The complex social structure and male-biased sex ratio of honeybee mating systems are analysed, followed by detailed treatments of panmictic drone congregation areas and species-specific daily mating flight periods. This is followed by an account of queen polyandry and drone monogamy and competition. Mating on the wing is a finely tuned technical tour de force involving initial docking, establishing the internal connection of drone and queen, the deposition and transfer of sperm and, finally, decoupling of the pair and deposition of a mating sign. Subsequent to mating, the problems of the ultimate storage and utilisation of sperm are discussed. Finally, the matter of reproductive isolation is considered.

Polyandry in queen honey bees (Apis mellifera) prolongs the duration of nuptial flights which increases costs and risks. Under conditions of limited drone numbers the hypothesis was tested whether or not there is a threshold for successful mating during mating flight. In 29 queens we found a significant negative correlation between mating flight duration and number of spermatozoa in the spermatheca (Pearson r = −0.38, P = 0.04). This negative correlation supports the idea that queens continuously get information on her mating success during flight and return to the colony as soon as they have met a sufficient number of drones. In case of normal availability of drones queens fly from 10 to 30 minutes, so we compared 2 groups of queens (flight duration less than 30 versus more than 30 minutes). Sperm numbers differed significantly between the two groups (3.0 ± 0.77 and 1.1 ± 1.04 million, Wilcoxon, P < 0.001). These results further indicate that queens monitor mating success during flight.

Queen honeybees (Apis mellifera) mate with a large number of drones on their nuptial flights. Not all drones contribute equally to the queen’s offspring and the queen’s utilization pattern of spermatozoa from different drones has an important impact on the genetic composition of the colony. Here we study the consequences of sperm use for the fitness of the queen’s mates with microsatellite DNA-fingerprinting. Eight queens were instrumentally inseminated with semen of six or seven drones. Each drone contributed either 0.5 µl or 1.0 µl semen, respectively, and we analyzed both the impact of the insemination sequence and the amount of semen on the sperm utilization. Our data show no significant effect of the insemination sequence but a strong impact of the semen volume of a drone on the frequency of his worker offspring in the colony. This effect was not linear and the patriline frequencies of the drones contributing larger semen volumes are disproportionately enhanced. If these observations are also valid for natural matings, drone honeybees should maximize the number of sperm but not apply specific mating tactics to be first or last male in a mating sequence.

Workers of six colonies of the giant honeybee Apis dorsata from Sabah, Malaysia (five colonies) and Java (one colony) were genotyped using single locus DNA fingerprinting. The colonies from Sabah nested in colony aggregations of 5 and 28 nests respectively on two trees. Three DNA microsatellite loci (A14, A76, A88) with a total of 27 alleles provided sufficient genetic variability to classify the workers into distinct sub-families revealing the degree of polyandry of the queens. Queens mated on average with 30.17 ± 5.98 drones with a range from 19 to 53. The average effective number of matings per queen was 25.56 ± 11.63. In the total sample of 192 workers, 22 individuals were found that were not offspring of the colony's queen. Three of these were potentially drifted offspring workers from genotyped queens of colonies nesting on the same tree.