Black spot disease caused by the hemibiotrophic ascomycete Diplocarpon rosae is the most devastating disease of field‐grown roses. Although resistance to black spot is an important trait for rose breeding, little information on the diversity of the pathogen is currently available. To date, a number of single‐spore isolates have been characterized based on a set of test genotypes of the host. In this study, six polymorphic simple sequence repeat (SSR) markers for D. rosae were developed and their potential application to single‐spore isolates collected from a wide range of geographic locations within and outside Europe discussed. Populations of the fungus were then analysed in different German rose collections/populations. Gene diversity was highest in older rose collections managed without fungicide application, and lowest in the two‐ to three‐year‐old testing sites of German rose breeders. Additional analyses of a global collection of single samples and populations revealed no unique alleles from any of the locations, indicating that global trading of rose varieties has led to an admixture of the pathogen.