Evidence for acoustic communication in the parasitoid wasp Glyptapanteles flavicoxis

Females of the parasitoid wasp Glyptapanteles flavicoxis (Marsh) (Hymenoptera: Braconidae) deposit sex pheromone on substrate that elicits attraction and wing fanning in conspecific males. We tested the hypothesis that wing fanning sound induces a behavioral response from females which, in turn, affects the males’ orientation toward them. Females exposed to playback of the males’ wing fanning sound engaged in short flights, with sound characteristics different from those of the males’ wing fanning sound. In two‐choice bioassays, playback of the females’ flight sound attracted significantly more males than a silent control stimulus, and in combination with pheromone‐containing body extract of females it attracted more males than female body extract alone. Our data support the conclusion that the males’ wing fanning induces sound and visual reply signals from females that help males orient toward them.     

Evidence for short-range sonic communication in lymantriine moths

Sexual communication of nun moth, Lymantria monacha (L.), pink gypsy moth, Lymantria mathura Moore, and fumida tussock moth, Lymantria fumida Butler (all Lepidoptera: Noctuidae: Lymantriinae), is known to be mediated by pheromones. We now show that males are attracted by the sounds of conspecific females over short distances and that wing fanning male and female L. monacha, L. mathura and L. fumida produce species- and sex-specific wing beat and associated click sounds that could contribute to reproductive isolation. Evidence for short-range communication in these lymantriines includes (i) scanning electron micrographs revealing metathoracic tympanate ears, (ii) laser interferometry showing particular sensitivity of tympana tuned to frequency components of sound signals from conspecifics, and (iii) phonotaxis of male L. monacha and L. fumida to speakers playing back sound signals from conspecific females. We conclude that tympanate ears of these moths have evolved in response not only to bat predation, but also for short-range mate finding and possibly recognition.     

Location,morphology and histology of sex pheromone glands of the female gypsy moth, Lymantria dispar (L.)

Scanning electron microscopy, histology and a male wing fanning bioassay were used in this study to locate the sex pheromone-producing glands of the female gypsy moth, Lymantria dispar. When exposed to female sex pheromone, adult males exhibit a strong wing fanning behaviour prior to take off. We found that adult males showed positive response to calling females and to tissue extract from both dorsal and ventral portions of the intersegmental membrane between 8th and 9th-abdominal segments. A typical male response usually starts with elevation of antennae, movement of head in different directions, walking, wing fanning and onset of search flight. Histological and scanning electron microscopic studies suggested that the sex pheromone glands are located on the dorsal and ventral aspects of the intersegmental membrane. The glands appear as two highly convoluted integumentary areas with hypertrophied glandular epidermal cells.     

Effects of pheromone plume structure and visual stimuli on the pheromone-modulated upwind flight of male gypsy moths (Lymantria dispar) in a Forest (Lepidoptera: Lymantriidae)

The pheromone-modulated upwind flight ofLymantria dispar males responding to different pheromone plume structures and visual stimuli designed to mimic trees was video recorded in a forest. Males flying upwind along pheromone plumes of similar structure generated tracks that were similar in appearance and quantitatively similar in almost all parameters measured, regardless of the experimentally manipulated visual stimuli associated with the pheromone source. Net velocities, ground speeds, and airspeeds of males flying in point-source plumes were slower than those of males flying in the wider, more diffuse plumes issuing from a cylindrical baffle. The mean track angle of males flying in plumes issuing from a point source was greater (oriented more across the wind) than that of males flying in plumes issuing from a transparent cylindrical baffle. Males flying in point-source plumes also turned more frequently and had narrower tracks overall than males responding to plumes from a cylindrical baffle. These data suggest thatL. dispar males orienting to pheromone sources (i.e., calling females) associated with visible vertical cylinders (i.e., trees) use predominantly olfactory cues to locate the source and that the structure of the pheromone plume markedly affects the flight orientation and the resultant track.     

Orientation of male gypsy moths,Lymantria Dispar (L.), to pheromone sources: The role of olfactory and visual cues

The role of olfaction and vision in the close-ranging flying and walking orientation of male gypsy moths, Lymantria dispar(L.), to females was studied in the forest and in the laboratory. In the forest, feral males found an isolated pheromone source as readily as one supplemented with female visual cues; dead, acetonerinsed females deployed without pheromone received virtually no visitations. In flight tunnel choice experiments using cylinders as surrogate trees and pheromone in different spatial configurations, visual attributes of the female did not influence either the males' choice of landing site or the efficiency with which they located the female. Rather, the presence of pheromone on the cylinder was necessary to elicit orientation as well as landing and walking on the cylinder. When a female visual model was placed in various positions around a pheromone source, walking males oriented primarily to the chemical stimulus. There were, however, indications that males would alter their walking paths in response to female visual cues over short distances (<5 cm), but only if they continued to receive pheromone stimulation. When visual and chemical cues were abruptly uncoupled by altering the trajectory of the pheromone plume, most males responded to the loss of the odor cue rather than to visual cues from the female. Temporal pheromone stimulation patterns affected male walking orientation. When stimulated by pheromone, males oriented toward the source; loss of the odor cue prompted an arearestricted local search characterized by primarily vertical and oblique movements with frequent reversals in direction. Presumably these maneuvers enhance the likelihood of recontacting the plume or serendipitously encountering the female. The apparent lack of visual response to the female is discussed in light of morphological and behavioral evidence suggesting that gypsy moths were formerly nocturnal.     

Disruption of the odour-mediated mating behaviour of Plodia interpunctella using high-frequency sound

Abstract Indian meal moths, Plodia interpunctella Hübner (Lepidoptera: Pyralidae), have ears which are sensitive to high‐frequency calls produced by echolocating, insectivorous bats. The influence of artificially generated, high‐intensity, ultrasound signals (25 kHz, 106 dB SPL at 1 m distance) on different parameters involved in the odour‐mediated mating behaviour of this species and its potential use in population control was investigated. All moths flying towards olfactory cues in flight tunnel experiments reacted strongly to a 1 s ultrasound pulse by cessation of flight and falling out of the odour plume. The source contact proportion of both male moths orienting towards the female‐produced sex pheromone and of mated female moths orienting towards an oviposition cue was reduced by 40%, compared to unexposed moths. Calling females responded to the sound by retraction of the ovipositor or by falling to the ground. Long‐term exposure to repetitive pulses of ultrasound suppressed female calling by up to 27%. Furthermore, mating in plastic tents was disrupted by up to 58% in ultrasound‐treated tents using different sound regimens, compared to control tents. The results are discussed in relation to the potential use of ultrasound technology for the population control of pyralid stored product pests.     

Pheromone‐binding proteins in the Asian gypsy moth females,Lymantria dispar,recognizing the sex pheromone and plant volatiles

Lepidopterans are known to have different pheromone‐binding proteins with differential expression patterns that facilitate specific signal transduction of semiochemicals. Two PBPs of the Asian gypsy moth, Lymantria dispar, were reported to express in both females and males, but their physiological functions were unknown. Results showed that LdisPBP1 and LdisPBP2 were expressed in the sensilla trichodea of males and the s. trichodea and s. basiconica of females. When LdisPBP1 gene was targeted by RNA interference (RNAi) in males, the expression of LdisPBP1 and LdisPBP2 decreased by 69 and 76%, respectively, and when LdisPBP2 gene was targeted by RNAi, they decreased by 60 and 42%, respectively. In females, after treatment with LdisPBP1 dsRNA, LdisPBP1 and LdisPBP2 levels were reduced by 26 and 69%, respectively, and LdisPBP2 dsRNA reduced the relative expression of them by 4 and 62%, respectively. The expression of LdisPBP1 and LdisPBP2 was interdependent. Electroantennogram (EAG) recordings showed that LdisPBPs participate in the recognition of the sex pheromone in males, and the sex pheromone and plant volatiles in females. The function of LdisPBPs represents the sex‐specific roles.     

Effects of light levels and plume structure on the orientation manoeuvres of male gypsy moths flying along pheromone plumes

The upwind zigzag flights of male gypsy moths (Lymantria dispar L.; Lepidoptera: Lymantriidae) along narrow, ribbon‐like and wide, turbulent plumes of pheromone were examined in a wind tunnel at light levels of 450 and 4 lux. Under all conditions tested males flew upwind zigzag paths. In 450 lux, males flying along turbulent plumes had the highest ground speeds and the widest crosswind excursions between counterturns, compared to slow flight and a narrow zigzag of males along a ribbon plume. In a turbulent plume, males flew more slowly and had narrower zigzags in 4 than in 450 lux. Across most treatments of plume structure and light level, the rate of transverse image flow and the frequency of counterturning remained relatively constant. The effects of light levels on orientation are not readily reconcilable with a model in which moths in low light levels would head more towards crosswind, thereby enhancing the rate of transverse image flow and the perception of wind‐induced drift.     

Conspecific females promote calling behavior in the noctuid moth,Pseudaletia adultera

Changes in female calling behavior in response to the presence of conspecific pheromones (pheromone autodetection) have been demonstrated in a number of moth species. However, the observed changes vary between species, and several ecological and adaptive explanations for autodetection have been proposed. We studied the effect of conspecific females on the calling behavior of the noctuid moth Pseudaletia adultera (Schaus) (Lepidoptera: Noctuidae, Hadenini), by comparing the age of first calling, as well as the onset and pattern of calling, when females were held individually or in the presence of conspecifics. Grouped females started calling at a lower age, a higher percentage of females called during the scotophase, and they called longer compared to females held in isolation. We also demonstrated that female antennae respond to each of the three main components of the sex pheromone – (Z)‐11‐hexadecen‐1‐ol, (Z)‐11‐hexadecen‐1‐yl acetate, and (Z)‐11‐hexadecenal – and that the response patterns differed from those of male antennae. By calling more and extending her calling window in presence of conspecific females, a female may increase her chances of accessing males. However, the potential benefits need to be considered within an ecological context, considering factors such as migration, oviposition, and foraging.     

Reproductive isolation inProkelisia planthoppers (Homoptera: Delphacidae): Acoustic differentiation and hybridization failure

Males and females of Prokelisia marginata (Van Duzee) and Prokelisia dolusWilson communicate through substrate-transmitted vibrations. The acoustic signals (attraction and courtship calls) of these planthoppers are effective in mate location, attraction, and mate choice. Attraction calls are structurally distinct for both species and differ in pulse type, pulse repetition rate, and pulse duration. Using playback of prerecorded calls, individuals discriminated between conspecific and heterospecific signals. Depending on the sex and species, response calls were produced three to eight times more frequently to conspecifics than to heterospecifics. However, acoustic signals alone did not explain reproductive isolation and hybridization failure in these two congeners. Some heterospecific pairs called, courted, and attempted to join genitalia, but no connections were successful and no progeny were produced. Thus, acoustic behavior is not a guaranteed premating isolating mechanism in no-choice situations. Other courtship behaviors and possibly morphological differences in genitalia also contributed to their isolation. Females displayed a variety of rejection behaviors to conspecific and heterospecific males, suggesting that sexual selection (female choice), in addition to species recognition, may be an important force in the evolution of the acoustic signals of planthoppers. Although signal structure was not dependent on wing form (planthoppers exhibit wing dimorphism), the age when females first began to call was related to wing form. Brachypterous (flightless) females of both species began calling early in adult life (day 2), whereas macropterous (migratory) females began calling later in adult life (day 6). This pattern is consistent with the oogenesis-flight syndrome, in which reproductive maturity is delayed until after migration occurs.     

Aggressive displays by male House Wrens are composed of multiple components that predict attack

Aggressive signals should predict whether the sender of the signal will attack the receiver, yet this criterion has been little studied. We conducted experiments with male House Wrens (Troglodytes aedon) in north‐central Illinois in 2009 to test the hypothesis that rates of song delivery and wing‐quivering (putative aggressive signals) signal aggressive intent. We simulated a conspecific territorial intrusion by combining playback of male song with a male taxidermic mount, predicting that these signals would be related to a male's likelihood of attacking a conspecific. All males (N = 37) sang in response to the intrusion. Males attacking the mount sang at significantly higher rates and performed significantly more wing quivers than males that did not attack. In addition, all males that attacked the mount performed wing quivers (9/9), whereas only 53.6% (15/28) of males that did not attack did so (P = 0.011). Our results are consistent with the hypothesis that both song and wing quivering indicate a signaler's aggressive intent and that these signals are likely components of a multi‐component, hierarchical display.     

Interspecific interactions and learning variability jointly drive geographic differences in mate preferences

Co‐occurrence of closely related species can cause behavioral interference in mating and increase hybridization risk. Theoretically, this could lead to the evolution of more species‐specific mate preferences and sexual signaling traits. Alternatively, females can learn to reject heterospecific males, to avoid male sexual interference from closely related species. Such learned mate discrimination could also affect conspecific mate preferences if females generalize from between species differences to prefer more species‐specific mating signals. Female damselflies of the banded demoiselle (Calopteryx splendens) learn to reject heterospecific males of the beautiful demoiselle (C. virgo) through direct premating interactions. These two species co‐occur in a geographic mosaic of sympatric and microallopatric populations. Whereas C. virgo males have fully melanized wings, male C. splendens wings are partly melanized. We show that C. splendens females in sympatry with C. virgo prefer smaller male wing patches in conspecific males after learning to reject heterospecific males. In contrast, allopatric C. splendens females with experimentally induced experience with C. virgo males did not discriminate against larger male wing patches. Wing patch size might indicate conspecific male quality in allopatry. Co‐occurrence with C. virgo therefore causes females to prefer conspecific male traits that are more species specific, contributing to population divergence and geographic variation in female mate preferences.     

Male moth songs tempt females to accept mating: the role of acoustic and pheromonal communication in the reproductive behaviour of Aphomia sociella

Background

Members of the subfamily Galleriinae have adapted to different selective environmental pressures by devising a unique mating process. Galleriinae males initiate mating by attracting females with either chemical or acoustic signals (or a combination of both modalities). Six compounds considered candidates for the sex pheromone have recently been identified in the wing gland extracts of Aphomia sociella males. Prior to the present study, acoustic communication had not been investigated. Signals mediating female attraction were likewise unknown.

Methodology/Principal Findings

Observations of A. sociella mating behaviour and recordings of male acoustic signals confirmed that males initiate the mating process. During calling behaviour (stationary wing fanning and pheromone release), males disperse pheromone from their wing glands. When a female approaches, males cease calling and begin to produce ultrasonic songs as part of the courtship behaviour. Replaying of recorded courting songs to virgin females and a comparison of the mating efficiency of intact males with males lacking tegullae proved that male ultrasonic signals stimulate females to accept mating. Greenhouse experiments with isolated pheromone glands confirmed that the male sex pheromone mediates long-range female attraction.

Conclusion/Significance

Female attraction in A. sociella is chemically mediated, but ultrasonic communication is also employed during courtship. Male ultrasonic songs stimulate female sexual display and significantly affect mating efficiency. Considerable inter-individual differences in song structure exist. These could play a role in female mate selection provided that the female''s ear is able to discern them. The A. sociella mating strategy described above is unique within the subfamily Galleriinae.     

Release mechanism of sex pheromone in the female gypsy moth <Emphasis Type="Italic">Lymantria dispar</Emphasis>: a morpho-functional approach

A morpho-functional investigation of the sex pheromone-producing area was correlated with the pheromone release mechanism in the female gypsy moth Lymantria dispar. As assessed by male electroantennograms (EAG) and morphological observations, the pheromone gland consists of a single-layered epithelium both in the dorsal and ventral halves of the intersegmental membrane between the 8th and 9th abdominal segments. By using the male EAG as a biosensor of real-time release of sex pheromone from whole calling females, we found this process time coupled with extension movements of the ovipositor. Nevertheless, in females in which normal calling behavior was prevented, pheromone release was detected neither in absence nor in presence of electrical stimulation of the ventral nerve cord/terminal abdominal ganglion (TAG) complex. Tetramethylrhodamine-conjugated dextran amine stainings also confirm the lack of any innervation of the gland from nerves IV to VI emerging from the TAG. These findings indicate that the release of sex pheromone from the glands in female gypsy moths is independent of any neural control exerted by the TAG on the glands, at least by way of its three most caudally located pairs of nerves, and appears as a consequence of a squeezing mechanism in the pheromone-producing area.     

ACOUSTIC COMMUNICATION AND MATING BEHAVIOUR IN WATER BUGS OF THE GENUS MICRONECTA

ABSTRACT

The role of male sounds in attracting females and in mating was investigated in the three most common species of Micronecta found in ponds 60 km NE of Melbourne, Australia: M. concordia, M. tasmanica and M. robusta. In playback experiments using recorded male signals, females were attracted to signals of conspecific males, in preference to signals of heterospecific sympatric males. Studies of mating behaviour, using video recording, showed that signals were obligatory for mating. These findings strongly suggest that acoustic signalling is important in reproductive isolation in Micronecta. Comparisons between matings and rejected matings showed that, within each species, copulation only occurred following a certain minimum number of acoustic signals with more pulse-trains than the mean for the species. Low amplitude sounds (after signals) were also important in courtship, immediately preceding mating. No sounds occurred during copulation.     

The significance of wing pattern diversity in the Lycaenidae: mate discrimination by two recently diverged species

Closely related species of lycaenid butterflies are determinable, in part, by subtle differences in wing pattern. We found that female wing patterns can act as an effective mate‐recognition signal in some populations of two recently diverged species. In field experiments, we observed that males from a Lycaeides idas population and an alpine population of L. melissa preferentially initiate courtship with conspecific females. A morphometric study indicated that at least two wing pattern elements were important for distinguishing the two species: hindwing spots and orange crescent‐shaped pattern elements called aurorae. We deceived male L. idas into initiating courtship with computer generated paper models of heterospecific females when these pattern elements were manipulated, indicating that the wing pattern elements that define the diversity of this group can be effective mate recognition signals.     

Pheromone‐trapping the nun moth,Lymantria monacha (Lepidoptera: Lymantriidae) in Inner Mongolia,China

The nun moth, Lymantria monacha L., is one of the most important defoliators of Eurasian coniferous forests. Outbreaks during 2011–2015 in the natural/planted larch, and larch‐birch mixed forests of the Greater Khingan Range in Inner Mongolia, China, caused tremendous timber losses from severe defoliation and tree mortality. A series of trapping experiments were conducted in these outbreak areas to evaluate the efficacy of a synthetic species‐specific pheromone lure based on the female pheromone blend of European nun moth populations. Our results clearly show that the nun moth in Inner Mongolia is highly and specifically attracted to this synthetic pheromone, with few gypsy moths (Lymantria dispar) captured. Flight activity monitoring of L. monacha male moths using pheromone‐baited Unitraps at 2 locations during the summer of 2015 indicated that the flight period started in mid‐July, peaking in early August at both locations. Based on male moth captures, there was a strong diurnal rhythm of flight activity throughout the entire scotophase, peaking between 22:00 and 24:00. Unitraps and wing traps had significantly and surprisingly higher catches than the gypsy moth traps. Unitraps fastened to tree trunks 2 m above ground caught significantly more male moths than those at the ground level or at 5 m height. Male L. monacha moths can be attracted to pheromone‐baited traps in open areas 150–200 m distant from the infested forest edge. Our data should allow improvement on the performance of pheromone‐baited traps for monitoring or mass‐trapping to combat outbreaks of this pest in northeastern China.     

Wing-Click Sounds of Heliconius cydno alithea (Nymphalidae: Heliconiinae) Butterflies

Field-collected Heliconius cydno Doubleday females were observed producing audible wing clicks during encounters between conspecifics in greenhouses in a large insectary during the day and at roosting time. Occasionally, these females also were observed producing sounds in aggressive encounters with females of a close relative, H. erato (L). However, the wing-clicks were not observed subsequently from first-generation adults born in the greenhouses. The sounds were produced in short trains of 3–10 wing-clicks at the rate of 10 clicks/s. The individual clicks had a mean duration of 1.48 ms and a broad frequency spectrum, with a peak near 1075 Hz. This peak lies near the 1200-Hz frequency of maximal sensitivity measured previously for auditory neurons of H. erato. The production of these previously unreported sounds suggests that wing clicks may play a role in both intra- and interspecific communication among Heliconius species.     

Avoiding Male Harassment: Wing‐Closing Reactions to Flying Individuals by Female Small Copper Butterflies

Males of many butterfly species persistently court and attempt to mate with females even if the females reject courtship. This male harassment almost certainly has negative effects on female fitness. Therefore, females have likely evolved strategies to avoid such encounters. To investigate the harassment avoidance strategy of females of the small copper butterfly, Lycaena phlaeas daimio, I observed the reactions of females to other individuals flying nearby in the field. In response to the conspecific butterflies, females closed their wings if they had previously been open and did not exhibit any action if the wings had been closed. Females that closed their open wings in response to a conspecific received fewer mating attempts than did females that held their wings open. These results indicate that the wing‐closing behaviour of L. phlaeas females functions to deter male mating attempts. The wing‐closing reaction occurred primarily in mated females. Because females of L. phlaeas copulate only once during their lives, this behaviour is not considered an indirect mate choice but rather an attempt to avoid persistent mating attempts (i.e. sexual harassment) by males.     

Sexual differentiation and seasonal variation in response to conspecific and heterospecific acoustic signals

Interspecific territoriality is frequently reported between closely related species; however, few studies have demonstrated interspecific territoriality between distantly related species living in sympatry. We conducted playback experiments to investigate territorial behaviour in male and female White‐bellied Wrens (Uropsila leucogastra) in response to simulated conspecific and heterospecific intruders during the breeding and non‐breeding seasons. We explored whether heterospecific songs of the Happy Wren (Pheugopedius felix), a distantly related species and ecological competitor, elicited antagonistic responses from focal White‐bellied Wrens, and whether such responses differed between the sexes. We also examined whether male and female responses to conspecific and heterospecific rivals varied with season. We found that male White‐bellied Wrens always responded to conspecific song, and responded significantly more to heterospecific song compared to a control stimulus (Tropical Parula, Setophaga pitiayumi). In contrast, although female White‐bellied Wrens responded strongly to conspecific song, their response to heterospecific song did not differ significantly from the control stimulus. The proportion of males that responded to heterospecific songs and the proportion of females that responded to conspecific songs varied seasonally, showing significantly lower responses during the breeding season. The intense responses of male White‐bellied Wrens to playback of heterospecific songs suggest that they recognise ecological competitors based on their vocal signals. Furthermore, the decrease in agonistic interactions during the breeding season is in line with the hypothesis that aggressive behaviour may be detrimental to reproductive and parental activity, and the hypothesis that heterospecific animals pose less of a threat during the breeding season.