Genebanks serve as a rich source of diversity that can be exploited for crop improvement. However, large numbers of accessions usually have to be evaluated to find material with the characters of interest, and therefore, enhanced trait information can facilitate the more efficient selection of accessions by users. In this study, we report on the distribution of resistances to 28 races of downy mildew among 1223 genebank accessions of cultivated lettuce (Lactuca sativa L.) and 14 related wild species. Due to modern plant breeding, the overall level of resistance of cultivars released after 1950 appears to have increased two- to three-fold compared with varieties from earlier periods. Although fully resistant reactions could be observed among the accessions of cultivated lettuce for each of the 28 investigated races, the resistance probability was more than two-fold higher on average for accessions from the wild gene pool. In general, species of the primary gene pool appeared less resistant than those of the secondary or tertiary gene pool. Probabilities for examined Lactuca species ranged from 0.29 for L. serriola to 1.00 for L. perennis compared with 0.19 for cultivated lettuce, with lower overall resistance probabilities observed only for L. altaica, L. dregeana and L. tenerrima. For L. serriola, the closest relative of cultivated lettuce and the wild species with the highest number of examined accessions, resistance probabilities to each of the investigated downy mildew races were relatively high for populations originating from Eastern Europe and Northern Asia.