Sven Steiner

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

SUMMARY 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.

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.

Host habitat assessment by a parasitoid using fungal volatiles

BackgroundThe 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.ResultsVolatiles 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.ConclusionWe 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.

Mating with sperm-depleted males does not increase female mating frequency in the parasitoid Lariophagus distinguendus

Sexual conflicts due to divergent male and female interests in reproduction are common in parasitic Hymenoptera. The majority of parasitoid females are monandrous, whereas males are able to mate repeatedly. Thus, accepting only a single mate might be costly when females mate with a sperm‐depleted male, which may not transfer a sufficient amount of sperm. In the present study, we investigated the reproductive performance in the parasitoid Lariophagus distinguendus Först. (Hymenoptera: Pteromalidae) and studied whether mating with experimentally sperm‐depleted males increases the tendency of females to remate. Males were able to mate with up to 17 females offered in rapid succession within a 10‐h test period. The resulting female offspring, as an indirect measure of sperm transfer, remained constant during the first six matings and then decreased successively with increasing number of copulations by the males. Experimentally sperm‐depleted males continued to mate even if they transferred only small amounts or no sperm at all. Unlike males, the majority of females mated only once during a 192‐h test period. A second copulation was observed only in a few cases (maximum 16%). The frequency of remating was not influenced by the mating status of the first male the females had copulated with, suggesting that these events are not controlled by sperm deficiency of the females. Furthermore, we investigated male courtship behaviour towards mated females. Male courtship intensity towards mated females decreased with increasing time. However, females that had mated with an experimentally sperm‐depleted male did not elicit stronger or longer‐lasting behavioural responses in courting males than those that had mated with a virgin male. As the observed behaviours in L. distinguendus are known to be elicited by a courtship pheromone, these results suggest that females no longer invest in pheromone biosynthesis after mating (as indicated by ceasing behavioural responses of courting males), irrespective of whether they have received a sufficient amount of sperm or not. We discuss the results with respect to a possible mating strategy of sperm‐depleted males.

Cuticular hydrocarbons as contact sex pheromone in the parasitoid Dibrachys cavus

Cuticular hydrocarbons (CHCs) cover the cuticle of virtually any insect and do not only protect them from desiccation, but also possess various communicative functions. Previous studies suggested a function as contact sex pheromones in the parasitic wasp family Pteromalidae, but further species need to be studied before more general conclusions are possible. Here, we demonstrate a contact sex pheromone in females of Dibrachys cavus (Walker) (Hymenoptera: Pteromalidae). Males of this parasitoid were arrested on cadavers of 1‐ to 2‐day‐old females, but neither on newly emerged females nor on males. Extracts from 1‐ to 2‐day‐old females and non‐polar fractions were also behaviorally active suggesting a non‐polar contact sex pheromone. Chemical analyses of bioactive and non‐active CHC profiles by gas chromatography–mass spectrometry and subsequent principal component analysis (PCA) revealed that the bioactive profiles from 1‐ to 2‐day‐old females were clearly distinguishable from all other profiles. Among the chemicals showing the strongest impact in the PCA were the two 3‐methylalkanes 3‐methylnonacosane and 3‐methylhentriacontane, which were absent or trace components in the inactive profiles and whose relative amounts increased in females when they became sexually attractive. The putative function of 3‐methylalkanes as key components of the contact sex pheromones in Pteromalidae is discussed.

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 C25 and C37 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.

Behavioural switch in the sex pheromone response of Nasonia vitripennis females is linked to receptivity signalling

The benefits for insects using chemical information may depend on their physiological state and the behavioural context in which the chemical stimuli are perceived. Hence, insect behaviour is often characterized by olfactory plasticity. Mechanisms triggering changes in the postmating behaviour of insect females are typically mediated by constituents of the male ejaculate. Here we show that the behavioural switch displayed by mated females of the jewel wasp Nasonia vitripennis in response to a male abdominal sex pheromone is independent of the transfer of a male ejaculate. Rather, our results suggest a pheromone interaction in that prior exposure of a female to one pheromone and her subsequent receptivity signal modulated her response to a second one. We tested the pheromone response of females that had been experimentally prevented from receiving sperm or from experiencing certain elements of the male courtship behaviour. We show that the behavioural switch in N. vitripennis females is linked to the receptivity signal shown by females in response to a male-derived oral aphrodisiac during precopulatory courtship.

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.