Fusarium oxysporum is a ubiquitous soilborne ascomycete responsible for vascular wilt in many plant species worldwide. This species comprises more than 120 putative host-specific formae speciales capable of causing marked economic losses. In summer 2009, wilt symptoms, including chlorosis and poor development of the root system, were observed on cultivars of chicory (Cichorium intybus) in northern Italy. The causal agent isolated from symptomatic tissues in this case was identified as F. oxysporum on the basis of both morphological features and molecular analyses. In this work, we attempted to characterize the isolates of F. oxysporum from C. intybus by both biological and molecular approaches. Pathogenicity trials performed on five species of the Asteraceae family with isolates of F. oxysporum from C. intybus indicated that the pathogen has a unique host range, infecting chicory only. Neither lettuce nor endive, lawn daisy or Paris daisy developed the disease. Five cultivars within C. intybus species were tested, and the cv. 'Clio' was the most susceptible. Phylogenetic analyses relative to the ribosomal intergenic spacer (IGS) and translation elongation factor 1-alpha (EF1-alpha) assigned isolates pathogenic to chicory to a single cluster, distinct from other pathogenic F. oxysporum. In light of these findings, we propose to designate this organism as Fusarium oxysporum f.sp. cichorii.