Collections of Puccinia triticina were made from rust-infected wheat leaves in Georgia, South Carolina, North Carolina, and Virginia in 1999 to examine if these states can be considered as a single epidemiological unit for virulence phenotypes of the wheat leaf rust pathogen. Singleuredinial isolates derived from the leaf rust collections were processed for identification of virulence phenotypes on seedling plants in greenhouse tests. Twenty-one virulence phenotypes from 253 isolates were described based on infection type to 16 Thatcher wheat lines near-isogenic for leaf rust resistance genes. Virulence phenotype MBRK (virulent to leaf rust resistance genes Lr1, Lr3, Lr3ka, Lr11, Lr30, Lr10, Lr14a, and Lr18) was the most common phenotype in the region, at 38.7% of all isolates. Phenotype TLGF (virulent to Lr1, Lr2a, Lr2c, Lr3, Lr9, Lr11, Lr14a, and Lr18) was the second most common phenotype overall, at 33.8% of isolates. Twenty-nine isolates selected on the basis of seedling virulence phenotypes also were tested for virulence to adult wheat plants with the resistance genes Lr12, Lr13, Lr22b, and Lr34. In all, 23 isolates were avirulent to Lr12 and 26 isolates were virulent to Lr13. All isolates had fewer and smaller uredinia on the Thatcher line with Lr34 compared with Thatcher. The widespread occurrence of the predominant P. triticina virulence phenotypes throughout the region indicated that the South Atlantic states should be considered as a single epidemiological area for wheat leaf rust. Some virulence phenotypes which occurred at lower frequencies were found primarily in the Coastal Plain and mountains of North Carolina or in breeding plots in southern Georgia. Localized populations of P. triticina may develop in the South Atlantic region due to overwintering of leaf rust infections or specific selection by leaf rust resistance genes in wheat cultivars.