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In this study, we successfully introgressed and validated <i>Yr59</i> into four elite wheat cultivars, Jimai 22, Chuanmai 42, Zhengmai 9023 and Xinmai 26 through marker-assisted backcross selection. Used as female parents, these four cultivars were crossed with wheat line PI 660061 (<i>Yr59</i>). After two backcrosses and marker-assisted selection, the progenies were selfed and advanced to the BC₂F₄ generation. A total of 123 BC₂F₄ lines were selected based on agronomic traits and stripe rust resistance, and their BC₂F₅ and BC₂F₆ progenies were further evaluated for stripe rust resistance and agronomic traits. Seven markers linked with relevant genes, including <i>Xbarc32</i>, <i>Xwgp5175</i>, <i>Xwmc557</i> and <i>Xcfa2040</i> linked with <i>Yr59</i>; <i>Xwmc658</i> with <i>YrJ22</i>; <i>WE173</i> and <i>Xbarc181</i> with <i>Yr26</i>, were used to genotype the breeding lines. A total of 109 introgression lines with positive markers for <i>Yr59</i> were identified for further stripe rust and agronomic trait evaluation. Finally, 16 lines had higher levels resistance to stripe rust, and similar or superior agronomic traits compared to their parents were obtained. These lines can be released as new cultivars for various regions after regional tests and also can be used as resistance stocks for regional breeding programs to develop new cultivars with adequate and durable resistance to stripe rust.