Uncoupling primer and releaser responses to pheromone in honey bees

Pheromones produce dramatic behavioral and physiological responses in a wide variety of species. Releaser pheromones elicit rapid responses within seconds or minutes, while primer pheromones produce long-term changes which may take days to manifest. Honeybee queen mandibular pheromone (QMP) elicits multiple distinct behavioral and physiological responses in worker bees, as both a releaser and primer, and thus produces responses on vastly different time scales. In this study, we demonstrate that releaser and primer responses to QMP can be uncoupled. First, treatment with the juvenile hormone analog methoprene leaves a releaser response (attraction to QMP) intact, but modulates QMP’s primer effects on sucrose responsiveness. Secondly, two components of QMP (9-ODA and 9-HDA) do not elicit a releaser response (attraction) but are as effective as QMP at modulating a primer response, downregulation of foraging-related brain gene expression. These results suggest that different responses to a single pheromone may be produced via distinct pathways.     

Selection on worker honeybee responses to queen pheromone (Apis mellifera L.)

 Disruptive selection for responsiveness to queen mandibular gland pheromone (QMP) in the retinue bioassay resulted in the production of high and low QMP responding strains of honeybees (Apis mellifera L.). Strains differed significantly in their retinue response to QMP after one generation of selection. By the third generation the high strain was on average at least nine times more responsive than the low strain. The strains showed seasonal phenotypic plasticity such that both strains were more responsive to the pheromone in the spring than in the fall. Directional selection for low seasonal variation indicated that phenotypic plasticity was an additional genetic component to retinue response to QMP. Selection for high and low retinue responsiveness to QMP was not an artifact of the synthetic blend because both strains were equally responsive or non-responsive to whole mandibular gland extracts compared with QMP. The use of these strains clearly pointed to an extra-mandibular source of retinue pheromones (Pankiw et al. 1995; Slessor et al. 1998; Keeling et al. 1999). Received: 2 March 2000 / Accepted in revised form: 4 September 2000     

Pheromonal regulation of starvation resistance in honey bee workers (<Emphasis Type="Italic">Apis mellifera</Emphasis>)

Most animals can modulate nutrient storage pathways according to changing environmental conditions, but in honey bees nutrient storage is also modulated according to changing behavioral tasks within a colony. Specifically, bees involved in brood care (nurses) have higher lipid stores in their abdominal fat bodies than forager bees. Pheromone communication plays an important role in regulating honey bee behavior and physiology. In particular, queen mandibular pheromone (QMP) slows the transition from nursing to foraging. We tested the effects of QMP exposure on starvation resistance, lipid storage, and gene expression in the fat bodies of worker bees. We found that indeed QMP-treated bees survived much longer compared to control bees when starved and also had higher lipid levels. Expression of vitellogenin RNA, which encodes a yolk protein that is found at higher levels in nurses than foragers, was also higher in the fat bodies of QMP-treated bees. No differences were observed in expression of genes involved in insulin signaling pathways, which are associated with nutrient storage and metabolism in a variety of species; thus, other mechanisms may be involved in increasing the lipid stores. These studies demonstrate that pheromone exposure can modify nutrient storage pathways and fat body gene expression in honey bees and suggest that chemical communication and social interactions play an important role in altering metabolic pathways.     

Chirality determines pheromone activity for flour beetles

Olfactory perception and orientation behaviour of female and male flour beetles (Tribolium castaneum, T. confusum) to single stereoisomers of their aggregation pheromone revealed maximal receptor potentials and optimal attraction in response to 4R,8R-(?)-dimethyldecanal, whereas its optical antipode 4S,8S-(+)-dimethyldecanal was found to be inactive in this respect. Female flour beetles of both species were ≈ 10³ times less attracted to 4R,8S-(+)- and 4S,8R-(?)-dimethyldecanal than to 4R,8R-(?)-dimethyldecanal, while male flour beetles failed to respond to the R,S-(+)- and S,R-(?)-stereoisomers. Pheromone extracts of prothoracic femora from unmated male flour beetles elicited higher receptor potentials in the antennae of females than in those of males. The results suggest that the aggregation pheromone emitted by maleT. castaneum as well as maleT. confusum has the stereochemical structure of 4R,8R-(?)-dimethyl-decanal, which acts as sex attractant for the females and as aggregant for the males of both species.     

A comparative study of an innate immune response in Lamprologine cichlid fishes

Social interactions facilitate pathogen transmission and increase virulence. Therefore, species that live in social groups are predicted to suffer a higher pathogen burden, to invest more heavily in immune defence against pathogens, or both. However, there are few empirical tests of whether social species indeed invest more heavily in immune defence than non-social species. In the current study, we conducted a phylogenetically controlled comparison of innate immune response in Lamprologine cichlid fishes. We focused on three species of highly social cichlids that live in permanent groups and exhibit cooperative breeding (Julidochromis ornatus, Neolamprologus pulcher and Neolamprologus savoryi) and three species of non-social cichlids that exhibit neither grouping nor cooperative behaviour (Telmatochromis temporalis, Neolamprologus tetracanthus and Neolamprologus modestus). We quantified the innate immune response by injecting wild fishes with phytohaemagglutinin (PHA), a lectin that causes a cell-mediated immune response. We predicted that the three highly social species would show a greater immune reaction to the PHA treatment, indicating higher investment in immune defence against parasites relative to the three non-social species. We found significant species-level variation in immune response, but contrary to our prediction, this variation did not correspond to social system. However, we found that immune response was correlated with territory size across the six species. Our results indicate that the common assumption of a positive relationship between social system and investment in immune function may be overly simplistic. We suggest that factors such as rates of both in-group and out-group social interactions are likely to be important mediators of the relationship between sociality and immune function.     

The effect of queen pheromones on worker honey bee ovary development

We report results that address a long-standing controversy in honey bee biology, the identity of the queen-produced compounds that inhibit worker honey bee ovary development. As the honey bee is the only organism for which identities have been proposed for any pheromone that regulates reproduction, the resolution of its identity is of broad significance. We examined the effects of synthetic honey bee queen mandibular pheromone (QMP), four newly identified queen retinue pheromone components, and whole-queen extracts on the ovary development of caged worker bees. The newly identified compounds did not inhibit worker ovary development alone, nor did they improve the efficacy of QMP when applied in combination. QMP was as effective as queen extracts at ovary regulation. Caged workers in the QMP and queen extract treatments had better developed ovaries than did workers remaining in queenright colonies. We conclude that QMP is responsible for the ovary-regulating pheromonal capability of queens from European-derived Apis mellifera subspecies.     

A new pheromone of the silkworm moth Bombyx mori

The female silkmoth Bombyx mori L. emits a second pheromone component bombykal (E-10, Z-12-hexade-cadien-1-al) in addition to the well-known sexual attractant bombykol (E-10, Z-12-hexadecadien-1-ol). Bombykal stimulates its own specialized and highly sensitive olfactory cells of the male moth. Surprisingly, the aldehyde inhibits the release of the male's wing-fluttering response to bombykol.     

Selective olfactory attention of a specialised predator to intraspecific chemical signals of its prey

Prey-specialised predators have evolved specific cognitive adaptations that increase their prey searching efficiency. In particular, when the prey is social, selection probably favours the use of prey intraspecific chemical signals by predatory arthropods. Using a specialised ant-eating zodariid spider, Zodarion rubidum, which is known to prey on several ant species and possesses capture and venom adaptations more effective on Formicinae ants, we tested its ability to recognise chemical cues produced by several ant species. Using an olfactometer, we tested the response of Z. rubidum towards air with chemical cues from six different ant species: Camponotus ligniperda, Lasius platythorax and Formica rufibarbis (all Formicinae); and Messor structor, Myrmica scabrinodis and Tetramorium caespitum (all Myrmicinae). Z. rubidum was attracted to air carrying chemical cues only from F. rufibarbis and L. platythorax. Then, we identified that the spiders were attracted to airborne cues coming from the F. rufibarbis gaster and Dufour's gland, in particular. Finally, we found that among several synthetic blends, the decyl acetate and undecane mixture produced significant attraction of spiders. These chemicals are produced only by three Formicine genera. Furthermore, we investigated the role of these chemical cues in the communication of F. rufibarbis and found that this blend reduces their movement. This study demonstrates the chemical cognitive capacity of Z. rubidum to locate its ant prey using chemical signals produced by the ants. The innate capacity of Z. rubidum to olfactory detect different ant species is narrow, as it includes only two ant genera, confirming trophic specialisation at lower than subfamily level. The olfactory cue detected by Zodarion spiders is probably a component of the recruitment or trail pheromone.     

Diel periodicity of pheromone release by females of Planococcus citri and Planococcus ficus and the temporal flight activity of their conspecific males

The diel periodicity of sex pheromone release was monitored in two mealybug species, Planococcus citri and Planococcus ficus (Hemiptera; Pseudococcidae), using sequential SPME/GCMS analysis. A maximal release of 2 ng/h pheromone by 9–12-day-old P. citri females occurred 1–2 h before the beginning of photophase. The highest release of pheromone by P. ficus females was 1–2 ng/2 h of 10–20-day-old females, approximately 2 h after the beginning of photophase. Mating resulted in termination of the pheromone release in both mealybug species. The temporal flight activity of the males was monitored in rearing chambers using pheromone baited delta traps. Males of both P. citri and P. ficus displayed the same flight pattern and began flying at 06:00 hours when the light was turned on, reaching a peak during the first and second hour of the photophase. Our results suggest that other biparental mealybug species display also diel periodicities of maximal pheromone release and response. Direct evaluation of the diel periodicity of the pheromone release by the automatic sequential analysis is convenient and will be very helpful in optimizing the airborne collection and identification of other unknown mealybug pheromones and to study the calling behavior of females. Considering this behavior pattern may help to develop more effective pheromone-based management strategies against mealybugs.     

Recognition of foreign oviposition-marking pheromone in a multi-trophic context

Both phytophagous and parasitic insects deposit oviposition-marking pheromones (OMPs) following oviposition that function to inform conspecifics of a previously utilized host of reduced suitability. The blueberry maggot fly, Rhagoletis mendax Curran (Diptera: Tephritidae), deposits eggs individually into blueberries and then marks the fruit surface with an OMP which reduces acceptance of fruit for oviposition by conspecifics. Diachasma alloeum (Muesebeck) (Hymenoptera: Braconidae) is a parasitic wasp attacking larval R. mendax which also deposits an OMP, signaling conspecifics of a wasp-occupied host. Behavioral studies were conducted testing the hypothesis that the OMP of the parasitic wasp modifies the oviposition behavior of its host fly. In this study, we show that the OMP of D. alloeum is recognized by R. mendax, and female flies will reject wasp-marked fruit for oviposition. Thus, we present a rare demonstration of pheromonal recognition between animals occupying different taxonomic orders and trophic levels. This chemical eavesdropping may enhance the ability of the fly to avoid fruit unsuitable for larval development.     

Sex pheromone component ratios and mating isolation among three Lygus plant bug species of North America

The plant bugs Lygus hesperus, Lygus lineolaris, and Lygus elisus (Hemiptera: Miridae) are major pests of many agricultural crops in North America. Previous studies suggested that females release a sex pheromone attractive to males. Other studies showed that males and females contain microgram amounts of (E)-4-oxo-2-hexenal, hexyl butyrate, and (E)-2-hexenyl butyrate that are emitted as a defense against predators. Using gas chromatography–mass spectrometry, we found that female L. lineolaris and L. elisus have a 4:10 ratio of hexyl butyrate to (E)-2-hexenyl butyrate that is reversed from the 10:1 ratio in female L. hesperus (males of the three species have ~10:1 ratio). These reversed ratios among females of the species suggest a behavioral role. Because both sexes have nearly equal amounts of the major volatiles, females should release more to attract males. This expectation was supported because L. hesperus females released more hexyl butyrate (mean of 86 ng/h) during the night (1800–0700 hours) than did males (<1 ng/h). We used slow-rotating pairs of traps to test the attraction of species to blends of the volatiles with a subtractive method to detect synergism. Each species’ major butyrate ester was released at 3 μg/h, the minor butyrate according to its ratio, and (E)-4-oxo-2-hexenal at 2 μg/h. The resulting catches of only Lygus males suggest that (E)-4-oxo-2-hexenal is an essential sex pheromone component for all three species, (E)-2-hexenyl butyrate is essential for L. elisus and L. lineolaris, and hexyl butyrate is essential for L. hesperus. However, all three components are recognized by each species since ratios of the butyrate esters are critical for conspecific attraction and heterospecific avoidance by males and thus play a role in reproductive isolation among the three species. Because L. hesperus males and females are known to emit these major volatiles for repelling ant predators, our study links defensive allomones in Lygus bugs with an additional use as sex pheromones.     

Codling moth males do not discriminate between pheromone and a pheromone/antagonist blend during upwind flight

Discrimination between conspecific and heterospecific signals is a key element in the evolution of pheromone-mediated communication in insects. Pheromone antagonists prevent heterospecific attraction. They are typically pheromone synergists in other species and enable specific communication in closely related species, using partly the same chemicals. In codling moth, Cydia pomonella, as in other moths, upwind flights to a pheromone/antagonist blend were slower and more convoluted than to pheromone. However, this deteriorated flight behaviour did not account for the strong decrease in male attraction. The pheromone/antagonist blend blocked instead the onset of upwind flight. This was corroborated by placing a separate source of pheromone close by, which restored male attraction. Males flying upwind did not discriminate between pheromone and the adjacent pheromone/antagonist blend, and landed on either source. This indicates differences in the neural regulation for initiating and maintaining a behavioural response to pheromone.     

Short- and long-range cues used by ground-dwelling parasitoids to find their host

Parasitoids of phytophagous insects face a detectability–reliability dilemma when foraging for hosts. Plant-related cues are easily detectable, but do not guarantee the presence of the host. Host-related cues are very reliable, but much harder to detect from a distance. Little is known in particular about the way coleopteran parasitoid females use these cues when foraging for a suitable place to lay their eggs. The question is of interest because, unlike hymenopteran larvae, coleopteran parasitoid larvae are highly mobile and able to forage for hosts on their own. We assessed whether females of the parasitoid rove beetle Aleochara bipustulata (L.) (Coleoptera: Staphylinidae) are attracted to plant (Swede roots, Brassica napus) and host-related cues [pupae of the cabbage root fly Delia radicum (L.) (Diptera: Anthomyiidae)]. In the field, A. bipustulata adult females were captured in selective pitfall traps containing pieces of roots damaged by D. radicum larvae, but not in traps containing pieces of healthy roots or D. radicum pupae. However, in the laboratory, the odour of D. radicum pupae attracted A. bipustulata females to mini-pitfalls. Video monitoring in the laboratory showed that foraging A. bipustulata females preferred a zone containing D. radicum pupae and larval tracks rather than one containing an extract of D. radicum-infested roots. Our results suggest a behavioural sequence where A. bipustulata females use plant-related cues at a distance, but then switch their preference to host-related cues at a close range. This would be the first observation of this behaviour in coleopteran parasitoids.     

Unusual pheromone chemistry in the navel orangeworm: novel sex attractants and a behavioral antagonist

Using molecular- and sensory physiology-based approaches, three novel natural products, a simple ester, and a behavioral antagonist have been identified from the pheromone gland of the navel orangeworm, Amyelois transitella Walker (Lepidoptera: Pyralidae). In addition to the previously identified (Z,Z)-11,13-hexadecadienal, the pheromone blend is composed of (Z,Z,Z,Z,Z)-3,6,9,12,15-tricosapentaene, (Z,Z,Z,Z,Z)-3,6,9,12,15-pentacosapentaene, ethyl palmitate, ethyl-(Z,Z)-11,13-hexadecadienoate, and (Z,Z)-11,13-hexadecadien-1-yl acetate. The C₂₃ and C₂₅ pentaenes are not only novel sex pheromones, but also new natural products. In field tests, catches of A. transitella males in traps baited with the full mixture of pheromones were as high as those in traps with virgin females, whereas control and traps baited only with the previously known constituent did not capture any moths at all. The navel orangeworm sex pheromone is also an attractant for the meal moth, Pyralis farinalis L. (Pyralidae), but (Z,Z)-11,13-hexadecadien-1-yl acetate is a behavioral antagonist. The new pheromone blend may be highly effective in mating disruption and monitoring programs.     

The effects of mating status and time since mating on female sex pheromone levels in the rice leaf bug,Trigonotylus caelestialium

Although mating status affects future mating opportunities, the biochemical changes that occur in response to mating are not well understood. This study investigated the effects of mating status on the quantities of sex pheromone components found in whole-body extracts and volatile emissions of females of the rice leaf bug, Trigonotylus caelestialium. When sampled at one of four time points within a 4-day postmating period, females that had copulated with a male had greater whole-body quantities of sex pheromone components than those of virgin females sampled at the same times. The quantities of sex pheromone components emitted by virgin females over a 24-h period were initially high but then steadily decreased, whereas 24-h emissions were persistently low among mated females when measured at three time points within the 4 days after mating. As a result, soon after mating, the mated females emitted less sex pheromones than virgin females, but there were no significant differences between mated and virgin females at the end of the experiment. Thus, postmating reduction in the rate of emission of sex pheromones could explain previously observed changes in female attractiveness to male T. caelestialium.     

Social spiders of the genus Anelosimus occur in wetter, more productive environments than non-social species

Latitude, rainfall, and productivity have been shown to influence social organisation and level of sociality in arthropods on large geographic scales. Social spiders form permanent group-living societies where they cooperate in brood care, web maintenance, and foraging. Sociality has evolved independently in a number of unrelated spider genera and may reflect convergent evolutionary responses to common environmental drivers. The genus Anelosimus contains a third of approximately 25 described permanently social spider species, eight to nine species that all occur in the Americas. To test for environmental correlates of sociality in Anelosimus across the Americas, we used logistic regression to detect effects of annual rainfall, productivity, and precipitation seasonality on the relative likelihood of occurrence of social and non-social Anelosimus spiders. Our analyses show that social species tend to occur at higher annual rainfall and productivity than non-social species, supporting the hypothesised effects of these environmental variables on the geographical distribution of social species. We did not find support for the hypothesis that permanently social species occur in areas with low precipitation seasonality. High annual precipitation and, to less extent, high productivity favour the occurrence of permanently group-living Anelosimus spiders relative to subsocial and solitary species. These results are partially consistent with previous findings for the Old World spider genus Stegodyphus, where a link between high habitat productivity and sociality was also found. Unlike Anelosimus, however, Stegodyphus typically occur in dry habitats negating a general importance of high precipitation for sociality. Sociality in spiders thus seems to be strongly linked to productivity, probably reflecting the need for relatively high availability of large prey to sustain social colonies.     

(<Emphasis Type="Italic">Z</Emphasis>)-9-tricosene identified in rectal gland extracts of <Emphasis Type="Italic">Bactrocera oleae</Emphasis> males: first evidence of a male-produced female attractant in olive fruit fly

It is well-known that Bactrocera oleae (olive fruit fly) females attract conspecific males by using 1,7-dioxaspiro[5,5]undecane (1) as the main component of their sex pheromone, and that 1 is produced in the female rectal gland. Although some authors have claimed that B. oleae males also attract females, to date no male-produced female attractants have been found in this species. In this paper, we report the first identification of a substance unique to males and able to attract females. The findings of the study include the following: (1) females responded in a bioassay to hexane extracts obtained from rectal glands of 15-day-old B. oleae males, (2) the presence of (Z)-9-tricosene (2) was consistently and unambiguously identified in these extracts using gas chromatography (GC) and GC-mass spectrometry methods, (3) in preliminary bioactivity tests, low doses (equivalent to a few males) of chemically and stereoisomerically pure synthetic (Z)-9-tricosene (2) attracted olive fruit fly females. Interestingly, compound 2, commonly called muscalure, is also a well-known component of the house fly (Musca domestica) sex pheromone.     

Female sex pheromone and male behavioral responses of the bombycid moth <Emphasis Type="Italic">Trilocha varians</Emphasis>: comparison with those of the domesticated silkmoth <Emphasis Type="Italic">Bombyx mori</Emphasis>

Analysis of female sex pheromone components and subsequent field trap experiments demonstrated that the bombycid moth Trilocha varians uses a mixture of (E,Z)-10,12-hexadecadienal (bombykal) and (E,Z)-10,12-hexadecadienyl acetate (bombykyl acetate) as a sex pheromone. Both of these components are derivatives of (E,Z)-10,12-hexadecadienol (bombykol), the sex pheromone of the domesticated silkmoth Bombyx mori. This finding prompted us to compare the antennal and behavioral responses of T. varians and B. mori to bombykol, bombykal, and bombykyl acetate in detail. The antennae of T. varians males responded to bombykal and bombykyl acetate but not to bombykol, and males were attracted only when lures contained both bombykal and bombykyl acetate. In contrast, the antennae of B. mori males responded to all the three components. Behavioral analysis showed that B. mori males responded to neither bombykal nor bombykyl acetate. Meanwhile, the wing fluttering response of B. mori males to bombykol was strongly inhibited by bombykal and bombykyl acetate, thereby indicating that bombykal and bombykyl acetate act as behavioral antagonists for B. mori males. T. varians would serve as a reference species for B. mori in future investigations into the molecular mechanisms underlying the evolution of sex pheromone communication systems in bombycid moths.     

A new C12 alcohol identified as a sex pheromone and a trail-following pheromone in termites: the diene (<Emphasis Type="Italic">Z</Emphasis>,<Emphasis Type="Italic">Z</Emphasis>)-dodeca-3,6-dien-1-ol

The diunsaturated C12 alcohol (Z,Z)-dodeca-3,6-dien-1-ol (dodecadienol) has been characterized by GC-MS and FTIR as a novel releaser pheromone in termites. This alcohol identified in Ancistrotermes pakistanicus (Termitidae, Macrotermitinae) possesses a double pheromonal function which again illustrates the chemical parsimony of termites compared with other social insects. In workers, dodecadienol elicits trail-following at a very low concentration (activity threshold at 0.1 pg/cm of trail); in male alates it induces trail-following at a low concentration (1–10 pg/cm) and sexual attraction at a higher concentration (about 1 ng). Traces of the monounsaturated C12 alcohol (Z)-dodec-3-en-1-ol (dodecenol), known as a trail pheromone of several Macrotermitinae, were also found in the sternal gland extracts of A. pakistanicus, although only dodecadienol was present at the surface of the sternal gland. Workers of A. pakistanicus are not sensitive to dodecenol, but they are as sensitive to dodecatrienol as to dodecadienol. However, in the study area (Vietnam), A. pakistanicus is living in sympatry only with those Macrotermitinae using dodecenol as a trail pheromone, the foraging populations therefore being well isolated through their respective trail pheromones. The presence of three types of unsaturated C12 alcohols as releaser pheromones in the only Macrotermitinae subfamily is discussed, and a possible biosynthetic pathway from linoleic acid is proposed for dodecadienol.     

A cost of alarm pheromone production in cotton aphids, Aphis gossypii

The sesquiterpene, (E)--farnesene, is used by many aphid species as an alarm pheromone to warn related individuals of predation. Disturbed cotton aphids, Aphis gossypii Glover, released (E)--farnesene into the air as detected by solid phase microextraction and gas chromatography mass spectrometry (GC–MS). Solvent extracts of cotton aphids of various life stages and weights also were analyzed by GC–MS for sums of ions 69 and 93, which discriminated (E)--farnesene from coeluting compounds. Aphids of all life stages and sizes reared on cotton plants in both an environmental chamber and glasshouse contained (E)--farnesene in amounts ranging from 0.1 to 1.5 ng per individual. The quantities of (E)--farnesene in aphids increased in relation to increasing body weight, and variation in individual weights explained about 82% of the variation in alarm pheromone. However, the concentrations (ng/mg fresh weight) declined exponentially with increasing body weight. These findings indicate that aphid nymphs try to compensate for their smaller size by producing relatively more pheromone per weight than adults but still cannot approach an evolutionary optimal load, as assumed in adults with the greatest total amounts. This suggests that young aphids need to balance costs of growth and maturation with costs of producing the alarm pheromone.