To determine stem rust resistance genes of wheat varieties in Yunnan province, 11 Chinese strains of Puccinia graminis f.sp. tritici with different virulence and 1 artificial mutant strain were used in 110 wheat varieties. The results indicated that among the 45 Sr genes, Sr31, Sr5, SrTmp, Sr30, Sr36, Sr8a, Sr11, Sr24, Sr29, Sr34, Sr9e, Sr26, Sr38, Sr47, and SrTt3 were characterised in 55 wheat varieties singly or in combination. Sr5, Sr31, SrTmp, Sr36, and Sr30 were contained in 17, 16, 5, 5, and 3 cultivars, respectively. Six cultivars contained Sr24 and/or Sr34. Three plant materials likely contained one or more of Sr8a, Sr11, Sr34, and Sr23 genes as well as other unknown genes. The 16 immune or highly resistant varieties contained one or more of Sr9e, Sr26, Sr38, Sr47, SrTt3, and other unknown resistance genes. The reaction types on 22 of the tested cultivars were different from those on Sr genes tested, and their resistance genes could not be analysed. Additionally, 17 varieties were susceptible to all the tested strains, having no postulation value. Our study provides a basis for improved breeding of stem rust resistant wheat in China.