【Objective】 To investigate the stripe rust resistance and resistance gene distribution in Xinjiang spring wheat germplasms, it’s necessary for identification stable resistant varieties (lines) carrying multiple resistance genes and providing reference information for the effective utilization of resistant resources and wheat disease-resistant breeding. 【Method】 Seedling-stage resistance evaluation was performed using the currently prevalent Puccinia striiformis f. sp. tritici races CYR32 and CYR34 for single-race inoculation assays. Adult-plant resistance assays were conducted during the 2023 and 2024 growing seasons at field sites in Qingshui, Gansu and Lugang, Xinjiang. A mixed inoculum of Puccinia striiformis f. sp. tritici races (CYR32, CYR33, CYR34, Su11-4, Su11-5, and G22-14) were used to simulate natural epidemic conditions. The presence of stripe rust resistance genes (Yr9, Yr15, Yr17, Yr18, Yr28, Yr29, Yr30, Yr80, Yr81, Yr82, Yr86, YrZH22 and YrZH84) was analyzed using tightly linked flanking markers or functional markers. 【Result】 Among the 155 tested wheat varieties (lines), 20 exhibited resistance to race CYR32, 26 showed resistance to CYR34, and 14 demonstrated resistance to both CYR32 and CYR34. Two wheat varieties (lines), Pirsabak-13 and Millat-11, exhibited immune responses (IT=0) against both CYR32 and CYR34 at the seedling stage. A total of 101 wheat varieties (lines) demonstrated stable resistance across both Xinjiang and Gansu field environments during the adult-plant stage, including 80 Xinjiang-bred varieties (lines) and 21 foreign-introduced varieties (lines). Eight wheat varieties (lines) (foreign varieties (lines): 10136, 10123, Xinjiang-bred varieties (lines): 9628, Hechun 137, Liangchun 1832, Liangchun 1773, Pin 9616 and Pin 9638) exhibited environmentally stable, high resistance (IT=0-3; DS≤20%) at the adult-plant stage. Molecular markers detection indicated that 14, 66, 6, 8, 17, 62, 17, 2, 1 and 11 varieties (lines) carried the resistance gene Yr9, Yr17, Yr18, Yr28, Yr29, Yr30, Yr81, Yr82, Yr86 and YrZH84, respectively. Yr15, Yr80 and YrZH22 were not detected in all wheat cultivars (lines). Pirsabak-13 (Yr9, Yr17, Yr29, Yr81), Borluag-16 (Yr17, Yr30, Yr86), Ufaq-2000 (Yr9, Yr18, Yr30), Bafeng 5 (Yr9, Yr17, Yr28, Yr30), 9628 (Yr30, Yr81, Yr82), Xinchun 5 (Yr17, Yr29, Yr30), Xinchun 50 (Yr9, Yr17, Yr30), Ning 33 (Yr17, Yr29, Yr81), and Ning 28 (Yr17, Yr28, Yr29) carry three to four resistance genes. These varieties exhibit stable resistance to the tested virulent races either throughout their entire growth period or during the adult plant stage. These varieties (lines) with multiple disease-resistant genes and exhibit stable disease resistance can be applied for breeding rust-resistant wheat in Xinjiang.【Conclusion】 The seedling resistance levels of spring wheat in Xinjiang to CYR32 and CYR34 are relatively low. However, the adult plant resistance was generally high. A total of 101 varieties (lines) exhibited stable resistance to the mixed pathogen races. Yr17 and Yr30 were the most widely distributed Yr genes among the tested spring wheat varieties. Wheat accessions carried unknown Yr genes or potentially novel genes/QTLs need further exploration.