Shifting range in a stingless bee leads to pre-mating reproductive interference between species

Human-induced shifts in species’ ranges can increase contact between closely related species and lead to reproductive interference. In Australia, climate change and trade in stingless bee colonies is increasing the range overlap of two cryptic species: Tetragonula carbonaria and T. hockingsi . To investigate reproductive interactions between these species, we validated a diagnostic-PCR test based on the mitochondrial gene COI to ID field specimens to species. We then assessed the likelihood of reproductive interference in four ways. First, we imaged the male genitalia of each species and found no evidence of reproductive character displacement. Second, we assessed species composition of mating aggregations in an area of sympatry (Southeast Queensland) and confirmed that some males join the mating aggregations of interspecific colonies. Third, we translocated T. hockingsi colonies into the southern range of T. carbonaria (Sydney) and tracked their ability to requeen. These translocated colonies attracted mating aggregations comprised almost entirely of interspecific males, but never formed hybrid colonies; instead, queens either mated with their brothers, or the colony failed to requeen at all. Finally, we presented T. carbonaria males with either conspecific or interspecific virgin queens and found that males attempted to mate only with their own species’ queens. In all, we conclude that reproductive barriers between these species are complete with respect to “short-range” mating cues, but not for “long-range” mate attraction cues. Our study highlights that hive movements can increase some forms of pre-mating reproductive interference between managed bee species, even where the species do not actually mate or hybridize.