We have isolated and characterized six chemically induced mutants of the filamentous fungus Aspergillus nidulans that are resistant to the experimental fungicide 8-chloro-4-(2-chloro-4-fluoro-phenoxy)quinoline (LY214352). The mutants are 13- to 430-fold more resistant to LY214352 than the parental strain, and one of the mutant strains requires LY214352 for maximal growth. The resistance trait is governed by a single dominant or partially dominant gene in each mutant, and it is likely that all of the mutations are allelic. The LY214352-resistant mutants were not cross-resistant to other compounds that inhibit the growth of A. nidulans. The implications of these findings on the potential for development of resistance to LY214352 are discussed.