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Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most devastating diseases in wheat. Due to the large-scale and widely-distributed planting pattern of wheat, the directional selection pressure of the pathogen is very strong. Therefore, it is urgent to pyramid more stripe rust resistance genes in wheat cultivars to enhance resistance durability and ensure wheat production safety. In this study, two quantitative trait loci (QTL) for adult plant resistance (APR) to stripe rust, QYr.nafu-2BL and QYr.nafu-3BS, were validated and introgressed from wheat line P9897 into three Chinese elite wheat cultivars, Chuanmai 42, Xiangmai 25, and Zhengmai 9023, through marker validation. The three Chinese elite varieties were used as the female parent to cross with wheat line P9897, and they were selfed to the F6 generation. A total of 114 lines were then selected based on field agronomic traits and stripe rust resistance. Four markers (Xcfd73, Xgwm120, Xbarc87 and Xbarc133) linked with the QTL's regions were employed to screen the 114 F6 lines. Subsequently, 27 lines combining two target QTL from P9897 were selected. The combination of agronomic traits and disease resistance results showed that 13 of these selected lines had favorable application prospects. The promising lines selected in this study could enrich the genetic resources of wheat stripe rust resistance genes, as well as provide material support and a theoretical basis for the prevention and control of wheat stripe rust in China. [ABSTRACT FROM AUTHOR]