In Ethiopia, stem rust caused by Puccinia graminis f. sp. tritici (Pgt) is one of the most serious wheat diseases. On vulnerable cultivars, it has the ability to cause yield losses of up to 100%. The infection responses of forty-two wheat lines at the seedling stage against eleven isolates of stem rust race TTKSK (Ug99 race), TTRTF, TKTTF (Digalu race), JRCQC, TKKTF, TRTTF, TTTTF, TTKTF, TTKTT, RRTTF, and TKPTF were evaluated in a study. Wheat lines were experimentally infected with urediniospores of the races under controlled conditions at Ambo Plant Protection Research Center in 2021. Against the Pgt races that were examined, different seedling resistance responses were seen. Thirteen wheat lines,