The role of the androconia in the mating behaviour of the European skipper, Thymelicus lineola, and evidence for a male sex pheromone

Abstract. Laboratory experiments were carried out to determine the role and characteristics of male scent scales (androconia), located in patches (stigmata) on the forewings of Thymelicus lineola (Ochsenheimer) (Lepidoptera: Hesperiidae). In behavioural tests, there were 30—40% fewer matings of virgin females by males with the stigmata removed or where females had their antennal sensilla covered with nail polish, when compared to sham-treated controls. These reductions occurred despite a large increase in male courtship activity. No physical contact was observed between male wings and female antennae during mating. A scanning electron microscope study of male wings and female antennae showed that: (1) lysis zones form over time on the androconia and pieces (osmo-phores) distal to these zones break off; (2) while few of these zones are present at emergence they are numerous in-day-old males; (3) during a single episode of courtship and mating approximately two-thirds of the available osmophores break off, yet the commonly observed courtship between males does not appear to result in osmophore release; and (4) osmophores were never observed on the female antennae. Electroantennogram recordings (EAGs) indicated that: (1) female antennae responded strongly to odours from male forewings but not from other parts of either males or females; (2) female antennae responded more strongly to young field-collected males than to newly-emerged or old, field-collected males; (3) female antennal response to male forewings was reduced if the androconia were scraped off, and was eliminated if the stigmata were removed; (4) male antennae gave weak EAG responses to both male and female forewings. We conclude that males release pheromone via the breakage of osmophores during courtship. We propose that the frequently observed refusals by females of courting males are at least partially dependent on the quantity or quality of male pheromone released, which are in turn correlated with the male's age and mating status.     

Pheromone production, male abundance, body size, and the evolution of elaborate antennae in moths

The males of some species of moths possess elaborate feathery antennae. It is widely assumed that these striking morphological features have evolved through selection for males with greater sensitivity to the female sex pheromone, which is typically released in minute quantities. Accordingly, females of species in which males have elaborate (i.e., pectinate, bipectinate, or quadripectinate) antennae should produce the smallest quantities of pheromone. Alternatively, antennal morphology may be associated with the chemical properties of the pheromone components, with elaborate antennae being associated with pheromones that diffuse more quickly (i.e., have lower molecular weights). Finally, antennal morphology may reflect population structure, with low population abundance selecting for higher sensitivity and hence more elaborate antennae. We conducted a phylogenetic comparative analysis to test these explanations using pheromone chemical data and trapping data for 152 moth species. Elaborate antennae are associated with larger body size (longer forewing length), which suggests a biological cost that smaller moth species cannot bear. Body size is also positively correlated with pheromone titre and negatively correlated with population abundance (estimated by male abundance). Removing the effects of body size revealed no association between the shape of antennae and either pheromone titre, male abundance, or mean molecular weight of the pheromone components. However, among species with elaborate antennae, longer antennae were typically associated with lower male abundances and pheromone compounds with lower molecular weight, suggesting that male distribution and a more rapidly diffusing female sex pheromone may influence the size but not the general shape of male antennae.     

草地贪夜蛾成虫触角感器超微结构观察

[目的]草地贪夜蛾是一种严重危害农业生产的迁飞性害虫,本研究旨在了解草地贪夜蛾成虫触角感器的类型、超微结构和分布.[方法]采用扫描电镜对其雌雄成虫触角感器类型及分布进行观察.[结果]草地贪夜蛾雌雄成虫触角由柄节、梗节及鞭节3部分组成,雄蛾触角稍长于雌蛾,且触角表面覆有大量鳞片.共观察到7种类型的感受器,分别为B?hm氏...     

Antennal glands in Psylliodes chrysocephala, and their possible role in reproductive behaviour

Abstract. The antennal morphology of Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae), an important pest of oilseed rape ( Brassica napus ), was studied. Two types of tricellular, integumentary glands were found. The common antennal glands are distributed under all sensilla-bearing parts of male and female antennae. The male-specific antennal glands are only located under a glabrous area found on antennomeres 6–10 of male beetles. The common antennal glands are synthetically active in both pre- and post-diapause adults, but the male specific antennal glands are only active in post-diapause (reproductively active) males. During studies of mating behaviour, the antennae of the males were highly active at the beginning and end of copulation, and in response to increased female activity. The male specific antennal glands may secrete a sex pheromone, and the glabrous area on the male antennae could be the release site for such a pheromone.     

Electrophysiological investigations of sex pheromone reception and release in Bruchidius atrolineatus

ABSTRACT. Male antennal sensitivity to female sex pheromone in Bruchidius atrolineatus was investigated electrophysiologically (EAG). The existence of receptors for this pheromone was shown in the antennae of males. (No such receptors exist in the antennae of females.) Young (age < 24h) males gave an EAG of amplitude 25–30% of the response of older males. Diapausing males several days old gave a much smaller response than active males of the same age.
EAGs were used to test the production/release of pheromone by groups of twenty females. Before they are 24 h old, active females do not produce/release the pheromone; older females release it at an almost constant rate. Sexually diapausing females do not produce and/or release the sex pheromone.
In sexually active females, mating inhibits pheromone release almost immediately. There is a correlation between reproductive status of the females (development of ovaries, oogenesis) and the production or release of the pheromone. A corresponding correlation also exists in the males whose antennae show a very low sensitivity when they are young or when they are in reproductive diapause.     

Electroantennographic resolution of pulsed pheromone plumes in two species of moths with bipectinate antennae

Trains of 20-ms-duration pulses of pheromone were deliveredat rates of 1–33 Hz to antennal preparations of malesof Bombyx mori and Lymantria dispar, two moth species with bipectinateantennae. Resolution of rapidly pulsed plumes of pheromone wasnot compromised by a complex antennal morphology or by moderatechanges in wind speed (25–50 cm/s). Fourier analysis ofthe electroantennograms resolved the temporal structure of thesignal at frequencies up to 25 Hz for B. mori and up to 5 Hzfor L. dispar. The ability of these sensory structures to identifythe original (unchanged) frequency of the pulse train is particularlynoteworthy because air is slowed by about an order of magnitudeas it passes through bipectinate antennae. Although an unchangingfrequency in slowed airflow may be counterintuitive, this flowpattern, and its effects on odorant patch shape and spacing,is explained from fluid mechanical principles (i.e., the principleof continuity). An unchanging frequency suggests that as deceleratingair passes through a bipectinate antenna, the slowed patchesof odorant are stretched, thinned, and brought closer togetherby the same factor with which they are slowed.     

Antennal responses of Cydia pomonella (L.) exposed to surfaces treated with methoxyfenozide

Electroantennogram (EAG) measurements were recorded from the antennae of male and female codling moth, Cydia pomonella L., to determine whether adult moths exposed to surfaces treated with the ecdysteroid agonist methoxyfenozide experience a decline in their antennal reception and thus olfactory sensitivity. Such a phenomenon would offer a possible mechanism for the previously reported decreased responsiveness from moths treated with methoxyfenozide to pheromone‐ and plant volatile‐based monitoring lures. Mean EAG data revealed that the antennae from methoxyfenozide‐treated male moths appear to be just as sensitive to various doses of synthetic codlemone as the antennae from the control and surfactant‐treated moths, but they appeared to be less sensitive to the pheromone component 12OH (collected from female effluvia) than the control male antennae. Mean male EAG responses to the pheromone components E8,E10‐12Al and codlemone collected from methoxyfenozide‐treated females were significantly less than the responses towards those two pheromone components collected from the control and surfactant‐treated females. Female moth exposure to methoxyfenozide did not negatively impact the sensitivity of female antennae to the plant volatile pear ester, but it did towards the apple volatile butyl hexanoate. Data from this study show that adult C. pomonella exposure to methoxyfenozide‐treated surfaces does appear to negatively impact, in a minor way, the (i) olfactory sensitivity (or detection) of male antennae towards some components of the female sex pheromone, (ii) the female antennal sensitivity towards a key apple volatile and (iii) the attractiveness of female pheromone effluvia.     

The effects of the antennal flight posture and orientation on the pheromonal stimulation patterns experienced by male Spodoptera littoralis (Lepidoptera: Noctuidae)

Heads of Spodoptera littoralis males with their antennae artificiallyelevated and stabilized to imitate their flight posture, wereexposed at different inclinations to a mobile beam of pulsedpheromones. The electroantennogram patterns recorded for variouslinear tracks simulated in this manner were closely linked tothe spatial pheromone density distribution. Thus, during in-flightmate detection, the normally elevated antennae appear to functionas a sensitive, instantaneous detector of conspecific femalesex pheromone, informing the insect of contact with, amount,loss or recovery of the stimulant.     

Olfactory attraction to aggregation pheromone is mediated by disti-flagellum of antennal segments in Riptortus pedestris

Riptortus pedestris (Hemiptera: Alydidae) is a serious pest of soybean and sweet persimmon and uses male produced aggregation pheromone, (E)-2-hexenyl (Z)-3-hexenoate, (E)-2-hexenyl (E)-2-hexenoate, and tetradecyl isobutyrate to facilitate food location and recognition by conspecifics. Using electroantennogram (EAG) and greenhouse bioassay, we determined which antennal segment is involved in the detection of their aggregation pheromone. In the first EAG test using individual antennal segments, significant EAG responses to 1:1:1 mixture of the aggregation pheromone were observed only from the disti-flagellum segments of both male and female antennae at both pheromone doses tested (1 µg and 100 µg). In the following EAG tests using gradually removed antennal segment(s), EAG response was still maintained when the distal half of a disti-flagellum was surgically removed, while EAG response was lost when whole segment of disti-flagellum or other whole segments were gradually removed from intact antenna of both sexes. In greenhouse experiment, removing one or both segment(s) of disti-flagellum from male or female antennae resulted in significant reduction in their attraction to the aggregation pheromone. Together, these findings support that the disti-flagellum of R. pedestris houses olfactory neurons associated with attraction to their aggregation pheromone.     

Male traits under cryptic female choice in the spotted cucumber beetle (Coleoptera: Chrysomelidae)

Males of the spotted cucumber beetle (Diabrotica undecimpunctata howardi) rhythmically stroke females with their antennae during copulation. Males that stroke quickly have a higher probabilityof being accepted as a mate. We determined (1) the mechanismby which females prevent unattractive males from passing spermatophores,(2) whether antennal stroking signals to females the likelihoodof receiving a nuptial gift, and (3) if other male traits inaddition to stroking are subjected to sexual selection fromfemale preference. Dissections of pairs flash-frozen in copuladuring and after antennal stroking showed musculature that,when contracted, folded the vaginal duct leading to the female'sbursa copulatrix in a way that prevented complete penetrationby the aedeagus. These muscles were always contracted whilemales were stroking and always relaxed after stroking had ceased.Males accepted as mates did not differ from males that failedto pass a spermatophore in either absolute or relative bodyweight, aedeagus length, or the amount of cucurbitacins (potentialnuptial gifts) sequestered in their spermatophores. Although99% of the beetles that came to cucurbitacin-rich Cucurbitafruits in the field were males, males that had sequestered cucurbitacins did not stroke females faster than males withno cucurbitacins, and fast-stroking males were not more likelyto find and sequester cucurbitacins than were males that strokedmore slowly. Males with a cucurbitacin slurry painted on theirantennae had no mating advantage over controls. We concludethat females discriminate among males after copulation hasbegun on the basis of antennal stroking displays (or some traitcorrelated with stroking speed) that males perform to enticefemales to relax their bursal sphincter.     

Sulfhydryl-reagent inhibition of olfaction in a moth and effects of exposure to pheromone compounds or a pheromone analogue

The effects on olfaction of N-ethylmaleimide (NEM), a specificreagent of free sulfhydryl groups, were studied in the mothMamestra brassicae. The antennae of male M.brassicae bear twotypes of specialist receptor neurons involved in pheromone communication.One type is tuned to (Z)-11-hexadecenyl acetate (Z11-16:Ac),the main pheromone component; the second type is tuned to (Z)-9-tetradecenylacetate (Z9-14:Ac), an interspecific inhibitor not producedby the females of this species. Vapours of NEM irreversiblyinhibited the electro-antennographic (EAG) responses to Z11-16:Acand Z9-14:Ac. When Zll-16:Ac was applied before and during NEMtreatment, the responses to Z9-14:Ac were preserved and someprotection was observed in the responses to Zll-16:Ac. In return,Z9-14:Ac partially prevented the disappearance of responsesto Zll-16:Ac but not to Z9-14:Ac. A third compound, hexadecylacetate (16:Ac), found in the pheromone gland, but not detectedby the antennal receptors, did not prevent the inhibition causedby NEM.     

Sexual Selection of Zorion guttigerum Westwood (Coleoptera: Cerambycidae: Cerambycinae) in Relation to Body Size and Color

Zorion guttigerum is a flower-visiting longhorned beetle endemic to New Zealand. Sexual selection of this species in relation to the body size and color form of different sexes was investigated in the field. The population sex ratio, based on censuses of feeding and mating sites (flowers), is male-biased. Females are significantly larger than males. Both sexes have antennae of similar length but the antennal length relative to the elytral length is greater in males than in females, and the antennal length of males increases more with an increase in body size than that of females. Both sexes have dark blue (DB) and yellowish-brown (YB) individuals. Both pair-bonded and solitary males are similar in elytral and antennal length. In pair-bonded males, DB individuals are significantly more numerous than YB ones, but in solitary males, the number of both color forms is similar. Males tend to have territory protection behavior, fighting with and chasing away rival males from feeding and mating sites. Larger males usually win the fight but the size-dependent fighting advantage does not translate into mating success. Male color plays an important role in mating success, with DB males having a significantly better chance to mate than YB males. Furthermore, male body size and color also have interactions in mating success: males of DB color morph obtain a greater mating advantage according to body size. Pair-bonded females are significantly larger and have longer antennae than solitary females, suggesting that males prefer larger females for mating. In addition, females of DB color morph with longer antennae are also preferred by males for mating. The significance of these findings is discussed.     

Interactions of pheromone with moth antennae: Adsorption,desorption and transport

Isolated antennae of the male moth Antheraea polyphemus adsorbed at least 32% of ³H-labelled pheromone molecules (E-6,Z-11 hexadecadienyl acetate) from an airstream passing the antenna. About 80% of the adsorbed molecules were caught by the long olfactory hairs (sensilla trichodea). The distal half of the hairs caught about twice as many molecules as the proximal half. About 40% of the molecules desorbed if the antennae were exposed to a clean airstream for 30 min. The adsorbed molecules were transported from the hairs towards the antennal branch. Transport due to diffusion would have a diffusion coefficient of 3 × 10^?7 cm²/s. Forty per cent of the total radioactivity per hair could be detected in receptor lymph extruded from the olfactory hairs, after an incubation time of 2 min. Dried antennae showed an increased desorption and an increased velocity of the transport along the hairs. One interpretation is that the molecules enter the receptor lymph of the intact antennae and diffuse more slowly than those on the cuticular surface. Fractional elution of fresh antennae revealed a diminishing elutability of pheromone from antennae in pentane (DEP-effect) and almost constant elutability in more polar solvents (chloroform-methanol toluene). The DEP-effect could be reversibly abolished by dehydration of the antennae. It could be shown that the DEP-effect occurs mainly on the antennal branch rather than on the hairs. Residual (uneluted) radioactivity builds up mainly on the branch.     

Adaptation of male moth antennal neurons in a pheromone plume is associated with cessation of pheromone-mediated flight

Recordings of the firing rates of single antennal neurons whenAgrotis segetum antennac were placed 70 cm downwind of a pheromonesource revealed that cells sensitive to the most volatile componentadapted rapidly in a plume from a high–concentration sourceknown from previous studies to cause in-flight arrestment ofprogress towards the source. No adaptation was found in responseto lower-concentration plumes known to promote high levels ofsustained flight to the source with little premature arrestment.Adaptation was not observed in antennal neurons of a secondspecies, Heliothis virescens, when they were placed in plumesof this species' sex pheromone blend, regardless of the concentration.In flight-tunnel tests these same pheromone sources evoked highlevels of source location with little arrestment. These resultsindicate that adaptation or attenuation of antennal neuronalburst frequencies in response to rapidly arriving pheromonefilaments in a plume may be important peripheral determinantsof whether or not prolonged upwind flight and successful pheromonesource location occurs.     

Pheromone-blend discrimination by European corn borer moths with inter-race and inter-sex antennal transplants

Transplantation of larval antennal imaginal disks between the pheromone races of the European corn borer moth produced males and females of one race with male antennal phenotypes of the race using the opposite pheromone blend. All of the adults with transplanted antennal disks that exhibited a pheromone-mediated response in the flight tunnel did so with the pheromone blend of the recipient's race, even though the respective males and females possessed male antennae of the opposite race. Our results support the conclusion that male antennal response type is not a determining factor in pheromone-blend discrimination in male moths. Accepted: 13 November 1998     

Sexual dimorphism and allometry in two seed beetles (Coleoptera: Bruchidae)

Male Callosobruchus chinensis (Coleoptera: Bruchidae) have elaborated, pectinate antennae, which are absent from conspecific females and both sexes of a congener, Callosobruchus maculatus. To begin to unravel the mechanisms producing this striking dimorphism, we examined which morphological traits best explain body size variation in bruchid beetles and quantified sexual dimorphism of antenna size through allometric analyses. Using principal component analyses, we found that elytron length and pronotum width were significantly correlated with the first principal component, which was interpreted as explaining variation in body size. Regressions of log‐transformed body size measures on log‐transformed antenna length revealed that males of both species had longer antennae than conspecific females for any given body size, although most of this effect was attributable to higher intercepts, rather than increased allometry, in males. Comparisons among heterospecific males revealed that C. maculatus males have noticeably longer antennae than C. chinensis males at large body sizes. Callosobruchus chinensis males, thus, appear to have increased the receptive area of their antennae by adding to the width of, rather than further elongating, their antennae. Finally, we found evidence for positive allometry between log‐transformed antenna length and log‐transformed antenna width in C. chinensis males. We discuss our results in the context of evidence supporting the presence of an additional, and potentially unique, sex pheromone in C. chinensis females.     

Electroantennogram responses to the sex pheromone and other odours in the American cockroach

Electroantennogram responses of adult American cockroaches were recorded for the sex pheromone and 30 kinds of monoterpenoids. Strong responses to the sex pheromone were obtained from the antennae of male adults, but not from females. The compounds possessing a hydroxyl or carbonyl group produced significant EAG responses in the antennae. Bornyl acetate, however, was an exception and gave strong responses in both male and female antennae. There was basically no difference between the antennal responsiveness to monoterpenoids tested in both sexes.     

Reidentification of pheromone composition of Sparganothis sulfureana (Clemens) and evidence of geographic variation in male responses from two US states

GC-EAD analyses of pheromone gland extracts of calling female Sparganothis sulfureana revealed at least 6 compounds that consistently elicited antennal responses from male antennae. In addition to the major pheromone compound, (E)-11-tetradecenyl acetate (E11–14:OAc), which was previously reported, the other compounds were found to be (E)-9-dodecenyl acetate (E9–12:OAc), (Z)-9-dodecenyl acetate (Z9–12:OAc), (Z)-9-tetradecenyl acetate (Z9–14:OAc), (Z)-11-tetradecenyl acetate (Z11–14:OAc), and (E)-11-tetradecenol (E11–14:OH). Tetradecyl acetate, hexadecyl acetate and hexadecenyl acetates were also present in the extracts, but elicited no EAG response from male antennae. Wind tunnel tests demonstrated that males from New Jersey responded equally well to a blend containing five pheromone components in relative to the pheromone glands of calling females. Different male-response profiles from field-trapping tests conducted in the states of Wisconsin and New Jersey were observed, respectively. Significantly higher numbers of male S. sulfureana were caught in New Jersey in traps baited with the binary blend of E11–14:OAc (30 μg) with 1% of Z11–14:OAc, but males from Wisconsin responded equally well to traps containing blends of E11–14:OAc with 0–10% of Z11–14:OAc. The addition of more than 10% of Z11–14:OAc to the primary pheromone compound reduced male captures significantly in both states. Male catches were doubled by adding E9–12:OAc and E11–14:OH to the most attractive binary blend in both states. The trapping test with caged live virgin female moths showed that males in Wisconsin preferred females from the local population than those from New Jersey. The differences in male responses observed may indicate the existence of pheromone polymorphism in this species.     

Discrimination of conspecific individuals via cuticular pheromones by males of the cricket Gryllus bimaculatus

Cuticular substances on the body surface of crickets serve as pheromones that elicit a variety of different behaviors in male crickets. Antennal contact between males and females resulted in courtship behavior, and that between two males resulted in aggressive displays. As a first step in elucidating how crickets recognize and discriminate individuals, behavioral responses of male individuals to cuticular substances of conspecific males or females were investigated. The behavioral responses of males to antennal or palpal stimulation with an isolated antenna from a male or a female were recorded. To both antennal and palpal stimulation with female antennae, the majority of males responded with courtship behavior; to stimulation with male antennae, males responded with aggressive displays. To gain insight into the chemical nature of the behaviorally relevant components, isolated antennae were washed in either n-hexane, acetone or ethanol before behavior assays. Washed antennae no longer elicited courtship or aggressive responses in males. Next, polypropylene fibers were smeared with substances from the body surface of females and used for antennal stimulation. This experiment showed that the quality and quantity of cuticular substances appear to be highly age-dependent. Significantly more males responded with courtship behavior to cuticular substances from younger females. Isolated males generally showed higher levels of aggression than males reared in groups. Grouped males also were more likely to display courtship behavior towards antennae from younger females, and aggressive behavior towards antennae from older females. These results suggest that male discrimination of mating partners depends on the nature of female cuticular substances.     

Transfer of pheromone from immature queen honeybees, Apis mellifera

ABSTRACT. Immediately after visiting cells containing immature queens, workers of Apis mellifera L. were observed to engage in prolonged cleaning, particularly of their tongues when they had visited larvae, and of their antennae when they had visited pupae. Thereafter other workers usually initiated and made antennal contacts with them. During such antennal contact the bee that had visited the queen larva often donated food. The implication of these findings on the distribution of pheromone produced by immature queens is discussed. Workers were stimulated to make antennal contact with the excised heads of bees from a queen's court, providing further evidence that queen pheromone is transferred between workers' antennae.