Fusarium oxysporum f. sp. cucumerinum is the fungal pathogen responsible for Fusarium vascular wilt of cucumber. In Australia, disease management in soilless greenhouse cucumbers is limited by the understanding of the disease cycle, in particular the risk associated with infection of pruning wounds by airborne propagules. Aerial dissemination of the pathogen in response to temperature and relative humidity was investigated using a specific and sensitive quantitative real-time PCR assay. Both macroconidia and microconidia were identified as airborne propagules and a potential relationship between fluctuation in relative humidity and spore release was found, however, it appears that crop disturbance may also influence conidia liberation. Experimental inoculation of stem wounds with conidia however, failed to establish infections. These results suggest that aerial inoculum propagates and disseminates the pathogen, however airborne spores are deposited on the substrate surface and infection occurs primarily through the root. We conclude that while airborne conidia are a risk to infection of cucumber plants in soilless greenhouse crops, resistant rootstocks are likely to provide good protection against this pathogen.