Fusarium disease of oats reduces yield quality due to decreasing germination that is caused by then contamination of grain with mycotoxins produced by Fusarium fungi. The aim of this study was to characterize the resistance of naked breeding lines of oats to fungal grain infection and to contamination with T-2 and HT-2 toxins. Thirteen naked oat breeding lines and two naked varieties, Nemchinovsky 61 and Vyatskiy, as well as a husked variety Yakov, were grown under natural conditions in the Nemchinovka Federal Research Center in 2019–2020. The contamination of grain with fungi was determined by the mycological method and real-time PCR. The analysis of mycotoxins was carried out by ELISA. In oats, Alternaria (the grain infection was 15–90 %), Cochliobolus (1–33 %), Cladosporium (1–19 %), Epicoccum (0–11 %), and Fusarium (3–17 %) fungi prevailed in the grain mycobiota. The predominant Fusarium species were F. poae (its proportion among Fusarium fungi was 49–68 %) and F. langsethiae (29–28 %). The highest amounts of F. langsethiae DNA ((27.9–71.9)×10–4 pg/ng) and T-2/HT-2 toxins (790–1230 μg/kg) were found in the grain of husked oat Yakov. Among the analysed naked oat lines, the amount of F. langsethiae DNA varied in the range of (1.2–42.7)×10–4 pg/ng, and the content of T-2/HT-2 toxins was in the range of 5–229 μg/kg. Two oat breeding lines, 54h2476 and 66h2618, as well as a new variety, Azil (57h2396), can be characterized as highly resistant to infection with Fusarium fungi and contamination with mycotoxins compared to the control variety Vyatskiy.