Honeybees (Apis mellifera L.) have an extreme polyandrous mating system. Worker offspring of 19 naturally mated queens was genotyped with DNA microsatellites, to estimate male reproductive success of 16 drone producing colonies. This allowed for estimating the male mating success on both the colony level and the level of individual drones. The experiment was conducted in a closed population on an isolated island to exclude interferences of drones from unknown colonies. Although all colonies had produced similar numbers of drones, differences among the colonies in male mating success exceeded one order of magnitude. These differences were enhanced by the siring success of individual drones within the offspring of mated queens. The siring success of individual drones was correlated with the mating frequency at the colony level. Thus more successful colonies not only produced drones with a higher chance of mating, but also with a significantly higher proportion of offspring sired than drones from less successful colonies. Although the life cycle of honeybee colonies is very female centred, the male reproductive success appears to be a major driver of natural selection in honeybees.