The aim of the present study was to identify Avena sterilis genotypes demonstrating a high level of resistance against oat powdery mildew, using host-pathogen tests. The study was conducted on 350 A. sterilis genotypes from different parts of the world. Six single-spore isolates of Blumeria graminis (DC.) f. sp. avenae, which demonstrated different levels of virulence to control lines and cultivars, were used in host-pathogen screening tests. To confirm the resistant response of selected genotypes, 13 other isolates were used. Reactions to the isolates were grouped into three classes: resistant, intermediate, and susceptible. Susceptible cultivars Sam and Fuchs were used as controls to estimate the degree of infection. The results of the screening test showed that 10 genotypes were classified as resistant. The second test based on 13 other isolates revealed that only four of the 10 genotypes were a valuable source of resistance against powdery mildew. The identified genotypes may be used in oat breeding programs to increase the level of resistance against powdery mildew. First, however, further studies aimed at identifying whether this resistance is conditioned by a single gene or combinations of different genes are required.