. In addition, isolates collected from fallen shoots of Pinussylvestris that were pruned by Tomicus sp. are described as Sporothrixtumidasp. nov. The new taxa were morphologically characterized and phylogenetically analyzed based on multi-loci sequence data (ITS, β-tubulin, calmodulin, and translation elongation factor 1-α genes). The Ophiostomatales species were especially abundant in soil under pine and oak stands. Leptographiumprocerum, S.silvicola, and S.roztoczensis were the most frequently isolated species from soil under pine stands, while S.brunneoviolacea was the most abundant in soil under oak stands. The results highlight that forest soil in Poland has a wide diversity of Ophiostomatales taxa, but further studies are required to uncover the molecular diversity and phylogenetic relationships of these fungi, as well as their roles in soil fungal communities.