Fusarium oxysporum f. sp. Vasinfectum (Fov) is considered the most destructive pathogen of cotton in Australia. In this study, BC3 progenies of chromosome addition lines between G. sturtianum (C genome), an Australian wild Gossypium species shown to be resistant to fusarium wilt, and G. hirsutum were genetically characterised to determine the number and identity of the G. sturtianum chromosomes in 47 G. hirsutum X G. sturtianum chromosome addition families. The 47 families were challenged with Fov (VCG 11) in glasshouse trials using root-dripping inoculations to determine their levels of fusarium wilt resistance. Overall, 20 of the BC3 families showed enhaced fusarium wilt resistance relative to their G. hirsutum parent. Logistic regression nominated five G. sturtianum linkage groups as having significant effect in a G. hirsutum background. Two linkage groups were associated with increased susceptibility.