Genetic variation within 34 Pythium irregulare isolates was analyzed using restriction fragment length polymorphisms (RFLPs) as genetic markers. Most isolates had two alleles at several codominant RFLP loci and were scored as heterozygous. Heterozygotes were detected in F1 progeny from an in vitro cross and segregation ratios of the F2 progeny were not significantly different from those expected for allelic variation in a diploid. This confirmed that outcrossing occurs and contributes to genetic variation within the species. Phenetic analysis showed that isolates formed genetically related groups due to their host species and not due to similarities in their geographical origins. All isolates originating from medic formed a discrete group and were highly differentiated from the cereal and sub-clover isolates. The allelic distributions between isolates from these host-groups were significantly different. Most isolates also showed significant differences in their pathogenicity between hosts, indicating that they varied in pathogenic fitness and were better adapted to parasitising some hosts relative to others. These isolates were however, not necessarily more pathogenic on their host of origin. This research provided estimates of the extent of genetic and pathogenic diversity within P. irregulare and qualitative evidence for the occurrence of host-mediated selection and sexual outcrossing in the field.