Pheromone communication in Nasonia vitripennis: abdominal sex attractant mediates site fidelity of releasing males.

Males of the parasitic wasp Nasonia vitripennis (Hymenoptera: Pteromalidae) use a substrate-borne sex pheromone to attract virgin females. The pheromone is synthesized in the rectal vesicle and deposited via the anus by dabbing movements of the abdominal tip. The chemicals attracting the females are composed of a mixture (4R,5R- and (4R,5 S)-5-hydroxy-4-decanolides (HDL) being synergized by the trace component 4-methylquinazoline (4-MeQ) which is not attractive for females when offered alone. Here we show that male pheromone deposits are not only attractive to virgin females but also for the releasing males themselves. In an olfactometer bioassay, males were strongly attracted by their own pheromone markings but were unable to discriminate between their own markings and those deposited by other males. Polar fractions of pheromone gland extracts containing the HDLs and 4-MeQ were also highly attractive for males. Bioassays using synthetic pheromones in natural doses revealed that combinations of HDL/4-MeQ and 4-MeQ alone attracted males whereas the HDLs alone were behaviorally inactive. Furthermore, males did not discriminate between HDL/4-MeQ and 4-MeQ alone. We conclude that the trace component 4-MeQ mediates site fidelity of N. vitripennis males at sites previously marked with the abdominal sex pheromone. The use of 4-MeQ to stay at and to return to scent-marked patches rather than marking new ones might be a strategy to economize semiochemical use in N. vitripennis males.

Quantity matters: male sex pheromone signals mate quality in the parasitic wasp Nasonia vitripennis.

Sexual selection theory asserts that females are well adapted to sense signals indicating the quality of potential mates. One crucial male quality parameter is functional fertility (i.e. the success of ejaculates in fertilizing eggs). The phenotype-linked fertility hypothesis (PLFH) predicts that functional fertility of males is reflected by phenotypic traits that influence female mate choice. Here, we show for Nasonia vitripennis, a parasitic wasp with haplodiploid sex determination and female-biased sex ratios, that females use olfactory cues to discriminate against sperm-limited males. We found sperm limitation in newly emerged and multiply mated males (seven or more previous matings) as indicated by a higher proportion of sons in the offspring fathered by these males. Sperm limitation correlated with clearly reduced pheromone titres. In behavioural bioassays, females oriented towards higher doses of the synthetic pheromone and were attracted more often to scent marks of males with a full sperm load than to those of sperm-limited males. Our data support the PLFH and suggest that N. vitripennis females are able to decrease the risk of getting constrained to produce suboptimal offspring sex ratios by orienting towards gradients of the male sex pheromone.

Mechanism and behavioral context of male sex pheromone release in Nasonia vitripennis.

Males of the parasitic wasp Nasonia vitripennis (Hymenoptera: Pteromalidae) attract virgin females by releasing a sex pheromone composed of (4R,5R)- and (4R,5S)-5-hydroxy-4-decanolide (HDL). The pheromone is biosynthesized in the rectal vesicle of males. In the present study, we investigated the mechanism and behavioral context of pheromone release, and determined the range of activity and the longevity of the chemical signal. Our data show that the sex pheromone of N. vitripennis is substrate-borne and is deposited on surfaces by dabbing movements of the abdominal tip, a behavior previously described in N. vitripennis males as 'abdomen dipping'. Chemical markings deposited by a single male were highly attractive to virgin females. Chemical analyses revealed the presence of HDL in surface washings of marked areas, and HDL amounts correlated with male marking activity. Pheromone deposition occurred spontaneously without any additional cues being present, but marking intensity increased greatly after copulation or after a single contact with a virgin female. In contrast, marking intensity was not influenced by the presence of host puparia. Male pheromone deposits were perceived by females in a still-air olfactometer at distances of up to 4.5 cm and remained attractive for at least 2 h. The function of the substrate-borne sex pheromone is discussed with respect to the mating system of N. vitripennis.

Costs of female odour in males of the parasitic wasp Lariophagus distinguendus (Hymenoptera: Pteromalidae).

The display of female traits by males is widespread in the animal kingdom. In several species, this phenomenon has been shown to function adaptively as a male mating strategy to deceive sexual rivals (female mimicry). Freshly emerged males of the parasitic wasp Lariophagus distinguendus (Hymenoptera: Pteromalidae) are perceived by other males as if they were females because of a very similar composition of cuticular hydrocarbons which function as a sex pheromone in this species inducing courtship behaviour in males. Within 32 h, however, males deactivate the pheromone and are no longer courted by other males. In this paper, behavioural experiments were performed to test hypotheses on potential costs and benefits associated with the female odour in young males. We did not find any benefits, but demonstrated that young males were significantly more often outrivaled in male-male contests when competing with two older males for a female. Also, young males were significantly more often mounted in homosexual courtship events during these contests. Thus, display of female traits by males is not necessarily beneficial, and in fact, can be disadvantageous. We suggest that these costs have favoured the evolution of the pheromone deactivation mechanism in L. distinguendus males. The function of cuticular hydrocarbons as a female courtship pheromone in L. distinguendus might have evolved secondarily from a primary function relevant for both genders, and the deactivation of the signal in males might have caused a shift of specificity of the chemical signal from the species level to the sex level.

4-methylquinazoline is a minor component of the male sex pheromone in Nasonia vitripennis.

We identified 4-methylquinazoline (4-MeQ) as a minor component of the male sex pheromone of the parasitoid Nasonia vitripennis. Like the major components (4R,5R)- and (4R,5S)-5-hydroxy-4-decanolide (HDL), 4-MeQ is synthesized in the abdomen of males. At doses of 6 or 1 ng, 4-MeQ synergized the response of virgin females to the HDL-diastereomers in a still-air olfactometer, but was not attractive as a single component. 4-MeQ is also responsible for the characteristic medicinal odor of N. vitripennis males.

A male sex pheromone in a parasitic wasp and control of the behavioral response by the female's mating status.

Male insects may increase their chance of successful reproduction by releasing pheromones that attract females or elicit sexual acceptance. In parasitic wasps, male pheromones have been suggested for a few species but no chemicals have been identified so far. Here we report the first identification of a male sex pheromone in parasitic Hymenoptera. In abdomens of male jewel wasps, Nasonia vitripennis Walker, we found a mixture of (4R,5R)- and (4R,5S)-5-hydroxy-4-decanolide (HDL), which was released intermittently and attracted virgin females, but no males, in an olfactometer bioassay. However, only a few minutes after copulation mated females avoided the male-derived pheromone. Neither preference nor avoidance was shown by mated females after 24 h and even after they had been allowed to oviposit for 6 days. Nasonia vitripennis females normally mate only once. Thus, their variable response to the sex attractant depending on the mating status makes sense from an evolutionary perspective. Firstly, it increases the chance of virgins to be inseminated. Secondly, by terminating the response or even avoiding the male pheromone, mated females decrease the probability of encountering males and being disturbed by their courtship activities when searching for new oviposition sites.

Courtship pheromones in parasitic wasps: comparison of bioactive and inactive hydrocarbon profiles by multivariate statistical methods.

Cuticular hydrocarbons play a significant role in the regulation of cuticular permeability and also in the chemical communication of insects. In the parasitoid Lariophagus distinguendus (Hymenoptera: Pteromalidae), male courtship behavior is mediated by a female-produced sex pheromone. Previous studies have shown that the chemicals involved are already present in the pupal stage of both males and females. However, pheromonal activity in males decreases shortly after emergence. This pheromonal deactivation occurs only in living males, suggesting an active process rather than simple evaporation of bioactive compounds. Here, we present evidence that the sex pheromone of L. distinguendus is composed of a series of cuticular hydrocarbons. Filter paper disks treated with nonpolar fractions of cuticular extracts of freshly emerged males and females, 72-hr-old females, and yellowish pupae caused arrestment and stimulated key elements of courtship behavior in males, whereas fractions of 72-hr-old males did not. Sixty-four hydrocarbons with chain length between C(25) and C(37) were identified in the fractions by gas chromatography-mass spectrometry (GC-MS). Methyl-branched alkanes with one to four methyl groups were major components, along with traces of n-alkanes and monoalkenes. Principal component analysis, based on the relative amounts of the compounds, revealed that cuticular hydrocarbon composition differed among all five groups. By using partial least squares-discriminant analysis, we determined a series of components that differentiate bioactive and bioinactive hydrocarbon profiles, and may be responsible for pheromonal activity of hydrocarbon fractions in L. distinguendus.

Host habitat assessment by a parasitoid using fungal volatiles.

BACKGROUND: The preference-performance hypothesis predicts that oviposition preference of insects should correlate with host suitability for offspring development. Therefore, insect females have to be able to assess not only the quality of a given host but also the environmental conditions of the respective host habitat. Chemical cues are a major source of information used by insects for this purpose. Primary infestation of stored grain by stored product pests often favors the intense growth of mold. This can lead to distinct sites of extreme environmental conditions (hot-spots) with increased insect mortality. We studied the influence of mold on chemical orientation, host recognition, and fitness of Lariophagus distinguendus, a parasitoid of beetle larvae developing in stored grain. RESULTS: Volatiles of wheat infested by Aspergillus sydowii and A. versicolor repelled female parasitoids in an olfactometer. Foraging L. distinguendus females are known to be strongly attracted to the odor of larval host feces from the granary weevil Sitophilus granarius, which may adhere in remarkable amounts to the surface of the grains. Feces from moldy weevil cultures elicited neutral responses but parasitoids clearly avoided moldy feces when non-moldy feces were offered simultaneously. The common fungal volatile 1-octen-3-ol was the major component of the odor of larval feces from moldy weevil cultures and repelled female parasitoids at naturally occurring doses. In bioassays investigating host recognition behavior of L. distinguendus, females spent less time on grains containing hosts from moldy weevil cultures and showed less drumming and drilling behavior than on non-moldy controls. L. distinguendus had a clearly reduced fitness on hosts from moldy weevil cultures. CONCLUSION: We conclude that L. distinguendus females use 1-octen-3-ol for host habitat assessment to avoid negative fitness consequences due to secondary mold infestation of host patches. The female response to fungal volatiles is innate, suggesting that host-associated fungi played a crucial role in the evolution of host finding strategies of L. distinguendus. Research on the role of host-associated microorganisms in the chemically mediated orientation of parasitoids is still at the beginning. We expect an increasing recognition of this issue in the future.

Characterization of a female-produced courtship pheromone in the parasitoid Nasonia vitripennis.

Males of the parasitoid Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) show a characteristic courtship behavior. We demonstrate that male arrestment and key behavioral elements of the courtship sequence are mediated by a female-derived contact sex pheromone. Males were arrested on paper disks treated with female extracts but not on those treated with male extracts. Male responsiveness was influenced by the surface to which female extracts were applied. Extracts applied to an extracted beetle elytron arrested males more strongly than those applied to filter paper of comparable size. However, more complex behavioral elements, such as head nodding and copulation attempts, were shown only when extracts were applied to extracted male cadavers, suggesting that tactile or visual cues synergize the male response. The chemicals involved are stable, of low volatility, and nonpolar. Dead females arrested males and elicited courtship behavior for at least 8 d. Males showed no sign of attraction to live females at a distance of 3 cm in an olfactometer. Fractionation of female extracts demonstrated that the activity was exclusively located in the nonpolar fraction. Analysis of the active fraction by gas chromatography-mass spectrometry revealed that cuticular hydrocarbons with chain lengths between 25 and 37 carbon units were present. Comparison of hydrocarbon profiles from males and females showed qualitative and quantitative differences. These results suggest that sex-specific cuticular hydrocarbons are the key signals mediating the male courtship behavior in N. vitripennis.