Stripe rust (yellow rust) of wheat, caused by Puccinia striiformis f. sp. tritici, has become more severe in eastern United States, Australia, and elsewhere since 2000. Recent research has shown that this coincided with a global spread of two closely related strains that were similar based on virulence phenotype and amplified fragment length polymorphism. The objective of this research was to quantify differences in aggressiveness among isolates representative of the pre-2000 and post-2000 populations. Representative isolates were evaluated at low (10 to 18°C) and high (12 to 28°C) temperature regimes for latent period, lesion length, lesion width, lesion area, and spore production on adult plants of a susceptible wheat cultivar with no known genes for resistance to stripe rust. “New” isolates (since 2000) were significantly more aggressive than “old” isolates (before 2000) for all variables. At the low temperature regime, new isolates sporulated 2.1 days (16%) sooner, grew 0.3 mm per day (18%) faster, produced 999 (140%) more spores per inoculation site per day, and produced 6.5 (71%) more spores per mm² of lesion per day compared with old isolates. At the high temperature regime, new isolates sporulated 3 days (26%) sooner, grew 0.2 mm per day (18%) and 2.2 mm² per day (88%) faster, grew 1.2 mm (50%) wider, produced 774 (370%) more spores per inoculation site per day, and produced 6.2 (159%) more spores per mm² of lesion per day than old isolates. New isolates showed significant adaptation to the warm temperature regime for all variables. Based on these results and previously published models for stripe rust epidemics, recent severe stripe rust epidemics were most likely enhanced by the pathogen's increased aggressiveness, especially at higher temperature. Furthermore, these results demonstrate that wheat rust fungi can adapt to warmer temperatures and cause severe disease in previously unfavorable environments.