The aim of this study was to identify the species composition and to evaluate the prevalence of genes involved in the synthesis of the following mycotoxins: deoxynivalenol, nivalenol and fumonisins in the population of Fusarium spp. isolated from ears of winter wheat in southern Poland. All fungal isolates were identified by the species-specific PCR or sequencing of the translation elongation factor. Significant differences were observed in both abundance and species composition of the collected strains between two years of studies. A total of 304 ear samples were processed and 107 Fusarium strains belonging to 10 species: F. graminearum, F. culmorum, F. sporotrichioides, F. poae, F. avenaceum, F. oxysporum, F. verticillioides, F. equiseti, F. tricinctum and F. cerealis were isolated in 2012 and 2013. Numerous presence of mycotoxin-biosynthesis pathway genes was detected in the examined material, which evidences the potential toxicity of the analyzed Fusarium fungi.