Tremulatory and Abdomen Vibration Signals Enable Communication through Air in the Stink Bug Euschistus heros

Communication by substrate-borne mechanical signals is widespread among animals but remains one of their least understood communication channels. Past studies of vibrational communication in insects have been oriented predominantly to communication during mating, showing that species- and sex-specific vibrational signals enable recognition and localization of potential mates on continuous solid substrates. No special attention has been paid to vibrational signals with less obvious specificity as well as to the possibility of vibrational communication across substrates that are not in physical contact. We aimed to reinvestigate emission of the aforementioned vibrational signals transmitted through a plant in the stink bug Euschistus heros (Pentatomidae: Pentatominae) and to check whether individuals are able to communicate across adjecent, physically separated substrates. We used laser vibrometry for registration of substrate-borne vibrational signals on a bean plant. Using two bean plants separated for 3 to 7 cm between two most adjacent leaves, we investigated the possibility of transmission of these signals through air. Our study showed that males and females of E. heros communicate using tremulatory, percussion and buzzing signals in addition to the previously described signals produced by vibrations of the abdomen. Contrary to the latter, the first three signal types did not differ between sexes or between pentatomid species. Experiments with two physically separated plants showed significant searching behaviour and localization of vibrational signals of an E. heros male or a female, in response to abdominal vibration produced signals of a pair duetting on the neighbouring plant, in comparison to control where no animals were on the neighbouring plant. We also confirmed that transmission through air causes amplitude and frequency decay of vibrational signals, which suggests high-amplitude, low-frequency tremulatory signals of these stink bugs their most plausible way of communication across discontinuous substrates.     

Exploitation of insect vibrational signals reveals a new method of pest management

Food production is considered to be the main source of human impact on the environment and the concerns about detrimental effects of pesticides on biodiversity and human health are likely to lead to an increasingly restricted use of chemicals in agriculture. Since the first successful field trial, pheromone based mating disruption enabled sustainable insect control, which resulted in reduced levels of pesticide use. Organic farming is one of the fastest growing segments of agriculture and with the continuously growing public concern about use of pesticides, the main remaining challenge in increasing the safety of the global food production is to identify appropriate alternative mating disruption approaches for the numerous insect pests that do not rely on chemical communication. In the present study, we show for the first time that effective mating disruption based on substrate-borne vibrational signals can be achieved in the field. When disruptive vibrational signals were applied to grapevine plants through a supporting wire, mating frequency of the leafhopper pest Scaphoideus titanus dropped to 9 % in semi-field conditions and to 4 % in a mature vineyard. The underlying mechanism of this environmentally friendly pest-control tactic is a masking of the vibrational signals used in mate recognition and location. Because vibrational communication is widespread in insects, mating disruption using substrate vibrations can transform many open field and greenhouse based farming systems.     

Substrate-Borne Vibration Mediates Intrasexual Agonism in the New Zealand Cook Strait Giant Weta (<Emphasis Type="Italic">Deinacrida rugosa</Emphasis>)

Substrate-borne vibrational communication is a common mode of information transfer in many invertebrate groups, with vibration serving as both primary and secondary signal channels in Orthopterans. The Cook Strait giant weta, Deinacrida rugosa (Orthoptera: Anostostomatidae), is an endangered New Zealand insect whose communication system has not been previously described. After field observations of intraspecific interactions in D. rugosa provided preliminary evidence for substrate-borne vibrational communication in the species, we sought to identify the following: vibrational signal structure, the mechanism of signal production, whether signal production is a sexually dimorphic trait, whether substrate-borne signals encode information regarding sender size, the primary social context in which vibration is utilized and finally, the function of vibrational signaling in the species. We used laser Doppler vibrometry to show that D. rugosa males produce low frequency (DF?=?37.00?±?1.63 Hz) substrate-borne vibrations through dorso-ventral tremulation. Rarely produced by females, male signals appear to target rivals while both are in the direct physical presence of a female. Tremulatory responses to playbacks were only produced by males in male-male-female trial contexts, and neither sex exhibited walking vibrotaxis to playback signals, indicating that substrate-borne vibrational signals are not likely a component of the courtship repertoire. While we found that vibrational signal structure was not closely related to signaler size, males that initiated male-male signaling bouts held a significant advantage in contests.     

Reproductive strategy of the Nearctic leafhopper Scaphoideus titanus Ball (Hemiptera: Cicadellidae)

Mating behaviour of Scaphoideus titanus Ball, the vector of the grapevine disease Flavescence dorée, was investigated in order to determine the role of substrate-borne vibrational signals in intra-specific communication and pair formation. Vibrational signals were recorded from grapevine leaves with a laser vibrometer. Signalling activity of single males changed throughout the day and the peak in activity was associated with twilight and early night when 'call and fly' behaviour was observed. Pair formation began with the spontaneous emission of male signals. The male calling signal consisted of a single series of pulses, partially accompanied with a 'rumble'. The male courtship phrase consisted of four consecutive sections characterized by two sound elements, pulse and 'buzz'. Female vibrational signals were emitted only in response to male signals. The female response was a single pulse that closely resembled male pulses and was inserted between pulses within the male signals. All recorded vibrational signals of S. titanus have a dominant frequency below 900 Hz. A unique feature of vibrational communication in S. titanus is well-developed intrasexual competition; males may use alternative tactics, in the form of disturbance signals, or silently approach duetting females (satellite behaviour). While the male-female duet appears to be essential for successful localization of females and copulation, it is also vulnerable to, and easily disrupted by, alternative tactics like masking.     

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.     

Mating behavior of Psammotettix alienus (Hemiptera: Cicadellidae)

The Wheat dwarf virus, the causal agent of the wheat dwarf disease, is transmitted by leafhoppers from the genus Psammotettix and currently the main protection strategy is based on the use of insecticide treatments. Sustainable management strategies for insect vectors should include methods that are targeted to disrupt reproductive behavior and here we investigated the mating behavior of Psammotettix alineus (Dahlbom 1850) in order to determine the role of vibrational signals in intra‐specific communication and pair formation. Both genders spontaneously emit species‐ and sex‐specific calling songs that consisted of regularly repeated pulse trains and differ primarily in pulse train duration and pulse repetition time. Females preferred the conspecific male calling song. After a coordinated exchange of pulse trains, the male approached the stationary female. During the close range courtship and also immediately prior to copulatory attempts distinct male vibrational signals associated with wing flapping and wing vibrations were recorded from the substrate. In the presence of a receptive female, competing males emitted vibrational signals most likely aimed to interfere with male‐female interaction. Mated females regained sexual receptivity after they laid eggs. Although results suggest that the viruliferous status of insects may have an effect on vibrational songs, our current results did not reveal a significant effect of virus on leafhopper performance in mating behavior. However, this study also suggests, that detailed understanding of plant–vector–virus interactions relevant for vector mating behavior is essential for trying new approaches in developing future control practices against plant viruses transmitted by insect vectors.     

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.     

Vibrational playbacks and microscopy to study the signalling behaviour and female physiology of Philaenus spumarius

The meadow spittlebug Philaenus spumarius, vector of the bacterium Xylella fastidiosa, relies on vibrational communication to accomplish mating: the female calls to establish a duet with a male. A deeper knowledge of the species’ reproductive biology and behaviour would provide useful information for developing control techniques based on principles of ‘biotremology’, which studies the vibrational behaviour of animals. Playback tests were conducted on single females and male–female pairs of P. spumarius from June to October 2018, and the features of the recorded calling signals were analysed using a wavelet decomposition. Dissections were performed on females to evaluate the relationship between calling activity and ovarioles development. From August onwards, females started to emit calling signals and to develop ovarioles. Female calling activity, duration of their chirps and their responsiveness to mating increased as the season progressed, and they were correlated with ovarioles’ development and presence of mature eggs. Hence, the ovarian maturation represents a key factor in association with the development of the sexual behaviour of P. spumarius females. Conversely, males produced advertisement signals soon after adult eclosion in May, but these signals were not involved in the pair formation process. Mating was achieved only when males produced courtship signals in response to female calling signals and established with them vibrational duets starting from August. Here, we provide new information regarding the P. spumarius’ ethology and hypothesize that potential mating disruption techniques should consider the insect physiology and be applied when both sexes are responsive to mating signals.     

Pure-tone vibrational signals in small Auchenorrhyncha (Homoptera)

Investigation of vibrational calling signals of about 500 species of small Auchenorrhyncha from Russia and adjacent territories has shown that more than 10% of the species studied produce signals fully or partially consisting of pure-tone components. Among these species, there are forms dwelling on various substrates including thick tree branches, slender twigs, grass stems, petioles, and leaves. Therefore, it is impossible to associate the presence of pure-tone signals in any species with the physical properties of the inhabited substrate. Pure-tone signals were recorded both in the largest and the smallest forms. Consequently, the type of signal frequency spectrum is not related to the insect size. Experiments under natural conditions confirm the assumption that pure-tone signals are more resistant to noise than wide-band ones. This property may compensate for the disadvantage of pure-tone signals arising from stronger attenuation in certain substrates. As a result, neither pure-tone nor noise signals give unambiguous advantages for vibrational communication. Since the carrier frequency of pure-tone signals of small Auchenorrhyncha increases with temperature, it seems to be determined by the contraction frequency of tymbal muscles, rather than by the resonance properties of any cavity or cuticular structure. Regular frequency modulations occur in the signals of many species; moreover, the calling signals of some species include both pure-tone and noise components. In most of the species studied, “frequency tuning” of their signals to the physical properties of a particular substrate is impossible because of the presence of frequency modulations, temperature-related variation in the carrier frequency, or the wide host range and the absence of strong preference for any particular plant organ. Sympatric species may differ in the carrier frequency of their pure-tone signals. If the signals occupy the same frequency range they sometimes differ in their frequency modulation pattern. Consequently, conspecific signal recognition not only by amplitude, but also by frequency pattern is possible in this case, which increases the efficiency of intraspecific communication.     

Vibrational communication and mating behaviour of Dichelops melacanthus (Hemiptera: Pentatomidae) recorded from loudspeaker membranes and plants

Vibrational communication is important for successful mating in various stink bugs species. The vibrational signals from males and females of Dichelops melacanthus Dallas (Hemiptera: Pentatomidae) are recorded from a nonresonant substrate (i.e. a loudspeaker membrane) to characterize the temporal and spectral properties of these vibrational signals, as well as on a resonant substrate (i.e. bean plants) to obtain information about how these signals are altered when they are transmitted through the plants. On the loudspeaker membrane, D. melacanthus males and females emit only one male or one female song, respectively. However, when the insects are placed on bean leaves, a more complex repertoire is recorded, with three different songs for each sex. The first female and male songs appear to have calling functions and the third male and female songs are emitted during courtship. The second female and male songs are emitted after the first song, although their functions in mating behaviour are not clear. The identified repertoire is similar to those of other Neotropical stink bugs, starting with songs 1 and 2 and developing into song 3. Frequency modulation is observed in the female songs recorded from the loudspeaker membrane and the plants. The signals recorded from plants present higher harmonic peaks compared with the signals recorded from the loudspeaker membrane. The presence of species and sex‐specific songs during mating confirms the important role of vibrational communication in mate location and recognition. The temporal and spectral characteristic signals are influenced by the substrate used to record the songs emitted by D. melacanthus.     

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.     

Historical use of substrate-borne acoustic production within the Hemiptera: first record for an Australian Lophopid (Hemiptera, Lophopidae)

Abstract  Here the first record of communication through substrate-borne vibrations for the Lophopidae family is reported. The signals from Magia subocellata that the authors recorded were short calls with a decreasing frequency modulation. Acoustic vibrations have been observed for other families within the Hemiptera and a scenario concerning the historical use of vibrational communication within the Hemiptera is tested using a phylogenetic inference. The most parsimonious hypothesis suggests that substrate-borne communication is ancestral for the hemipteran order and highlights the groups for which future acoustic research should be undertaken.     

A signal-substrate match in the substrate-borne component of a multimodal courtship display

The environment can impose strong limitations on the efficacy of signal transmission. In particular, for vibratory communication, the signaling environment is often extremely heterogeneous at small scales. Nevertheless, natural selection is expected to select for signals well-suited for effective transmission. Here, we test for substrate-dependent signal efficacy in the wolf spider Schizocosa stridulans Stratton 1991. We first explore the transmission characteristics of this important signaling mo-dality by playing recorded substrate-bome signals through three different substrates (leaf Utter, pine litter, and red clay) and measuring the propagated signal. We found that the substrate-borne signal of S stridulans attenuates the least on leaf litter, the substrate upon which the species is naturally found. Next, by assessing mating success with artificially muted and non-muted males across different signaling substrates (leaf litter, pine litter, and sand), we explored the relationship between substrate-bome signaling and signaling substrate for mating success. We found that muted males were unsuccessful in obtaining copulations re-gardless of substrate, while mating success was dependent on the signaling substrate for non-muted males. For non-muted males, more males copulated on leaf litter than any other substrate. Taken together, these results confirm the importance of sub-strate-borne signaling in & stridulans and suggest a match between signal properties and signal efficacy - leaf litter transmits the signal most effectively and males are most successful in obtaining copulations on leaf litter.     

Substrate Vibrations during Courtship in Three Drosophila species

While a plethora of studies have focused on the role of visual, chemical and near-field airborne signals in courtship of Drosophila fruit flies, the existence of substrate-borne vibrational signals has been almost completely overlooked. Here we describe substrate vibrations generated during courtship in three species of the D. melanogaster group, from the allegedly mute species D. suzukii, its sister species D. biarmipes, and from D. melanogaster. In all species, we recorded several types of substrate vibrations which were generated by locomotion, abdominal vibrations and most likely through the activity of thoracic wing muscles. In D. melanogaster and D. suzukii, all substrate vibrations described in intact males were also recorded in males with amputated wings. Evidence suggests that vibrational signalling may be widespread among Drosophila species, and fruit flies may provide an ideal model to study various aspects of this widespread form of animal communication.     

Orientation of a hemipteran predator to vibrations produced by feeding caterpillars

Observations of search behavior in the predatory stinkbugPodisus maculiventris (Say) suggested that vibrations produced by prey as they chew on leaves may be an important cue used by this predator to locate prey. To test this hypothesis, studies were conducted to determine ifP. maculiventris search and make directional choices in response to vibrational stimuli produced by feeding green cloverworms,Plathypena scabra (F.), and to recordings of chewing vibrations. Modified soybean plants [Glycine max (L.)] were used in Y-choice tests. Individuals exposed to vibrational signals finished trials significantly more often on branches through which vibrations entered the plants than on no-stimulus branches. Also, a significantly higher proportion of individuals that initially moved onto branches with no stimulus reversed course than did those moving up branches with vibrational stimuli. The response ofP. maculiventris individuals to vibrational signals produced by a common prey species demonstrates that these predators are capable of using substrate-borne vibrations as cues for prey location.     

Vibrational Communication and the Ecology of Group-Living, Herbivorous Insects

Communication among members of a colony is a key feature ofthe success of eusocial insects. The same may be true in otherforms of insect sociality. I suggest that substrate-borne vibrationalcommunication is important in the success of group-living, herbivorousinsects. I examine three challenges encountered by herbivorousinsects: locating and remaining in a group of conspecifics;locating food resources; and avoiding predation. Studies ofgroups of immature treehoppers, sawflies and butterflies suggestthat vibrational communication can be important in each of thesecontexts, enhancing the ability of these group-living herbivoresto exploit the resources of their host plants.     

Pea leafminer Liriomyza huidobrensis (Diptera: Agromyzidae) uses vibrational duets for efficient sexual communication

The pea leafminer (Liriomyza huidobrensis) is a notorious pest of vegetables and ornamental plants worldwide. Despite a large number of studies on its biology and ecology, the courtship behavior and sexual communication of this species remain unclear. Here, we studied vibrational communication in the sexual interaction of the pea leafminer. On host plant leaves, females and males behaviorally displayed the bobbing-quivering alternation, which finally led to copulation. Moreover, records of laser vibrometry revealed threesignal duets underlying the behavioral alternation. Sexually mature males spontaneously emitted calls (MCs) to initiate the duets. The females rapidly responded to MCs by emitting replies (FRs) that are longer in duration. The FRs further triggered male replies (MRs) in their search for potential partners. Leafminer-produced vibrational signals convey efficient information to partners and generate pair formation on stretched substrates, such as plant leaves and nylon mesh, but cannot elicit responses on dense substrates, such as glass and plastic. Vibrational playbacks of both MCs and FRs can elicit replies in females and males, respectively. This study completely characterizes substrate-borne vibrational duets in a dipteran insect. The discovery of vibrational sex signals in the pea leafminer provides new insights for the development of novel approaches to control the pest and its relative species.     

Mate searching in Ennya maculicornis (Membracidae: Polyglyptini) initiated by females: behavioural and acoustic descriptions

1. In treehoppers in which courtship has been studied, males initiate the search for females by periodically emitting a vibrational signal. The responses by the female are used by males as a beacon and give rise to a duet. 2. Courtship and mating of the treehopper Ennya maculicornis were characterised through the simultaneous recording of vibrational signals and the behaviour of males and females in an arena. 3. In E. maculicornis, female initiated mate searching. Females produced two types of signals during the this process: (i) a signal that preceded the approach by the male and (ii) a signal that preceded mating. Males emitted two signals associated with two stereotyped body movements: (i) a signal produced as a response to the first signal emitted by the female, involving a change in the male's locomotory mode and the approach to the female, and (ii) a signal produced after finding and holding on to the female, involving simultaneous abdomen raising and wing fluttering. These signals were repeated several times before the female emitted the second signal. The four signalling patterns were observed in all recordings in which mating was observed. When any of the signals was missing, mating did not occur. 4. Female‐biased sex ratios in E. maculicornis, along with iteroparity, are suggested to explain the initiation of mate searching behaviour by females. A comparison of data with that from other treehoppers indicates that vibrational signals and their associated behaviour are more diverse among treehoppers than has been appreciated previously.     

Acoustics, context and function of vibrational signalling in a lycaenid butterfly-ant mutualism

Juveniles of the Australian common imperial blue butterfly, Jalmenus evagoras, produce substrate-borne vibrational signals in the form of two kinds of pupal calls and three larval calls. Pupae stridulate in the presence of conspecific larvae, when attended by an ant guard, and as a reaction against perturbation. Using pupal pairs in which one member was experimentally muted, pupal calls were shown to be important in ant attraction and the maintenance of an ant guard. A pupa may use calls to regulate levels of its attendant ants and to signal its potential value in these mutualistic interactions. Therefore substrate-borne vibrations play a significant role in the communication between J. evagoras and its attendant ants and pupal calls appear to be more than just signals acting as a predator deterrent. Similarly, caterpillars make more sound when attended by Iridomyrmex anceps, suggesting that larval calls may be important in mediating ant symbioses. One larval call has the same mean dominant frequency, pulse rate, bandwidth and pulse length as the primary signal of a pupa, suggesting a similarity in function. Copyright 2000 The Association for the Study of Animal Behaviour.