Fusarium culmorum and Fusarium graminearum are the major agents of Fusarium head blight (FHB) in Poland. Both pathogens produce trichothecene B mycotoxins, such as deoxynivalenol (DON), nivalenol (NIV), and their acetylated derivatives. Over one hundred isolates of F. culmorum and F. graminearum were collected from symptomatic wheat, triticale and barley spikes from fields at several locations in Poland. A PCR assay with selective primers was used to identify the strains of F. culmorum and F. graminearum producing nivalenol (NIV), 3-acetyl-deoxynivalenol (3Ac-DON) and 15-acetyl-deoxynivalenol (15Ac-DON). The results indicated that DON- and NIV-producing strains were present among the tested isolates of these species. However, most of the isolates under study were classified as DON-producing strains (84 of 97 isolates of F. culmorum and 23 of 26 isolates of F. graminearum). According to the data presented, the DON chemotype of F. culmorum and F. graminearum is predominant in Poland. Additionally, all F. culmorum isolates were identified as 3Ac-DON-producers, while F. graminearum isolates were identified as 15Ac-DON-producers only.