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.     

Vibration‐mediated territoriality in the warty birch caterpillar Drepana bilineata

Abstract The warty birch caterpillar Drepana bilineata produces two distinct types of vibrational signals (mandible drumming and anal scraping) during interactions with conspecifics. Vibrational signalling is characterized using standard and high‐speed videography synchronized with laser‐doppler vibrometry, and behavioural experiments test the hypothesis that signalling functions to advertise occupancy of birch (Betula) leaves. Drumming involves raising the head and striking the leaf with the sharp edges of the open mandibles. Anal scraping involves dragging a pair of specialized oar‐shaped setae against the leaf surface. Staged encounters between leaf residents and conspecific intruders result in the resident signalling, with rates increasing as the intruder moves closer. Intruders signal significantly less often than residents. Conflicts are typically resolved within a few minutes, with the resident winning in 61% of the trials, and the intruder winning in 6%. Contests that last more than 30 min are deemed ‘ties’ and comprise the remaining 33% of trials. The results support the hypothesis that vibrational signals function to advertise leaf occupancy. Vibrational communication is believed to be widespread in Drepanoidea caterpillars, but has only been described in two species to date: D. bilineata (present study) and Drepana arcuata. It is proposed that differences in territorial behaviour and signalling between these species are related to their relative investments in silk leaf mats and shelters. The proximate and ultimate bases for the evolution of vibrational communication in caterpillars are discussed.     

Chemical information transfer between wounded and unwounded plants: backing up the future

This special issue on “Chemical information transfer between wounded and unwounded plants” provides an overview of past and ongoing experiments on plant-to-plant communication. Since the studies on plant responses to single gaseous compounds were not particularly emphasised, the actual number of studies relevant to the subject is underestimated. All in all, we think the amount of data on damage-induced plant-to-plant information transfer makes that the phenomenon can no longer be denied and deserves intensified attention by the scientific community. In this epilogue we highlight a couple of issues which received little attention and present some speculative ideas. First we concentrate on functional aspects of plant-plant communication we stress the concept of damage-induced signalling as an ecological cost to the signal-sending plant and we discuss the theoretical development on interplant signalling, which is still in its infancy. With respect to mechanisms, we compare above- to belowground signalling, discuss potential cues and stress the possibility that responses in signal-exposed plants may be hidden. Finally, we address some future prospects which may help in the further development of the still underexposed phenomenon of damage-induced plant-to-plant information transfer.     

Acoustic alarm signalling facilitates predator protection of treehoppers by mutualist ant bodyguards

Mutualism is a net positive interaction that includes varying degrees of both costs and benefits. Because tension between the costs and benefits of mutualism can lead to evolutionary instability, identifying mechanisms that regulate investment between partners is critical to understanding the evolution and maintenance of mutualism. Recently, studies have highlighted the importance of interspecific signalling as one mechanism for regulating investment between mutualist partners. Here, we provide evidence for interspecific alarm signalling in an insect protection mutualism and we demonstrate a functional link between this acoustic signalling and efficacy of protection. The treehopper Publilia concava Say (Hemiptera: Membracidae) is an insect that provides ants with a carbohydrate-rich excretion called honeydew in return for protection from predators. Adults of this species produce distinct vibrational signals in the context of predator encounters. In laboratory trials, putative alarm signal production significantly increased following initial contact with ladybeetle predators (primarily Harmonia axyridis Pallas, Coleoptera: Coccinellidae), but not following initial contact with ants. In field trials, playback of a recorded treehopper alarm signal resulted in a significant increase in both ant activity and the probability of ladybeetle discovery by ants relative to both silence and treehopper courtship signal controls. Our results show that P. concava treehoppers produce alarm signals in response to predator threat and that this signalling can increase effectiveness of predator protection by ants.     

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.     

Palomena prasina (Hemiptera: Pentatomidae) vibratory signals and their tuning with plant substrates

Palomena prasina is interesting for the study of vibrational communication within the Pentatomid subfamily Pentatominae, because its host range is limited to woody plants, unlike the better known Nezara viridula, whose vibrational communication is commonly used as a model for the whole family. The vibrational repertoire of P. prasina was described several decades ago and is redescribed in this paper using modern methods for non-contact vibration recording. Additionally, we hypothesized that this species has retained the capacity for signal frequency variation necessary for tuning to resonance properties of various host plants of Pentatominae, but if the signals are emited in the absence of mechanical feedback, they are tuned more specifically to their native acoustic environment — woody plants. By recording live bugs signalling on different substrates and comparing spectral properties of their signals among substrates, we found that there is a match between the signals emitted on a woody branch and those emitted on a non-resonant surface, while spectral properties of signals emitted on herbaceous plants differ. Our findings provide evidence in support of the signal tuning hypothesis and shed further light on the crucial role of substrate in vibrational communication of insects.     

Vibrational communication facilitates cooperative foraging in a phloem-feeding insect

Insects are the dominant herbivores in tropical forests, with a range of mechanisms for exploiting plant resources. For group-living species, such mechanisms may involve communication. The Neotropical treehopper Calloconophora pinguis (Hemiptera: Membracidae) is a sap-feeding species in which groups of siblings feed on new leaves during the brief period of leaf expansion. Using an experimental approach, a process of cooperative foraging among siblings was documented, in which a few individuals in a group behave as scouts, locating a new feeding site and advertising it using plant-borne vibrational signals. Signalling leads to a period of positive feedback in which newly recruited individuals signal in concert with those already there. The food signalling system of C. pinguis is unique in its use of synchronized group displays and in the tight coordination of receiver responses with collective signals. Examples from a number of taxonomic groups show that vibrational communication can allow group-living insects to solve the challenges of feeding on plants, such as remaining in a foraging group or avoiding predation. While most research has focused on leaf-feeding species, sap-feeding species may remove just as much biomass. This study shows that cooperative vibrational communication underlies the ability of a sap-feeding species to exploit plant resources during a narrow window of availability.     

G protein-coupled receptor signalling and cross-talk: achieving rapidity and specificity

Activation of a given type of G protein-coupled receptor (GPCR) triggers a limited set of signalling events in a very rapid and specific manner. The classical paradigm of GPCR signalling was rather linear and sequential. Emerging evidence, however, has revealed that this is only a part of the complex signalling mediated by GPCR. Propagation of GPCR signalling involves cross-regulation of many but specific pathways, including cross-talks between different GPCRs as well as with other signalling pathways. Moreover, it is increasingly apparent that GPCRs can activate both heterotrimeric G protein-dependent and G protein-independent signalling pathways. In this review, we discuss how the signallings initiated by GPCRs achieve rapidity as well as specificity, and how the GPCRs can cross-regulate other specific signalling pathways at the same time. New concepts regarding GPCR signalling have been arising to address this issue, which include multiprotein signalling complex and signalling compartment in microdomain concepts that enable close colocalization or even contact among the proteins engaged in the specific signal transduction. The final outcome of a stimulation of GPCR will thus be the sum of its own specific set of intracellular signalling pathways it regulates.     

Visual signalling by asymmetry: a review of perceptual processes.

Individual levels of asymmetry in traits that display fluctuating asymmetry could be used as visual signals of phenotypic (and perhaps genotypic) quality, as asymmetry can often be negatively related to fitness parameters. There are some data to support this hypothesis but the experimental protocols employed have commonly resulted in asymmetries far larger than those observed in nature. To date, there has been little consideration of the ability of animals to accurately discriminate small asymmetries (of the magnitude observed in the wild) from perfect symmetry. This is key to assessing the plausibility of the asymmetry-signalling hypothesis. Here, I review the perceptual processes that may lead to the discrimination of asymmetry and discuss a number of ecologically relevant factors that may influence asymmetry signalling. These include: signal orientation, distance of trait elements from the axis of symmetry, trait complexity, trait contrast and colour, and the behaviour of both signaller and receiver. I also discuss the evolution of symmetry preferences and make suggestions as to where researchers should focus attention to examine the generality of asymmetry-signalling theory. In highly developmentally stable signalling systems the magnitude of asymmetry may be too small to be detected accurately and reliably, hence asymmetry signalling is unlikely to have evolved in these situations.     

Characteristics of the Signals of Male <Emphasis Type="Italic">Anoeconeossa bundoorensis</Emphasis> Taylor and Burckhardt (Hemiptera: Aphalaridae) Associated with Female Responsiveness

Sexual selection requires intra-specific variation in the characteristics mediating mate choice. In species reliant on substrate-borne vibrational signalling (SBVS), differences in the attractiveness of individual signallers’ calls can influence relative mating success since they can indicate the quality of the sender. We used laser vibrometry and playback experiments to study duet signalling in a psyllid (Anoeconeossa bundoorensis Taylor and Burckhardt), in particular to identify characteristics linked to female responsiveness. Signals were sex-specific with the syllables (or calls) of the smaller males attaining higher frequencies than the syllables of the larger females. Male syllables build over time with more energy in the second half of the call while those of females have a more uniform energy distribution. Male syllables vary in the timing of the halfway point of energy production. We used playback to examine species recognition and female responsiveness to the first and second halves of male calls. We demonstrated that females responded only to calls of male A. bundoorensis and that they were more likely to respond to the first half of male calls. Based on the results of our second playback experiment, we found that a gradation in energy production is more important for eliciting a female reply than a uniform distribution of energy. Our findings show that the rate of energy production facilitates differentiation between syllables which could be used to indicate the quality of individual males as mates.     

Signalling shutdown strategies in aging immune cells

It is important for the resolution of inflammation that the number and activity of immune cells are reduced. Clearance of immune cells may be achieved by apoptosis and phagocytosis of cell fragments by macrophages. However, signalling shutdown by immune cells committed to apoptosis occurs early in the progression of these cells towards fragmentation and, it could be argued, is a key feature of apoptosis. There is surprisingly little known about the mechanisms that underlie this signalling shutdown, in particular the shutdown of Ca(2+) influx. The consequences and the potential mechanisms by which Ca(2+) influx shutdown is achieved are discussed. In addition, the potential consequences for cell signalling of cytochrome c release from mitochondria and of phosphatidyl-serine externalization are discussed. The aim of the review is therefore to highlight the evidence for various signalling shutdown strategies in immune cells that may limit their activity during progression towards apoptosis.     

HSPG synthesis by zebrafish Ext2 and Extl3 is required for Fgf10 signalling during limb development

Heparan sulphate proteoglycans (HSPGs) are known to be crucial for signalling by the secreted Wnt, Hedgehog, Bmp and Fgf proteins during invertebrate development. However, relatively little is known about their effect on developmental signalling in vertebrates. Here, we report the analysis of daedalus, a novel zebrafish pectoral fin mutant. Positional cloning identified fgf10 as the gene disrupted in daedalus. We find that fgf10 mutants strongly resemble zebrafish ext2 and extl3 mutants, which encode glycosyltransferases required for heparan sulphate biosynthesis. This suggests that HSPGs are crucial for Fgf10 signalling during limb development. Consistent with this proposal, we observe a strong genetic interaction between fgf10 and extl3 mutants. Furthermore, application of Fgf10 protein can rescue target gene activation in fgf10, but not in ext2 or extl3 mutants. By contrast, application of Fgf4 protein can activate target genes in both ext2 and extl3 mutants, indicating that ext2 and extl3 are differentially required for Fgf10, but not Fgf4, signalling during limb development. This reveals an unexpected specificity of HSPGs in regulating distinct vertebrate Fgfs.     

Vibrational communication during spawning behavior in the himé salmon (landlocked red salmon,Oncorhynchus nerka)

Summary Sexual behaviors of the salmon are composed of a stimulus-reaction chain, which ensures synchronous spawning between the sexes and successful fertilization. To characterize the signals involved in such a stimulusreaction chain, the body vibration and electromyographic activity of the trunk muscles during spawning were simultaneously recorded from freely behaving male and female pairs of himé salmon (landlocked red salmon,Oncorhynchus nerkd) and were analyzed in combination with a videographic analysis of behavior sequences. The results showed that the himé salmon have an elaborate communication system in which characteristic vibrational signals are exchanged. These are produced by body vibration due to trunk muscle activity related to spawning and are transmitted between the sexes with an accurate timing through the stimulus-reaction chain. They act as timing cues to synchronize gamete release and are thought to be shared among a wide variety of fishes. It was hypothesized that the lateral line sense is involved in the detection of these vibrational signals. Furthermore, based on the sequence matrix analysis as well as on information theory, intersexual behavioral sequences during spawning were analyzed statistically. The results showed that statistically significant interactions occur between the sexes and statistically significant amounts of information are transmitted through the interactions, supporting the results from recording experiments mentioned above. Characters of the signalling system and possible origins of the vibrational signals are also discussed.     

Nematode parasites reduce carotenoid-based signalling in male red grouse

Carotenoids determine the yellow-red colours of many ornaments, which often function as signals of quality. Carotenoid-based signalling may reliably advertise health and should be particularly sensitive to parasite infections. Nematodes are among the commonest parasites of vertebrates, with well-documented negative effects on their hosts. However, to date, little is known about the effects that these parasites may have on carotenoid-based signalling. Tetraonid birds (grouse) exhibit supra-orbital combs, which are bright integumentary ornaments pigmented by carotenoids. We tested the effect of the nematode parasite Trichostrongylus tenuis on signalling in free-living male red grouse Lagopus lagopus scoticus. We show that experimentally reduced nematode infection increases plasma carotenoid concentration and comb redness, demonstrating for the first time that nematodes can influence carotenoid-based signals.     

Wounding increases salt tolerance in tomato plants: evidence on the participation of calmodulin-like activities in cross-tolerance signalling

Cross-tolerance is the phenomenon by which a plant resistance to a stress results in resistance to another form of stress. It has previously been shown that salt stress causes the accumulation of proteinase inhibitors and the activation of other wound-related genes in tomato plants (Solanum lycopersicum). However, very little is known about how different stresses interact with one another, and which are the signalling components that interrelate the responses triggered by different stress types. In the present work, it is shown that mechanical wounding increases salt-stress tolerance in tomato plants through a mechanism that involves the signalling peptide systemin and the synthesis of JA. Data are also provided indicating that calmodulin-like activities are necessary for the downstream signalling events that lead to cross-tolerance between wounding and salt stress. Finally, evidence was gathered supporting the hypothesis that LeCDPK1, a Ca2+ -dependent protein kinase from tomato previously described in our laboratory, could participate in this cross-tolerance mechanism interrelating the signalling responses to wounding and salt stress.