Fusarium wilt, caused by the soilborne pathogen Fusarium oxysporum f. sp. vasinfectum race 1, is a vascular disease in cotton (Gossypium spp.), and is a component of a disease complex with root-knot nematodes (Meloidogyne incognita). Genetic analysis of two interspecific crosses (G. barbadense Pima S-7 × G. hirsutum Acala NemX and Pima S-7 × Acala SJ-2) showed that one major gene (designated Fov1) with allele dosage effect conferred resistance to F. oxysporum f. sp. vasinfectum race 1 in Pima S-7. Two amplified fragment length polymorphism (AFLP) markers were linked to Fov1 in Pima S-7, with genetic distance from the gene of 9.3 and 14.6 centimorgans. Less severe wilt symptoms in Acala NemX than Acala SJ-2 indicated that Acala NemX possesses one or more minor genes contributing to delay of wilt symptoms. Highly resistant plants in F2 and F3 (Pima S-7 × NemX) families indicated transgressive segregation effects of minor genes in Acala NemX combined with Fov1 from Pima S-7. The effects of wilt and nematode resistance on the nematode-wilt disease complex were assayed with two inoculation methods. In the presence of both pathogens, wilt damage measured as shoot and root weight reductions was greatest on wilt- and nematode-susceptible Acala SJ-2 and least in root-knot nematode-resistant and wilt-susceptible Acala NemX. Intermediate damage occurred in wilt-resistant and root-knot nematode-susceptible Pima S-7. The results indicated that nematode resistance was more effective than wilt resistance in suppressing wilt symptoms when either resistance was present alone. Nematode resistance combined with intermediate wilt resistance, as in the F1 (Pima S-7 × NemX), was highly effective in protecting plants from root-knot nematodes and race 1 of Fusarium wilt as a disease complex.