Seed samples of maize (Zea mays ssp. mays) from Mexico and of teosintes (Zea spp.), the nearest wild relatives of maize, from Mexico, Guatemala, and Nicaragua were assessed for infection with Fusarium species. Strains similar in morphology to Fusarium moniliforme and F. subglutinans were the most frequent isolates from maize and from teosinte species including Z. diploperennis, Z. luxurians, Z. mays ssp. mexicana, and Z. mays ssp. parviglumis. Analysis of fertility, vegetative compatibility and mycotoxin production identified 63% of the 70 F. moniliforme strains from teosinte as genetically diverse members of Gibberella fujikuroi mating population A, a common pathogen of maize. The F. subglutinans strains from maize and teosinte were similarly genetically diverse, but were not fertile with standard testers of G. fujikuroi mating populations B and E, common pathogens of Poaceae, or of mating population H, which causes pitch canker disease of pine. Fifty-four percent of the 80 F. subglutinans strains were fertile when crossed with female tester strains from teosinte and maize collected in a field at Netzahualcoyotyl in the state of Mexico. These strains from Mexico and Central America may comprise a new and distinct G. fujikuroi mating population, but a strain from the Netzahualcoyotyl field site was fertile with a strain of G. fujikuroi mating population H from California. Thus, F. subglutinans from teosinte and maize may have a close relationship to mating population H from pine.