The distribution of genetic diversity between Oryza sativa L. ssp. indica and O. sativa L. ssp. japonica covering different ecological zones in Yunnan was studied, and specific markers of indica/japonica subspecies, paddy/upland rice and different ecological zones were screened, using 36 microsatellite primers and 113 accessions in the Yunnan landrace rice core collection. The genetic diversity of japonica was higher than that of indica, and the ecological zone with the highest and smallest genetic diversity lay in south-east and north-east Yunnan, respectively. This distribution was consistent at morphological and isozyme levels with studies on the entire Yunnan rice resources and core collection. In addition, the results showed that, among 416 markers, there were six indica/japonica-specific markers, 15 specific markers for paddy/upland rice and three specific markers in different ecological zones. The main conclusions are that the landrace rice core collection in Yunnan genetically represents the entire landrace rice resources in Yunnan, the centre of genetic diversity at DNA level lies in south-east Yunnan, and the DNA differentiation between indica and japonica is small. Furthermore, microsatellite markers were useful for studying the genetic diversity, classification and ecotype of germplasm resources and their core collection.