The tomato Fusarium resistance gene 1-2 confers resistance to F. oxysporum f. sp. lycopersici race 2, which expresses the corresponding avirulence gene avrl-2. To elucidate the molecular basis of this gene-for-gene interaction, we initiated a search for the avrl-2 gene. Gamma irradiation mutagenesis, using 137Cs, was performed to generate an avrI-2 mutant of E oxysporum f. sp. lycopersici. To this end, a race 2 isolate was first transformed with a phleomycine resistance gene and a GUS marker gene in order to distinguish mutants from contaminating isolates. A total of 21,712 mutagenized colonies was tested for loss of avirulence on I-2-containing tomato seedlings. One mutant was selected that showed the expected loss of avirulence but, surprisingly, also showed reduced pathogenicity toward susceptible tomato plants. DNA analysis was subsequently used to visualize genomic changes in the mutant. Southern analysis on contour-clamped homogeneous electrophoretic field blots demonstrated a translocation of a 3.75-Mb chromosome in the mutant. Random amplified polymorphic DNA and amplified fragment length polymorphism analysis identified at least nine polymorphisms between the wild-type and mutant isolates. Most of these polymorphisms appeared as extra fragments in the mutant and contained repetitive DNA sequences.