The random amplified polymorphic DNA (RAPD) technique was used to determine the mating groups of several members of the Fusarium section Liseola recovered from maize, rice and sorghum collected from different locations in Ghana. Three mating groups were identified, A, D and F, of which all A and F isolates were confirmed by mating studies. Fertile crosses were also obtained in crosses involving two of the isolates identified as belonging to the D population. Variability within the A population isolated from seeds and stem‐bases of maize was investigated to determine whether the sub‐structuring of this population was related to the host tissue from which the isolates were obtained. The relative merits of the RAPD procedure, compared to the mating procedure, for determining the mating affiliations of isolates and for more detailed analyses of isolates within a population, as well as its possible advantages over established RFLP methodologies are discussed.