Urediniospore isolates of Puccinia triticina were obtained from wheat leaf collections made in three wheat-growing regions in Nebraska in 1997 and in four regions in 1998. Using 16 Thatcher lines that are near-isogenic for leaf rust resistance, 17 virulence phenotypes were found among 121 single uredinial isolates in 1997, and 42 virulence phenotypes were found among 178 isolates in 1998. The most prevalent phenotype in 1997 was MDRR (virulent on Lr1, 3, 3ka, 10, 11, 18, 23, 24, and 30). In 1998, virulence phenotypes MDRR and MDRM (virulent on Lr1, 3, 3ka, 10, 11, 23, 24, and 30) were the most prevalent. In both years, virulence frequency was above 80% to genes Lr1, 3, 3ka, 10, 11, 23, 24, and 30 and below 21% to Lr2a, 17, and 26. Virulence frequency to Lr2c was 37% in 1997 and 22% in 1998. No virulence was found to Lr9, 16, or 21 in either year. New virulence phenotypes were detected in 1998 that were not found in 1997. In 1998, virulence was less frequent on Lr2a, 2c, 3ka, 18, 24, and 26 and more frequent on Lr17 than in 1997.