Summary • Australia's savannas typically are burnt every 1-3 years. Although there are concerns about the effect of frequent fire on recruitment of Australian savanna species, there is a lack of information. This research aimed to determine whether seed or microsite availability limits seedling recruitment of the overstorey tree Eucalyptus miniata and the midstorey shrub Acacia oncinocarpa, whether seed or microsite availability is affected by frequent fire, and the consequent effect on seedling recruitment. • Quadrats were established in unburnt areas. Experimental manipulations were addition of seed and/or disturbance of the soil surface to increase the number of microsites suitable for germination. In a second experiment, seed was added to quadrats established in three fire regimes after the annual burning event (unburnt, burnt early in the dry season, burnt late in the dry season). • In unburnt areas, seedling regeneration was limited by both seed supply and microsite availability. Both burning regimes reduced seedling emergence, possibly because the reduced canopy cover caused unfavourable microclimate, the increased grass and forb ground cover increased competition for resources, and there was increased loss of seed to seed harvesters. • The results indicate that sexual regeneration of these common species is disadvantaged by current burning practices because both seed supply and the number of microsites are reduced. Thus, long-term changes in savanna floristic structure seem likely unless fire managers aim to increase the fire-free intervals. The relative abundance of species able to reproduce vegetatively may increase under frequent fire regimes. Such a change may take a long time to detect given the fire-resistant and long-lived nature of overstorey species, and the capacity for vegetative regeneration in many savanna species. The impacts of fire on limiting seedling regeneration will require savanna managers to consider fire-free intervals of several years for effective recruitment of dominant woody species.