Stink Bug Communication and Signal Detection in a Plant Environment.

Plants influenced the evolution of plant-dwelling stink bugs' systems underlying communication with chemical and substrate-borne vibratory signals. Plant volatiles provides cues that increase attractiveness or interfere with the probability of finding a mate in the field. Mechanical properties of herbaceous hosts and associated plants alter the frequency, amplitude, and temporal characteristics of stink bug species and sex-specific vibratory signals. The specificity of pheromone odor tuning has evolved through highly specific odorant receptors located within the receptor membrane. The narrow-band low-frequency characteristics of the signals produced by abdomen vibration and the frequency tuning of the highly sensitive subgenual organ vibration receptors match with filtering properties of the plants enabling optimized communication. A range of less sensitive mechanoreceptors, tuned to lower vibration frequencies, detect signals produced by other mechanisms used at less species-specific levels of communication in a plant environment. Whereas the encoding of frequency-intensity and temporal parameters of stink bug vibratory signals is relatively well investigated at low levels of processing in the ventral nerve cord, processing of this information and its integration with other modalities at higher neuronal levels still needs research attention.

Inhibitory Copulation Effect of Vibrational Rival Female Signals of Three Stink Bug Species as a Tool for Mating Disruption.

Stink bugs are major pests in diverse crops around the world. Pest management strategies based on insect behavioral manipulation could help to develop biorational management strategies of stink bugs. Insect mating disruption using vibratory signals is an approach with high potential for pest management. The objective of this work was to investigate the effect of conspecific female rival signals on the mating behavior and copulation of three stink bug species to establish their potential for mating disruption. Previously recorded female rival signals were played back to bean plants where pairs of the Neotropical brown stink bug, Euschistus heros, and two green stink bugs, Chinavia ubica and Chinavia impicticornis were placed. Vibratory communication and mating behavior were recorded for each pair throughout the experimental time (20 min). Female rival signals show a disrupting effect on the reproductive behavior of three conspecific investigated stink bug species. This effect was more clearly expressed in E. heros and C. ubica than in C. impicticornis. The likelihood of copulating in pairs placed on control plants, without rival signals, increased 29.41 times in E. heros, 4.6 times in C. ubica and 1.71 times in C. impicticornis. However, in the last case, the effect of female rivalry signals in copulation was not significant. The effect of mating disruption of female rival signals of the three stink bug species may originate from the observed reduction in specific vibratory communication signals emitted, which influences the duet formation and further development of different phases of mating behavior. Our results suggest that female rival signals have potential for application in manipulation and disruption of mating behavior of stink bugs. Further work needs to focus on the effects of female rival signals used in long duration experiments and also their interactions with chemical communication of stink bugs.

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.

Interference of Overlapping Insect Vibratory Communication Signals: An Eushistus heros Model.

Plants limit the range of insect substrate-borne vibratory communication by their architecture and mechanical properties that change transmitted signal time, amplitude and frequency characteristics. Stinkbugs gain higher signal-to-noise ratio and increase communication distance by emitting narrowband low frequency vibratory signals that are tuned with transmission properties of plants. The objective of the present study was to investigate hitherto overlooked consequences of duetting with mutually overlapped narrowband vibratory signals. The overlapped vibrations of the model stinkbug species Eushistus heros, produced naturally or induced artificially on different plants, have been analysed. They represent female and male strategies to preserve information within a complex masked signal. The brown stinkbugs E. heros communicate with species and gender specific vibratory signals that constitute characteristic duets in the calling, courtship and rivalry phases of mating behaviour. The calling female pulse overlaps the male vibratory response when the latency of the latter is shorter than the duration of the female triggering signal or when the male response does not inhibit the following female pulse. Overlapping of signals induces interference that changes their amplitude pattern to a sequence of regularly repeated pulses in which their duration and the difference between frequencies of overlapped vibrations are related inversely. Interference does not occur in overlapped narrow band female calling pulses and broadband male courtship pulse trains. In a duet with overlapped signals females and males change time parameters and increase the frequency difference between signals by changing the frequency level and frequency modulation pattern of their calls.

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.

Mating behaviour and vibratory signalling in non-hearing cave crickets reflect primitive communication of Ensifera.

In Ensifera, the lack of well-supported phylogeny and the focus on acoustic communication of the terminal taxa hinders understanding of the evolutionary history of their signalling behaviour and the related sensory structures. For Rhaphidophoridae, the most relic of ensiferans following morphology-based phylogenies, the signalling modes are still unknown. Together with a detailed description of their mating process, we provide evidence on vibratory signalling for the sympatric European species Troglophilus neglectus and T. cavicola. Despite their temporal shift in reproduction, the species' behaviours differ significantly. Signalling by abdominal vibration constitutes an obligatory part of courtship in T. neglectus, while it is absent in T. cavicola. Whole-body vibration is expressed after copulation in both species. While courtship signalling appears to stimulate females for mating, the function of post-copulation signals remains unclear. Mating and signalling of both species were found to take place in most cases on bark, and less frequently on other available substrates, like moss and rock. The signals' frequency spectra were substrate dependent, but with the dominant peak always expressed below 120 Hz. On rock, the intensity of T. neglectus courtship signals was below the species' physiological detection range, presumably constraining the evolution of such signalling in caves. The species' behavioural divergence appears to reflect their divergent mating habitats, in and outside caves. We propose that short-range tremulation signalling in courtship, such as is expressed by T. neglectus, represents the primitive mode and context of mechanical signalling in Ensifera. The absence of high-frequency components in the signals may be related to the absence of the crista acoustica homologue (CAH) in the vibratory tibial organ of Rhaphidophoridae. This indirectly supports the hypothesis proposing that the CAH, as an evolutionary precursor of the ear, evolved in Ensifera along the (more) complex vibratory communication, also associated with signals of higher carrier frequency.

Resonance in herbaceous plant stems as a factor in vibrational communication of pentatomid bugs (Heteroptera: Pentatomidae).

Pentatomid bugs communicate using substrate-borne vibrational signals that are transmitted along herbaceous plant stems in the form of bending waves with a regular pattern of minimal and maximal amplitude values with distance. We tested the prediction that amplitude variation is caused by resonance, by measuring amplitude profiles of different vibrational pulses transmitted along the stem of a Cyperus alternifolius plant, and comparing their patterns with calculated spatial profiles of corresponding eigenfrequencies of a model system. The measured distance between nodes of the amplitude pattern for pulses with different frequencies matches the calculated values, confirming the prediction that resonance is indeed the cause of amplitude variation in the studied system. This confirmation is supported by the resonance profile obtained by a frequency sweep, which matches theoretical predictions of the eigenfrequencies of the studied system. Signal bandwidth influences the amount of amplitude variation. The effect of both parameters on signal propagation is discussed in the context of insect vibrational communication.

Basidiomycete Clitocybe nebularis is rich in lectins with insecticidal activities.

Basidiomycete mushrooms are a rich source of unique substances, including lectins, that could potentially be useful in biotechnology or biomedical applications. Lectins are a group of carbohydrate-binding proteins with diverse biological activities and functions. Here, we demonstrate the presence of a number of lectins in the basidiomycete mushroom Clitocybe nebularis. Glucose-, galactose-, sucrose-, lactose-, and Sepharose-binding lectins were isolated from fruiting bodies using affinity chromatography on Sepharose-immobilized sugars or on Sepharose. The lectins were characterized biochemically and their binding specificities examined by agglutination and agglutination inhibition assays. In addition, insecticidal and anti-nutritional properties of the lectins were studied against a model organism, fruit fly (Drosophila melanogaster), and Colorado potato beetle (Leptinotarsa decemlineata). Of the several basidiomycete mushrooms screened, C. nebularis extract showed the most potent insecticidal activity. Sucrose-binding lectin showed the strongest activity against D. melanogaster, followed by lactose- and galactose-binding lectins. Feeding bioassays with Colorado potato beetle revealed that C. nebularis extract exhibited high anti-nutritional activity against the insect; and of those tested, only lactose-binding lectin, named CNL showed the effect. Mushroom C. nebularis is shown to be rich in a variety of lectins with versatile biological activities, including insecticidal and anti-nutritional effects. C. nebularis lectins could thus have potential for use as natural insecticides.

Effects of heterospecific and conspecific vibrational signal overlap and signal-to-noise ratio on male responsiveness in Nezara viridula (L.).

Animals often communicate in environments with high levels of biotic noise that arises from the signals of other individuals. Although effects of background biotic noise on mate recognition and discrimination have been widely studied in air-born sound communication, little is known about incidental interference between signallers that use substrate-borne vibrational signals. In this study we investigated the ability of males of the southern green stink bug Nezara viridula (L.) (Heteroptera: Pentatomidae) to recognize conspecific female song in the presence of biotic noise originating from conspecific and heterospecific vibrational signals. We tested male responsiveness on a bean plant in playback experiments. One leaf was vibrated with conspecific female song, while to the other one we simultaneously applied either heterospecific female signal or various altered conspecific signals with different temporal parameters. We tested males in three levels of biotic noise, ranging from +6 dB to -6 dB and we compared male responsiveness in each treatment with response obtained in unilateral treatment with unaltered conspecific female calling song. Male responsiveness was reduced in the presence of heterospecific signals or when background noise from conspecific signals obscured the species-specific temporal pattern of conspecific female song. By contrast, the presence of two sources of conspecific female songs had a positive effect on male responsiveness, for as long as the signal repetition rate of perceived song did not differ from the species-specific value. In the presence of interfering background signals, searching activity was less affected than male signalling. Increased signal-to-noise ratio restored male responsiveness to the level expressed in unilateral stimulation with conspecific female song. The results are discussed with regard to male behavioural strategies for vibrational communication in a noisy environment.

Sound is involved in multimodal communication of Loxosceles intermedia Mello-Leitão, 1934 (Araneae; Sicariidae).

Signals of different modalities are involved during courtship of the brown spider Loxosceles intermedia. A spine on the pedipalp is rubbed against the grooves on the retrolateral region of the chelicerae producing stridulatory signals, which have a dominant frequency of the airborne component range around 770 Hz for females and around 170 Hz for males. These values are significantly lower for the substrate-borne component. The sound pressure level of stridulatory signals lies below 50 dB and the velocity values below 1mm/s. The copulation frequency does not depend on the presence of pedipalps in females; however, in males the removal of pedipalps decreases the courtship frequency. During courtship, females vibrate their abdomens after being touched by the courting male, producing tremulatory signals with the dominant frequency below 100 Hz, sound pressure level below 60 dB and velocity below 3mm/s. This vibration may function as a sign of the akinesia state since it precedes the introduction of the embolus. Cuticular compounds probably determine the recognition of the male by the female. Data from the present study corroborate the generalist nature of L. intermedia in which signals of different modalities are used during courtship.

Stink bug interaction with host plants during communication.

In solitary plant-dwelling stink bug species, success depends crucially on efficient mate location and recognition, mediated by signals transmitted through the plant. All stink bugs investigated so far communicate with species and sex-specific narrow-band calling and courtship song signals produced by abdomen vibration. Calling songs of lower specificity are characterized by readily repeated units emitted with regular repetition rate from the same place on a plant, while courtship songs take place at shorter distances in the process of species and sex recognition, together with signals of other modalities. Signal spectra with about 100Hz fundamental frequency and harmonics below 1000Hz are tuned to the resonant properties of their green host plants. The majority of the identified leg vibratory receptor cells and the underlying ventral cord interneurons respond best in the frequency range below 500Hz. Green plants with low pass filtering properties transmit optimally signals with a dominant frequency around 100Hz and strongly attenuate vibrations above 600Hz. Accurate tuning of signal spectral properties with the plant's mechanical characteristics enables communication over several meter distances, with dispersive bending waves running through the plant's rod-like structures under standing wave conditions.

Morphology and physiology of vibratory interneurons in the thoracic ganglia of the southern green stinkbug Nezara viridula (L.).

The central processing mechanisms of vibratory signals in small plant-dwelling insects that rely primarily on substrate-borne vibratory communication are still largely unknown. To elucidate the neural mechanisms involved in vibratory signaling, the vibration-sensitive interneurons in thoracic ganglia of the southern green stinkbug, Nezara viridula, were investigated electrophysiologically by single-cell recordings and staining. Ten types of interneurons were described and divided into four categories, based on their gross morphology. The cell body of the L-shaped CG-AC neurons is located in the metathoracic neuromere of the central ganglion, and the axon ascends contralaterally. This group comprises five types of neurons differing in their fine structure and functional properties. CG-AB neurons are dorsal unpaired median (DUM) neurons with cell bodies in the mesothoracic neuromere of the central ganglion and two axons that ascend bilaterally into the prothoracic ganglion. Group CG-L includes three types of local neurons limited to the central ganglion. With ipsilateral dendritic arborizations and contralateral axonal branching, their gross morphology is similar to that of cricket omega cells. Interneuron PTG-DC, with the cell body in the prothoracic ganglion (PTG) and a contralaterally descending axon, conveys information received by the sensory organs of the front contralateral leg to the neuropil regions of the ipsilateral middle and hind legs. Based on their frequency tuning and acceleration sensitivity, the vibratory interneurons fall into two groups: the low-frequency units are tuned to 50 Hz and the middle frequency units to 200 Hz, with their acceleration thresholds at 10(-1) m/s(2) and 5 x 10(-3) m/s(2), respectively. Their function is discussed with relevance to the vibratory communication of N. viridula.

Vibrational communication along plants by the stink bugs Nezara viridula and Murgantia histrionica.

The velocity and spectral characteristics of vibrational signals of Nezara viridula (L.) and Murgantia histrionica (Hahn) (Heteroptera: Pentatomidae) were analyzed as the signals were transmitted through different plants. The velocity parameter of the body vibrations ranges from 0.1 to 1 mm/s. According to the mechanical properties of different substrates, the signal is attenuated or amplified during transmission from the insect's body to the substrate. Attenuation of up to 20 dB occurs during transmission of signals from leaves to stalks or stems. The velocity decrease with distance is below 0.5 dB/cm during transmission through less dense green stems, whereas it ranges between 0.6 and 1.6 dB/cm during transmission through more dense, woody stems. Signal velocity decreases non-linearly with increasing distance from the signal source. Regularly repeated velocity minima (nodes) and maxima (internodes) spaced 10-15 cm apart are characteristic of signal transmission through green plants but not woody stems. The signal velocity at some internodes exceeds the input value for N. viridula but not M. histrionica signals. The relative amplitude of the dominant frequency spectral peak varies with distance, along with overall signal velocity. Variable ratios of spectral peak amplitudes are characteristic for signals recorded at different distances from the source.

Tuning of host plants with vibratory songs of Nezara viridula L (Heteroptera: Pentatomidae).

Songs of the southern green stink bug Nezara viridula L, recorded on a non-resonant loudspeaker membrane, reflect frequency characteristics of body vibrations. The song dominant frequency directly depends on the repetition rate of potentials recorded from synchronously contracting muscles that vibrate the abdomen during singing. Spectra of naturally emitted signals recorded on the pronotum of a singing bug or on a plant contain peaks characteristic of plant resonant spectra. The dominant resonant frequency of sound-induced vibrations in bean and other stink bug host plants ranges between 160 and 215 Hz and subdominant peaks do not exceed 600 Hz. There is no correlation between spectral peak positions and recording points on a plant. The dominant resonance peak corresponds to the best frequency sensitivity of N. viridula middle frequency subgenual receptor cell. Subdominant peaks around and below 100 Hz lie close to the dominant frequency of body vibrations during singing and to the range of best frequency sensitivity of low frequency receptor cells. Tuning of plant resonant frequencies with spectral properties of songs and frequency sensitivity of sensory organs is discussed in the context of stink bug substrate-borne communication.

Vibrational communication in insects / A comunicação entre insetos através de vibrações

Communication through substrate-borne vibrations has for long been recognized but in comparison with air-borne sound it has received very little attention. However, in recent years it has become increasingly clear that vibrational signals play a crucial role in communication in many insect groups and we provide a short overview. Vibrational signals are related to sexual behavior, alarm and defensive behavior and are often used to mediate coordinated group actions and complex social interactions. For small insects they are probably the least costly and most far-reaching signals for intraspecific communication and also not easily perceived by a potential predator or parasitoid. Substrate-borne signals are produced by diversed methods and detected by sensitive receptors in all six legs. The courtship behavior of the southern green stink bug Nezara viridula (L.) (Hemiptera: Pentatomidae) is taken as a representative model in illustrating some principal mechanisms of vibrational communication in insects. Species and sex specific vibrational signals produced during the courtship are well suited for propagation through plants and to transmit the relevant information about the species and sex of the sender as well as provide the directional cue for locating the mate. The role of substrate-borne signals as a part of the specific mate recognition systems which are unique for each species makes studies of vibrational signals a very useful tool for resolving taxonomic problems.

A comunicação através de vibrações originárias do substrato tem sido reconhecida há muito tempo, mas tem recebido muito menos atenção que a comunicação através do som propagado pelo ar. Recentemente, entretanto, tem se tornado cada vez mais claro o papel crucial que sinais vibratórios desempenham na comunicação em muitos grupos de insetos, o que motivou essa revisão suscinta. Sinais vibracionais estão relacionados ao comportamento sexual, de alarme e de defesa, e são em geral usados para mediar ações coordenadas em grupo e interações sociais complexas. Para insetos pequenos esses sinais representam um custo energético mínimo, com alcance máximo, e que não são percebidos facilmente por predadores e por parasitóides em potencial. Sinais originários dos substrato são produzidos por diversos métodos e detectados por receptores presentes nas seis pernas. O comportamento de corte do percevejo verde, Nezara viridula (L.) (Hemiptera: Pentatomidae) é analisado como um modelo representativo na ilustração de alguns mecanismos importantes na comunicação vibracional em insetos. Sinais vibratórios são específicos das espécies e de sexos, os quais são produzidos durante a corte e que são adequados para serem transmitidos pelas plantas, contendo informações relevantes sobre a espécie e o sexo do organismo emissor, bem como informações de onde localizar o parceiro. O papel de sinais originados do substrato, os quais são únicos para cada espécie, faz com que os estudos em sinais vibracionais sejam uma importante ferramenta para resolver problemas taxonômicos.

Communication with substrate-borne signals in small plant-dwelling insects.

Vibratory signals of plant-dwelling insects, such as land bugs of the families Cydnidae and Pentatomidae, are produced mainly by stridulation and/or vibration of some body part. Signals emitted by the vibratory mechanisms have low-frequency characteristics with a relatively narrow frequency peak dominant around 100 Hz and differently expressed frequency modulation and higher harmonics. Such spectral characteristics are well tuned to the transmission properties of plants, and the low attenuation enables long-range communication on the same plant under standing wave conditions. Frequencies of stridulatory signals extend up to 10 kHz. In some groups, vibratory and stridulatory mechanisms may be used simultaneously to produce broadband signals. The subgenual organ, joint chordotonal organs, campaniform sensilla and mechanoreceptors, such as the Johnston's organ in antennae, are used to detect these vibratory signals. Species-specific songs facilitate mate location and recognition, and less species-specific signals provide information about enemies or rival mates.

Temporal and spectral properties of the songs of the southern green stink bug Nezara viridula (L.) from Slovenia.

Substrate born songs of the southern green stinkbug Nezara viridula (L.) from Slovenia were recorded and analysed. The male calling song is composed of narrow-band regularly repeated single pulses and of broad-band frequency modulated pulses grouped into pulse trains. The female calling song is characterised by broad-band pulsed and narrow-band non-pulsed pulse trains. A frequency modulated pre-pulse precedes the narrow-band pulse train. A frequency-modulated post-pulse usually follows the pulse train of the male courtship song. The male calling song triggers broad-band pulse trains of the female courtship song. The female also produces a repelling low-frequency vibration that inhibits male calling and courtship. The male rival song is characterised by prolonged pulses with a typical frequency modulation.

Vibratory songs of hybrids from Brazilian and Slovenian populations of the green stink bug Nezara viridula.

Males and females of the green stink bug Nezara viridula produce species and sex specific vibrational signals. The songs of bugs from geographically isolated population differ in their temporal characteristics. Hybrids were produced between the bugs from Brazilian and Slovenian populations in order to examine the levels of genetic inheritance of their vibratory songs. Hybrid males and females produced songs which are distinctly different from parental songs and these differences can be attributable to genetic factor. The results show that in some parameters the hybrid songs are intermediate between the parental types. Several song parameters are apparently sex-linked. It remains to be established whether observed genetically determined differences in vibratory songs also indicate that cryptic species exist within the taxon N. viridula.