Wheat leaf rust (Puccinia triticina) is becoming a serious concern in Spanish wheat, especially on durum wheat where acreage has enormously increased. Host resistance is the preferred method of disease control, but the virulence spectrum of the leaf rust population in Spain is currently unknown. In order to deploy effective Lr genes, this study was conducted to characterize the virulence spectrum of leaf rust in Andalusia (Spain). Isolates were obtained from surveys of wheat fields across Andalusia from 1998 to 2000. From 56 isolates phenotyped, 35 pathotypes were identified. Virulence to Lr10, Lr11, Lr14a, Lr14b and Lr18 was high (>96%), while virulence to Lr9 and Lr24 were not found. None of the isolates collected from durum wheat were virulent to Lr1, Lr3, Lr3ka, Lr3bg, Lr15, Lr16 and Lr17, while many of the isolates collected on bread wheat showed virulence on these genes, indicating a certain specialization in the leaf rust infecting durum wheat. Population dynamics of current wheat leaf rust pathotypes in terms of mutation and migration are discussed.