Andrew F. G. Bourke

Genetic analysis of spatial foraging patterns and resource sharing in bumble bee pollinators.

Conservation biologists, evolutionary ecologists and agricultural biologists require an improved understanding of how pollinators utilize space and share resources. Using microsatellite markers, we conducted a genetic analysis of space use and resource sharing at several spatial scales among workers of two ecologically dissimilar bumble bee species (Bombus terrestris and B. pascuorum) foraging in an urban landscape (London, UK). At fine scales, the relatedness of workers visiting small patches of flowers did not differ significantly from zero. Therefore, colonies shared flower patches randomly with other colonies, suggesting that worker scent‐marks deterring visits to unrewarding flowers have not evolved as signals benefiting nestmates. To investigate space use at intermediate scales, we developed a program based on Thomas & Hills maximum likelihood sibship reconstruction method to estimate the number of colonies utilizing single sites. The average number of colonies (95% confidence limits) sending workers to forage at sites of ≈ 1 ha in area was 96 colonies (84–118) in B. terrestris and 66 colonies (61–76) in B. pascuorum. These values are surprisingly high and suggested that workers travelled far from their colonies to visit the sites. At the landscape scale, there was little or no genetic differentiation between sites. We conclude that urban habitats support large bumble bee populations and are potentially valuable in terms of bumble bee conservation. In addition, bumble bee‐mediated gene flow in plants is likely to occur over large distances and plant–bumble bee conservation requires landscape‐scale action.

Parentage, reproductive skew and queen turnover in a multiple-queen ant analysed with microsatellites

We investigated the fine genetic structure of colonies of the ant, Leptothorax acervorum, to examine how queens share parentage (skew) in a social insect with multiple queens (polygyny). Overall, 494 individuals from eight polygynous field colonies were typed at up to seven microsatellite loci each. The first main finding was that surprisingly many sexual progeny (60% of young queens and 49% of young males) were not the offspring of the extant queens within their colonies. This implies that a high turnover (brief reproductive lifespan) of queens within colonies could be an important feature of polygyny. The second main result was that in most colonies relatedness among sexual progeny fell significantly below that expected among full siblings, proving that these progeny were produced by more than one singly–mated queen, but that skew in two colonies where the data permitted its calculation was moderate to high. However, relative to a German population, the study population is characterized by low queen–queen relatedness and low skew in female production, which is in line with the predictions of skew theory.

Landscape context not patch size determines bumble-bee density on flower mixtures sown for agri-environment schemes

Bumble-bee declines across Europe have been linked to loss of habitat and forage availability due to agricultural intensification. These declines may have severe ecological and commercial consequences since bumble-bees pollinate a range of wildflowers and crops. In England, attempts are being made to reintroduce forage resources through agri-environment schemes, yet there are few data on how the area of forage, or the landscape context in which it is provided, affects their success. We investigated the effects of sown forage patches on bumble-bees across sites varying in landscape characteristics. Bumble-bee densities were higher on sown patches compared with control habitats but did not vary with patch size, i.e. total forager numbers were proportional to patch area. Importantly, the relative response to sown forage patches varied widely across a landscape gradient such that their impact in terms of attracting foraging bumble-bees was greatest where the proportion of arable land was highest.

Non-lethal sampling of DNA from bumble bees for conservation genetics

SummaryNon-lethal sampling of DNA from individuals in wild populations will often be required for studies of the conservation genetics of social insects, since it avoids destroying members of scarce or declining species. We investigated the effectiveness and consequences of methods of non-lethal sampling of DNA from bumble bee workers. In an experiment with two captive and confined Bombus terrestris colonies, we found that, unlike sampling haemolymph, sampling the terminal portion of the tarsus of a mid-leg of a worker reliably yielded amplifiable microsatellite DNA and did not significantly reduce worker survivorship. In a further experiment with four B. terrestris colonies whose workers were allowed to forage freely at flowers in the external environment, tarsal sampling of either a mid-leg or a hind-leg had no significant effects on worker survivorship, the mean body mass of foraging workers, the frequency or duration of foraging trips, mass of pollen loads or mass of nectar loads. We therefore suggest that tarsal sampling of either a mid-leg or a hind-leg is an effective and acceptable means of non-lethally sampling DNA from workers in wild populations of bumble bees, because effects on individual and colony performance are likely to be absent or minimal.

Differential gene expression in queen–worker caste determination in bumble-bees

Investigating how differential gene expression underlies caste determination in the social Hymenoptera is central to understanding how variation in gene expression underlies adaptive phenotypic diversity. We investigated for the first time the association between differential gene expression and queen–worker caste determination in the bumble-bee Bombus terrestris. Using suppression subtractive hybridization we isolated 12 genes that were differentially expressed in queen- and worker-destined larvae. We found that the sets of genes underlying caste differences in larvae and adults failed to overlap greatly. We also found that B. terrestris shares some of the genes whose differential expression is associated with caste determination in the honeybee, Apis mellifera, but their expression patterns were not identical. Instead, we found B. terrestris to exhibit a novel pattern, whereby most genes upregulated (i.e. showing relatively higher levels of expression) in queen-destined larvae early in development were upregulated in worker-destined larvae late in development. Overall, our results suggest that caste determination in B. terrestris involves a difference not so much in the identity of genes expressed by queen- and worker-destined larvae, but primarily in the relative timing of their expression. This conclusion is of potential importance in the further study of phenotypic diversification via differential gene expression.

Hamilton's rule and the causes of social evolution

Hamiltons rule is a central theorem of inclusive fitness (kin selection) theory and predicts that social behaviour evolves under specific combinations of relatedness, benefit and cost. This review provides evidence for Hamiltons rule by presenting novel syntheses of results from two kinds of study in diverse taxa, including cooperatively breeding birds and mammals and eusocial insects. These are, first, studies that empirically parametrize Hamiltons rule in natural populations and, second, comparative phylogenetic analyses of the genetic, life-history and ecological correlates of sociality. Studies parametrizing Hamiltons rule are not rare and demonstrate quantitatively that (i) altruism (net loss of direct fitness) occurs even when sociality is facultative, (ii) in most cases, altruism is under positive selection via indirect fitness benefits that exceed direct fitness costs and (iii) social behaviour commonly generates indirect benefits by enhancing the productivity or survivorship of kin. Comparative phylogenetic analyses show that cooperative breeding and eusociality are promoted by (i) high relatedness and monogamy and, potentially, by (ii) life-history factors facilitating family structure and high benefits of helping and (iii) ecological factors generating low costs of social behaviour. Overall, the focal studies strongly confirm the predictions of Hamiltons rule regarding conditions for social evolution and their causes.

Split sex ratios in a multiple-queen ant population

Split sex ratio theory is an important extension of sex allocation theory. It suggests that colony sex ratios in social insects vary because workers control sex allocation and respond to variations in their comparative relatedness with females and males (relatedness asymmetry). In a population of the ant Leptothorax acervorum, 21 monogynous (single-queen) colonies produced a female-biased sex ratio (62% females), and 24 polygynous (multiple-queen) colonies produced a male-biased sex ratio (28% females). Within the polygynous colonies, queen number did not affect sex ratio (with colony productivity statistically controlled). As colony productivity rose, the sex ratio either did not change (monogynous colonies) or became more male-biased (polygynous colonies). The fraction invested in sexuals rose with increasing colony size and productivity in monogynous but not in polygynous colonies, which invested less in sexuals. These findings suggest that split sex ratios in L. acervorum stem from two processes. The first is workers’ response to the variation in relatedness asymmetry caused by variable queen numbers. The second is sex-ratio compensation by monogynous colonies for male-biased production in polygynous colonies. This arises because polygynous colonies reproduce, partly, by colony budding and so have daughter colonies subject to local resource competition.

Mating frequency and mating system of the polygynous ant, Leptothorax acervorum.

Multiple mating by queens (polyandry) and the occurrence of multiple queens in the same colony (polygyny) alter patterns of relatedness within societies of eusocial insects. This is predicted to influence kin‐selected conflicts over reproduction. We investigated the mating system of a facultatively polygynous UK population of the ant Leptothorax acervorum using up to six microsatellite loci. We estimated mating frequency by genotyping 79 dealate (colony) queens and the contents of their sperm receptacles and by detailed genetic analysis of 11 monogynous (single‐queen) and nine polygynous colonies. Results indicated that 95% of queens were singly mated and 5% of queens were doubly mated. The corrected population mean mating frequency was 1.06. Parentage analysis of adults and brood in 17 colonies (10 monogynous, 7 polygynous) showed that female offspring attributable to each of 31 queens were full sisters, confirming that queens typically mate once. Inbreeding coefficients, queen–mate relatedness of zero and the low incidence of diploid males provided evidence that L. acervorum sexuals mate entirely or almost entirely at random. Males mated to queens in the same polygynous colony were not related to one another. Our data also confirmed that polygynous colonies contain queens that are related on average and that their workers had a mixed maternity. We conclude that the mating system of L. acervorum involves queens that mate near nests with unrelated males and then seek readoption by those nests, and queens that mate in mating aggregations away from nests, also with unrelated males.

Reproductive conflict in bumblebees and the evolution of worker policing

Worker policing (mutual repression of reproduction) in the eusocial Hymenoptera represents a leading example of how coercion can facilitate cooperation. The occurrence of worker policing in “primitively” eusocial species with low mating frequencies, which lack relatedness differences conducive to policing, suggests that separate factors may underlie the origin and maintenance of worker policing. We tested this hypothesis by investigating conflict over male parentage in the primitively eusocial, monandrous bumblebee, Bombus terrestris. Using observations, experiments, and microsatellite genotyping, we found that: (a) worker‐ but not queen‐laid male eggs are nearly all eaten (by queens, reproductive, and nonreproductive workers) soon after being laid, so accounting for low observed frequencies of larval and adult worker‐produced males; (b) queen‐ and worker‐laid male eggs have equal viabilities; (c) workers discriminate between queen‐ and worker‐laid eggs using cues on eggs and egg cells that almost certainly originate from queens. The cooccurrence in B. terrestris of these three key elements of “classical” worker policing as found in the highly eusocial, polyandrous honeybees provides novel support for the hypothesis that worker policing can originate in the absence of relatedness differences maintaining it. Worker policing in B. terrestris almost certainly arose via reproductive competition among workers, that is, as “selfish” policing.

Lifetime reproductive success and longevity of queens in an annual social insect.

Although central to understanding life‐history evolution, the relationship between lifetime reproductive success and longevity remains uncertain in many organisms. In social insects, no studies have reported estimates of queens’ lifetime reproductive success and longevity within populations, despite the importance of understanding how sociality and associated within‐group conflict affect life‐history traits. To address this issue, we studied two samples of colonies of the annual bumblebee, Bombus terrestris audax, reared from wild‐caught queens from a single population. In both samples, queens’ lifetime reproductive success, measured as either queens’ inclusive fitness or as total biomass of queen‐produced sexuals (new queens and males), was significantly positively associated with queen longevity, measured from the day the first worker was produced. We suggest that a positive relationship between reproductive success and longevity was inherited from nonsocial ancestors showing parental care and maintained, at least in part, because the presence of workers buffers queens against extrinsic mortality.