Exploring new sources of resistance and widening genetic resources of disease resistance are important strategic reserves to ensure the sustainable development of agricultural production. In order to get it clear about the resistance to stripe rust and the distribution of stripe rust resistance genes in 400 wheat varieties (lines), the resistance to stripe rust was identified on adult plants in the field by using the dominant mixed races (CYR31,CYR32 and Su11-14) in Guanzhong area, and Yr5, Yr9, Yr10, Yr15, Yr17, Yr18 and Yr26 which are known as stripe rust resistance genes were detected and screened by molecular markers in this study for comprehensive analysis of resistance genes that may be carried by tested materials. The results showed that out of the 400 materials, 177 (44.25%) were highly resistance to immunity (IT=0~1) to mixed races at adult stage, 62 (15.5%) were moderate resistance (IT=2) and 162 (40.25%) were medium or high susceptible (IT=3/4). Combined with disease resistance performance and molecular analysis of known Yr gene, the results showed that Yr5 was carried by 121 (32.5%) wheat materials, Yr9 was carried by 96 (24%) materials,Yr10 was carried by 10 (2.5%) materials, Yr15 was carried by 19 (4.75%) materials, Yr17 was carried by 150 (37.5%) materials, Yr18 was detected in 15(3.75%) materials, and Yr26 was detected in 127 (31.75%) materials, respectively. But Yr5, Yr9, Yr10, Yr15, Yr17, Yr18 and Yr26 were not found in 25 materials such as Dingxi 24 and N7187 among the 400, they might contain other unknown resistance genes or new genes. The results of the study established the identification and evaluation system of resistance to wheat stripe rust, and 177 resistant materials with different resistance characteristics were selected out, of which 25 might contain new resistance sources, which laid a foundation for further cultivation of new varieties resistant to stripe rust