Reproductive Ground Plan Hypothesis Research Articles

Mating status and the evolution of eusociality: Oogenesis is independent of mating status in the solitary bee Osmia bicornis

The fundamental trait underlying eusociality is the reproductive division of labour. In honeybees (Apis mellifera), queens lay eggs while workers forage, defend and care for brood. The division of labour is maintained by pheromones including queen mandibular pheromone (QMP) produced by the queen. QMP constrains reproduction in adult honeybee workers, but in the absence of their queen workers can activate their ovaries and, although they cannot mate, they lay haploid male eggs. The reproductive ground plan hypothesis suggests that reproductive constraint may have evolved by co-opting mechanisms of reproductive control in solitary ancestors. In many insects mating is required to activate or accelerate oogenesis. Here, we use the solitary bee Osmia bicornis (Megachilidae) to test whether reproductive constraint evolved from ancestral control of reproduction by mating status. We present a structural study of the O. bicornis ovary, and compare key stages of oogenesis with honeybee workers. Importantly, we show that mating does not affect any aspect of the reproductive physiology of O. bicornis. We therefore conclude that mechanisms governing reproductive constraint in honeybees were unlikely to have been co-opted from mechanisms pertaining to mating status.

Caste-based differential transcriptional expression of hexamerins in response to a juvenile hormone analog in the red imported fire ant (Solenopsis invicta).

The reproductive ground plan hypothesis proposes that gene networks regulating foraging behavior and reproductive female physiology in social insects emerged from ancestral gene and endocrine factor networks. Expression of storage proteins such as vitellogenins and hexamerins is an example of this co-option. Hexamerins, through their role modulating juvenile hormone availability, are involved in caste determination in termites. The genome of the fire ant (Solenopsis invicta) encodes four hexamerin genes, hexamerin-like (LOC105192919, hereafter called hexamerin 1), hexamerin (LOC105204474, hereafter called hexamerin 2), arylphorin subunit alpha-like, and arylphorin subunit beta. In this study, a phylogenetic analysis of the S. invicta hexamerins determined that each predicted protein clustered with one of the orthologous Apis mellifera hexamerins. Gene expression analyses by RT-qPCR revealed differential expression of the hexamerins between queens and workers, and between specific task-allocated workers (nurses and foragers). Queens and nurses had significantly higher expression of all genes when compared to foragers. Hexamerin 1 was expressed at higher levels in queens, while hexamerin 2 and arylphorin subunit beta were expressed at significantly higher levels in nurses. Arylphorin subunit alpha-like showed no significant difference in expression between virgin queens and nurses. Additionally, we analyzed the relationship between the expression of hexamerin genes and S-hydroprene, a juvenile hormone analog. Significant changes in hexamerin expression were recorded in nurses, virgin queens, and foragers 12 h after application of the analog. Hexamerin 1 and arylphorin subunit alpha-like expression were significantly lower after analog application in virgin queens. In foragers, hexamerin 2 and arylphorin subunit beta were significantly lower after analog application, while in nurses expression of all genes were significantly lower after analog application. Our results suggest that in S. invicta hexamerin genes could be associated with reproductive division of labor and task-allocation of workers.

Ovary activation does not correlate with pollen and nectar foraging specialization in the bumblebee Bombus impatiens.

Social insect foragers may specialize on certain resource types. Specialization on pollen or nectar among honeybee foragers is hypothesized to result from associations between reproductive physiology and sensory tuning that evolved in ancestral solitary bees (the Reproductive Ground-Plan Hypothesis; RGPH). However, the two non-honeybee species studied showed no association between specialization and ovary activation. Here we investigate the bumblebee B. impatiens because it has the most extensively studied pollen/nectar specialization of any bumblebee. We show that ovary size does not differ between pollen specialist, nectar specialist, and generalist foragers, contrary to the predictions of the RGPH. However, we also found mixed support for the second prediction of the RGPH, that sensory sensitivity, measured through proboscis extension response (PER), is greater among pollen foragers. We also found a correlation between foraging activity and ovary size, and foraging activity and relative nectar preference, but no correlation between ovary size and nectar preference. In one colony non-foragers had larger ovaries than foragers, supporting the reproductive conflict and work hypothesis, but in the other colony they did not.

Support for the reproductive ground plan hypothesis in a solitary bee: links between sucrose response and reproductive status.

In social bees, foraging behaviour is correlated with reproductive status and sucrose sensitivity via endocrine pathways. This association led to the hypothesis that division of labour in social insect societies is derived from an ancestral ground plan that functions to synchronize dietary preferences with reproductive needs in solitary insects. However, the relationship between these traits is unknown for solitary bees, which represent the ancestral state of social bees. We used the proboscis extension response assay to measure sucrose response among reproductive females of the solitary alkali bee (Nomia melanderi) as a function of acute juvenile hormone (JH) treatments and reproductive physiology. We also tested long-term effects of JH on reproductive development in newly emerged females. JH did not have short-term effects on reproductive physiology or sucrose response, but did have significant long-term effects on ovary and Dufour's gland development. Dufour's gland size, not ovary development, was a significant predictor of sucrose response. This provides support for the reproductive ground plan hypothesis, because the Dufour's gland has conserved reproductive functions in bees. Differing results from this study and honeybees suggest independent origins of division of labour may have evolved via co-option of different components of a conserved ground plan.

Testing the reproductive groundplan hypothesis in ants (Hymenoptera: Formicidae).

The evolution of complex societies with obligate reproductive division of labor represents one of the major transitions in evolution. In such societies, functionally sterile individuals (workers) perform many of fitness-relevant behaviors including allomaternal ones, without getting any direct fitness benefits. The question of how such worker division of labor has evolved remains controversial. The reproductive groundplan hypothesis (RGPH) offers a powerful proximate explanation for this evolutionary leap. The RGPH argues that the conserved genetic and endocrinological networks regulating fitness-relevant behavior (e g. foraging and brood care) in their solitary ancestors have become decoupled from actual reproduction in the worker caste and now generate worker behavioral phenotypes. However, the empirical support for this hypothesis remains limited to a handful of species making its general validity uncertain. In this study, we combine data from the literature with targeted sampling of key species and apply phylogenetically controlled comparative analysis to investigate if the key prediction of the RGPH, namely an association between allomaternal behavior and an allomaternal physiological state holds in the largest and most species-rich clade of social insects, the ants. Our findings clearly support the RPGH as a general framework to understand the evolution of the worker caste and shed light on one of the major transition in evolutionary history.

Specialization on pollen or nectar in bumblebee foragers is not associated with ovary size, lipid reserves or sensory tuning.

Foraging specialization allows social insects to more efficiently exploit resources in their environment. Recent research on honeybees suggests that specialization on pollen or nectar among foragers is linked to reproductive physiology and sensory tuning (the Reproductive Ground-Plan Hypothesis; RGPH). However, our understanding of the underlying physiological relationships in non-Apis bees is still limited. Here we show that the bumblebee Bombus terrestris has specialist pollen and nectar foragers, and test whether foraging specialization in B. terrestris is linked to reproductive physiology, measured as ovarian activation. We show that neither ovary size, sensory sensitivity, measured through proboscis extension response (PER), or whole-body lipid stores differed between pollen foragers, nectar foragers, or generalist foragers. Body size also did not differ between any of these three forager groups. Non-foragers had significantly larger ovaries than foragers. This suggests that potentially reproductive individuals avoid foraging.

Vitellogenin and vitellogenin receptor gene expression is associated with male and female parenting in a subsocial insect.

Complex social behaviour in Hymenoptera has been hypothesized to evolve by co-opting reproductive pathways (the ovarian ground plan hypothesis, OGPH) and gene networks (the reproductive ground plan hypothesis, RGPH). In support of these hypotheses, in eusocial Hymenoptera where there is reproductive division of labour, the yolk precursor protein vitellogenin (Vg) influences the expression of worker social behaviour. We suggest that co-opting genes involved in reproduction may occur more generally than just in the evolution of eusociality; i.e. underlie earlier stages of social evolution such as the evolution of parental care, given that reproduction and parental care rarely overlap. We therefore examined vitellogenin (vg) gene expression associated with parental care in the subsocial beetle Nicrophorus vespilloides. We found a significant reduction in the expression of vg and its receptor, vgr, in head tissue during active parental care, and confirmed that the receptor is expressed in the brains of both sexes. Ours is the first study to show that vgr is expressed in the brain of a non-eusocial insect. Given the association between behaviour and gene expression in both sexes, and the presence of vitellogenin receptors in the brain, we suggest that Vg was co-opted early in the evolution of sociality to have a regulatory function. This extends the association of Vg in parenting to subsocial species and outside of the Hymenoptera, and supports the hypothesis that the OGPH is general and that heterochrony in gene expression is important in the evolution of social behaviour and precedes subsequent evolutionary specialization of social roles.

Queen and young larval pheromones impact nursing and reproductive physiology of honey bee (Apis mellifera) workers.

Several insect pheromones are multifunctional and have both releaser and primer effects. In honey bees (Apis mellifera), the queen mandibular pheromone (QMP) and e-beta-ocimene (eβ), emitted by young worker larvae, have such dual effects. There is increasing evidence that these multifunctional pheromones profoundly shape honey bee colony dynamics by influencing cooperative brood care, a fundamental aspect of eusocial insect behavior. Both QMP and eβ have been shown to affect worker physiology and behavior, but it has not yet been determined if these two key pheromones have interactive effects on hypopharyngeal gland (HPG) development, actively used in caring of larvae, and ovary activation, a component of worker reproductive physiology. Experimental results demonstrate that both QMP and eβ significantly suppress ovary activation compared to controls but that the larval pheromone is more effective than QMP. The underlying reproductive anatomy (total ovarioles) of workers influenced HPG development and ovary activation, so that worker bees with more ovarioles were less responsive to suppression of ovary activation by QMP. These bees were more likely to develop their HPG and have activated ovaries in the presence of eβ, providing additional links between nursing and reproductive physiology in support of the reproductive ground plan hypothesis.Electronic supplementary materialThe online version of this article (doi:10.1007/s00265-014-1811-y) contains supplementary material, which is available to authorized users.

The Architecture of the Pollen Hoarding Syndrome in Honey Bees: Implications for Understanding Social Evolution, Behavioral Syndromes, and Selective Breeding.

Social evolution has influenced every aspect of contemporary honey bee biology, but the details are difficult to reconstruct. The reproductive ground plan hypothesis of social evolution proposes that central regulators of the gonotropic cycle of solitary insects have been coopted to coordinate social complexity in honey bees, such as the division of labor among workers. The predicted trait associations between reproductive physiology and social behavior have been identified in the context of the pollen hoarding syndrome, a larger suite of interrelated traits. The genetic architecture of this syndrome is characterized by a partially overlapping genetic architecture with several consistent, pleiotropic QTL. Despite these central QTL and an integrated hormonal regulation, separate aspects of the pollen hoarding syndrome may evolve independently due to peripheral QTL and additionally segregating genetic variance. The characterization of the pollen hoarding syndrome has also demonstrated that this syndrome involves many non-behavioral traits, which may be the case for numerous "behavioral" syndromes. Furthermore, the genetic architecture of the pollen hoarding syndrome has implications for breeding programs for improving honey health and other desirable traits: If these traits are comparable to the pollen hoarding syndrome, consistent pleiotropic QTL will enable marker assisted selection, while sufficient additional genetic variation may permit the dissociation of trade-offs for efficient multiple trait selection.

Vitellogenin Underwent Subfunctionalization to Acquire Caste and Behavioral Specific Expression in the Harvester Ant Pogonomyrmex barbatus

The reproductive ground plan hypothesis (RGPH) proposes that the physiological pathways regulating reproduction were co-opted to regulate worker division of labor. Support for this hypothesis in honeybees is provided by studies demonstrating that the reproductive potential of workers, assessed by the levels of vitellogenin (Vg), is linked to task performance. Interestingly, contrary to honeybees that have a single Vg ortholog and potentially fertile nurses, the genome of the harvester ant Pogonomyrmex barbatus harbors two Vg genes (Pb_Vg1 and Pb_Vg2) and nurses produce infertile trophic eggs. P. barbatus, thus, provides a unique model to investigate whether Vg duplication in ants was followed by subfunctionalization to acquire reproductive and non-reproductive functions and whether Vg reproductive function was co-opted to regulate behavior in sterile workers. To investigate these questions, we compared the expression patterns of P. barbatus Vg genes and analyzed the phylogenetic relationships and molecular evolution of Vg genes in ants. qRT-PCRs revealed that Pb_Vg1 is more highly expressed in queens compared to workers and in nurses compared to foragers. By contrast, the level of expression of Pb_Vg2 was higher in foragers than in nurses and queens. Phylogenetic analyses show that a first duplication of the ancestral Vg gene occurred after the divergence between the poneroid and formicoid clades and subsequent duplications occurred in the lineages leading to Solenopsis invicta, Linepithema humile and Acromyrmex echinatior. The initial duplication resulted in two Vg gene subfamilies preferentially expressed in queens and nurses (subfamily A) or in foraging workers (subfamily B). Finally, molecular evolution analyses show that the subfamily A experienced positive selection, while the subfamily B showed overall relaxation of purifying selection. Our results suggest that in P. barbatus the Vg gene underwent subfunctionalization after duplication to acquire caste- and behavior- specific expression associated with reproductive and non-reproductive functions, supporting the validity of the RGPH in ants.

Nutrition-dependent fertility response to juvenile hormone in non-social Euodynerus foraminatus wasps and the evolutionary origin of sociality

The reproductive ground plan hypothesis (RGPH) proposes that the ovarian cycle in solitary insects provides the basis for social evolution, so similar mechanisms are predicted to influence reproductive plasticity in social and solitary species. Specifically, reproductive plasticity in social species originated via modification of nutrition-dependent fertility response to juvenile hormone (JH) in solitary insects. Testing this prediction requires information about the factors that influence fertility in non-social relatives of the eusocial hymenoptera. However, no previous studies have examined how JH or nutritional condition influence fertility in Eumenines, the non-social group most closely related to social wasps. Here, we find support for the RGPH, as JH increases Euodynerus foraminatus fertility. Fertility is also condition-dependent, as heavy E. foraminatus are more fertile than light E. foraminatus. In addition, we measure the factors associated with mating success in E. foraminatus, finding that multiple factors influence mating success, including male weight, male mating experience, and female age. There is also higher variance in male than female reproductive success, suggesting that males may experience substantial sexual selection in this species. Overall, the relationships between JH, body weight, and fertility in E. foraminatus support the RGPH for the origin of sociality by demonstrating that there are strong parallels in the mechanisms that mediate fertility of social and non-social wasps.

Ovarian Control of Nectar Collection in the Honey Bee (Apis mellifera)

Honey bees are a model system for the study of division of labor. Worker bees demonstrate a foraging division of labor (DOL) by biasing collection towards carbohydrates (nectar) or protein (pollen). The Reproductive ground-plan hypothesis of Amdam et al. proposes that foraging DOL is regulated by the networks that controlled foraging behavior during the reproductive life cycle of honey bee ancestors. Here we test a proposed mechanism through which the ovary of the facultatively sterile worker impacts foraging bias. The proposed mechanism suggests that the ovary has a regulatory effect on sucrose sensitivity, and sucrose sensitivity impacts nectar loading. We tested this mechanism by measuring worker ovary size (ovariole number), sucrose sensitivity, and sucrose solution load size collected from a rate-controlled artificial feeder. We found a significant interaction between ovariole number and sucrose sensitivity on sucrose solution load size when using low concentration nectar. This supports our proposed mechanism. As nectar and pollen loading are not independent, a mechanism impacting nectar load size would also impact pollen load size.

Worker division of labor and endocrine physiology are associated in the harvester ant,Pogonomyrmex californicus

In Pogonomyrmex californicus harvester ants, an age-associated division of labor occurs in the worker caste, in which young workers perform in-nest tasks and older workers forage for food. Here, we tested whether this behavioral division is age based or age flexible, and whether it coincides with differential expression of systemic hormones with known roles in behavioral regulation. Whole-body content of juvenile hormone (JH) and ecdysteroids was determined in workers from (1) age-typical colonies, in which a typical age structure is maintained and workers transition across behaviors naturally, and (2) single-cohort colonies, which are entirely composed of same-aged workers, facilitating the establishment of age-independent division of labor. Foragers from both colony types had higher JH and lower ecdysteroid content than workers performing in-nest tasks, suggesting that age is not the sole determinant of worker behavior. This association between hormone content and behavior of P. californicus workers is similar to that previously observed in founding queens of this species. Because these hormones are key regulators of development and reproductive behavior, our data are consistent with the reproductive ground plan hypothesis (RGPH), which posits that the reproductive regulatory mechanisms of solitary ancestors were co-opted to regulate worker behavior.

Ovarian regulation of foraging division of labour in Africanized backcross and pollen-hoarding honeybees

Division of labour (i.e. differential task performance) is the hallmark of social insects. One highly studied component of honeybee division of labour is foraging specialization through collection of protein (pollen) or carbohydrates (nectar). The reproductive ground plan hypothesis proposes that this foraging division of labour is regulated by the same physiological control networks that controlled foraging behaviour during the reproductive life cycle of solitary honeybee ancestors. Based on observed differences in foraging behaviour and reproductive anatomy between bees selected for storing high and low quantities of pollen (high and low pollen-hoarding strains) and observed differences between wild-type pollen and nectar foragers, the reproductive ground plan hypothesis suggests that ovary size in facultatively sterile worker honeybees has a regulatory effect on variation in foraging division of labour, with increased protein bias correlated with increased ovary size (measured as ovariole number). An alternative explanation for the link between ovary size and foraging behaviour is that genes for ovarian development and foraging are inherited together because of genetic linkage or chance gene associations in the selected and natural populations. To address this alternative explanation, we investigated the relationship between ovary size and foraging bias in honeybees where genetic linkage was broken using a backcross mating design. We also studied high- and low-strain bees where ovary size effects were partitioned from other potential linkage effects. We found that ovariole number was related to foraging bias in the backcross bees and in the selected strains, as predicted by the reproductive ground plan hypothesis. An interaction effect between nectar sugar concentration and ovariole number was observed in the backcross and high-strain bees. This result suggests a potential mechanism by which the ovary affects sucrose perception, which in turn influences foraging behaviour.

Support for the reproductive ground plan hypothesis of social evolution and major QTL for ovary traits of Africanized worker honey bees (Apis melliferaL.)

BackgroundThe reproductive ground plan hypothesis of social evolution suggests that reproductive controls of a solitary ancestor have been co-opted during social evolution, facilitating the division of labor among social insect workers. Despite substantial empirical support, the generality of this hypothesis is not universally accepted. Thus, we investigated the prediction of particular genes with pleiotropic effects on ovarian traits and social behavior in worker honey bees as a stringent test of the reproductive ground plan hypothesis. We complemented these tests with a comprehensive genome scan for additional quantitative trait loci (QTL) to gain a better understanding of the genetic architecture of the ovary size of honey bee workers, a morphological trait that is significant for understanding social insect caste evolution and general insect biology.ResultsBack-crossing hybrid European x Africanized honey bee queens to the Africanized parent colony generated two study populations with extraordinarily large worker ovaries. Despite the transgressive ovary phenotypes, several previously mapped QTL for social foraging behavior demonstrated ovary size effects, confirming the prediction of pleiotropic genetic effects on reproductive traits and social behavior. One major QTL for ovary size was detected in each backcross, along with several smaller effects and two QTL for ovary asymmetry. One of the main ovary size QTL coincided with a major QTL for ovary activation, explaining 3/4 of the phenotypic variance, although no simple positive correlation between ovary size and activation was observed.ConclusionsOur results provide strong support for the reproductive ground plan hypothesis of evolution in study populations that are independent of the genetic stocks that originally led to the formulation of this hypothesis. As predicted, worker ovary size is genetically linked to multiple correlated traits of the complex division of labor in worker honey bees, known as the pollen hoarding syndrome. The genetic architecture of worker ovary size presumably consists of a combination of trait-specific loci and general regulators that affect the whole behavioral syndrome and may even play a role in caste determination. Several promising candidate genes in the QTL intervals await further study to clarify their potential role in social insect evolution and the regulation of insect fertility in general.

Genotype effect on regulation of behaviour by vitellogenin supports reproductive origin of honeybee foraging bias

In honeybee colonies, food collection is performed by a group of mostly sterile females called workers. After an initial nest phase, workers begin foraging for nectar and pollen, but tend to bias their collection towards one or the other. The foraging choice of honeybees is influenced by vitellogenin (vg), an egg-yolk precursor protein that is expressed although workers typically do not lay eggs. The forager reproductive ground plan hypothesis (RGPH) proposes an evolutionary path in which the behavioural bias toward collecting nectar or pollen on foraging trips is influenced by variation in reproductive physiology, such as hormone levels and vg gene expression. Recently, the connections between vg and foraging behaviour were challenged by Oldroyd and Beekman (2008), who concluded from their study that the ovary, and especially vg, played no role in foraging behaviour of bees. We address their challenge directly by manipulating vg expression by RNA interference- (RNAi) mediated gene knockdown in two honeybee genotypes with different foraging behaviour and reproductive physiology. We show that the effect of vg on the food-loading decisions of the workers occurs only in the genotype where timing of foraging onset (by age) is also sensitive to vg levels. In the second genotype, changing vg levels do not affect foraging onset or bias. The effect of vg on workers' age at foraging onset is explained by the well-supported double repressor hypothesis (DHR), which describes a mutually inhibitory relationship between vg and juvenile hormone (JH) - an endocrine factor that influences development, reproduction, and behaviour in many insects. These results support the RGPH and demonstrate how it intersects with an established mechanism of honeybee behavioural control.

The developmental genetics and physiology of honeybee societies

Eusocial animal societies, as diverse as those found in the ants, bees, wasps, shrimp and naked mole-rats, are structured around one or few reproductive females. The remaining females are helpers called 'workers' that are mostly sterile. A paradigm in studies of eusociality is that worker sterility is a key to societal functions because advanced sociality cannot be achieved when there is conflict over reproduction. Yet, traits such as sensory responsiveness, foraging and hoarding behaviour that change between female reproductive life stages also vary between workers. This variation is central to worker division of labour, a complex social trait believed to be instrumental for the ecological success of animal societies. Thus, we took a step back from established views on worker sterility and societal functions, and hypothesized that division of labour can be better understood if adaptive variation in worker behaviour is seen as emerging from pre-existing mechanisms associated with female reproduction. In exploring this reproductive ground plan hypothesis (RGPH) in honeybee workers, we established that variation in foraging division of labour correlates with ovary size and is affected by expression changes in vitellogenin, an egg yolk protein precursor. Here, we explain and reconcile the RGPH with data on honeybee sensory sensitivity, genomic mapping, transcript and endocrine profiling, and link our discussion with Ihle et al. (2010, this issue, pp. xx-xx). The findings bring together mechanistic and evolutionary explanations of honeybee worker behaviour. This essay suggests that a broader view on worker reproductive traits can increase the understanding of animal social behaviour.

Association between Larger Ovaries and Pollen Foraging in Queenless Apis cerana Workers Supports the Reproductive Ground-plan Hypothesis of Social Evolution

Based on the ovarian ground-plan hypothesis (West-Eberhard 1987; West-Eberhard 1996) the reproductive ground-plan hypothesis of social evolution has recently been proposed as a theoretical framework to understand social evolution in honeybees (Amdam et al. 2004, 2006). The reproductive ground-plan hypothesis seeks to explain the evolution of honeybee social complexity from solitary ancestors by co-option of control elements of the female, gonadotropic reproductive cycle (Amdam et al. 2004). One of its central predictions, therefore, is that reproductive physiology and social behavior, in particular division of labor, be linked in particular ways (Amdam et al. 2006). Ovary size influences the level of reproductive hormones (Goodman and Granger 2005) and is therefore predicted to be correlated with differences in social behavior. Honey bee workers exhibit age-dependent division of labor between in-hive tasks and foraging (Beshers and Fewell 2001) but intrinsic worker differences affect the timing of behavioral transitions and further aspects of the division of labor, such as the specialization during foraging on nectar or pollen collection (Page and Erber 2002).

Effects of selection for honey bee worker reproduction on foraging traits.

The “reproductive ground plan” hypothesis (RGPH) proposes that reproductive division of labour in social insects had its antecedents in the ancient gene regulatory networks that evolved to regulate the foraging and reproductive phases of their solitary ancestors. Thus, queens express traits that are characteristic of the reproductive phase of solitary insects, whereas workers express traits characteristic of the foraging phase. The RGPH has also been extended to help understand the regulation of age polyethism within the worker caste and more recently to explain differences in the foraging specialisations of individual honey bee workers. Foragers that specialise in collecting proteinaceous pollen are hypothesised to have higher reproductive potential than individuals that preferentially forage for nectar because genes that were ancestrally associated with the reproductive phase are active. We investigated the links between honey bee worker foraging behaviour and reproductive traits by comparing the foraging preferences of a line of workers that has been selected for high rates of worker reproduction with the preferences of wild-type bees. We show that while selection for reproductive behaviour in workers has not altered foraging preferences, the age at onset of foraging of our selected line has been increased. Our findings therefore support the hypothesis that age polyethism is related to the reproductive ground plan, but they cast doubt on recent suggestions that foraging preferences and reproductive traits are pleiotropically linked.