The introduction of Phakopsora pachyrhizi, the cause of soybean rust, into the United States is a classic case of a pathogen introduction that became established in a new geographical region overwintering on a perennial host (kudzu, Pueraria lobata). The objective of our study was to classify the pathogenic variation of P. pachyrhizi isolates collected in the United States, and to determine the spatial and temporal associations. In total, 72 isolates of P. pachyrhizi collected from infected kudzu and soybean leaves in the United States were purified, then established and increased on detached soybean leaves. These isolates were tested for virulence and aggressiveness on a differential set of soybean genotypes that included six genotypes with known resistance genes (Rpp), one resistant genotype without any known characterized resistance gene, and a susceptible genotype. Three pathotypes were identified among the 72 U.S. P. pachyrhizi isolates based on the virulence of these isolates on the genotypes in the differential set. Six aggressiveness groups were established based on sporulating-uredinia production recorded for each isolate on each soybean genotype. All three pathotypes and all six aggressiveness groups were found in isolates collected from the southern region and from both hosts (kudzu or soybean) in 2008. Shannon's index based on the number of pathotypes indicated that isolates from the South region were more diverse (H = 0.83) compared with the isolates collected in other regions. This study establishes a baseline of pathogenic variation of P. pachyrhizi in the United States that can be further compared with variation reported in other regions of the world and in future studies that monitor P. pachyrhizi virulence in association to deployment of rust resistance genes.