Repetitive DNA elements discovered in the basidiomycete Chondrostereum purpureum were characterized and validated for use as genetic markers. Regions of these marker sequences were similar to retrotransposon and retrotransposon-like sequences, as indicated by BLAST searches of NCBI databases. These sequences occur in multiple DNA fragments of variable length in a given C. purpureum isolate, and thus can serve as strain-specific genetic markers. The segregation of the markers within a progeny set demonstrated their stability through meiosis. The population structure of C. purpureum was assessed using the markers. There was no evidence of a barrier to gene flow between C. purpureum populations separated by 1400 km and no indication of population sub-structuring based on host or geographical source of isolate. Repetitive fragments were amplified from four other species, suggesting the occurrence of these retrotransposon-like elements in other basidiomycetes and the potential utility of these markers for other fungi.