A comparative study of genome organization and inferences for the systematics of two large bushcricket genera of the tribe Barbitistini (Orthoptera: Tettigoniidae: Phaneropterinae).

BACKGROUND: Poecilimon and Isophya are the largest genera of the tribe Barbitistini and among the most systematically complicated and evolutionarily intriguing groups of Palearctic tettigoniids. We examined the genomic organization of 79 taxa with a stable chromosome number using classical (C-banding, silver and fluorochrome staining) and molecular (fluorescence in situ hybridization with 18S rDNA and (TTAGG)n telomeric probes) cytogenetic techniques. These tools were employed to establish genetic organization and differences or similarities between genera or species within the same genus and determine if cytogenetic markers can be used for identifying some taxonomic groups of species. RESULTS: Differences between the karyotypes of the studied genera include some general changes in the morphology of the X chromosome in Isophya (in contrast to Poecilimon). The number of major rDNA clusters per haploid genome divided Poecilimon into two main almost equal groups (with either one or two clusters), while two rDNA clusters predominated in Isophya. In both genera, rDNA loci were preferentially located in the paracentromeric region of the autosomes and rarely in the sex chromosomes. Our results demonstrate a coincidence between the location of rDNA loci and active NORs and GC-rich heterochromatin regions. The C/DAPI/CMA3 bands observed in most Poecilimon chromosomes suggest the presence of more families of repetitive DNA sequences as compared to the heterochromatin patterns in Isophya. CONCLUSIONS: The results show both differences and similarities in genome organization among species of the same genus and between genera. Previous views on the systematics and phylogenetic grouping of certain lineages are discussed in light of the present cytogenetic results. In some cases, variation of chromosome markers was observed to correspond with variation in other evolutionary traits, which is related to the processes of ongoing speciation and hybridization in zones of secondary contact. It was concluded that the physical mapping of rDNA sequences and heterochromatin may be used as an additional marker for understanding interspecific relationships in these groups and their routes of speciation.

Review of the Balkan Isophya (Orthoptera: Phaneropteridae) with particular emphasis on the Isophya modesta group and remarks on the systematics of the genus based on morphological and acoustic data.

A critical review of the taxonomy and systematics of the genus Isophya from the Balkan Peninsula, together with a revision of the I. modesta group, including its representatives outside the Balkans, has been made using morphological, bioacoustic and karyological data. As a result, the following taxonomic considerations have been proposed: (1) the status of two taxa has been reconsidered: Isophya rhodopensis leonorae Kaltenbach, stat.n., I. rhodopensis petkovi Peshev, stat.n.; (2) five synonymies have been established: I. hospodar (Saussure) = I. hospodar medimontana Nedelkov, syn.n.; I. plevnensis Peshev, sensu novo = I. pravdini pravdini Peshev, syn.n.; I. rhodopensis leonorae Kaltenbach = I. kisi Peshev, syn.n.; I. obtusa = I. pravdini bazyluki Peshev, syn.n.; I. modesta Frivaldszky = I. modesta intermedia Kis syn.n. The descriptions of the taxa under consideration are supplemented with unpublished morphological and/or bioacoustic data. An updated list of the taxa occurring on the Balkans includes 24 taxa (21 species, including the doubtful data on I. camptoxypha). A dichotomic and tabulated key for recognition and maps of distribution of the established taxa are presented.

A review of Eupholidoptera (Orthoptera, Tettigoniidae) from Crete, Gavdos, Gavdopoula, and Andikithira.

Being nocturnal, hiding in prickly bushes and shrubs during the day, Eupholidoptera species in Crete and its neighbouring islands are easily overlooked, and until now our knowledge about their distribution was based on some thirty sightings across 11 species. In this paper results are presented of a study of Eupholidoptera specimens collected between 1987 and 2020 by hand-catches and pitfall and fermenting traps on the Greek islands of Crete, Gavdos, Gavdopoula, and Andikithira. Diagnostic features of all known species are presented and illustrated with stacked images. An updated key to all species is provided. Eupholidopterafrancisae Tilmans & Odé, sp. nov. from Andikithira and southwestern Crete and Eupholidopteramarietheresae Willemse & Kotitsa, sp. nov. from Mt. Dikti are described. Female E.cretica, E.gemellata, and E.mariannae are described, and the female of E.astyla is redescribed. Bioacoustics for E.francisae Tilmans & Odé, sp. nov., E.giuliae, and E.jacquelinae are presented for the first time. Eupholidopterasmyrnensis is reported for the first time from Crete. A substantial amount of new distribution data for Eupholidoptera species on Crete is presented. The current distribution pattern and first analyses of phylogeny based on molecular data of Eupholidoptera species on Crete are discussed in relation to paleogeographical events.

A multi-taxon analysis of European Red Lists reveals major threats to biodiversity.

Biodiversity loss is a major global challenge and minimizing extinction rates is the goal of several multilateral environmental agreements. Policy decisions require comprehensive, spatially explicit information on species' distributions and threats. We present an analysis of the conservation status of 14,669 European terrestrial, freshwater and marine species (ca. 10% of the continental fauna and flora), including all vertebrates and selected groups of invertebrates and plants. Our results reveal that 19% of European species are threatened with extinction, with higher extinction risks for plants (27%) and invertebrates (24%) compared to vertebrates (18%). These numbers exceed recent IPBES (Intergovernmental Platform on Biodiversity and Ecosystem Services) assumptions of extinction risk. Changes in agricultural practices and associated habitat loss, overharvesting, pollution and development are major threats to biodiversity. Maintaining and restoring sustainable land and water use practices is crucial to minimize future biodiversity declines.

On some Moroccan Pamphagidae, with the description of a new species of Euryparyphes (Orthoptera: Caelifera).

An update to the distribution records of the Moroccan Pamphagidae is presented. One new species to science, Euryparyphes minor sp. n., is described, and one new combination is proposed.

Review of song patterns and sound production in armoured ground crickets (Orthoptera: Tettigoniidae: Hetrodini) with karyological data and taxonomic notes.

In the mate finding system of bush-crickets, acoustical signals play a central role. Here we review and describe the bioacoustics of Hetrodini, a morphologically uniform group of Tettigonioidea with a distribution centered in Africa. The male calling songs are produced by tegmino-tegminal stridulation. In all species, these fore wings are completely covered by the pronotum (invisible in intact specimens) and lack the glossy mirror cells which are well-known from many other singing Ensifera. Concerning spectral composition, the broad frequency peak and the parts with the most energy lie in the high audio or low ultrasonic range. In amplitude modulation, the songs are relatively simply structured and contain only one type of syllables each. These syllables consist of heavily dampened impulses (non-resonant song) and are arranged in long series of echemes (chirps) or trills. As far as it is known, syntopic species differ in syllable and/or echeme repetition rates. In contrast to the uniform morphology, the karyotypes are surprisingly diverse with chromosome numbers ranging from 2n = 29 to 17 and one of two sex determination systems: X0 and neo-XY. In our opinion, taxonomically the group is at the moment at best considered as a tribe incertae sedis (without subfamily classification) within Tettigoniidae (not Tettigoniinae sensu OSF).

Geographic differentiation in male calling song of Isophyamodestior (Orthoptera, Tettigoniidae, Phaneropterinae).

We studied the songs and morphology of the stridulatory file of Isophyamodestior across its complete geographic range, in order to test our hypothesis that the male calling song of the species shows strong differentiation between the northern (Pannonian) and southern (Balkan) parts of its distribution range, reflecting its disjunct distribution. Our analyses confirm this hypothesis, separating analyzed specimens of I.modestior into two main groups - one present in the central part of the Balkan Peninsula (representing Isophyamodestior sensu stricto), with the second group occurring in the Pannonian Basin, Dinarides, Slovenia and NE Italy. The most reliable difference between the groups is the duration of the main syllable, the number of stridulatory teeth and number of pulses in the main syllable, where all values are higher in specimens from the Balkan Peninsula. Additional analyses showed that within the second group, there are differences in analyzed characters between specimens from the Pannonian Basin and specimens from the Dinaric area, the latter ones having intermediate song characteristics, closer to the group from the Balkan Peninsula. Our study shows that detailed bioacoustic analyses can help to unravel patterns of intraspecific differentiation and thus provide a useful tool for taxonomic studies.

A taxonomic outline of the Poecilimon affinis complex (Orthoptera) using the geometric morphometric approach.

The genus Poecilimon contains 145 species, widely distributed in the Palaearctic, among which the Poecilimon ornatus group has the greatest diversity in the Balkans. Despite several revisions of the genus, the systematics of the species group, and in particular, of the taxa associated with the species Poecilimon affinis, is still unsolved. Due to morphological similarity, P. affinis with its subspecies, P. nonveilleri and P. pseudornatus form the Poecilimon affinis complex. The aim of this study is to test the hypotheses of an outlined species complex, namely the P. affinis complex, within the P. ornatus group using morphological data. Geometric analysis was conducted to explore variation in the structure of the male tegmen, ovipositor, male cercus, and male pronotum. The number of teeth and stridulatory file measurements provided additional information on morphological variation within the complex. A phylogenetic tree based on the cytochrome c oxidase subunit I gene (COI) was used for comparison with the morphological data. Canonical variate analysis showed that male tegmen and male cercus are good morphostructures to distinguish the taxa belonging to the P. affinis complex from other species in the P. ornatus group. This may confirm our assumption for the designation of the P. affinis complex. The results of the principal component analysis of stridulatory file measurements, molecular data, and CVA of the ovipositor suggest adding two additional species to the complex: P. ornatus and P. hoelzeli.

Songs in the genus Uromenus (Orthoptera: Tettigoniidae: Ephippigerini): A review with new information about some species.

With nearly 60 described forms Uromenus is the most species-rich genus of the tribe Ephippigerini, a west Palearctic group of flightless bush-crickets (katydids). As it is typical for bush-crickets, Uromenus males produce species-specific calling songs to attract females prepared to mate. These insects are relatively large and their songs have always components audible to humans. Nevertheless, acoustical information are available only for few species. Also the phylogenetic relationships of species and species groups are poorly known. In this paper we present new data on the song and morphology of several species (U. dyrrhachiacus, U. elegans, U. finoti, U. galvagnii, U. robustus, U. tobboganensis and U. innocentii) and give a review of all published acoustical data. Judging from the still few data, as in other Ephippigerini the song patterns often seem to contain species-group characteristics making them candidates to be used in phylogenetic studies. Additionally, we have studied the female stridulatory organs which are unique in structure in Ephippigerini and completely different from those of the males. Despite in Uromenus used only for defensive stridulation, possibly species-specific similarities in structure between males and females exist.

Bioacoustics of poorly known Poecilimon taxa (Insecta: Orthoptera: Tettigoniidae) with redescriptions of P. pechevi and P. stschelkanovzevi.

The genus Poecilimon has been object of several studies dealing with its systematics, sound communication and evolution. Yet, published data contradict in the classification, while many taxa are still insufficiently known. In the present study we supplement the knowledge of 13 poorly known species and one additional subspecies with data on their sound communication and/or morphology. Most species concerned here are classified within two acoustic groups. First group includes P. celebi, P. obtusicercus and P. toros, showing typical characteristics of the P. syriacus group. Second group is more heterogeneous. Among that, P. pechevi, P. armeniacus, P. harveyi, P. guichardi, P. haydari, P. doga, P. davisi and P. excisus present a song pattern similar to that of P. ampliatus. In addition, P. ataturki is also quite similar in basic song structure to the latter but differs in its fine song structure. Poecilimon stschelkanovzevi, with unclear relationships, is described morphologically in detail. The present study provides hints for solving some disagreement between recent revisions, proposing unification of the Poecilimon celebi with P. syriacus group and opening discussion based on close relationships of the groups ampliatus, armeniacus, davisi, luschani and the species P. guichardi on one hand, while the complex P. ataturki/glandifer shows significant differences on the other. The acoustic peculiarities of the ampliatus group allies are discussed in an evolutionary light.

Ceraeocercus Uvarov, a genus recorded in China for the first time (Orthoptera, Tettigoniidae; Tettigoniinae; Drymadusini).

Ceraeocercus Uvarov, 1910 of Tettigoniinae; Drymadusini is receded in China for the first time. Ceraeocercus fuscipennis fuscipennis Uvarov, 1910 is documented. Morphology and male calling songs are described and illustrations are provided.

Comparative analysis of C-heterochromatin, ribosomal and telomeric DNA markers in chromosomes of Pamphagidae grasshoppers from Morocco.

The karyotypes and the localization of C-bands, clusters of ribosomal DNA and telomeric repeats of 10 species of the family Pamphagidae from Morocco are described for the first time. The species studied belong to the subfamilies Pamphaginae and Thrinchinae. All species have karyotypes consisting of 19 and 20 acrocentric chromosomes and X0/XX sex chromosome system in males and females, respectively (2n♂=19, NF=19; 2n♀=20, NF=20). Despite the karyotype conservatism, we revealed differences in the location and size of C-heterochromatin blocks and ribosomal DNA clusters. A comparative analysis of these differences shows that karyotype divergences in this group is connected not to structural chromosome rearrangements, but to the evolution of repetitive DNA.

The first record of genus Gryllomorpha Fieber in Romania (Orthoptera: Gryllidae: Gryllomorphinae).

Although the fauna of Dobrogea (south-eastern Romania and north-eastern Bulgaria) is relatively well studied, the discovery of a crevice cricket was quite unexpected. Described from Ukraine and known to occur in Bulgaria, Greece, Turkey, Kazakhstan and possibly Uzbekistan, Gryllomorpha miramae is now recorded for the first time in Romania.

Taxonomy of the genus Isophya (Orthoptera, Phaneropteridae, Barbitistinae): comparison of karyological and morphological data.

The genus Isophya represents one of the largest orthopteran genera with about 45 species occurring in Europe. All over its range the genus includes groups of sibling species, the recognition of which is hampered by their morphological similarity. In part, some species were grouped according to their morphology: the Isophya straubei-group, the I. amplipennis-group and the I. major-group. Nevertheless, many species have not been studied using new methods. This is particularly true for 15 taxa described from the Balkans, more or less resembling I. modesta, known mostly from old, unsatisfactorily informative descriptions. Chromosomal analysis of 25 species/subspecies of the genus Isophya showed the karyotypic evolution among species. The karyotype of 18 species/subspecies was investigated for the first time. The most remarkable changes in the sex chromosomes were the ancient acrocentric X chromosome inversions in 19 species and an X chromosome/autosome mutual tandem translocation, from which the sex determination system neo-XY originated in I. hemiptera. Karyotype differentiation has been less rapid in autosomes than in sex chromosomes, in this case interspecific autosomal differentiation has involved the distribution and quantity of C-heterochromatin as well as the number of NORs. Cytogenetic analysis revealed some changes reflecting the level of genomic organization, and the results suggest some association of related taxa from biogeographic regions.

Review of Poecilimon species with inflated pronotum: description of four new taxa within an acoustically diverse group.

Poecilimon Fischer (Orthoptera, Tettigoniidae) consists of about 130 species and a functional taxonomy of the genus requires arranging species into phylogenetic groups. However, this is a task that requires detailed empirical testing. This paper documents data on Poecilimon species with an inflated pronotum. The Poecilimon (Poecilimon) inflatus group is a lineage ranging along a narrow zone in the south-west corner of Anatolia and in Crete plus some other Aegean islands. By producing data of morphology and calling songs of males and responding songs of females the composition and intra-group diversity of P. inflatus group are studied. Both morphology and male calling and female responding songs, exhibited a considerable intra-group diversity. In the light of data obtained the following nomenclatural changes were made. The genus Parapoecilimon Karabag, 1975 syn. n. is synonymised with Poecilimon Fischer and Parapoecilimon antalyaensis Karabag, 1975 is given a new combination, Poecilimon antalyaensis (Karabag, 1975) comb. n. Four new taxa are described in the group: (i) Poecilimon isopterus sp.n.; (ii) Poecilimon inflatus lyciae subsp. n.; (iii) Poecilimon antalyaensis myrae subsp. n., and (iv) Poecilimon antalyaensis anemurium subsp. n. The Poecilimon (P.) inflatus species group consists of the following species: (i) Poecilimon inflatus Brunner von Wattenwyl, 1891; (ii) Poecilimon martinae Heller, 2004; (iii) Poecilimon cretensis Werner, 1903; (iv) Poecilimon antalyaensis (Karabag, 1975) comb. n.; (v) Poecilimon bilgeri Karabag, 1953 and (vi) Poecilimon isopterus sp.n. The species group is defined by a combination of characters including the pronotum inflated in metazona and a male cerci mostly black in their apical 1/2-1/3. The male calling song has typically two types of short syllables and the females respond acoustically with very short latency times. By using combination of morphology and song data we suggest three main lineages in the group: (i) P. bilgeri, (ii) P. antalyaensis and (iii) the other four species.

New insights into the karyotype evolution of the genus Gampsocleis (Orthoptera, Tettigoniinae, Gampsocleidini).

Five species belonging to the genus Gampsocleis Fieber, 1852 were analyzed using fluorescence in situ hybridization (FISH) with 18S rDNA and telomeric probes, as well as C-banding, DAPI/CMA3 staining and silver impregnation. The studied species showed two distinct karyotypes, with 2n = 31 (male) and 2n = 23 (male) chromosomes. The drastic reduction in chromosome number observed in the latter case suggests multiple translocations and fusions as the main responsible that occurred during chromosome evolution. Two groups of rDNA distribution were found in Gampsocleis representatives analyzed. Group 1, with a single large rDNA cluster on the medium-sized autosome found in four species, carried in the haploid karyotype. Group 2, represented only by G.abbreviata, was characterized by the presence of two rDNA signals. TTAGG telomeric repeats were found at the ends of chromosome arms as expected. The rDNA clusters coincided with active NORs and GC-rich segments.

Comparative analysis of chromosomes in the Palaearctic bush-crickets of tribe Pholidopterini (Orthoptera, Tettigoniinae).

The present study focused on the evolution of the karyotype in four genera of the tribe Pholidopterini: Eupholidoptera Maran, 1953, Parapholidoptera Maran, 1953, Pholidoptera Wesmaël, 1838, Uvarovistia Maran, 1953. Chromosomes were analyzed using fluorescence in situ hybridization (FISH) with 18S rDNA and (TTAGG) n telomeric probes, and classical techniques, such as C-banding, silver impregnation and fluorochrome DAPI/CMA3 staining. Most species retained the ancestral diploid chromosome number 2n = 31 (male) or 32 (female), while some of the taxa, especially a group of species within genus Pholidoptera, evolved a reduced chromosome number 2n = 29. All species show the same sex determination system X0/XX. In some taxa, a pericentric inversion has changed the morphology of the ancestral acrocentric X chromosome to the biarmed X. The rDNA loci coincided with active NORs and C-band/CG-rich segments. A comparison of the location of the single rDNA/NOR in the genus Pholidoptera suggests that reduced chromosome number results from Robertsonian translocation between two pairs of autosomes, one carrying the rDNA/NOR. The results constitute a step towards better understanding of the chromosomal reorganization and evolution within the tribe Phaneropterini and the whole subfamily Tettigoniinae.

The unexpected finding of Parapholidoptera castaneoviridis in south-eastern Romania (Insecta, Orthoptera, Tettigoniidae).

The Balkano-Anatolian genus Parapholidoptera comprises 21 species and the westernmost one, Parapholidoptera castaneoviridis, previously recognized to occur in western Turkey, north-eastern Greece and south-eastern Bulgaria is recorded for the first time from south-eastern Romania, almost 300 km away from the closest known locality. Illustrations and measurements of morphological characters are given and the male calling song from this new, northernmost population is described.

New data on the bush-cricket Montana medvedevi (Orthoptera: Tettigoniidae), critically endangered in Europe (EU 28), and a comparison of its song with all known song patterns within the genus.

Montana medvedevi is reported for the first time from Serbia. New information about the distribution, morphology and song of this species is discussed. The song of M. medvedevi is different from that of all other members of the genus, all figured for comparison. Montana is quite diverse regarding the amplitude pattern of the calling song of its members (known in 15 species). Surprisingly, some Montana species seem to have two song patterns, one produced during the day and one at night.

Corrigenda: Iorgu IȘ, Iorgu EI, Puskás G, Ivkovic S, Borisov S, Gavril VD, Chobanov DP (2016) Geographic distribution of Gryllotalpa stepposa (Insecta, Orthoptera, Gryllotalpidae) in South-eastern Europe, with first records for Romania, Hungary and Serbia. ZooKeys 605: 73-82, doi: 10.3897/zookeys.605.8804.

[This corrects the article DOI: 10.3897/zookeys.605.8804.].