Species Recognition During Substrate-Borne Communication in <Emphasis Type="Italic">Nezara viridula</Emphasis> (L.) (Pentatomidae: Heteroptera)

The information code in the temporal and spectral characteristics of the substrate-borne communication signals produced by insects has been primarily studied in insects in the suborder Auchenorrhyncha. In the present study we investigated which of the female calling song (FCS) parameters in Nezara viridula (L.) (Heteroptera, Pentatomidae) are essential for recognition by conspecific males. In playback experiments we measured male vibrational responsiveness to FCS signals varying in the durations of pulse trains and inter-pulse train intervals, repetition times, duty cycles, and dominant frequencies, and determined the preference range for each specific parameter. Males were able to distinguish songs of different temporal and frequency parameters and responded best to values characteristic of the song of conspecific females. Signal recognition is achieved on the basis of two temporal filters tuned to the durations of the pulse train and inter-pulse train interval. Males responded best to the dominant frequency characteristic of conspecific songs, which are tuned to the resonant properties of the herbaceous plants used for intraspecific signal transmission during communication.     

Male–male interactions and male mating success in the leafhopper Aphrodes makarovi

In mating systems based on substrate‐borne vibrations, sexual communication often involves a reciprocal exchange of species‐ and sex‐specific vibrational signals and male is searching for a stationary female. In the leafhopper Aphrodes makarovi, female reply is essential for successful location of the female and its variable duration directly affects male's costs associated with signalling and searching. We studied male and female behaviour in a trio situation (two males and one female), and our results show that male–male competition had important effects on male mating success. Females replied equally to advertisement calls emitted by the winning and losing males and mated with the first male that located them, regardless of his investment in calling effort. Males eavesdropped to male–female duet maintained by the rival, and the winners were better at exploiting female replies to the rival's advertisement calls by silently approaching the female. To interfere with the ongoing male–female duet, males also emitted masking signals overlapping the latter part of the female reply. More overlapped female replies were registered in response to the losers and masking signals most likely delay the rival in reaching the female. Our study shows that a comprehensive understanding of male mating success and female preferences in vibrational duetting systems requires also investigations in more complex settings that more realistically represent the situation in nature.     

The Effect of Timing of Female Vibrational Reply on Male Signalling and Searching Behaviour in the Leafhopper Aphrodes makarovi

Sexual communication in animals often involves duetting characterized by a coordinated reciprocal exchange of acoustic signals. We used playback experiments to study the role of timing of a female reply in the species-specific duet structure in the leafhopper Aphrodes makarovi (Hemiptera: Cicadellidae). In leafhoppers, mate recognition and location is mediated exclusively by species- and sex-specific substrate-borne vibrational signals and a female signal emitted in reply to male advertisement calls is essential for recognition and successful location of the female. In A. makarovi, males have to initiate each exchange of vibrational signals between partners, and in a duet the beginning of a female reply overlaps the end of the male advertisement call. Results of playback treatments in which female replies were delayed and did not overlap with the male call revealed that in order to trigger an appropriate behavioural response of the male, female reply has to appear in a period less than 400 ms after the end of the initiating male call. Results also suggest that males are not able to detect a female reply while calling, since female reply that did not continue after the end of male call triggered male behaviour similar to behaviour observed in the absence of female reply. Together, our results show that vibrational duets are tightly coordinated and that the species-specific duet structure plays an important role in mate recognition in location processes.     

Acoustic behaviour and phonotaxis in the duetting ephippigerines,Steropleurus nobrei* and Steropleurus stali (Tettigoniidae)

Both sexes of the ephippigerines Steropleurus stall and S. nobrei can stridulate and produce multisyllabic calls which are described. Female stridulation is in response to the conspecific male call. In both species cither sex can perform phonotaxis on the call of the conspecific member of the opposite sex, but ignore the calls of other species. The parameters of the calls are examined with the conclusion that the only reliably distinctive feature is the modal or carrier frequency generated during stridulation. There are frequency differences between male and female calls in both species. Males only perform phonotaxis on female replies generated in response to their own call, implying that there is also some time window involved. Male phonotaxis was faster and more accurate than that of the female. Acoustic rivalry and aggression were also noted, particularly in females.     

Evolution and Diversity of Vibrational Signals in Mantophasmatodea (Insecta)

Vibrational communication for species identification and mate location is widespread among insects. We investigated the vibrational communication signals of 13 species of the insect order Mantophasmatodea (Heelwalkers). Males and females produce percussive signals by tapping their abdomens on the substrate to locate conspecific mates. We show that male and female calls are of similar general structure but differ in temporal characteristics. Using a principal component analysis, we demonstrate that most species can be distinguished by their calls only. Mapping the calls onto an existing molecular phylogenetic tree reveals a slow diverging drift of male call pattern but no specific trend. For females, a trend from faster towards slower pulse repetition times is indicated. Two sympatric species, Karoophasma biedouwense and Viridiphasma clanwilliamense (Austrophasmatidae), exhibit very different call parameters. The latter species produces calls rather different from all other investigated species, which might hint at reproductive character displacement.     

Duetting Behaviour in the Leafhopper Aphrodes makarovi (Hemiptera: Cicadellidae)

Mate recognition and location in Cicadellidae is mediated exclusively via substrate-borne vibrational signals. In the present study we investigated vibrational signals and mate searching behaviour of the leafhopper Aphrodes makarovi. We studied mating behaviour and exchange of vibrational signals between live insects and in playback experiments. Males emitted long and complex calling signals composed of several sections. Female reply was long and always overlapped the end of the male call. The exchange of male and female vibrational signals was a complex and dynamic interaction during which both partners modified their signals according to partner’s reply. The duration of female reply was influenced by the duration of the male call to which she was responding, while the duration of male call was influenced by the duration of the previous female reply. Such relationship suggests the role of sexual selection in the evolution of male vibrational signals.     

The effect of vibratory disturbance on sexual behaviour of the southern green stink bug Nezara viridula (Heteroptera, Pentatomidae)

The effect of vibratory disturbance on sexual behaviour and substrate-borne sound communication of the southern green stink bug, Nezara viridula L. was studied. Disturbance signals do not change the time N. viridula males need to locate the source of vibratory signals, but decrease the number of males responding with the calling and courtship song to calling females. Female N. viridula proceed calling during stimulation with disturbance signals but some of them change the song rhythm by skipping one or more signal intervals or emitting the repelling signals. The number of females which change the dominant frequency of the calling song decreases proportionally with increasing differences between the dominant frequency of the disturbance signals and the emitted female calling song. Variation of the song dominant frequency probably serves females to avoid interference by increasing the signal to noise ratio. Signal duration and repetition rate do not change significantly when the female is stimulated with the disturbance signals. This indicates that frequency shift by calling females is the main strategy for reducing interference by competitive signalers in N. viridula vibrational communication.     

Species-Specificity of Chemical Signals: Silk Source Affects Discrimination in a Wolf Spider (Araneae: Lycosidae)

Female spiders deposit chemical cues that elicit male courtship behavior with silk. These cues are often assumed to be species-specific although male spiders may court in response to chemical cues of closely-related species. We used behavioral assays to test the extent of species discrimination of female chemical cues by male Schizocosa ocreata, a wolf spider (Lycosidae). Discrimination, expressed as relative courtship intensity of males, varied significantly with phylogenetic distance. Males did not discriminate between female cues of conspecifics and a sibling species, S. rovneri. Courtship response was intermediate for another species within the ocreata clade and not different from control for spiders outside the clade. These findings support the sibling species status of S. ocreata and S. rovneri, and also suggest the composition of female chemical signals is conserved across closely related wolf spider species.     

Molecular diagnostics reveal spiders that exploit prey vibrational signals used in sexual communication

Vibrational signalling is a widespread form of animal communication and, in the form of sexual communication, has been generally regarded as inherently short‐range and a private communication channel, free from eavesdropping by generalist predators. A combination of fieldwork and laboratory experiments was used to test the hypothesis that predators can intercept and exploit such signals. First, we developed and characterized PCR primers specific for leafhoppers of the genus Aphrodes and specifically for the species Aphrodes makarovi. Spiders were collected from sites where leafhoppers were present and screened with these primers to establish which spider species were significant predators of this species during the mating period of these leafhoppers. Analysis using PCR of the gut contents of tangle‐web spiders, Enoplognatha ovata (Theridiidae), showed that they consume leafhoppers in the field at a greater rate when signalling adults were present than when nymphs were dominant, suggesting that the spiders were using these vibrations signals to find their prey. Playback and microcosm experiments then showed that E. ovata can use the vibrational signals of male leafhoppers as a cue during foraging and, as a result, killed significantly more male than female A. makarovi. Our results show, for the first time, that arthropod predators can exploit prey vibrational communication to obtain information about prey availability and use this information to locate and capture prey. This may be a widespread mechanism for prey location, one that is likely to be a major unrecognized driver of evolution in both predators and prey.     

Vibrational directionality in the southern green stink bug, Nezara viridula (L.), is mediated by female song

We tested the hypothesis that male southern green stink bugs, Nezara viridula (L.), use substrate-borne songs to locate females. We recorded the responses of bugs on plants to the vibrations caused by a prerecorded female song and by an artificial sound. The female song caused males to walk, to respond with the calling and courtship songs and to approach the source of the song with characteristic search behaviour at junctions between branches on the plants. At a junction, a searching male stopped, stretched his legs and antennae and compared the vibratory signals on the two branches, with different combinations of legs and antennae. The males then left the junction and approached the source of the vibration. Males located the loudspeaker significantly more frequently in the presence than in the absence of vibratory stimuli on cyperus, Cyperus alternifolius L., and beans, Phaseolus vulgaris L. Vibrational directionality was also elicited by artificial pure tones whose spectral and temporal parameters were similar to those of natural female song. Females showed no reaction to vibratory stimulation and no vibrational directionality. We discuss possible mechanisms underlying vibrational directionality in the light of expected signal changes during transmission through plants. Copyright 1999 The Association for the Study of Animal Behaviour.     

Differences in preference for species-specific female calls between acoustically experienced and acoustically naive maleRibautodelphax planthoppers (Homoptera: Delphacidae)

Males of the planthopper Ribautodelphax imitanswere exposed to playbacks of either conspecific or heterospecific (R. imitantoides)female calls during their development from egg to adult, and thereafter these, as well as naive males,were offered a two- way choice between these calls. Males of all treatments approached the conspecific call significantly more often. However, males primed by the conspecific call chose the heterospecific call almost four times less often than did males primed by heterospecific calls or naive males, thus showing that the preference for conspecific calls can be partly learned. Males primed by heterospecific calls performed very similarly to completely naive males, suggesting that the signal recognition mechanism is much less sensitive to heterospecific calls than to conspecific calls. Males with experience of the conspecific female call tended to take more time to reach the call source in the trials than both other types of males. The evolutionary implications of these findings are discussed.     

Dermacentor variabilis and Dermacentor andersoni: Genital sex pheromones

There is evidence for the existence of a previously undescribed sex pheromone (or pheromones) in the ticks Dermacentor variabilis and D. andersoni. In addition to 2,6-dichlorophenol, which attracts mate-seeking males, a pheromone released on the cuticle of the female genital area enables the sexually excited male to locate the gonopore. The compound (or compounds) appears to act as a contact pheromone; male copulation responses are greatly reduced when the female genital surface is washed with solvents, especially hexane. It is also a potent excitant; males will puncture or dislodge barriers placed over the gonopore to copulate. However, the response is eliminated if the genital area is washed (hexane or acetone) prior to sealing the gonopore, suggesting the reproductive system as the source of the pheromone. A species specific copulation-eliciting pheromone appears necessary to excite the male to form and implant its spermatophore in the vulva of a conspecific female. Males encountering trans-specific females probe their gonopores, but mating attempts are almost always aborted within 5–10 min. The copulation-eliciting pheromone may be the same, or similar, to that used to locate the gonopore. Physical differences in the shape of the female gonopore in the two species, although slight, may contribute to the male's ability to identify conspecific females. Using this pheromone-guided process of attraction and identification, females present in mixed species populations will almost always be distinguished and inseminated by conspecific mate-seeking males.     

The role of signals of different modalities in initiating vibratory communication in Nezara viridula

Signals of different modalities are involved in the behaviour of the green stink bug, Nezara viridula (L.) (Pentatomidae, Heteroptera). Long range attraction is mediated by male pheromones, resulting in aggregation of bugs on the same plant where vibratory signals, vision and various chemical signals become important. Both males and females sing spontaneously. When both are on the plant, males start vibratory communication as often as females. Females induce the exchange of vibratory signals spontaneously or triggered by the male pheromone while males initiate the duet either spontaneously or after seeing the female. Males and females sing spontaneously and respond to signals of different modalities more often in the daylight than in the dark. Long lasting autonomous emission of the female calling song is present when triggered by the male pheromone and males respond to female calling predominantly by the emission of the courtship song.     

Vibrational Communication in Two Sympatric Species of Mantophasmatodea (Heelwalkers)

The communication via percussion of the abdomen on the substrate for species recognition and mate location of males and females of two sympatric species of the recently described insect order Mantophasmatodea (Heelwalkers) was investigated. Each sex produced a single and distinctive call. The female call consisted of repeated single pulses, whereas the more complex male call comprised repeated pulse trains. The calls of males and females of the two species were of similar general structure, but differed in most temporal characters such as pulse and pulse train repetition time. In behavioral playback experiments females reacted to the call of conspecific males by calling and decreasing locomotion. When stimulated with the call of the heterospecific, sympatric male, females showed no reaction. Males exhibited abdominal rubbing, high tapping rates, increased activity (both movement and active searching) as well as characteristic searching behavior at branch nodes, when presented with the conspecific female call. Being stimulated with the playback of the heterospecific female call and the conspecific male call respectively, males responded with less intense locomotor and searching behavior. The drumming behavior in the control situation (no playback) suggests that males sometimes call in the absence of other individuals.     

Responses of Second-Instar Male Nymphs of Four Mealybug Species (Hemiptera: Pseudococcidae) to Conspecific and Heterospecific Female Sex Pheromones

The response of the late second-instar male nymphs of the mealybug species (Hemiptera, Pseudococcidae), Planococcus citri (Risso), Planococcus ficus (Signoret), Pseudococcus cryptus Hempel Nipaecoccus viridis (Newstead), to their conspecific and heterospecific female pheromone was studied. Males that exhibited the typical appearance of late-second-instar nymphs were tested. The male behavior was monitored soon after their exposure to the tested female sex pheromone in glass Petri dish arenas. Male nymph behavior toward the pheromone source was characterized based on their aggregation on the disks in the arena. Males of all four tested mealybug species were attracted to their conspecific female pheromone. By contrast, almost no interceptions of male nymphs with disks impregnated with a heterospecific female pheromone were observed. The mode of attraction of each of male nymphs of P. ficus, among most of the tested individuals (>80%), to the conspecific female sex pheromone, (S)-lavandulyl senecioate and or (S)-lavandulyl isovalerate, was the same as the mode of attraction latter on as adult. We suggest that by being attracted to the conspecific pheromone these males may direct themselves to a suitable pupation site near conspecific non-sibling mature females, thus preventing inbreeding. The repellency of heterospecific sex pheromone to males that are looking for a pupation site suggests that the latter try to avoid close contact with heterospecific females.     

Male adaptations to minimize sexual cannibalism during reproduction in the funnel‐web spider Hololena curta

Males of many spider species risk being attacked and cannibalized while searching for, courting, and mating with conspecific females. However, there are exceptions. We show that the funnel‐web spider, Hololena curta, has 3 adaptations that minimize risk to males during courtship and mating, and enhance reproductive success. First, males detected chemical or tactile signals associated with webs of virgin females, and differentiated them from webs of mated females, enabling males to increase encounter rates with virgin females and avoid aggressive mated females. Second, males produced stereotyped vibrational signals during courting which induced female quiescence and suppressed female aggression. Third, when touched by males, sexually receptive females entered a cataleptic state, allowing males to safely approach and copulate. Because males can mate multiple times and the sex ratio in natural populations of H. curta is female biased, overall reproductive output is likely increased by males of this species avoiding sexual cannibalism.     

Vibratory signals of the harlequin bug and their transmission through plants

Abstract. Males of the harlequin bug, Murgantia histrionica (Hahn), produce five different vibrational songs, whereas females produce one song. Songs differ from those of other stink bugs primarily in their species‐specific temporal characteristics. The broad band male courtship songs of M. histrionica are achieved by a combination of different frequency modulated and/or narrow band subunits, with several higher harmonic frequencies. Males rather than females initiate substrate‐borne vibrational communication, and the longer‐range calling songs found typically in other pentatomid species are lacking. Interindividual differences in song temporal and spectral characteristics are discussed. Transmission of vibrational songs through a cabbage head is more efficient along veins than along lamina. Attenuation of signals transmitted through veins is low and similar to that reported previously for plant stalks. On the leaf vein, distances between peak amplitude minima and maxima are different for the dominant and subdominant frequencies. At any distance from the vibration source, a different relationship between spectral peak amplitudes can be recorded. Resolution of these differences, together with velocity differences between signals recorded on the vein and lamina, may help small stink bugs to estimate distance and to locate each other on a plant.     

Predatory bug <Emphasis Type="Italic">Picromerus bidens</Emphasis> communicates at different frequency levels

The Asopinae (Heteroptera: Pentatomidae) are a subfamily of stinkbugs with predaceous feeding habits and poorly understood communication systems. In this study we recorded vibratory signals emitted by Picromerus bidens L. on a non-resonant substrate and investigated their frequency characteristics. Males and females produced signals by vibration of the abdomen and tremulation. The female and male songs produced by abdominal vibrations showed gender-specific time structure. There were no differences in the temporal patterns of male or female tremulatory signals. The signals produced by abdominal vibrations were emitted below 600 Hz whereas tremulatory signals had frequency ranges extending up to 4 kHz. Spectra of male vibratory signals produced by abdominal vibrations contained different peaks, each of which may be dominant within the same song sequence. Males alternated with each other during production of rivalry signals, using different dominant frequency levels. We show that the vibratory song repertoire of P. bidens is broader than those of other predatory stinkbugs that have been investigated. The emission of vibrational signals with different dominant frequencies but the same production mechanism has not yet been described in heteropteran insects, and may facilitate location of individual sources of vibration within a group.     

Substrate-borne vibrational signals and stridulatory organs for sexual communication in leafminer,Liriomyza sativae (Diptera: Agromyzidae)

The vegetable leafminer (Liriomyza sativae [Burgess]) is a highly polyphagous pest that threatens vegetables and horticultural plants. Although sexual communication is a key component of the animal behavioral repertoire, the mechanism underlying sexual communication in L. sativae remains to be elucidated. Here, we used laser vibrometry to characterize the vibrational signals emitted by L. sativae during pair formation. By emitting trains of vibrational pulses (male calling) the male initiated communication on the host plant. The female then became immobile and responded to the male calling by emitting replies (female replies), which in turn triggered male replies consisting of a rapid series of chirps and trills. If the female replied, a continuous exchange of male and female replies ensued, representing a duet. In playback trials, a playback signal caused responses from the opposite sex. Moreover, scanning electron microscopy revealed vibration-producing stridulatory organs in both male and female individuals. The files in males were more developed than those in females, and older male specimens had more signs of abrasion. The results provide new insight into the mating biology of L. sativae.     

Female competition for availability of males in insects: the Nezara viridula (Linnaeus, 1758) model

Multimodal communication in solitary stinkbugs enables them to meet, mate and copulate. Many plant‐dwelling species exchange information during the calling phase of mating behavior using substrate‐borne vibratory signals. A female‐biased gender ratio induces rivalry and competition for a sexual partner. Female competition for males, first described among Heteroptera in three stinkbug species, revealed species specific differences and opened the question of plasticity in individually emitted temporal and frequency signal characteristics during calling and rival alternation. To address this question and gain an insight into the mechanisms underlying stinkbug female rivalry, we compared the characteristics of alternated signals in the southern green stinkbug Nezara viridula (Linnaeus, 1758) (Hemiptera: Pentatomidae). Compared to male rivalry, female rivalry is more complex, lasts longer and runs through successive phases by a combination of different song types. The male pheromone triggers alternation between females, producing song pulses that occasionally overlap each other. One female initiates the rivalry by changing individual pulses into pulse trains of three different types. The competing female alternates with pulses of changed temporal characteristics at lower levels of rivalry and by varying the frequency characteristics of pulse trains at higher levels. During female rivalry, the male either stops responding or occasionally emits calling and courtship signals in response to the female that has produced signals of steady temporal characteristics. Female rivalry shows complex and species specific patterns of information exchange at different levels with a broad‐range variation of temporal and frequency characteristics of, until now, unidentified vibratory emissions.