High acoustic diversity and behavioral complexity of katydids in the Mesozoic soundscape.

Acoustic communication has played a key role in the evolution of a wide variety of vertebrates and insects. However, the reconstruction of ancient acoustic signals is challenging due to the extreme rarity of fossilized organs. Here, we report the earliest tympanal ears and sound-producing system (stridulatory apparatus) found in exceptionally preserved Mesozoic katydids. We present a database of the stridulatory apparatus and wing morphology of Mesozoic katydids and further calculate their probable singing frequencies and analyze the evolution of their acoustic communication. Our suite of analyses demonstrates that katydids evolved complex acoustic communication including mating signals, intermale communication, and directional hearing, at least by the Middle Jurassic. Additionally, katydids evolved a high diversity of singing frequencies including high-frequency musical calls, accompanied by acoustic niche partitioning at least by the Late Triassic, suggesting that acoustic communication might have been an important driver in the early radiation of these insects. The Early-Middle Jurassic katydid transition from Haglidae- to Prophalangopsidae-dominated faunas coincided with the diversification of derived mammalian clades and improvement of hearing in early mammals, supporting the hypothesis of the acoustic coevolution of mammals and katydids. Our findings not only highlight the ecological significance of insects in the Mesozoic soundscape but also contribute to our understanding of how acoustic communication has influenced animal evolution.

Evolutionary rates of and selective constraints on the mitochondrial genomes of Orthoptera insects with different wing types.

Orthoptera is the most diverse order of polyneopterans, and the forewing and hindwing of its members exhibit extremely variability from full length to complete loss in many groups; thus, this order provides a good model for studying the effects of insect flight ability on the evolutionary constraints on and evolutionary rate of the mitochondrial genome. Based on a data set of mitochondrial genomes from 171 species, including 43 newly determined, we reconstructed Orthoptera phylogenetic relationships and estimated the divergence times of this group. The results supported Caelifera and Ensifera as two monophyletic groups, and revealed that Orthoptera originated in the Carboniferous (298.997 Mya). The date of divergence between the suborders Caelifera and Ensifera was 255.705 Mya, in the late Permian. The major lineages of Acrididae seemed to have radiated in the Cenozoic, and the six patterns of rearrangement of 171 Orthoptera mitogenomes mostly occurred in the Cretaceous and Cenozoic. Based on phylogenetic relationships and ancestral state reconstruction, we analysed the evolutionary selection pressure on and evolutionary rate of mitochondrial protein-coding genes (mPCGs). The results indicated that during approximately 300 Mya of evolution, these genes experienced purifying selection to maintain their function. Flightless orthopteran insects accumulated more non-synonymous mutations than flying species and experienced more relaxed evolutionary constraints. The different wing types had different evolutionary rates, and the mean evolutionary rate of Orthoptera mitochondrial mPCGs was 13.554 × 10-9 subs/s/y. The differences in selection pressures and evolutionary rates observed between the mitochondrial genomes suggested that functional constraints due to locomotion play an important role in the evolution of mitochondrial DNA in orthopteran insects with different wing types.

Material heterogeneity of male genitalia reduces genital damage in a bushcricket during sperm removal behaviour.

Sperm removal behaviour (SRB) is known in many animals, and male genital structures are often involved in the SRB, e.g. rubbing female genitalia vigorously. However, it remains unclear how those male genital structures function properly without severe genital damage during SRB. In the present study, we focused on the bushcricket Metaplastes ornatus and examined the biomechanics of male and female genital structures, involved in their SRB as a model case. During an initial phase of mating, males of this species thrust their subgenital plate with hook-like spurs and many microscopic spines into the female genital chamber. By moving the subgenital plate back-and-forth, males stimulate females, and this stimulation induces the ejection of sperm previously stored in females. We aimed to uncover the mechanics of the interaction between the subgenital plate and genital chamber during SRB. The genital morphology and its material composition were investigated using modern imaging and microscopy techniques. The obtained results showed a pronounced material heterogeneity in the subgenital plate and the genital chamber. The material heterogeneity was completely absent in that of a second bushcricket species, Poecilimon veluchianus, which does not exhibit SRB. Finite element simulations showed that the specific material heterogeneity can redistribute the stress in the subgenital plate of M. ornatus and, thereby, reduces stress concentration during SRB. This may explain why only a few examined males had a broken spur. We suggest that the observed structural features and material heterogeneity in M. ornatus are adaptations to their SRB.

Exaggerated male legs increase mating success by reducing disturbance to females in the cave weta Pachyrhamma waitomoensis.

Mate guarding is a widespread behaviour resulting from sperm competition and conflict over optimal remating rates. It is a key way in which males exhibit differential mating investment, and represents a complex interplay between mating effort, intrasexual competition, opportunity costs and sexual conflict. Nevertheless, although there are many examples of exaggerated male structures used to fight rivals, few animals have developed specialized male morphological adaptations for directly sheltering females from disturbance by non-rivals. Here we report on the use of sexually dimorphic, elongated male hind legs, which are used to guard females in the New Zealand cave weta Pachyrhamma waitomoensis (Orthoptera: Rhaphidophoridae). We found that male hind legs alongside the female failed to deter rivals from accessing her or disrupting copulation. However, they did reduce the disturbance to females from other, non-rival animals such as juveniles and heterospecifics. Males with longer hind legs were more effective in reducing disturbance, and remained with females for longer. Longer guarding periods also led to higher numbers of matings between pairs. Models of males with artificially altered hind leg dimensions also showed a benefit to greater leg length, and artificially altering the disturbance rate to females also had a significant effect on pair duration. Our results indicate that nuisance disturbance to females may play an important role in driving sexual selection on male leg length and its exaggeration in this species.

Are Anatomical Measurements Useful for Interspecific and Sexual Differentiation of Temnomastax (Orthoptera: Eumastacidae) Species?

Species of Temnomastax have wide morphological similarities, and differentiation is usually based on male and female genitalia. In this study, we tested whether morphometric differences contribute to differentiation of species of Temnomastax, proposed an identification key for males, and morphometrically studied the sexual dimorphism of Temnomastax hamus and Temnomastax ricardoi. Analysis was performed using 204 specimens belonging to six species; fifteen morphometric variables were used. We used MANOVA and ANOVA to test the morphometric differences among species and Discriminant Analysis for sexual distinction. Temnomastax sp. nov. 1 had the highest values in ⅓ of all analyzed variables used for morphometric distinction of males, followed by Temnomastax latens, T. ricardoi and Temnomastax sp. nov. 2, and T. hamus and Temnomastax tigris. Two groups were formed by the MANOVA, with length of body, length of hind femur, and length of tegmen as the main variables that distinguished them. Overall, females of T. hamus and T. ricardoi presented higher averages for measurement values used in morphometric distinction between sexes than conspecific males and based on discriminant analysis we found significant differences between sexes. There are significant differences in morphometric variations and the ratio between body length and tegmen length provided reliable evidence for differentiation among species, which can be used as a valid tool that complements the identification of Temnomastax species.

Functional basis of the sexual dimorphism in the auditory fovea of the duetting bushcricket Ancylecha fenestrata.

From mammals to insects, acoustic communication is in many species crucial for successful reproduction. In the duetting bushcricket Ancylecha fenestrata, the mutual acoustic communication between males and females is asymmetrical. We investigated how those signalling disparities are reflected by sexual dimorphism of their ears. Both sexes have tympanic ears in their forelegs, but male ears possess a significantly longer crista acustica containing 35% more scolopidia. With more sensory cells to cover a similar hearing range, the male hearing organ shows a significantly expanded auditory fovea that is tuned to the dominant frequency of the female reply to facilitate phonotactic mate finding. This sex-specific auditory fovea is demonstrated in the mechanical and neuronal responses along the tonotopically organized crista acustica by laservibrometric and electrophysiological frequency mapping, respectively. Morphometric analysis of the crista acustica revealed an interrupted gradient in organ height solely within this auditory fovea region, whereas all other anatomical parameters decrease continuously from proximal to distal. Combining behavioural, anatomical, biomechanical and neurophysiological information, we demonstrate evidence of a pronounced auditory fovea as a sex-specific adaptation of an insect hearing organ for intraspecific acoustic communication.

Morphological determinants of signal carrier frequency in katydids (Orthoptera): a comparative analysis using biophysical evidence of wing vibration.

Male katydids produce mating calls by stridulation using specialized structures on the forewings. The right wing (RW) bears a scraper connected to a drum-like cell known as the mirror and a left wing (LW) that overlaps the RW and bears a serrated vein on the ventral side, the stridulatory file. Sound is generated with the scraper sweeping across the file, producing vibrations that are amplified by the mirror. Using this sound generator, katydids exploit a range of song carrier frequencies (CF) unsurpassed by any other insect group, with species singing as low as 600 Hz and others as high as 150 kHz. Sound generator size has been shown to scale negatively with CF, but such observations derive from studies based on few species, without phylogenetic control, and/or using only the RW mirror length. We carried out a phylogenetic comparative analysis involving 94 species of katydids to study the relationship between LW and RW components of the sound generator and the CF of the male's mating call, while taking into account body size and phylogenetic relationships. The results showed that CF negatively scaled with all morphological measures, but was most strongly related to components of the sound generation system (file, LW and RW mirrors). Interestingly, the LW mirror (reduced and nonfunctional) predicted CF more accurately than the RW mirror, and body size is not a reliable CF predictor. Mathematical models were verified on known species for predicting CF in species for which sound is unknown (e.g. fossils or museum specimens).

Evolutionary diversification of the auditory organ sensilla in Neoconocephalus katydids (Orthoptera: Tettigoniidae) correlates with acoustic signal diversification over phylogenetic relatedness and life history.

Neoconocephalus Tettigoniidae are a model for the evolution of acoustic signals as male calls have diversified in temporal structure during the radiation of the genus. The call divergence and phylogeny in Neoconocephalus are established, but in tettigoniids in general, accompanying evolutionary changes in hearing organs are not studied. We investigated anatomical changes of the tympanal hearing organs during the evolutionary radiation and divergence of intraspecific acoustic signals. We compared the neuroanatomy of auditory sensilla (crista acustica) from nine Neoconocephalus species for the number of auditory sensilla and the crista acustica length. These parameters were correlated with differences in temporal call features, body size, life histories and different phylogenetic positions. By this, adaptive responses to shifting frequencies of male calls and changes in their temporal patterns can be evaluated against phylogenetic constraints and allometry. All species showed well-developed auditory sensilla, on average 32-35 between species. Crista acustica length and sensillum numbers correlated with body size, but not with phylogenetic position or life history. Statistically significant correlations existed also with specific call patterns: a higher number of auditory sensilla occurred in species with continuous calls or slow pulse rates, and a longer crista acustica occurred in species with double pulses or slow pulse rates. The auditory sensilla show significant differences between species despite their recent radiation, and morphological and ecological similarities. This indicates the responses to natural and sexual selection, including divergence of temporal and spectral signal properties. Phylogenetic constraints are unlikely to limit these changes of the auditory systems.

Chamber music: an unusual Helmholtz resonator for song amplification in a Neotropical bush-cricket (Orthoptera, Tettigoniidae).

Animals use sound for communication, with high-amplitude signals being selected for attracting mates or deterring rivals. High amplitudes are attained by employing primary resonators in sound-producing structures to amplify the signal (e.g. avian syrinx). Some species actively exploit acoustic properties of natural structures to enhance signal transmission by using these as secondary resonators (e.g. tree-hole frogs). Male bush-crickets produce sound by tegminal stridulation and often use specialised wing areas as primary resonators. Interestingly, Acanthacara acuta, a Neotropical bush-cricket, exhibits an unusual pronotal inflation, forming a chamber covering the wings. It has been suggested that such pronotal chambers enhance amplitude and tuning of the signal by constituting a (secondary) Helmholtz resonator. If true, the intact system - when stimulated sympathetically with broadband sound - should show clear resonance around the song carrier frequency which should be largely independent of pronotum material, and change when the system is destroyed. Using laser Doppler vibrometry on living and preserved specimens, microcomputed tomography, 3D-printed models and finite element modelling, we show that the pronotal chamber not only functions as a Helmholtz resonator owing to its intact morphology but also resonates at frequencies of the calling song on itself, making song production a three-resonator system.

The thorax of the cave cricket Troglophilus neglectus: anatomical adaptations in an ancient wingless insect lineage (Orthoptera: Rhaphidophoridae).

BACKGROUND: Secondary winglessness is a common phenomenon found among neopteran insects. With an estimated age of at least 140 million years, the cave crickets (Rhaphidophoridae) form the oldest exclusively wingless lineage within the long-horned grasshoppers (Ensifera). With respect to their morphology, cave crickets are generally considered to represent a `primitive' group of Ensifera, for which no apomorphic character has been reported so far. RESULTS: We present the first detailed investigation and description of the thoracic skeletal and muscular anatomy of the East Mediterranean cave cricket Troglophilus neglectus (Ensifera: Rhaphidophoridae). T. neglectus possesses sternopleural muscles that are not yet reported from other neopteran insects. Cave crickets in general exhibit some unique features with respect to their thoracic skeletal anatomy: an externally reduced prospinasternum, a narrow median sclerite situated between the meso- and metathorax, a star-shaped prospina, and a triramous metafurca. The thoracic muscle equipment of T. neglectus compared to that of the bush cricket Conocephalus maculatus (Ensifera: Tettigoniidae) and the house cricket Acheta domesticus (Ensifera: Gryllidae) reveals a number of potentially synapomorphic characters between these lineages. CONCLUSIONS: Based on the observed morphology we favor a closer relationship of Rhaphidophoridae to Tettigoniidae rather than to Gryllidae. In addition, the comparison of the thoracic morphology of T. neglectus to that of other wingless Polyneoptera allows reliable conclusions about anatomical adaptations correlated with secondary winglessness. The anatomy in apterous Ensifera, viz. the reduction of discrete direct and indirect flight muscles as well as the strengthening of specific leg muscles, largely resembles the condition found in wingless stick insects (Euphasmatodea), but is strikingly different from that of other related wingless insects, e.g. heel walkers (Mantophasmatodea), ice crawlers (Grylloblattodea), and certain grasshoppers (Caelifera). The composition of direct flight muscles largely follows similar patterns in winged respectively wingless species within major polyneopteran lineages, but it is highly heterogeneous between those lineages.

Biomechanics of hearing in katydids.

Animals have evolved a vast diversity of mechanisms to detect sounds. Auditory organs are thus used to detect intraspecific communicative signals and environmental sounds relevant to survival. To hear, terrestrial animals must convert the acoustic energy contained in the airborne sound pressure waves into neural signals. In mammals, spectral quality is assessed by the decomposition of incoming sound waves into elementary frequency components using a sophisticated cochlear system. Some insects like katydids (or bushcrickets) have evolved biophysical mechanisms for auditory processing that are remarkably equivalent to those of mammals. Located on their front legs, katydid ears are small, yet are capable of performing several of the tasks usually associated with mammalian hearing. These tasks include air-to-liquid impedance conversion, signal amplification, and frequency analysis. Impedance conversion is achieved by a lever system, a mechanism functionally analogous to the mammalian middle ear ossicles, yet morphologically distinct. In katydids, the exact mechanisms supporting frequency analysis seem diverse, yet are seen to result in dispersive wave propagation phenomenologically similar to that of cochlear systems. Phylogenetically unrelated katydids and tetrapods have evolved remarkably different structural solutions to common biophysical problems. Here, we discuss the biophysics of hearing in katydids and the variations observed across different species.

Wing stridulation in a Jurassic katydid (Insecta, Orthoptera) produced low-pitched musical calls to attract females.

Behaviors are challenging to reconstruct for extinct species, particularly the nature and origins of acoustic communication. Here we unravel the song of Archaboilus musicus Gu, Engel and Ren sp. nov., a 165 million year old stridulating katydid. From the exceptionally preserved morphology of its stridulatory apparatus in the forewings and phylogenetic comparison with extant species, we reveal that A. musicus radiated pure-tone (musical) songs using a resonant mechanism tuned at a frequency of 6.4 kHz. Contrary to previous scenarios, musical songs were an early innovation, preceding the broad-bandwidth songs of extant katydids. Providing an accurate insight into paleoacoustic ecology, the low-frequency musical song of A. musicus was well-adapted to communication in the lightly cluttered environment of the mid-Jurassic forest produced by coniferous trees and giant ferns, suggesting that reptilian, amphibian, and mammalian insectivores could have also heard A. musicus' song.

First record of the subfamily Anoplophilinae (Orthoptera: Rhaphidophoridae) from Russia with description of a new species of the genus Alpinanoplophilus Ishikawa, 1993.

The subfamily Anoplophilinae (Rhaphidophopridae) is recorded from Russia for the first time. Alpinanoplophilus kurilensis Storozhenko, sp. nov. is described from Kunashir Island. The holotype of the new species is deposited in the Zoological Institute of Russian Academy of Science (St. Petersburg, Russia). A revised key to the species of the genus Alpinanoplophilus Ishikawa, 1993 is provided.

Notes on the genus Conanalus Tinkham, 1943 (Orthoptera: Tettigoniidae: Conocephalinae) with one new species from Yunnan, China.

The paper describes one new species, i.e. Conanalus bilobus sp. nov., and provides additional information on Conanalus brevicaudus Shi, Mao & Ou, 2008 (including female description for the first time), and Conanalus robustus Shi, Mao & Ou, 2008 from Yunnan, as well as illustrations of important morphological characters of Conanalus axinus Shi, Wang & Fu, 2005. A distribution map of the species of Conanalus from Yunnan, China is presented. The type material is deposited in the Museum of Hebei University.

The tribe Dysoniini part IV: New species of Quiva Hebard, 1927 (Orthoptera: Tettigoniidae: Phaneropterinae) from Brazilian rainforest and some clarifications.

Two new species of the genus Quiva: Quiva buhrnheimi n. sp. and Quiva gutjahrae n. sp. from Brazilian Amazon are described. Determinations for distributional data previously published by Sovano & Mendes (2013) are clarified and the synonymy of Ituana dorisae under Q. abacata is confirmed. In this paper, an updated key to subgenus Quiva is provided.

Sensory evolution of hearing in tettigoniids with differing communication systems.

In Tettigoniidae (Orthoptera: Ensifera), hearing organs are essential in mate detection. Male tettigoniids usually produce calling songs by tegminal stridulation, whereas females approach the males phonotactically. This unidirectional communication system is the most common one among tettigoniids. In several tettigoniid lineages, females have evolved acoustic replies to the male calling song which constitutes a bidirectional communication system. The genus Poecilimon (Tettigoniidae: Phaneropterinae) is of special interest because the ancestral state of bidirectional communication, with calling males and responding females, has been reversed repeatedly to unidirectional communication. Acoustic communication is mediated by hearing organs that are adapted to the conspecific signals. Therefore, we analyse the auditory system in the Tettigoniidae genus Poecilimon for functional adaptations in three characteristics: (i) dimension of sound-receiving structures (tympanum and acoustic spiracle), (ii) number of auditory sensilla and (iii) hearing sensitivity. Profound differences in the auditory system correlate with uni- or bidirectional communication. Among the sound-receiving structures, the tympana scale with body size, whereas the acoustic spiracle, the major sound input structure, was drastically reduced in unidirectional communicating species. In the unidirectional P. ampliatus group, auditory sensilla are severely reduced in numbers, but not in the unidirectional P. propinquus group. Within the P. ampliatus group, the number of auditory sensilla is further reduced in P. intermedius which lost acoustic signalling due to parthenogenesis. The auditory sensitivity correlated with the size of the acoustic spiracle, as hearing sensitivity was better with larger spiracles, especially in the ultrasonic range. Our results show a significant reduction in auditory structures, shaped by the differing sex roles during mate detection.

Notes on the genus Acosmetura Liu, 2000 (Orthoptera: Tettigoniidae: Meconematinae).

This paper deals with one new species, Acosmetura mabianensis Bian & Shi sp. nov., one new combination, Acosmetura jinyunensis (Shi & Zheng, 1994) com. nov., and the first record of female of Acometura nigrogeniculata (Liu & Wang, 1998). Photographs of two known species, a key to the species, male stridulatory files and a distribution map are provided. We also observe the male stridulatory files and left tegmina for four known species of the genus. The material is deposited in the Museum of Hebei University.

The phallus in Tettigoniidae (Insecta: Orthoptera: Ensifera): revision of morphology and terminology, and discussion on its taxonomic importance and evolution.

The phallus in Tettigoniidae (katydids) is a structure informative relative to the systematics of the group. Despite this, it is often not considered in descriptions of taxa. The lack of adequate descriptions of phalli is not only a gap for sytematic and morphological studies, but postpones works on the evolution of copula. Here we study the exoskeletal morphology of the phallus in katydids, its components, and revised the terminology for them. We carried out dissections for morphological comparisons, and complement the observational information with published data. We stained phalli of katydids with chlorazol black, to better contrast membranous versus sclerotized components. We demonstrate that phallic components vary at specific, generic and suprageneric levels, and that internal and external components vary in number, shape, size and position. Currently there is little comparative data to support hypotheses on the evolution of this structure, but possibly the possession of a titillator is an ancestral condition. We identify additional sclerotized components, the sclerites of the ventral fold of the dorsal lobe, which can modify the shape and function of the titillator, being also important for understanding the evolution of the phallus. Potential functional relationships based on hypothetical morphological correlations between the shape of titillator and cerci are proposed, categorized in three main groups: (i) phallus devoid of titillator and cerci simple, (ii) titillator with bifurcated or paired sclerites, and cerci adapted for grasping, and (iii) titillator with single process and/or sclerite and cerci simple, sometimes with a pointed tip. Two explanations for these hypothetical morphological correlations and morphological variation are proposed: first, species with similar structures at the postabdomen would share similar copulatory behaviour, and second, more than one selective pressure would have acted over the structures of the postabdomen.

A new genus of Phaneropterinae (Orthoptera: Ensifera: Tettigoniidae) from Central Africa.

The new genus Pseudoplangia is described for Plangia laminifera Karsch, 1896. Its general aspect is similar to that of the genus Plangia Stål, 1873, but it differs remarkably in the vertex width, in the shape of eyes, that are oval and elongate, in the length of mid femora, that are shorter than pronotum length, in the shape of fore and mid legs that are very much laterally compressed, and in the presence of broad-based spines on the hind tibiae.

Description of Dociostaurus biskrensis sp. nov. and male allotypes of four species: Pamphagulus bodenheimeri dumonti, P. uvarovi, Sphingonotus ebneri and Notopleura pygmaea (Orthoptera: Acridoidea) in the region of Biskra, Algeria.

The new species Dociostaurus biskrensis Moussi & Petit 2013 was collected in the region of Biskra, at the arido-Saharan limit of Eastern Algerian. We also describe the males of four species for which only females were known: Pamphagulus bodenheimeri dumonti, P. uvarovi, Notopleura pygmaea and Sphingonotus ebneri. The key to the genus Dociostaurus in North Africa is given. The species P. uvarovi and Notopleura pygmaea are new for the Algeria. The diets and life cycles of D. biskrensis and of the two species of Pamphagulus are defined.