Queen Mandibular Pheromone Research Articles

The effects of queen mandibular pheromone on Nosema (Vairimorpha) ceranae infections in caged honey bees

Honey bees utilize queen mandibular pheromone (QMP) for maintaining social hierarchy and colony development. In controlled cage studies, synthetic QMP is often introduced to mimic natural conditions. However, questions have arisen about the effects of QMP on nosema disease studies. This short report identifies significant early-stage suppression effects of QMP on Nosema (Vairimorpha) ceranae infections. QMP was found to significantly lower infection rates below the reported infectious dose for 50 % infectivity (ID50) and to slow disease development in a dose-independent manner. These effects diminished at doses exceeding ID100. We recommend that studies investigating treatment effects using caged bees avoid QMP to ensure unambiguous results. Additionally, employing multiple infectious doses with shorter incubation times would be useful for evaluating other treatments that may have subtle effects. Furthermore, our findings support previous field studies suggesting that queen replacement reduces nosema disease at levels similar to treatment with fumagillin.

Early life exposure to queen mandibular pheromone mediates persistent transcriptional changes in the brain of honey bee foragers.

Behavioural regulation in insect societies remains a fundamental question in sociobiology. In hymenopteran societies, the queen plays a crucial role in regulating group behaviour by affecting individual behaviour and physiology through modulation of worker gene expression. Honey bee (Apis mellifera) queens signal their presence via queen mandibular pheromone (QMP). While QMP has been shown to influence behaviour and gene expression of young workers, we know little about how these changes translate in older workers. The effects of the queen pheromone could have prolonged molecular impacts on workers that depend on an early sensitive period. We demonstrate that removal of QMP impacts long-term gene expression in the brain and antennae in foragers that were treated early in life (1day post emergence), but not when treated later in life. Genes important for division of labour, learning, chemosensory perception and ageing were among those differentially expressed in the antennae and brain tissues, suggesting that QMP influences diverse physiological and behavioural processes in workers. Surprisingly, removal of QMP did not have an impact on foraging behaviour. Overall, our study suggests a sensitive period early in the life of workers, where the presence or absence of a queen has potentially life-long effects on transcriptional activity.

Effects of microRNA-305 knockdown on brain gene expression associated with division of labor in honey bee colonies (Apis mellifera).

Division of labor in honey bee colonies is based on the behavioral maturation of adult workers that involves a transition from working in the hive to foraging. This behavioral maturation is associated with distinct task-related transcriptomic profiles in the brain and abdominal fat body that are related to multiple regulatory factors including juvenile hormone (JH) and queen mandibular pheromone (QMP). A prominent physiological feature associated with behavioral maturation is a loss of abdominal lipid mass as bees transition to foraging. We used transcriptomic and physiological analyses to study whether microRNAs (miRNAs) are involved in the regulation of division of labor. We first identified two miRNAs that showed patterns of expression associated with behavioral maturation, ame-miR-305-5p and ame-miR-375-3p. We then downregulated the expression of these two miRNAs with sequence-specific antagomirs. Neither ame-miR-305-5p nor ame-miR-375-3p knockdown in the abdomen affected abdominal lipid mass on their own. Similarly, knockdown of ame-miR-305-5p in combination with JH or QMP also did not affect lipid mass. By contrast, ame-miR-305-5p knockdown in the abdomen caused substantial changes in gene expression in the brain. Brain gene expression changes included genes encoding transcription factors previously implicated in behavioral maturation. The results of these functional genomic experiments extend previous correlative associations of microRNAs with honey bee division of labor and point to specific roles for ame-miR-305-5p.

Sublethal effects of imidacloprid-contaminated honey stores on colony performance, queens, and worker activities in fall and early winter colonies.

Neonicotinoid-contaminated sugar stores can have both near term and long term effects on honey bees due to their persistence in honey stores. Effects of imidacloprid food stores contaminants were examined in subtropical colonies that experience reduced brood rearing and foraging during overwintering. Colonies were given treatment sugar syrup containing 0 ppb (control), 20 ppb (field relevant), or 100 ppb (above field relevant) imidacloprid over six weeks to simulate contaminated fall nectar. Colonies were evaluated immediately (post-treatment) and 10 weeks (mid-winter) after treatment to compare proximal and latent effects. Post-treatment 0 ppb and 20 ppb colonies had more workers than 100 ppb colonies while 0 ppb colonies more brood than 20 ppb or 100 ppb colonies. Mid-winter 0 ppb and 20 ppb colonies had more workers than 100 ppb colonies and 0 ppb colonies more brood than 100 ppb colonies. Colonies experienced seasonal declines in stored pollen but no treatment effects. Lower 100 ppb colony performance was associated with reduced effort rather than lifespan. RFID (Radio Frequency Identification) tracking revealed that workers had similar adult lifespans across treatments; however, 100 ppb workers engaged in activities outside the colony for less time than 0 ppb workers. Imidacloprid exposure affected queen but not worker nutritional physiology. Nurses retained well-developed hypopharyngeal glands (as indicated by head protein) across treatments. Mid-winter queens from 0 ppb colonies had marginally higher ovary protein than queens from 100 ppb colonies and more ovary lipids than queens from 20 ppb colonies. However, queen nutrient stores in non-reproductive tissues (fat bodies) did not differ across treatments. Queens from different treatments were attended by comparable numbers of retinue workers and had similar gland contents of four QMP (Queen Mandibular Pheromone) components essential to queen care. High levels of imidacloprid in sugar stores can negatively affect colony performance months after initial storage.

Honey bee retinue workers respond similarly to queens despite seasonal differences in Queen Mandibular Pheromone (QMP) signaling.

Honey bee colonies maintain viable queens in part through communication with Queen Mandibular Pheromone (QMP), a mixture that signals the queen's presence and reproductive quality to workers. In turn, workers are thought to provide retinue queen care or replace queens partially based on QMP profiles. We examined the effects of seasonal dearth (overwintering in a warm subtropical location) on queen-worker interactions. Retinue worker responses to continuously ovipositing queens were considered in view of QMP signaling and queen reproductive quality. QMP signaling was estimated from QMP residues recovered from nest worker bodies, which is the primary mode of QMP transfer from the queen to the colony at large. QMP residues varied seasonally but not at all with queen reproductive quality (spermatheca sperm storage, ovary protein and lipid contents). 9-HDA and 9-ODA were lower in January than other months. HOB decreased from July to January, while HVA, a component associated with mated queens, increased sharply in January. Despite these seasonal signaling differences, retinue workers attended queens at similar levels through the months. In terms of reproductive quality, queens did not differ over the months in matedness (spermatheca sperm storage) or physiological age (protein carbonyl content), but varied in nutrient allocation to reproductive and non-reproductive tissues. Queen ovaries contained more protein in September than in November, and more lipid in July and September than in November and January. Queen fat bodies had more protein in July than September or November, but less lipid in July and September than November or January. Retinue worker responses did not vary with seasonal QMP changes, but reflected overall continuous brood rearing efforts and queen matedness throughout the year. The absence of seasonal differences in worker responses to QMP should be considered in the broader context of continuous reproductive efforts in warm subtropical colonies.

Differences in ASP1 expression and binding dynamics to queen mandibular pheromone HOB between Apis mellifera and Apis cerana workers reveal olfactory adaptation to colony organization

The eastern Apis cerana (Ac) and the western Apis mellifera (Am) are two closely related and most economically valuable honeybee species managed extensively worldwide. However, how worker bees of Ac and Am are adapted to their colony organization remains to be uncovered. Here, we found that the expression level of gene encoding antennae-specific proteins 1 (ASP1, a key regulator in recognizing queen mandibular pheromone) was positively correlated with the colony sizes in both bee species, and the expression level in Am was higher than that in Ac, suggesting that ASP1 may play an important role in maintaining colony homeostasis. Using competitive binding assay, molecular docking, and site-directed mutagenesis, we then confirmed the good binding affinities of both Ac-ASP1 and Am-ASP1 to methyl p-hydroxy benzoate (HOB), and Val115 was the key amino acid. However, the affinity of Am-ASP1 was stronger than that of Ac-ASP1. EAG analysis further demonstrated that antennae of Am worker bees had faster depolarization and repolarization in response to HOB stimulation. Taken together, these findings indicate that the differences in expression levels and binding dynamics allow ASP1 recognizing HOB to potentially serve as a specific regulator of colony organization in Ac and Am.

Methoxyfenozide has minimal effects on replacement queens but may negatively affect sperm storage

Honey bees are incidentally exposed to pesticides such as the insect growth regulator methoxyfenozide (MEOF) during crop pollination, exposures that extend into the hive via contaminated stored food. We examined the sublethal effects of MEOF-contaminated pollen and queen cell wax on replacement queen development. MEOF-exposed colonies were largely able to produce replacement queens of similar physiological and reproductive quality as unexposed colonies. Newly established queens did not differ in their body mass, ovariole development, or protein and fatty acid contents in their ovaries and fat bodies. MEOF and control queens had similar glandular contents of queen mandibular pheromone (QMP) and queen retinue pheromone (QRP) compounds. However, MEOF queens stored less sperm in their spermathecae than control queens. Given that queen productivity is ultimately limited by sperm availability, MEOF contamination might shorten the functional lifespan of exposed queens.

Morphology of Nasonov and Tergal Glands in Apis mellifera Rebels.

Simple SummaryCommunication in a colony of social insects, such as the honeybee, is possible thanks to the pheromones secreted by all individuals. Pheromones are produced and secreted by the glands. Examples of such structures are Nasonov and tergal glands. Nasonov glands are characteristic of worker bees, while tergal glands are primarily found in queens. There are situations in the colony in which the queen and her pheromones are missing. In these instances, the larvae develop into rebels, which are reproductive workers. We therefore assumed that the rebels would have a reduced Nasonov gland and developed tergal glands. Our assumption turned out to be correct. These discoveries bring us closer to explaining the evolutionary formation of different castes of honeybees.Social insect societies are characterized by a high level of organization. This is made possible through a remarkably complex array of pheromonal signals produced by all members of the colony. The queen’s pheromones signal the presence of a fertile female and induce daughter workers to remain sterile. However, the lack of the queen mandibular pheromone leads to the emergence of rebels, i.e., workers with increased reproductive potential. We suggested that the rebels would have developed tergal glands and reduced Nasonov glands, much like the queen but contrary to normal workers. Our guess turned out to be correct and may suggest that the rebels are more queen-like than previously thought. The tergal gland cells found in the rebels were numerous but they did not adhere as closely to one another as they did in queens. In the rebels, the number of Nasonov gland cells was very limited (from 38 to 53) and there were fat body trophocytes between the glandular cells. The diameters of the Nasonov gland cell nuclei were smaller in the rebels than in the normal workers. These results are important for understanding the formation of the different castes of Apis mellifera females, as well as the division of labor in social insect societies.

Impact of Chronic Exposure to Two Neonicotinoids on Honey Bee Antennal Responses to Flower Volatiles and Pheromonal Compounds.

Volatile compounds provide important olfactory cues for honey bees (Apis mellifera L.), which are essential for their ecology, behavior, and social communication. In the external environment bees locate food sources by the use of floral scents, while inside the hive, pheromones such as the queen mandibular pheromone (QMP) and alarm pheromones serve important functions in regulating colony life and inducing aggressive responses against intruders and parasites. Widely reported alterations of various behaviors in- and outside the hive following exposure to pesticides could therefore be associated with a disturbance of odor sensitivity. In the present study, we tested the effects of neonicotinoid pesticides at field concentrations on the ability of honey bees to perceive volatiles at the very periphery of the olfactory system. Bee colonies were subjected to treatments during the summer with either Imidacloprid or Thiacloprid at sublethal concentrations. Antennal responses to apple (Malus domestica L.) flower volatiles were studied by GC-coupled electro-antennographic detection (GC-EAD), and a range of volatiles, a substitute of the QMP, and the alarm pheromone 2-heptanone were tested by electroantennography (EAG). Short-term and long-term effects of the neonicotinoid treatments were investigated on bees collected in the autumn and again in the following spring. Treatment with Thiacloprid induced changes in antennal responses to specific flower VOCs, with differing short- and long-term effects. In the short term, increased antennal responses were observed for benzyl-alcohol and 1-hexanol, which are common flower volatiles but also constituents of the honey bee sting gland secretions. The treatment with Thiacloprid also affected antennal responses to the QMP and the mandibular alarm pheromone 2-heptanone. In the short term, a faster signal degeneration of the response signal to the positive control citral was recorded in the antennae of bees exposed to Thiacloprid or Imidacloprid. Finally, we observed season-related differences in the antennal responses to multiple VOCs. Altogether, our results suggest that volatile-specific alterations of antennal responses may contribute to explaining several behavioral changes previously observed in neonicotinoid-exposed bees. Treatment effects were generally more prominent in the short term, suggesting that adverse effects of neonicotinoid exposure may not persist across generations.

Honeybee queen mandibular pheromone fails to regulate ovary activation in the common wasp.

The queen mandibular pheromone (QMP) identified from the honeybee is responsible for maintaining reproductive division of labour in the colony, and affects multiple behaviours. Interestingly, QMP inhibits reproduction not only in honeybee workers, but also in distantly related insect species such as fruit flies and bumblebees. This study examines whether QMP also affects worker reproduction in the common wasp Vespula vulgaris. Wasp workers were exposed to one of the following treatments: QMP, wasp queen pheromone (the hydrocarbon heptacosane n-C27), or acetone (solvent-only control). After dissecting the workers, no evidence that QMP inhibits development in V. vulgaris could be found. However, this study could confirm the inhibitory effect of the hydrocarbon heptacosane on ovary activation. The reason why non-social species such as the fruit fly and social species such as bumblebees and ants respond to the QMP, while the social wasp V. vulgaris does not, is unclear. The investigation of whether olfaction is key to sensing QMP in other insect species, and the detailed study of odorant receptors in other social insects, may provide insights into the mechanisms of response to this pheromone.

Pesticide Exposure During Development Does Not Affect the Larval Pheromones, Feeding Rates, or Morphology of Adult Honey Bee (Apis mellifera) Queens

Recent work demonstrated that honey bee (Apis mellifera L.) queens reared in pesticide-laden beeswax exhibit significant changes in the composition of the chemicals produced by their mandibular glands including those that comprise queen mandibular pheromone, which is a critical signal used in mating as well as queen tending behavior. For the present study, we hypothesized that pesticide exposure during development would alter other queen-produced chemicals, including brood pheromone in immature queens, thus resulting in differential feeding of queen larvae by nurse workers, ultimately impacting adult queen morphology. We tested these hypotheses by rearing queens in beeswax containing field-relevant concentrations of (1) a combination of tau-fluvalinate and coumaphos, (2) amitraz, or (3) a combination of chlorothalonil and chlorpyrifos. These pesticides are ubiquitous in most commercial beekeeping operations in North America. We observed nurse feeding rates of queen larvae grafted into pesticide-laden beeswax, analyzed the chemical composition of larval queen pheromones and measured morphological markers in adult queens. Neither the nurse feeding rates, nor the chemical profiles of immature queen pheromones, differed significantly between queens reared in pesticide-laden wax compared to queens reared in pesticide-free wax. Moreover, pesticide exposure during development did not cause virgin or mated adult queens to exhibit differences in morphological markers (i.e., body weight, head width, or thorax width). These results were unexpected given our previous research and indicate that future work is needed to fully understand how pesticide exposure during development affects honey bee queen physiology, as well as how various adult queen quality metrics relate to each other.

Queen pheromone modulates the expression of epigenetic modifier genes in the brain of honeybee workers.

Pheromones are used by many insects to mediate social interactions. In the highly eusocial honeybee (Apis mellifera), queen mandibular pheromone (QMP) is involved in the regulation of the reproductive and other behaviour of workers. The molecular mechanisms by which QMP acts are largely unknown. Here, we investigate how genes responsible for epigenetic modifications to DNA, RNA and histones respond to the presence of QMP in the environment. We show that several of these genes are upregulated in the honeybee brain when workers are exposed to artificial QMP. We propose that pheromonal communication systems, such as those used by social insects, evolved to respond to environmental signals by making use of existing epigenomic machineries.

Drosophila melanogaster and worker honeybees (Apis mellifera) do not require olfaction to be susceptible to honeybee queen mandibular pheromone

Eusociality is characterised by the reproductive division of labour; a dominant female (queen) or females are responsible for the majority of reproduction, and subordinate females are reproductively constrained. Reproductive constraint can be due to behavioural aggression and/or chemical cues, so-called queen pheromones, produced by the dominant females. In the honeybee, Apis mellifera, this repressive queen pheromone is queen mandibular pheromone (QMP). The mechanism by which honeybee workers are susceptible to QMP is not yet completely understood, however it is thought to be through olfaction via the antennae and/or gustation via trophallaxis.We have investigated whether olfaction is key to sensing of QMP, using both Drosophila melanogaster- a tractable non-eusocial insect which is also reproductively repressed by QMP- and the target species, A. mellifera worker honeybees. D. melanogaster are still capable of sensing and responding to QMP without their antenna and maxillary palps, and therefore without olfactory receptors. When worker honeybees were exposed to QMP but unable to physically interact with it, therefore required to use olfaction, they were similarly not reproductively repressed. Combined, these findings support either a non-olfactory based mechanism for the repression of reproduction via QMP, or redundancy via non-olfactory mechanisms in both D. melanogaster and A. mellifera. This study furthers our understanding of how species are susceptible to QMP, and provides insight into the mechanisms governing QMP responsiveness in these diverse species.

Influence of RNA interference-mediated reduction of Or11 on the expression of transcription factor Kr-h1 in Apis mellifera drones

9-Oxo-2-decenoic acid (9-ODA, the predominant component of honeybee queen mandibular pheromones) acts as a sex pheromone attracting drones during mating flights in midair. Odorant receptor 11 (Or11), which is located on the membrane of antennal olfaction receptor neurons in bees, can specifically recognize 9-ODA. At present, the molecular pathway of honeybee drones responding to 9-ODA is still unclear. Studies have demonstrated that 9-ODA could downregulate the expression of Kruppel-homolog1 (Kr-h1, a transcription factor related to the regulation of reproduction and division of labor mediated by juvenile hormone) gene in the mushroom of honeybee brain. We speculate that Kr-h1 may be the downstream gene of Or11, which is involved in the pathway of drones responding to 9-ODA. Therefore, we analyzed the influence of 9-ODA on the expression of Or11 and Kr-h1 in the antennae of sexually immature (4 days) and mature (14 days) male honeybees (Apis mellifera) by quantitative polymerase chain reaction (qPCR). The results demonstrated that 9-ODA significantly downregulated the expression of Or11 and Kr-h1 in the antennae of sexually immature and mature drones. Additionally, siRNA-Or11 was injected into the antennae and brain tissues of 8-day-old drone pupae, and the expression patterns of Or11, Kr-h1 and Broad-Complex (Br–c, downstream gene of Kr-h1) were determined by qPCR at 72 h. The RNAi-induced knockdown of Or11 significantly decreased the expression of Or11, Kr-h1 and Br–c in the antennae and brains of drones. This study suggests that the transcription factor Kr-h1 is downstream of Or11, Kr-h1, which may play an important role in the signal transduction process of drones responding to 9-ODA.

Queen honey bee (Apis mellifera) pheromone and reproductive behavior are affected by pesticide exposure during development

Pollinator diversity and abundance in North America have been at a steep decline over the last two decades due to the combinatorial effects of several environmental and anthropogenic stressors. In particular, managed honey bees (Apis mellifera) face multiple health risks including nutritional stress, exposure to pests and pathogens, poor queen quality, and pesticide contamination, which cause problems at the individual and colony levels. One of the gravest problems faced by honey bees is parasitization by the mite Varroa destructor, which is typically controlled through the application of miticides such as tau-fluvalinate, coumaphos, and amitraz. In addition to miticides, colonies are also exposed to pesticides brought by foragers from agricultural settings, including the fungicide chlorothalonil and the insecticide chlorpyrifos. Here, we explored whether exposure of wax to combinations of these pesticides during development affects honey bee queen physiology and worker behavior. To do this, we reared queens in plastic cups coated with molten beeswax that was either pesticide-free or containing field-relevant concentrations of tau-fluvalinate and coumaphos, amitraz, or chlorothalonil and chlorpyrifos. Once queens mated naturally, we placed them in observation hives to measure egg-laying rate and worker retinue size. We then dissected the queens and used the contents of their mandibular glands to measure worker attractiveness in caged bioassays and to analyze their chemical components using GC-MS. Exposure of wax to field-relevant concentrations of the tested pesticides during queen development significantly lowered the adult queens’ egg-laying rate and worker retinue size. Miticide exposure during development also lowered the attractiveness of queen mandibular gland contents to workers and affected the relative amounts of the glands’ chemical components. Our results support the ideas that mandibular gland pheromones act as honest indicators of queen reproductive fitness and that pesticide exposure of wax during bee development is an important and concerning factor impairing honey bee health. Pollinator health is currently being threatened by multiple environmental factors, including poor nutrition, habitat fragmentation, and pesticide contamination. In particular, honey bee health is facing challenges caused by combinations of these factors, leading to an annual mortality of over 30% of all managed colonies in the USA. The conditions in which honey bee queens are reared can cause severe physiological and behavioral effects in adults, which in turn can affect the behavior of workers that tend to those queens, thus affecting colony productivity, growth, and survival. In this study, we show that queens reared in beeswax contaminated with miticides or agrochemicals lay fewer eggs over time and exhibit mandibular gland pheromone profiles that differ from those of queens reared in pesticide-free wax. Workers discern these differences, preferentially investigating the gland contents and forming larger retinues (thus performing more tending behaviors) for queens reared in pesticide-free wax. These results indicate that pesticide contamination of the queen-rearing wax directly affects queen physiology and behavior and further affirm that queen mandibular pheromones seem to act as honest indicators to workers of a queen’s reproductive capacity and overall health.

Dopamine regulation of reproduction in insects

Dopamine (DA) regulates reproductive behavior and promotes maturation of the reproductive system in insects. In Drosophila , male courtship behavior is regulated by the protocerebral posteriolateral dopaminergic cluster neuron 2ab (PPL2ab). The ability to synthesize DA in the PPL2ab affects male-male and male-female courtship behavior. The DA level in the brain of adult male Drosophila influences sexual orientation; although, male-male courtship is induced when the DA level is too high or too low. DA also regulates changes in reproductive behavior and post-copulation behavior. As a neurohormone, DA regulates taste and smell perception in insects. In the central nervous system, DA regulation of reproductive behavior involves the DA/MH (molting hormone) receptor (DopEcR) and synergy between DA and ecdysone, with the tritocerebrum P1 neurons playing an important role in the male courtship information loop. DA can promote development in the eggs and regulate the reproductive state in social insects. The promotional effect of DA on egg development involves developmental stage differences in DA receptor expression and interactions among DA, juvenile hormone (JH), MH, and octopamine (OA). For example, the expression level of dopamine D2-like receptor (DD2R) in the corpora allata of D. melanogaster young females is far lower than that in sexually mature females. Meanwhile, the expression level of dopamine D1-like receptor (DopR) in young females is much higher than that in mature females. By contrast, the expression of DopR and DD2R in the fat body is the opposite to that in the corpora allata. The interactions among DA, JH, MH, and OA depend on the expression of certain genes and regulation of metabolic enzyme activity. In addition, the interactions among different hormones also involve the insulin-like signaling (ILPs) pathway. As to reproductive state regulation in social insects, it has been found that the reproductive state of worker bees is regulated by queen mandibular pheromone (QMP), larvae pheromones, and the ovarian DA D2 receptor gene. Research on DA regulation of insect reproduction will provide us with references for the development of new drugs, and for research on mechanisms in human brain science and nervous system diseases.

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.

Cross-activity of honeybee queen mandibular pheromone in bumblebees provides evidence for sensory exploitation

AbstractThe evolutionary origin of queen pheromones (QPs), which regulate reproductive division of labor in insect societies, has been explained by two evolutionary scenarios: the sender-precursor hypothesis and the sensory exploitation hypothesis. These scenarios differ in terms of whether the signaling system was built on preadaptations on the part of either the sender queens or the receiver workers. While some social insect QPs—such as cuticular hydrocarbons—were likely derived from ancestral fertility cues and evolved according to the former theory, the honeybee’s queen mandibular pheromone (QMP) has been suggested to act directly on preexisting gene-regulatory networks linked with reproduction. This is evidenced by the fact that QMP has been shown to also inhibit ovary activation in fruit flies, thereby implying exploitation of conserved physiological pathways. To verify whether QMP has similar effects on more closely related eusocial species, we here tested for QMP cross-activity in the bumblebee Bombus terrestris. Interestingly, we found that the non-native QMP blend significantly inhibited egg laying in both worker and queen bumblebees and caused accompanying shifts in ovary activation. The native bumblebee QP pentacosane, by contrast, only inhibited the reproduction of the workers. Overall, these findings support the hypothesis that honeybee QMP likely evolved via a route of sensory exploitation. We argue that such exploitation could allow social insect queens to produce compounds that manipulate the workers to remain sterile, but that a major hurdle would be that the queens themselves would have to be immune to such compounds.

Ancestral hymenopteran queen pheromones do not share the broad phylogenetic repressive effects of honeybee queen mandibular pheromone

Queen pheromones effect the reproductive division of labour, a defining feature of eusociality. Reproductive division of labour ensures that one, or a small number of, females are responsible for the majority of reproduction within a colony. Much work on the evolution and function of these pheromones has focussed on Queen Mandibular Pheromone (QMP) which is produced by the Western or European honeybee (Apis mellifera). QMP has phylogenetically broad effects, repressing reproduction in a variety of arthropods, including those distantly related to the honeybee such as the fruit fly Drosophila melanogaster. QMP is highly derived and has little chemical similarity to the majority of hymenopteran queen pheromones which are derived from cuticular hydrocarbons. This raises the question of whether the phylogenetically widespread repression of reproduction by QMP also occurs with more basal saturated hydrocarbon-based queen-pheromones. Using D. melanogaster we show that saturated hydrocarbons are incapable of repressing reproduction, unlike QMP. We also show no interaction between the four saturated hydrocarbons tested or between the saturated hydrocarbons and QMP, implying that there is no conservation in the mechanism of detection or action between these compounds. We propose that the phylogenetically broad reproductive repression seen in response to QMP is not a feature of all queen pheromones, but unique to QMP itself, which has implications for our understanding of how queen pheromones act and evolve.

Honeybees possess a structurally diverse and functionally redundant set of queen pheromones.

Queen pheromones, which signal the presence of a fertile queen and induce workers to remain sterile, play a key role in regulating reproductive division of labour in insect societies. In the honeybee, volatiles produced by the queen's mandibular glands have been argued to act as the primary sterility-inducing pheromones. This contrasts with evidence from other groups of social insects, where specific queen-characteristic hydrocarbons present on the cuticle act as conserved queen signals. This led us to hypothesize that honeybee queens might also employ cuticular pheromones to stop workers from reproducing. Here, we support this hypothesis with the results of bioassays with synthetic blends of queen-characteristic alkenes, esters and carboxylic acids. We show that all these compound classes suppress worker ovary development, and that one of the blends of esters that we used was as effective as the queen mandibular pheromone (QMP) mix. Furthermore, we demonstrate that the two main QMP compounds 9-ODA and 9-HDA tested individually were as effective as the blend of all four major QMP compounds, suggesting considerable signal redundancy. Possible adaptive reasons for the observed complexity of the honeybee queen signal mix are discussed.