Anatolian Short-Horned Grasshoppers Unveiled: Integrating Biogeography and Pest Potential.

Biogeographically, Anatolia harbours a rich diversity of short-horned grasshoppers (Orthoptera, Caelifera). The number of species recorded from Anatolia so far stands at 300. They inhabit diverse habitats ranging from arid Eremial to Euro-Siberian-like montane meadows, aligning with the topographical and climatological heterogeneity of Anatolia. Alongside some swarming species, the pest potential of several pullulating species needs attention. This is especially important concerning global warming, a scenario expected to be more severe in the Northern Mediterranean Basin in general and Anatolia specifically. A faunal list of biogeographic Anatolia, the area extending from the Aegean Sea in the west to the intermountain basin of the Caucasus in the northeast, the lowlands of Lake Urmia in the east, and Mesopotamia in the southeast, was developed. The recorded species were classified according to the phytogeographical provinces of Anatolia. Distributions of the species with the potential for pullulating were modelled using ecological-niche-modelling approaches for the present and future. The results have the potential to lead to the development of a concept that merges biogeography and the pest potential of certain Anatolian grasshopper species. Our results reveal the following: (i) Acrididae and Pamphagidae are the most diverse families represented in Anatolia; (ii) roughly 40% of Caelifera and 71% of Pamphagidae are endemics, suggesting Anatolia is a biodiversity hotspot; (iii) according to Caelifera diversity, the phytogeographical provinces of Anatolia follow an order of Irano-Anatolia, Euro-Siberia, Mediterranean, and Mesopotamia; and (iv) based on ecological modelling and personal observations, Dociostaurus maroccanus, Locusta migratoria, Calliptamus italicus, Heteracris pterosticha, Notostaurus anatolicus, Oedipoda miniata, and O. schochii should be monitored regarding their pest potential.

Microgeographic speciation in a complex of Anatolian bush crickets facilitated by fast evolution of reproductive isolation.

Identifying the drivers of microgeographic speciation (i.e., speciation over small, local geographic scales) is key to understand the origin of speciose groups. Here, we use genomic data to infer the demographic processes underlying diversification in Poecilimon luschani (Orthoptera: Tettigoniidae), a species complex belonging to the most diverse genus of bush crickets from the Mediterranean region (>170 taxa) that comprises three recognized subspecies with small allopatric distributions in the topographically complex Teke Peninsula, southwestern Anatolia. Phylogenomic reconstructions that include all other taxa within the species group confirmed that subspecies of P. luschani originated from a common ancestor during the Pleistocene, supporting recent (<1 Ma) diversification within a small geographical area (ca. 120 × 80 km). Genetic clustering analyses corroborated the distinctiveness of each subspecies and the cohesiveness of their respective populations, with abrupt genetic discontinuities coinciding with contemporary range boundaries. Indeed, our analyses uncovered the presence of two sympatric cryptic sister lineages that diverged <300 ka ago and do not admix despite being co-distributed. Collectively, these results support that all lineages within the complex represent independently evolving entities corresponding to full-fledged species. Statistical evaluation of alternative models of speciation strongly supports a scenario of divergence in isolation followed by a period of limited gene flow during the last glacial period, when all lineages experienced marked expansions according to demographic reconstructions. Our study exemplifies how localized allopatric divergence and fast evolution of reproductive isolation can promote microgeographic speciation and explain the high rates of endemism characterizing biodiversity hotspots.

Mitochondrial genome of Poecilimon cretensis (Orthoptera: Tettigoniidae: Phaneropterinae): Strong phylogenetic signals in gene overlapping regions.

We report the complete mitochondrial genome of the Cretan bush cricket Poecilimon cretensis. The mitogenome consists of 13 protein-coding regions, 22 tRNAs, two rRNAs, and one control region. The length of mitogenome in P. cretensis varies between15477 and 15631 bp, mainly due to variability in control region. The start and stop codons of protein coding genes exhibit the general pattern in Phaneropterinae. Phylogenetic tree constructed with the mitogenome obtained during this study and 12 mitogenomes of Phaneropterinae downloaded from GenBank, placed P. cretensis in Barbitistini as sister group to Poecilimon luschani. Data indicate that the gene overlapping pattern exhibit strong phylogenetic signals.

Anatolian endemic genus Bolua (Orthoptera: Tettigoniidae: Tettigoniinae): genetic and phenotypic data indicate inconsistent diversity and evolutionary patterns.

Anatolia harbours diverse and unique biodiversity. Nearly half of the genera and more than 80% of the species belonging to the tettigoniid subfamily Tettigoniinae are endemic to Anatolia. Although the Anatolian Tettigoniinae have been well studied, proper documentation of Tettigoniinae biodiversity requires studies using contemporary approaches and techniques. We attempted to study the taxonomy, phylogeny, and phylogeography of the Bolua, a monotypic genus endemic to northwest Anatolia. Species delimitation tests applied to three mitochondrial and two nuclear gene segments indicated three putative species in the genus. Analyses suggested inversely related genetic and phenotypic divergence. By integrating these results, we reached the following conclusions. (i) The plausible taxonomic composition recognizes three species in the genus; the type species Bolua turkiyae and two new species B. balikesirensis sp. n. and B. bursaensis sp. n., (ii) the radiation steps in Bolua correspond to the starting of the Pliocene, Mid Pleistocene Transition (MPT) and end of cold periods of Günz and Riss glacial. By considering intra generic divergence steps and habitat preference plus the distribution pattern of the species, ecological speciation seems to be the mode of generic radiation, (iii) possibly due to habitat conservatism, the isolated relict populations on highlands evolved under similar selection pressures, and this led to a conserved phenotype, consequently morphologically poorly diverged species, though genetic divergence is deep and (iv) although morphological diagnoses is poor, genetic divergence between species is deep, and each species fulfils the criteria of several species concepts.

Review of emUvarovistia/em (Orthoptera: Tettigoniidae; Tettigoniinae): genetic data indicate a new cryptic species, emUvarovistia/em emmunzurensis/em sp.n.

Genus Uvarovistia ranged along the Zagros Mountain belt. We presented first genetic data from three species of the genus and a taxonomic rectification indicated by these data. Sequences of three mitochondrial and two nuclear gene segments were obtained from different populations. Phylogenetic and automatic species delimitation analyses consistently suggested three distinct phylogroups as U. zebra, U. satunini and the third a new species, U. munzurensis Uluar Yahyaoglu sp. n. Time estimation and population genetic analyses supported consistent results. The following conclusions were reached: (i) five species in the genus constitute two distinct species groups named as Zebra Group and Satunini Group, (ii) data suggest intra generic relationships as U. zebra + ((U. satunini + U. munzurensis) + (U. bakhtiara +U. iraka)), (iii) molecular clock estimations indicated a deep divergence time and no gene flow between U. satunini and U. munzurensis, (iv) although these two species cannot be clearly distinguished by morphology, range of U. munzurensis seems to be isolated from other by lowlands of Euphrates Valley, and (v) age of generic ancestor is around five million years a time corresponding to connection of Anatolian and Zagros plates, and genus radiated along Zagros belt after dispersal of ancestral stock here.

Evolution, characterization and phylogenetic utility of ITS2 gene in Orthoptera and some Polyneoptera: Highly variable at the order level and highly conserved at the species level.

ITS2 is often suggested as a potential marker for evolutionary studies and species barcoding. However, there are many lineages have not been studied. This study focuses on ITS2 in Polyneoptera at the order and species levels. ITS2 sequences representing six polyneopteran orders and 15 species in the genus Anterastes are studied. We arrived at the following conclusions: (i) ITS2 is highly variable and contains little phylogenetic information in Polyneoptera, (ii) the shortest length and the highest GC content of ITS2 is found in Orthoptera among insects, (iii) the secondary structure exhibits general characteristics of eukaryotes especially in helices II and III, and with no order-specific architecture, (iv) ITS2 is highly conserved at the species level, both in linear sequences and secondary structures, (v) helices I, IA, II, IIA and III almost invariable in nucleotide sequence shared by all species in the genus. At the generic level, the most conspicuous result is the variable pattern in ITS2. It is highly conserved in helical sequences, but highly variable in non/peri-helical regions which we considered to be mutation islands. These frequently mutated regions contain a significant amount of molecular homoplasy, thus, the utility of ITS2 in phylogenetic analyses and species barcoding is low, at least in Polyneoptera.

Complete mitogenome of Psorodonotus venosus (Orthoptera, Tettigoniidae; Tettigoniinae): Short intergenic spacers shorten the total genome.

Mitogenomes are popular sources of data in evolutionary studies. By development of next generation sequencing the number of total mitogenome in data bases rapidly increased. However, there is still a limited number of total mitogenome known from species of Tettigoniinae. This paper aims to describe the total mitogenome of Psorodonotus venosus (Orthoptera, Tettigoniidae; Tettigoniinae) obtained by NGS reads. The total mitogenome is 15836-15845 bp and consists of 13 protein coding genes (PCG), 22 tRNA genes, two rRNA genes and an AT rich control region as in other metazozans. The mitogenome is AT skewed with 69.5% AT percentage. The genes are ordered as in pancrustacean. Total length of PCGs is 11229 bp, the start codon for all fits ATN pattern and stop codons are incomplete T-- / TA- and rarely complete TAA. Total length of 22 tRNA genes is 1447 bp and their anticodons are identical to other members of Tettigonioidea. The mitogenome contains 12 overlapping regions constituting 41 bp in total. Of these 12 overlapping regions those between trnW-trnC, atp6-atp8, nad4-nad4L, nad6 -cytb and atp6-cox3 gene pairs seem to be conserved. The total length of seven noncoding intergenic spacers is 46 bp. We concluded that P. venosus is one of the species with short mitogenome amongst Tettigonioidea because of limited number and length of noncoding intergenic spacers.

Mitogenome of Anterastes babadaghi (Orthoptera: Tettigoniinae; Platycleidini): Frequent conserved overlapping regions within Tettigoniinae.

Development of next generation sequencing rapidly increased the number of total mitogenome in data bases. However, the documented number of total mitogenome from species of Tettigoniinae is still limited and a comparison at subfamily level has not been made sufficiently. This paper aims to describe total mitogenome of A. babadaghi (Orthoptera, Tettigoniidae) by comparing to the known mitogenomes of other Tettigoniinae. The total mitogenome of A. babadaghi is 15882-15883 bp, AT skewed with 70.2% AT percentage, and consists of 13 protein coding genes (PCG), 22 tRNA genes, two rRNA genes and an AT rich control region. The genes are ordered as in pancrustacean. The comparative description of mitogenomes in Tettigoniinae showed that total length varies between 15766-16788 bp, the start codon for protein coding genes almost always fits to the ATN pattern, the stop codons are incomplete T-- / TA- and rarely complete TAA, intergenic spacers (IGS) and overlapping regions (OR) in species of the subfamily are similar in number, location, length and nucleotide sequence. We arrived to following conclusion from comparative data: (i) A. babadaghi has a typical orthopteran mitogenome by general features; (ii) this generalisation seems valid for Tettigoniinae as gene content, gene location, gene order, average AT content, anticodons and secondary structure of the tRNA genes, the start and stop codons of the protein coding genes, and several IGSs/ORs are similar to other orthopteran and hexapopods, (iii) variation range in total mitogenome length is narrow in Tettigoniinae and mainly determined by the lengths of control region and total IGSs, (iv) mitogenome of the subfamily exhibits conserved patterns especially in overlapping regions, but conserved features are mostly plesiomorphic.

Possibility of numt co-amplification from gigantic genome of Orthoptera: testing efficiency of standard PCR protocol in producing orthologous COI sequences.

Mitochondrial DNA has been the preferential genome biodiversity studies. However, several factors contribute to its inadequacy. Numts constitute one of the main complications that prevent obtaining orthologous mitochondrial sequences. Orthoptera have been a model group in numt studies because of their huge genome size. In this study we aimed to; (i) test efficiency of standard PCR protocol in producing orthologous sequences of cytochrome C oxidase, (ii) study presence/absence of numts in several unstudied Orthoptera species, (iii) test if there is a threshold between the length of mtDNA targeted for amplification and possibility of encountering numts, and (iv) estimate reliability of the sequences in databases in light of these findings. For these aims we studied 38 species of Orthoptera representing different sublineages and genome sizes. DNA extracted from each sample was used to amplify five different fragments of COI region by standard PCR protocol. Sequenced PCR amplicons were checked for numt possibility by several different numt criteria. No sequences without numt signs were obtained for the first fragment. The number of samples with numt signs for the other four fragments differed between the suborders Ensifera and Caelifera. The percentage of samples with numt signs was higher in Caelifera than Ensifera for all fragments. The numt percentage considerably decreased for the longest two fragments. Numts are more prevalent in families with larger genome size. We arrived at the following conclusions: (i) numts are common in all members of Orthoptera, but, their prevalence differs among intra-lineages, especially more prevalent in Caelifera, (ii) there seems a correlation between numt rate and genome size, (iii) there is no threshold to avoid numt co-amplification, but, a 1,000 bp length may be a threshold for Ensifera, (iv) Folmer region of COI doesn't seem an appropriate marker for animal barcoding. Additionally, a phylogenetic tree produced from the numt sequences of fragment four detected in genus Anterastes suggested a paleonumt gained in generic ancestor a 3.5-4 times slower divergence rate for numt sequences.

Contribution to the taxonomy of Poecilimon bosphoricus species group (Orthoptera: Phaneropteridae): two new species from its core range.

The Poecilimon bosphoricus species group, representing the most diverse lineage of the bushcricket genus Poecilimon, has been studied in the Basin of Marmara Sea. Two new species, P. warchalowskae sp. n. and P. canakkale sp. n., are described morphologically and acoustically in comparison to their related taxa of the P. bosphoricus species group. Song type and morphology of the new species suggests that they are related to P. turcicus, P. turciae and P. athos and each of these five species can be distinguished by their specific male cerci. The diagnoses are supplemented with comparative figures of morphology and song. Additionally, a short account is given on the synonymy of P. anatolicus with P. sureyanus.

Review of Psorodonotus Specularis Group (Orthoptera, Tettigoniidae, Tettigoniinae): two new species from North-east Anatolia.

The Anatolio-Caucasio-Balkan genus Psorodonotus (Orthoptera, Tettigoniidae) presently includes 13 species belonging to three species groups. In the present study we review the Specularis group and describe two new species-P. rize Kaya & Ciplak sp. n. and P. giresun Kaya & Ciplak sp. n.. Data obtained of 21 different populations from the North-eastern Anatolia and the Caucasus are used for descriptive and analytical purposes. Morphology of this group has been studied both qualitatively and quantitatively using linear metric data of the hind femur and geometric data of male and female pronotum, male cerci and ovipositor. Male calling songs are described and statistically analysed. Qualitative and/or quantitative morphology and male calling song suggested seven taxonomic units: P. specularis specularis, P. specularis inermis, P. inflatus, P. davisi, P. soganli, P. rize sp. n., P. giresun sp. n.. Our results suggest closest relationships of P. rize sp. n. with P. davisi and of P. giresun sp. n. with P. soganli. 

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.

Psorodonotus venosus group (Orthoptera, Tettigoniidae; Tettigoniinae): geometric morphometry revealed two new species in the group.

Psorodonotus (Orthoptera, Tettigoniidae) includes 11 species distributed in Caucasus, Anatolia and Balkans. Although its present taxonomy is problematic, mainly three species groups can be distinguished; (i) The Specularis Group, (ii) The Caucasicus Group and (iii) The Venosus Group. Our recent studies on the genus have revealed presence of two new species in the last species group. Morphology of the species group studied both qualitatively and quantitatively using linear metric data of pronotum, tegmina and hind femur, and geometric data of male cerci and ovipositor. Morphological data were accompanied by data obtained from male calling song. Morphological and song data were produced from six different populations from North and Eastern part of Turkey: (1) Hakkari, (2) Tendürek, (3) Giresun, (4) Artvin, (5) Kars and (6) Agri. Qualitative and quantitative morphology, either linear-metric or geometric, suggest last three population as members of the same unit, but each of other three as different units. Song data are also largely in support of the morphological results. Necessary illustrations were provided to document results visually. Following conclusions were made: (1) the Artvin, Kars and Agri populations represent typical P. venosus and the Giresun population P. rugulosus, (2) each of the Hakkari and Tendürek populations represents a new species and P. hakkari sp. n. and P. tendurek sp. n. described by comparing with other members of P. venosus group, (3) P. rugulosus, P. hakkari sp. n. and P. tendurek sp. n. differ from P. venosus mainly by the longer cerci (extend to or beyond end of abdomen) and indistinct tubercles on surface of pronotal disc in female. P. rugulosus and P. tendurek sp. n. are also similar by sharing presence of two loud elements in a syllable (one in P. venosus, song of P. hakkari sp. n. is not available). But, similarities in phenotype are in conflict with relationships suggested by genetic data.