The aim of the study was to monitor the mycobiota in grape samples at 3 maturation stages (pea berry, early veraison and ripe berry) and to test the ability of selected Penicillium strains to produce mycotoxins in year 2014. Grapes came from Nitra wine growing region, Šintava subregion and were analyzed by plating methods. From Devín grape variety the 339 strains were detected and identified. The most abundant genera found by descending order were Alternaria (42.8%), Cladosporium (24.2%) and Epicoccum (10.0%). Penicillium expansum was isolated from all 3 maturation stages but in low average relative density (1.2%). From Frankovka modrá (Frankish) grape variety the 562 strains were detected and identified. The most abundant genera found by descending order were Alternaria (30.6%), Cladosporium (21.3%) and Penicillium (20.3%). Totally were identified 8 species of the genus Penicillium. The most important species, on the basis of the isolation from all maturation stages and relative density, was P. expansum. Selected isolates of Penicillium species were tested for their toxigenity, according to thin layer chromatography method. In total 14 Penicillium strains representing 3 potentially toxigenic species were tested for their toxigenic ability, namely P. expansum, P. griseofulvum and P. chrysogenum. Out of 14 strains, 64% produced et least one mycotoxin as revealed by the method used here.