For the present study, samples of cleistothecia were collected in Central Lithuania and Southern Ukraine. To characterize the virulence, complexity and diversity of powdery mildew (Blumeria graminis f. sp. tritici) populations, 80 isolates were derived from single ascospores, 40 isolates from each population. Pathotype analysis was conducted on 16 differentials with known Pm genes. According to the proposed nomenclature, 32 pathotypes were identified in the Lithuanian powdery mildew population and 30-in the Ukrainian population. The most frequent phenotype in Lithuania was NGDE (7.5%), and in Ukraine it was NGKE (15%). The Ukrainian powdery mildew population was more complex and contained more virulence genes per isolate. The virulence test was carried out by inoculation on detached leaves of 26 common wheat (Triticum aestivum L.) differentials with known Pm genes and 12 resistant winter wheat breeding lines and cultivars. The frequencies of virulence to these differentials ranged from 0% to 100%, and most of them had high level of virulence. No virulence was found to Pm20 and Pm25+3a genes, cultivar 'Lastivka odeska' and breeding lines CN 89/16 and PI 170911 in both populations, whereas the frequencies of virulence were more than 50% to Pm1a,