(Orthoptera, Tettigoniidae): morphology, song and

Zootaxa 3401: 49–59 (2012) www.mapress.com / zootaxa/ Copyright © 2012 · Magnolia Press

ISSN 1175-5326 (print edition)

Article

ZOOTAXA ISSN 1175-5334 (online edition)

Anterastes davrazensis sp. n. (Orthoptera, Tettigoniidae): morphology, song and 16S rDNA phylogeny SARP KAYA1, DRAGAN CHOBANOV2 & BATTAL ÇIPLAK3, 4 1

Department of Biology, Graduate School of Applied and Natural Sciences, Antalya, Turkey, E-mail: [email protected] Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, Tsar Osvoboditel boulevard 1, 1000 Sofia, Bulgaria, E-mail: [email protected] 3 Department of Biology, Faculty of Science, Akdeniz University 07058 Antalya, Turkey, Tel: +90 242 310 23 56, Fax: +90 242 227 89 11. E-mail: [email protected] 4 Corresponding Author 2

Abastract The new species Anterastes davrazensis sp. n. (Orthoptera, Tettigoniidae) is described from south-eastern Turkey. Description, diagnosis and relationships of the new species were studied utilizing morphology, male calling songs and 16S rDNA sequence data from all species in the genus. Morphology and song syllable structure indicate A. davrazensis sp. n. is related to A. uludaghensis. Phylogenetic analyses based on representative haplotypes of 16S rDNA, using Sureyaella bella, Parapholidoptera distincta and Bolua turkiyae as outgroups, also suggested strong support to the relationship of these two species. A. davrazensis sp. n. differs from its closest relative A. uludaghensis by the higher number of stridulatory pegs and the song, consisting of irregular syllable groups. Key words: Anterastes, Anterastes davrazensis sp. n., Anatolia, taxonomy, song, phylogeny

Introduction The genus Anterastes was established in 1882 to include A. serbicus by Brunner von Wattenwyl (1882). Karabağ (1951) made the first revision of Anterastes listing eight species, all occurring in Anatolia, and A. serbicus also in the Balkans. With the description of A. niger (Ünal 2000), the species number in the genus rose to nine. In the second revision of the group Çıplak (2004) included 10 species. A recent paper by Kaya & Çıplak (2011) increased the species number to 12 by transferring Koroglus disparalatus Ünal, 2002 to Anterastes and synonymising its monotypic genus and by describing an additional new species, A. antecessor. The recent increase in the species number might be a result of extensive field studies and the application of contemporary and comprehensive taxonomic approaches (Kaya & Çıplak 2011). Curiously, nearly all of the recently described species seem to be restricted to mountain summits in Anatolia: A. niger from Çamlıbel Mountain-range between Tokat and Sivas provinces, A. disparalatus from the summit of Koroglu Mt. in Bolu province and A. antecessor from the summit of Akdağ Mt. between Antalya and Muğla provinces (Çıplak 2004, Ünal 2002, Kaya & Çıplak 2011). The recently described A. antecessor and A. disparalatus, in addition to the previously known A. uludaghensis, occupy the most basal branches in a phylogenetic tree obtained using sequences of 16S rDNA (Kaya & Çıplak 2011). Their restricted distribution and the phylogenetic relationships indicate that these species are possibly relicts of an ancestral stock exhibiting similar ecological preference. One of the authors (DC) occasionally met another population representing a new species again from a summit in south-west Anatolia that has phylogenetic affinities with the primitive species of genus. This study aims to describe this new species and to discuss its relationships and distribution by combining its sequences to that presented in Çıplak et al. (2010) and Kaya & Çıplak (2011).

Accepted by D. Rentz: 7 May 2012; published: 27 Jul. 2012

49

Material and methods Morphology and bioacoustics Specimens were preserved in 96% alcohol kept at -20oC. Morphological structures were examined and measured using Leica M6 stereomicroscope. Leica DC200 digital imaging system was used to obtain morphological images. Specimens examined during this study are preserved in Akdeniz University, Department of Biology, Zoological Museum, Antalya, Turkey (AUZM) and the collection of D. Chobanov, Bulgaria. We also benefited from the data presented in Çıplak (2004) (obtained from The Natural History Museum, London and Museum für Naturkunde, Berlin), Çıplak et al. (2010) and Kaya & Çıplak (2011). Male calling songs were recorded in laboratory. For sound recording a FOSTEX FR-2 digital recorder was used with a G.R.A.S. Type 40BF microphone (frequency response 10 hz—40 Khz ± 1.0 db, 4 hz—100 Khz ± 2.0 dB) and the G.R.A.S. Power Module Type 12AK. Oscillograms and sound measuring were made using a PC with the software programs Turbolab (Stemmer AG) and CoolEdit Pro. V. 2.0 (Syntrillium Software Corporation). In song descriptions the following bioacoustic terminology by Heller (1988) and Ragge & Reynolds (1998) is used: Calling song—spontaneous song produced by an isolated male, echeme—a first-order assemblage of syllables, syllable—the song produced by one opening + closing movement cycle of the tegmina, hemisyllable—the song produced by an opening or closing movement of the tegmina, impulse—a simple, undivided transient train of sound waves, syllable duration—the time interval starting from beginning of one syllable to its end, s—second, ms—millisecond.

TABLE 1. Data relating to the haplotypes used in phylogenetic analyses (* haplotypes published in Çıplak et al. (2010), ** those in Kaya & Çıplak (2011) - all obtained from Genebank; *** haplotypes given in this study). Location Rize

Coordinates

Altitude (m)

Genus/Species (and authors) Parapholidoptera Maran, 1953 Parapholidoptera distincta (Uvarov, 1921) Bolua Ünal, 1999 Bolua turkiyae Ünal, 1999 Sureyaella Uvarov, 1934 Sureyaella bella Uvarov, 1934 Anterastes Brunner von Wattenwyl, 1882

40o41'84"N

40o41'10"E

1627

Balıkesir

39o41'38"N

26o54'56"E

1428

Konya

38°08'25"N

32°51'21"E

1600

Antalya (AnA)

36o34'56"N

29o34'99"E

2238

A. antecessor Kaya & Çıplak 2011

Bolu (BO)

40o35'47"N

31o46'92"E

2020

A. disparalatus (Ünal, 2002)

Isparta (IS)

37o46'18"N

30o44'34"E

1950

A. davrazenssis sp.n.

Bursa (BU) İzmir (IZ) Afyon (AF) Konya (KO) Bolu (BO) Bursa (BU) Giresun (GI) Kastamonu (KS) Sivas (SI)

40o07'18"N 38o21'15"N 38o28'08"N 41o03'24"N 40o35'47"N 40o07'19"N 40o27'96"N 41o03'24"N 40o09'95"N

29o08'57"E 28o06'14"E 30o22'69"E 33o43'05"E 31o46'92"E 29o08'57"E 38o42'48"E 33o43'05"E 37o49'73"E

2014 1550 1802 1560 2020 1734 2294 2006 1907

A. uludaghensis Karabağ, 1950 A. tolunayi Karabağ, 1951 A. antitauricus Çıplak, 2004 A. antitauricus A. burri Karabağ, 1951 A. burri A. serbicus Brunner von Wattenwyl, 1882 A. serbicus A. serbicus

Bulgaria (BUL) Antalya (AnZ) Denizli (DB) Antalya (AnC) Sivas (SI) Antalya (AnT)

Stara Mts 36o56'21"N 37 o47'45"N 36°36'11"N 40o09'79"N 36o49'78"N

29o47'63"E 28o46'83"E 29°37'49"E 37o51'02"E 30o17'62"E

50 · Zootaxa 3401 © 2012 Magnolia Press

2100-2850 1580 1572 1750 1809 1790

A. serbicus A. babadaghi Uvarov, 1939 A. babadaghi A. ucari Çıplak, 2004 A. niger Ünal, 2000 A. turcicus Karabağ, 1951

Genbank accesion no JX024230***

HQ848285** HQ848286** HQ850974** JX024233*** HQ850975** HQ850976** JX024231*** JX024232*** HM856139* HM856144* HM856160* HM856140* HM856169* HM856166* HM856147* HM856146* HM856145* HM856162* HM856167* HQ850980** HQ850978** HQ850979** HQ850977** HQ850981**

KAYA ET AL.

Sequence data and phylogenetic analyses In this study 16S rDNA region of mitochondrial genome was amplified. Procedures for DNA extraction and amplification of the gene region, and the primers used, are given in Çıplak et al. (2010). Previously published DNA sequences of Anterastes were obtained from GenBank (Çıplak et al. 2010, Kaya & Çıplak 2011). Three different out groups were chosen for phylogenetic analyses: Parapholidoptera distincta, Bolua turkiyae and Sureyaella bella. The sequences downloaded from GenBank, accession numbers for the published and unpublished haplotypes and the haplotype abbreviations are shown in Table 1. The sequences were aligned manually in Sequencher v. 4.1 sequence analysis software (Gene Codes Corporation). DnaSP v. 5 (Librado & Rozas 2009) was used to determine unique haplotypes. Haplotype matrix was prepared using MEGA v. 5 (Tamura et al. 2011). In the matrix, gaps were coded as the fifth character state in maximum parsimony (MP) analysis. The MP was carried out 100 random addition with the heuristic search approach using the TBR algorithm. The confidence of branching was assessed using 10000 nonparametric bootstrap resampling (Felsenstein 1985). The parameters and the bestfit model were estimated using MODELTEST version 3.06 (Posada & Crandall 1998) for a gap-loaded positions excluded matrix. The selected model was implemented in the maximum likelihood (ML) analysis. A nonparametric bootstrapping (Felsenstein 1985) was used to evaluate the support of nodes based on 1000 pseudoreplicates analysed by ML. Bayesian search was carried out using four simulations of Markov Chains, three million generations and sampling every 100 generation. The software tool TRACER v. 1.5 (Rambout & Drummond 2003) was used to examine the parameters and determine the number of trees needed to reach stationary (burn-in). Bayesian posterior branch probabilities were obtained by taking the majority rule consensus of the sampled trees, excluding the first 3000 trees as burn-in. MP and ML analyses were conducted using PAUP v. 4.0b10 (Swofford 2000) and Bayesian inference of phylogeny (BI) by MrBayes v. 3.1.2 (Ronquist & Huelsenbeck 2003). Branches that received a bootstrap or posterior probability support higher than 50% from each of MP, ML and BI analyses were indicated by their support value.

Results Anterastes davrazensis sp. n. (Figs 1–11) Material examined: Holotype, male, TURKEY: Isparta, Davraz Mt. (N 37o46'18", E 30o44'34"), 1950 m, 22.07.2011 (Leg. D. Chobanov) (in alcohol). Paratypes—8 males, 5 females from the same place. Depositories: holotype and 4 male paratypes—Department of Biology, Zoological Museum, Akdeniz University, Antalya, Turkey (AZUM); 4 male and 5 female paratypes— collection of D. Chobanov, Bulgaria. Diagnosis: A. davrazensis sp. n. is similar to A. uludaghensis and A. antecessor in morphology. The male left tegmen not strongly narrowing toward apex on its right margin is a typical synapomorphy of these three species that distinguishes them from the other members of Anterastes. However, A. antecessor differs from its two relatives by the short and robust male cerci with a huge tooth and the apically not or weakly curved apical arms of the titillators. Prominent morphological character, diagnosing A. davrazensis from A. uludaghensis, is the higher number of stridulatory pegs (72–89 in the new species and 55–60 in the second; Çıplak 2004). Apart from the morphology, the most prominent synapomorphy of A. davrazensisis sp. n. and A. uludaghensis is the typical syllable with four elements (Fig. 9C, F). In all other species of Anterastes (see Kaya & Çıplak 2010) each syllable contains two elements. Although their syllable structure is similar, A. uludaghensis and A. davrazensis differ from each other in song pattern. Males of A. davrazensis produce irregular echemes with short or long-lasting syllable groups, while those of A. uludaghensis produce regular echemes (Fig. 9A, D). The relationship pattern suggested by morphology and syllable type is congruent with that suggested by phylogenetic trees produced from 16S rDNA data. Description (holotype, male): Fastigium of vertex rounded, twice or more the width of scapus, not sulcate. Pronotum relatively short, cylindrical or slightly depressed dorsally in metazona; disc with a trace of median carina, lateral carinae absent, hind margin widely obtuse (Fig. 1A). Micropterous, tegmina equal to pronotum in length (Fig. 1A, B); right tegmen slightly longer than left one, weakly narrowing distal ward on its right margin; stridulatory file prominently curved at distal part, with 72–89 teeth; teeth in the medial part wider than those on

ANTERASTES DAVRAZENSIS SP. N.

Zootaxa 3401 © 2012 Magnolia Press ·

51

FIGURES 1–5. Anterastes davrazensis sp. n., male (Scale: 1mm): 1—Pronotum and tegmina from above (A) and lateral view (B), 2—Cerci, 3—Tenth tergite (A, B different individuals), 4—Subgenital plate (A, B different individuals), 5—Titillators (A, B different individuals).


KAYA ET AL.

FIGURES 6–8. Anterastes davrazensis sp. n. female (Scale: 1mm): 6—Pronotum from above (A) and lateral view (B), 7—Subgenital plate and 5–7 sterna, 8—Ovipositor in whole (A) and apical view (B).

the proximal and distal ends; distal teeth small and conical. All three pairs of femora without ventral spines/ spinules; fore tibiae spineless on dorso-anterior margin and with two or three spines on dorso-posterior margin. Mid tibiae with three spines on each of dorso-anterior and dorso-posterior margins. Each of the ventro-anterior and ventro-posterior margins of fore and mid tibiae with six spines. Hind femur as long as or barely extends beyond the end of abdomen, swollen in the basal 2/3; 3.6–3.8 times as long as its maximal width; hind tibiae with one pair of spurs apico-ventrally; platulae two-thirds of the length of metatarsus. Tenth abdominal tergite with two short, incurved triangular processes, and with a wide shallow incision between them, setose medially and marginally (Fig. 3A, B). Cerci short and thick, with a huge pre-apical tooth, base of the tooth depressed and its apex slightly downcurved; width of cercus along the tooth 1/3 or more of the total cercal length (Fig. 2). Male subgenital plate equal or wider than long, with small styli and a deep incision at posterior margin (Fig. 4A, B). Titillators slender; basal arms longer than apical arms; basal arms smooth and obtusely curved; apical arms almost straight in proximal 3/4 and strongly curved in apical 1/4, their outer surface rough or sometimes with small tubercles (Fig. 5A, B). Paratypes are similar to the holotype.



53

FIGURE 9. Male calling song. A–C, A. davrazensis sp. n.: A—irregular syllable groups, B—a group of syllables, C—opening and closing hemisyllable; D–F, A. uludaghensis: D—regular echemes, E—groups of echemes, F—opening and closing hemisyllable.

Description of female (paratype): Similar to male in general characteristics. Tegmina strongly abbreviated, not overlapping dorsally (Fig. 6A, B); hind femur ca. 3.0 times as long as its maximal width. Sixth and seventh abdominal sterna not modified (Fig. 7). Subgenital plate wider than long, with a semi-elliptical incision and two obtuse lobes apically (Fig. 7). Ovipositor short, robust and slightly up curved, 4–5 times as long as its maximal width; ventral valves with 7–9 tubercles in the apical one-sixth (Fig. 8A, B).


KAYA ET AL.

FIGURE 10. Map showing localities and haplotypes used in phylogenetic analyses. Triangle represents the locality of new species.

Coloration: Black or blackish brown. A yellow pattern with a narrow light stripe is situated behind the eyes; pronotal disc dark brown, paranota with a large black spot or black pattern, a bright creamish posterio-ventral band; hind femur with a black spot dorso-anteriorly and indistinct black pattern dorso-externally; tegmina brownish (Fig. 1A, B). Abdominal terga black, with irregular brownish pattern; ovipositor light brown with dark-brown pattern. Measurements (in mm, mean in the brackets): Body, male 16.5–18.0 (17.4), female 22.2–26.0 (24.1); length of pronotum, male 4.16–4.46 (4.3), female 4.2–4.6 (4.5); tegmina, male 2.96–3.83 (3.4); number of stridulatory pegs 72–96 (80); length of hind femur, male 10.38–10.79 (10.6), female 10.6–12.0 (11.4); maximum width of hind femur, male 2.72–3.08 (3.0), female 2.93–3.23 (3.03); length of ovipositor 9.6–10.8 (10.2); maximum width of ovipositor 1.7–1.86 (1.8). Song (n= 24 for each parameter, otherwise stated): The male calling song of type specimens (the holotype and a paratype male) was recorded in laboratory at 27.5 °C (Fig. 9A–C). The calling song contains irregular long and short syllable groups. The duration of syllable groups ranges between 0.559 –15 s (mean: 2.96 s) and the syllable number per syllable group ranges between 15–377 (mean; 67.54). Syllable period has a length of 0.037–0.040 ms (mean 0.038) and includes a quiet opening- and a loud closing-hemisyllable, each of which consisting of two different elements (Fig 9C). The first elements of opening and closing hemisyllables are longer than the seconds and contain prominent impulses. The opening hemisyllable includes 7–10 (mean 9; n=35) inseparable impulses in a period of 0.014–0.016 ms (mean 0.015; n=35). The closing hemisyllable contains 8–16 (mean 12) impulses and lasts 0.019–0.024 ms (mean 0.021). Etymology: The species is named after the name of the summit it occupies, the Davraz Mountain, Isparta, Turkey.



55

FIGURE 11. Phylogenetic tree obtained from MP analyses of the 16S rDNA haplotypes given in Table 1. In the phylogenetic analyses S. bella, P. distincta and B. turkiyae used to as outgroups. Bootstrap (MP, ML) and posterior probability (BI) support (those lower than 50% indicated by an *) to branches are shown as MP/ML/BI respectively.

Phylogenetic relationships of the new species Localities of specimens sampled for sequencing are shown in Figure 10 and Table 1. After alignment and trimming, the final length of the used sequences was 530 bp (with five gaps). The MP phylogenetic analysis was accomplished using 530 bp of 25 haplotypes (including five gaps). Of the 530 sites, 416 are constant and 114 are variable (84 of these are parsimony informative). MP analyses resulted with eight equally parsimonious trees (tree length=262, CI=0.588, RI=0.690, RC=0.405) and the strict consensus tree is presented in Figure 11. In the MP tree representative haplotypes of P. distincta, B. turkiyae and S. bella branch off successively leading to in-group haplotypes. At the next step, a clade containing two haplotypes of A. antecessor occurs in the base of all others.


KAYA ET AL.

The upper next step is a dichotomy; the first phylogroup contains haplotypes of A. disparalatus + A. uludaghensis + A. davrazensis sp. n. and the second – those of the A. babadaghi + A. serbicus species groups. The MP bootstrap support values to the most basal branches are low, but that to the A. uludaghensis + A. davrazensis sp. n. is higher than 50% (Fig. 11). The ML and BI analyses were made using 525 bp of 24 haplotypes (excluding five gaps). Of the 525 sites, 416 are constant and 109 are variable. MODELTEST, applied to the dataset (Table 1), selected the TrN+I+Γ model (Tamura & Nei 1993) according to hLRT and AIC with proportion of invariable sites I of 0.7044, gamma shape parameter Γ of 1.1183 and ti/tv ratio of 3.29. The BI and ML analyses under the selected model and parameters did not suggest the same topology with that of MP. In both of ML and BI the P. distincta and B. turkiyae branch off basally. The next step on the both trees is a polytomy. BI tree suggested a polytomy with eight clades; (i) S. bella, (ii) A. antecessor, (iii) A. koroglus, (iv) A. davrazensis sp. n. + A. uludaghensis, (v) A. turcicus, (vi) A. niger, (vii) A. serbicus group + A. tolunayi and (viii) A. babadaghi + A. ucari. The topology of ML tree is similar to that of BI tree, but A. tolunayi occurs as the ninth clade in the basal polytomy (see Fig. 11). Sister group relation of A. davrazensis and A. uludaghensis is supported by both of ML and BI trees with high bootstrap (ML) and posterior probability (BI) values.

Discussion All of the three data sources, morphology, song, and DNA, are in agreement regarding A. davrazensis sp. n., either for its distinct species entity or for its relationships with the other taxa concerned in this study. The male left tegmen not or indistinctly narrowing toward apex on its right margin is a typical synapomorphy of A. uludaghensis (see Çıplak 2004), A. antecessor (see Kaya & Çıplak 2011) and A. davrazensis sp. n. However, the latter is more closely related to A. uludaghensis than A. antecessor by similarities in other morphological characters. A. antecessor differs from other two especially by its unique structure of male cerci. Although A. uludaghensis and A. davrazensis share several morphological similarities, they are easily distinguishable, especially by the number of stridulatory pegs. The syllable structure of A. davrazensis sp. n. and A. uludaghensis is even more important than the morphology for their recognition. Their song contains a syllable of four elements (Fig. 9C, F), which seems to be a unique synapomorphy of these two species (in other species of the genus the syllable contains only two elements; see Kaya & Çıplak 2011; Çıplak et al. unpublished data). Contrary to the syllable structure, the song patterns are different in both species – A. davrazensis produces irregular syllable groups, while A. uludaghensis produces regular echemes. Apart of the song pattern both species also differ in some temporal parameters of the song. In synopsis, A. davrazensis is a distinct species according to its phenotypical traits and it may be regarded most closely related to A. uludaghensis. Phylogenetic analyses partly supported the relationship pattern suggested from phenotypical features (Fig. 11). The sister group relationship of A. uludaghensis and A. davrazensis sp. n. was supported by all of MP, ML and BI analyses and received high bootstrap or posterior probability values. Yet, the relationships of this clade with other members of the genus remained unresolved. Strict consensus tree of eight MP trees suggested the topology A. disparalatus + (A. uludaghensis and A. davrazensis), however, ML and BI trees did not support this relationship. The possible reasons for this incongruency may be due to either small sample size in the analyses and/or poor phylogenetic information of 16S rDNA. Presently the sample number per species is restricted. Additionally, there are five indel positions which were coded as fifth character state, but deleted in the matrix used in ML and BI analyses. Another reason, reported for some lineages, might be that this marker gene is poor for revealing long time divergences (Simon et al. 1994, Muraji, 2000). Solving this requires studying larger samples size and further marker genes. Distribution of the new species alone or in relation to other members of the genus, especially its sister group A. uludaghensis, exhibits some specific aspects to be mentioned. The new species is restricted to a summit in southwest Anatolia. A. ucari was recorded from the same mountain, usually occurring at lower altitude. Yet, at the type locality of A. davrazensis, both species occurred in a close neighbourhood, A. ucari preferring sites of higher humidity and denser vegetation, while A. davrazensis kept on dry stony ground with very sparse vegetation cover and subjected to strong erosion. Additionally, A. ucari becomes adult earlier than the new species. A. uludaghensis, being most closely related to A. davrazensis, occurs in Uludağ Mts., ca. 400 km away from the type locality of the



57

new species. Thus, there are two important inferences to be concluded. First, according to the phylogenetic relationships, we assume that the new species is a remnant of the generic ancestral stock. Second, there is a growing evidence that the southern Anatolian highlands are extremely important biodiversity hotspots preserving rich fauna and serving as refugia as suggested by studies on several lineages (e.g. Çıplak 2003, 2008, Şirin et. al. 2010, 2011, Gündüz et al. 2007, Médail & Quezel 1997). Therefore, the concerned ancestral lineages, presently isolated in so-called sky-islands offer good examples for outlining biogeographic hypotheses to be further tested using comprehensive genetic data.

Acknowledgement We thank Dr. İslam Gündüz (Samsun) for guiding us in the early stage of the molecular studies and to the young orthopterist Zehra Boztepe (Antalya) for her help during laboratory studies. Our research was supported by the Scientific and Technical Research Council of Turkey (TUBITAK, TBAG110T462). Collecting the material was partly financed by grant N N303 611738 of the Ministry of Science and Higher Education of Poland. Studies were carried out in laboratories of the Department of Biology, Akdeniz University, and the paper was supported by the Akdeniz University Research Found. Specimens collected by D. Chobanov, data produced by S. Kaya, D. Chobanov and B. Çıplak, and B. Çıplak lead the writing.

References Brunner von Wattenwyl, C. (1882) Prodromus der europäischen Orthopteren. Engelmann, Leipzig, 1–456. Çıplak, B. (2003) Distribution of Tettigoniinae (Orthoptera, Tettigoniidae) bush-crickets in Turkey: the importance of the Anatolian Taurus Mountains in biodiversity and implications for conservation. Biodiversity and Conservation, 12, 47–64. Çıplak, B. (2004) Systematics, phylogeny and biogeography of Anterastes (Orthoptera, Tettigoniidae, Tettigoniinae): evolution within a refugium. Zoologica Scripta, 33, 19–44. Çıplak, B. (2008) The analogy between glacial cycles and global warming for the glacial relicts in a refugium: a biogeographic perspective for conservation of Anatolian Orthoptera. In: Fattorini, S.(Ed): Insect Ecology and Conservation, Research Signpost, Kerela. pp 135–163. Çıplak, B., Kaya, S. & Gündüz, İ. (2010) Phylogeography of Anterastes serbicus species group (Orthoptera, Tettigoniidae): phylogroups correlate with mountain belts, but not with the morphospecies. Journal of Orthoptera Research, 19, 89–100. Felsenstein, J. (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution, 39, 783–791. Gündüz, İ., Jaarola, M., Tez, C., Yeniyurt, C., Polly, P.D., Searle, J.B. (2007) Multigenic and morphometric differentiation of ground squirrels (Spermophilus, Sciuridae, Rodentia) in Turkey. Molecular Genetics and Evolution, 43, 916–935. Heller, K.G. (1988) Bioakustik der europäischen Laubheuschrecken. Weikersheim, Verlag Josef Margraf, 358 pp. Karabağ, T. (1951) Revision of the genus Anterastes Brunner (Orthoptera, Tettigoniidae). Annual Magazine of Natural History, 4, 1043–1051. Kaya, S., & Çıplak, B. (2011) Taxonomy of Anterastes and related genera: a new synonym and a new species of Anterastes. Zootaxa, 2771, 41–52. Librado, P. & Rozas, J. (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25, 1451–1452. Médail, F. & Quézel, P. (1997) Hot-spots analysis for conservation of plant biodiversity in the Mediterranean Basin. Annals of the Missouri Botanical Garden, 84, 112–127. Muraji, M., Kawasaki, K. & Simizu, T. (2000) Phylogenetic utility of nuclear sequences of mitochondrial 16S ribosomal RNA and cytochrom b gene in anthocorid bugs (Heteroptera: Anthocoridae). Entomological Zoology, 35, 293–300. Posada, D. & Crandal, K.A. (1998) MODELTEST: testing the model of DNA substitution. Bioinformatics, 14, 817–818. Ragge, D.R. & Reynolds, W.J. (1998) The songs of the grasshoppers and crickets of western Europe. Harley Books, London. 221 pp. Rambout, A. & Drummond, A.J. (2003) Tracer v1.3. Available from: http: //evolve.zoo.ox.ac.uk/. Ronquist, F. & Huelsenbeck, J.P. (2003) MRBAYES 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19, 1572–1574. Swofford, D.L. (2002) PAUP* Phylogenetic Analysis Using Parsimony (* and other methods). v. 4.0 beta. M.A. Sinauer Associates, Sunderland. Uvarov, B.P. (1921) The geographical distribution of orthopterous insects in the Caucasus and in Western Asia. Proceeding of Zoological Society of London, 31, 447–472.


KAYA ET AL.

Uvarov, B.P. (1934) Studies in the Orthoptera of Turkey, Iraq and Syria. Revista Espanola de Entomologia, Madrid, 10, 21–119. Ünal, M. (1999) Notes on Orthoptera of western Turkey, with description of a new genus and four new species. Journal of Orthoptera Research, 8, 243–255. Ünal, M. (2000) A new Anterastes Brunner von Wattenwyl from Turkey (Orthoptera: Tettigoniidae). Centre for Entomological Studies, Miscellaneous Papers, 65–66, 5–7. Ünal, M. (2002) First data on Orthoptera of Mount Köroğlu, N.W. Anatolia, with description of three new taxa. Entomological News, 113(4), 275–288. Simon, C., Frati, F., Beckenbach, A., Crespi, B., Liu, H. & Flook, P. (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved PCR primers. Annals of the Entomological Society of America, 87, 651–701. Şirin, D., Mol A. & Çıplak, B. (2011) Myrmeleotettix Bolivar (Orthoptera, Gomphocerinae) in Anatolia on the basis of morphological and behavioural characters: data suggest a new species from southern end of the Anatolian refugium. Zootaxa, 2917, 29–47. Şirin, D., Von Helversen, O. & Çıplak, B. (2010) Chorthippus brunneus subgroup (Orthoptera, Gomphocerinae) in Anatolia with description of two new species: data suggest an Anatolian origin for the lineage. Zootaxa, 2410, 1–28. Tamura, K. & Nei, M. (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolution, 10, 512–526. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution, 28, 2731–2739.



59