We report the characterization of ten microsatellite markers in the fungus Puccinia striiformis f.sp. tritici, responsible for yellow rust disease on wheat. A published EST library was scanned for microsatellite motives, and over 15 selected EST sequences, 13 were successfully amplified and ten exhibited polymorphism over an international collection of 43 isolates. These new microsatellites, added to the few previously published ones, provide a sufficient set of markers to perform population genetic studies.