Isolates of Alternaria solani, cause of potato early blight, collected in 1998 through 2001 from various potato growing areas across the midwestern United States, were tested for sensitivity to azoxystrobin. Isolates collected in 1998, prior to the introduction of azoxystrobin, were tested to establish the baseline sensitivity of the fungus to this fungicide. Isolates collected in subsequent years, not necessarily from the same sites as baseline isolates, were tested to determine if populations of A. solani had become less sensitive to azoxystrobin. Azoxystrobin sensitivity was determined utilizing an in vitro spore germination assay. The effective fungicide concentration that inhibited spore germination by 50% (EC50) was determined for each isolate. There was no significant difference in mean EC50 values between baseline isolates and all other isolates collected through 1999. Mean azoxystrobin EC50 values of A. solani isolates collected in 2000 and 2001 were significantly higher compared with means from previous years, and mean azoxystrobin EC50 values from 2001 were significantly higher than means from isolates collected in 2000. A subset of 54 A. solani isolates was evaluated in vitro for cross-sensitivity to pyraclostrobin and trifloxystrobin. A highly significant and strong correlation among the isolates tested for fungicide cross-sensitivity was detected between azoxystrobin and pyraclostrobin; however, the correlation between azoxystrobin and trifloxystrobin, and between trifloxystrobin and pyraclostrobin, was significant but weak. A second subset of five isolates was chosen for in vivo assessment of azoxystrobin, pyraclostrobin, and trifloxystrobin sensitivity. Disease severity on plants treated with azoxystrobin and pyraclostrobin was significantly greater with reduced-sensitive A. solani isolates compared with sensitive isolates. Disease severity was not statistically different between azoxystrobin reduced-sensitive and sensitive A. solani isolates on plants treated with trifloxystrobin. This is the first report of a shift in sensitivity to QoI fungicides in a fungus possessing only an anamorphic stage.