Sharp eyespot on wheat is caused by Rhizoctonia cerealis anastomosis group D subgroup I (AG-DI) and is an economically important stem-base disease of wheat in temperate regions worldwide. However， the understanding about the field population structure of R. cerealis is limited. In this study， the genetic structure of four wheat-infecting populations in China was investigated using six microsatellite markers characterized from the transcriptome data of R. cerealis AG-DI. A total of 173 unique genotypes were identified among 235 fungal isolates. Departure from Hardy-Weinberg equilibrium， a significant degree of inbreeding， and a significant deficit in heterozygotes indicated a nonrandom mating pattern. Combining the low to intermediate degrees of gametic disequilibrium， although with high genotypic diversity and low to moderate clonal fractions， sexual reproduction probably existed， but the asexual reproduction should be the predominant reproductive mode. Structural analysis showed three gene pools among the four populations， which indicated the existence of three evolutionary origins of R. cerealis AG-DI. The long-distance movement of contaminated material， especially the infected seed， might have caused the moderate gene flow among these populations， which was consistent with the high differentiation among these populations. Overall， the genetic characteristics of the populations suggested a moderate evolutionary potential for R. cerealis AG-DI in China.