Chorthippus (Glyptobothrus) Bornhalmi Harz, 1971 Karyotype Analysis†

Chorthippus (Glyptobothrus) bornhalmi Harz, 1971 Karyotype Analysis†

Fatih Çakmak and Serdar Koca*

Department of Biology, Science and Art Faculty, Adnan Menderes University, 09010 Aydin, Turkey

Received April 12, 2014; accepted July 20, 2014

Summary In this study, the karyotype of the Chorthippus (Glyptobothrus) bornhalmi species (chromosome number, chromosome morphology and chromosome lengths) belonging to the Gomphocerinae subfamily of the Acrididae family was examined. As a result of these examinations, the species’ number of chromosome was determined as 2n♂= 17 (X0). It was found that one pair of

autosomal chromosome 3 (L1–L3) was submetacentric whereas ﬁve pairs (M4–S8) and X chromosome had an acrocentric structure. As a result of the counting carried out on ﬁve individuals, the mean chiasma frequency was found to be 15.36.

Key words Chorthippus (Glyptobothrus) bornhalmi, Orthoptera, Karyotype, Chromosome, Chiasma frequency.

Orthoptera is one of the insect order which has been systematically well-studied in Turkey (Karabağ 1983, Demirsoy 1975, Ünal 2000, Çıplak 2003, Sevgili and Heller 2003). According to recent studies, 649 species and subspecies belonging to Orthoptera together with 90 species and subspecies belonging to the Gomphocerinae subfamily of the Acrididae family have been listed in Turkey (Ünal 2012). Hundreds of species systematically exist in the Acrididae family. Among these species, Chorthippus Fiber,1852 genus, has more than 250 species and subspecies. It has been stated that 27 species and subspecies belonging to the Chorthippus genus exist in Turkey (Ünal 2012). Even though many studies have been carried out under systematic conditions on the Chorthippus (Glyptobothrus) bornhalmi genus in the name of determining diversity in Turkey, no survey on the cytogenetic features of Ch. (Glyptobothrus) bornhalmi genus (Karaca et al. 2006, Ünal 2008, Sevgili et al. 2011) has yet been conducted. However, a relatively large number of studies have been carried out on cytogenetic features related to chromosome number and structure of many species from the Orthoptera order (Lopez-Fernandes et al. 1984, Gusachenko et al. 1992, Carvalho et al. 2011, Rocha et al. 2011). There have been few studies conducted with Orthopteras cytogenetically in Turkey (Koca 1993, Turkoglu 2001, Turkoglu and Koca 2002a, 2002b, Turkoglu et al. 2003, Koca and Tunçbaş 2006). In chromosome studies conducted with Chorthippus genus grasshoppers belonging to Gomphocerinae subfamily, it was generally determined that the chromosome number was 2n=17,

XO ♂ and 2n=18, XX ♀ and that the chromosome structure consisted of three pairs of long (L1– L3) meta or submetacentric submetasentric and acrocentric or ﬁve pairs of (M4–S8) subacrocentric chromosomes ranging from medium to short length. It was also reported that the X chromosome was acrocentric in structure (Cano and Santos 1990, Gusachenko et al. 1992, Bugrov 1996, Li et al. 2008).

* Corresponding author, e-mail: [email protected] † This study is a part of the summary of an MSc thesis of the ﬁrst researcher. DOI: 10.1508/cytologia.79.509 510 F. Çakmak and S. Koca Cytologia 79(4)

Chiasmata are the points in which two of the four chromatids in bivalent are joined together in one or more than one place when homologous chromosomes separate from each other (John 1990). The presence of a chiasma indicates the event of genetical crossing over (part replacement). This study intends to contribute to the literature by ﬁrstly determining the chromosome number, structure and chiasma distribution of the Ch. (Glyptobothrus) bornhalmi Harz, 1971 genus belonging to the Orthoptera order from in the Aydın region.

Materials and methods

After grasshoppers collected from various places in Aydin were brought into the laboratory, their testes were removed and they were stored in a colchicine-hypotonic solution for two hours at room temperature. At the end of this period, the testes were held in ethyl alcohol–glacial acetic acid (3 : 1) for 24 h at 4°C. Later, the testes were preserved in 70% alcohol at 4°C and an examination was eventually made, where testes were stained with 2% aceto-orcein and a crushed preparation was made with 45% acetic acid. Photographs of cells which showed good distribution in preparations, whose chromosome morphologies were clearly visible and whose chromosomes were located on a same plane were taken using a Olympus BX51 microscope. The measurement of chromosome lengths was micrometrically performed with the IM50 measurement module computer program. Chromosome lengths of 10 cells measured with the program were seperately micrometrically recorded and the classiﬁcation of chromosomes was performed according to Levan et al. (1964). Among the examined samples prepared, chiasmata were counted in 25 diplotene cells of ﬁve individuals in bivalents and the mean chiasma frequency of the species was determined.

Results

As a result of cytogenetic examinations made on the Ch. (Glyptobothrus) bornhalmi genus, the chromosome number was found to be 2n♂=17, X0 (NF=23) (Fig. 1). Three pairs of autosomes

(L1–L3) were submetacentric, ﬁve pairs (M4–S8) ranged from medium to short length and the X chromosome was acrocentric. Chromosome lengths varied between 1.82–11.08 μm. The relative lengths were between 3.26–19.93. The length of the X chromosome was determined to be 6.49 μm. It was the ﬁfth largest chromosome of the karyotype and covered 11.64% of the genome. Karyogram belonging to Ch. (Glyptobothrus) bornhalmi is shown in Fig. 2 while the idiogram is provided in

Fig. 1. Metaphase chromosomes of Ch. (Glyptobothrus) bornhalmi (2n♂=17). Fig. 2. Karyotype of Ch. (Glyptobothrus) bornhalmi. 2014 Karyotype of Chorthippus (Glyptobothrus) bornhalmi 511

Fig. 3. Idiogram of Ch. (Glyptobothrus) bornhalmi.

Table 1. Morphometric characteristics of the chromosomes of Ch. (Glyptobothrus) bornhalmi.

Number of Chromosome length Relative length Centromeric index Arm ratio Chromosome chromosome pair (μm) Mean±S.E. (% of 2n set) Mean+S.E. Mean+S.E. morphology

I 11.08+0.29 19.93+0.55 37.12+1.12 1.70+0.08 sm II 9.77+0.26 17.55+0.22 38.20+1.89 1.74+0.10 sm III 8.53+0.27 15.32+0.46 37.97+1.96 1.72+0.09 sm IV 7.36+0.13 13.25+0.35 ̶ ∞ a V 5.16+0.27 9.24+0.33 ̶ ∞ a VI 3.18+0.13 5.70+0.13 ̶ ∞ a VII 2.26+0.07 4.06+0.11 ̶ ∞ a VIII 1.82+0.09 3.26+0.12 ̶ ∞ a IX (X) 6.49+0.22 11.64+0.20 ̶ ∞ a T.C.L. 55.65 sm: submetacentric, a: acrocentric, T.C.L.: Total Chromosome Length.

Table 2. Mean chiasma frequency and distribution of Ch. (Glyptobothrus) bornhalmi (S.D.; Standart deviation; Chi: Chiasma).

Mean chiasma Bivalent Individual Counted cell 5 Chi 4 Chi 3 Chi 2 Chi 1 Chi frequency+S.D. number

1 25 15.56+1.35 1 27 39 24 109 200 2 25 14.36+0.81 ̶ 7 50 38 105 200 3 25 16.20+1.19 ̶ 37 29 35 99 200 4 25 15.40+1.11 ̶ 27 43 20 110 200 5 25 15.32+1.14 6 13 48 25 108 200 X¯=15.36 7 111 209 142 531 1000

Fig. 3. The measurements of chromosome morphology are given in Table 1. The mechanism of sex determination is of XX (♀)/X0 (♂) type. Mean chiasma frequency and distribution of the genus are given in Table 2. As can be seen in this table, long bivalents (L1–L3) have three chiasmata in general. However, bivalents with two chiasmata and a few with four chiasmata were also observed

(Fig. 4). Although rare, ﬁve chiasmata were encountered in long bivalents (L1–L3). Generally, only one chiasma occured in short bivalents. 512 F. Çakmak and S. Koca Cytologia 79(4)

Fig. 4. (A, B) Diplotene in Ch. (Glyptobothrus) bornhalmi. Eight bivalents and the univalent X-chromosome (arrowed) are present.

Discussion

Determination of differences and similarities between chromosome’s differences and similarities is of great importance in determining both the proximity and distance between genuses. Karyotype is an important part of a genus. While karyology and genetical structure provide an identity to a genus, these identities provide a better understanding of evolutionary relationship and separation between these species. Although chromosome numbers vary from genus to genus and even between species, it is fixed for each species. However it is a common occurrence that this number changes by itself or is changed due to external factors (Oraler Temizkan 1994). The species in the Acrididae family generally show a fixed situation in terms of their number and morphology (John and Hewitt 1968). It is accepted that the chromosome number of many members of this family is 2n♂=23, X0 with 2n♀=24, XX and consists of acro- or subacrocentric chromosomes. However, some karyological changes in some species of the Acrididae family have been observed in recent studies. It is thought that the observed karyotype changes result from some chromosal arrangements arising from small changes in their chromosome numbers and morphologies (Hewitt 1979, Camacho 1980, Cabrero and Camacho 1982). The Gomphocerinae subfamily members of the Acrididae family have 2n♂=17, X0 and 2n♀=18, XX chromosome numbers and it is thought that three long (L1–L3) chromosome pairs occur as a result of centric fusion (Cabrero and Camacho 1985). It is also known that the Chorthippus species belonging to the Gomphocerinae subfamily have 2n♂=17(X0) and 2n♀=18(XX) chromosome numbers. As for chromosome morphologies, they consist of three long pairs (L1–L3) which are metacentric or submetacentric, five pairs (M4–S8) which are acrocentric or subacrocentric autosomes ranging from medium to small length and also X chromosome which, again, possesses an acrocentric structure (Gusachenko et al. 1992, Bugrov 1996, Li et al. 2008). The fact that the chromosome number of Ch. (Glyptobothrus) bornhalmi is 2n♂=17(X0) was determined for the first time in this study. The chromosome number and morphology of this species are given in Fig. 2 and Table 1, respectively. In a study carried out with Ch. brunneus huabeiensis and Ch. minutus of the Gomphocerinae subfamily, Li et al. (2008) stated that the chromosome number of the species was 2n=17(X0), and that chromosome morphologies are three pairs of long metacentric (L1–L3), X chromosome and five pairs of chromosomes ranging from medium to small length (M4–S8) that are acrocentric in structure. Cano and Santos (1990), found that the chromosome number was 2n=17(X0) in their study carried out on Omocestus panteli, Euchorthippus pulvinatus, Euchorthippus chopardi, Ch. vagans, Ch. 2014 Karyotype of Chorthippus (Glyptobothrus) bornhalmi 513 parallelus and Ch. jucundus. It was also detected that chromosome morphologies were three pairs of long submetacentric (L1–L3), five pairs of autosomes ranging from medium to small length (M4– S8) and that the X chromosome were acrocentric in type. Gusachenko et al. (1992) determined that the chromosome number of the species was 2n=17(X0) in males and 2n=18(XX) in females according to their study carried out on Ch. albomarginatus. Among these, the morphology was three pairs of long (L1–L3) submetacentric chromosomes, five pairs of autosomes ranging from medium to small length (M4–S8) and an X chromosome, both of which were observed to be acrocentric in structure. Bugrov (1996) stated that the chromosome number of Ch. macrocerus, Ch. vicinus, Ch. ferganensis, Ch. biguttulus, Ch. jacobsoni, Ch. intermedius, Ch. montanus, Ch. lorarus, Ch. dichrous, Ch. albomarginatus, Ch. saxatilis, Ch. angulatus, Ch. parallelus and Ch. fallax species was

2n=16+X0/XX, that chromosome morphologies were three pairs of long (L1–L3) and metacentric, and that X chromosome and ﬁve pairs of autosomes ranging from medium to small length (M4–S8) were acrocentric in structure within the scope of the collection made of Chorthippus by various researchers, while all chromosomes of the Ch. schmidti species whose chromosome number is 2n=23(X0) are acrocentric in structure. It was found that the Ch. hammarstroemi species has

2n=21(X0) chromosomes; the largest chromosome pair was (L1) metacentric while the X chromosome and other chromosome pairs (M2–S10) were acrocentric in structure. In this study, it was determined that the chromosome number of the Ch. bornhalmi species was 2n=17(X0), and that the chromosome morphology was three pairs of long (L1–L3) submetacentric, with five pairs of autosomes ranging from medium to small length (M4–S8) and X chromosomes which were acrocentric in structure. The XX♀/X0 ♂ and XX♀/neo-XY♂ sex-determination mechanisms have been observed in grasshoppers. The XX♀/X0 ♂ sex determination mechanism has been detected in many species belonging to the Acrididae and Tettigoniidae families (White 1968, 1973, Warchalowska-Sliwa 1984, Warchalowska-Sliwa et al. 1993, Bugrov 1996). However, the XX♀/neo-XY♂ sex development mechanism has also been observed in some Tettigoniidae species and it has been widely accepted that it developed from the ancestral XX♀/X0 ♂ mechanism (Alicata et al. 1974, Messina et al. 1975). That X chromosomes of species belonging to Chorthippus genus are generally acrocentric in structure has been repeatedly demonstrated in studies made by Cano and Santos (1990) (Ch. vagans, Ch. parallelus ve Ch. jucundus), Gusachenko et al. (1992) (Ch. albomarginatus), Bugrov (1996) (Ch. macrocerus, Ch. vicinus, Ch. ferganensis, Ch. biguttulus, Ch. jacobsoni, Ch. intermedius, Ch. montanus, Ch. lorarus, Ch. dichrous, Ch. albomarginatus, Ch. saxatilis, Ch. angulatus, Ch. parallelus, Ch. fallax, Ch. schmidti and Ch. hammarstroemi), Turkoglu (2001) (Ch. brunneus), Bridle et al. (2002) (Ch. brunneus and Ch. jacobsi) and Li et al. (2008) (Ch. brunneus huabeiensis and Ch. minutus). The fact that the X chromosome of the Ch. bornhalmi species has an acrocentric structure has also been detected in this study. The chromosome arm number, also know as NF (Fundamental Number) value, is of great importance in cytogenetic studies as it gives the genetical index of a chromosome. Even if the chromosome number may change by means of events such as centric fission and fusion, the branch number remains unchanged. In our study, it was determined that the chromosome number of the Ch. (Glyptobothrus) bornhalmi species was 2n=17, X0 (NF=23) and the chromosome morphology was three submetacentric pairs and five pairs and an X chromosome with an acrocentric structure. However, the Ch. schimidti species was reported to have 2n=23, X0 (NF=23) acrocentric chromosomes in Bugrov’s (1996) study. Although the Ch. schimidti species has chromosome numbers greater than those of Ch. (Glyptobothrus) bornhalmi, it has been seen that the arm number was not changed and remained fixed. Chiasma frequency is of great importance in terms of reflecting genetic exchange ratio 514 F. Çakmak and S. Koca Cytologia 79(4)

(Sybenga 1975). It is known that many internal and external factors affect chiasma frequency. Chromosome size and the organism’s genotype influence chiasma frequency (Henderson 1963, Fox 1973, Jones 1974, Koca 1993). Chiasma frequencies in 25 diplotene cells of five individuals of Ch. (Glyptobothrus) bornhalmi collected from Aydın province vary between 14.36 and 16.20 in our study. The average chiasma frequency of this species was determined to be 15.36. It can be seen that there were significant differences between chiasma frequencies of individuals assembled from the same region and this may result from genetic differences between individuals. Indeed, some chromosome aberrations such as inversion and fusion affected chiasma frequency in both positive and negative ways (Teoh and Yong 1983, Viseras and Camacho 1984, Goni et al. 1985). Although differences were detected in terms of karyotype between individuals in our study, it is possible that some small changes may result in differences in chiasma frequency. Gusachenko et al. (1992) found that average chiasma frequencies were 14.69 in the first region, 15.18 in the second region and 14.80 in the third region in the scope of their study performed on Ch. albomarginatus individuals cellected from three different locations (Ojcow, Novosibirsk and Kosh-Agach). The mean chiasma frequencies of Ch. loratus individuals collected from three different regions (Sinop, Tokat and İzmir) were detected as being 14.23 in the Sinop region, 14.20 in the Tokat region and 14.91 in the İzmir region (Koca 1993). As a result of statistical calculations that were performed, there was no difference in terms of chiasma frequency between Sinop and Tokat, while there were significant differences between Sinop-İzmir and Tokat-İzmir. This difference may be explained in geographical terms. Furthermore, differences in bivalent size may affect chiasma number. Short bivalents have one or two chiasmata whereas long bivalents have three or more chiasmata. As a result of measurements, it was pointed out that bivalent sizes in Ch. loratus ranged between 15.63–

2.70 μm (L1–S8) in the Sinop population, between 17.73–2.95 μm (L1–S8) in Tokat and between 25.95–2.65 μm (L1–S8) in İzmir. The Ch. loratus collected from İzmir were detected as being bigger than the Ch. loratus collected from Tokat and Sinop in terms of body size. Furthermore, in our survey, one or two chiasmata were seen in short bivalents while three or more chiasmata were encountered in long bivalents. Our results show similarity with those of the study carried out on Ch. loratus (Koca 1993). Consequently, the chromosomes of the Ch. (Glyptobothrus) bornhalmi species have been defined for the first time in this study. As the study of only morphological differences remains insufficient in systematic studies, studies at cytogenetic, biochemical and molecular levels are also needed.

Acknowledgments

We thank Dr. Hasan Sevgili from Science and Art Faculty, Department of Biology, Ordu University, for the taxonomic identiﬁcation of the species. This study was supported by Adnan Menderes University Scientiﬁc Resarches Unit (FEF-12024).

References

Alicata, P., Messina, A. and Oliveri, S. 1974. Determinismo cromosomico del sesso in Odontura stenoxipha (Orth., Phaneropteridae) un nuovo caso di Neo-XY. Animalia 1: 109–122. Bridle, J. R., Torre, J., Bella, J. L., Butlin, R. K. and Gosalvez, J. 2002. Low levels of chromosomal differentiation between the grasshopper Chorthippus jacobsi (Orthoptera; Acrididae) in northen Spain. Genetica 114: 121–127. Bugrov, A. G. 1996. Karyotypes of the short-horned Orthopteran insects (Orthoptera, Caelifera) from Russia, Kazakhstan, Central Asia and Caucasus. Folia Biol. (Krakow) 44: 15–25. Cabrero, J. and Camacho, J. P. M. 1982. Perisentric inversion polymorphism in Aiolopus strepens (Orthoptera: Acrididae): 2014 Karyotype of Chorthippus (Glyptobothrus) bornhalmi 515

effects of chiasma formation. Caryologia 35: 411–424. Cabrero, J. and Camacho, J. P. M. 1985. A Spontaneous interchange heterozygote mosaic in the grasshopper Stauroderus scalaris: interchromosamal chiasma effects. Heredity 54: 235–243. Camacho, J. P. M. 1980. Variabilidad Cromosomica en Poblaciones Naturales de Tettigoniidae, Pamphagoidea and Acridoidea. Tesis Doctoral, Universidad de Granada, Granada. Cano, M. I. and Santos, J. L. 1990. Chiasma frequencies and distributions in gomphocerine grasshoppers: A comparative study between sexes. Heredity 64: 17–23. Carvalho, D. B., Rocha, M. F., Loreto, V., Silva A. E. B. and Souza, M. J. 2011. Ommexecha virens (Thunberg 1824) and Descampsacris serrulatum (Serville 1831) (Orthoptera Ommexechidae): Karyotypes, constitutive heterochromatin and nucleolar organizing regions. Comp. Cytogenet. 5: 123–132. 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. Biodivers. Conserv. 12: 47–64. Demirsoy, A. 1975. Erzurum Bölgesi Orthoptera (Insecta) faunasının tespiti ve taksonomik incelenmesi. Atatürk Üniversitesi Yayınları. 39: 1–114. Fox, D. P. 1973. The Control of chiasma distribution in the locust, Schistocerca gregaria (Forskal). Chromosoma 43: 289– 328. Goni, B., De Vaio, E. S., Beltrami, M., Leira, M. S., Crivel, M., Panzera, F., Castellanos, P. and Basso, A. 1985. Geographic patterns of chromosomal variation in the populations of the grasshopper (Trimetropis pallidipennis) from southern Argentina. Can. J. Genet. Cytol. 27: 254–271. Gusachenko, A. M., Warchalowska-Sliwa, E., Maryanska-Nadachowska, A., Bugrov, A. G. and Vysotskaya, V. 1992. Cytogenetic analysis of populations of Chorthippus albomarginatus (DE GEER) (Acrididae: Orthoptera). Folia Biol. (Krakow) 40: 27–31. Henderson, S. A. 1963. Chiasma distribution at diplotene in a locust. Heredity 18: 173–190. Hewitt, G. M. 1979. Orthoptera: Grasshoppers and Crickets. Insecta I. Vol. 3. In: John, B. (ed.). Animal Cytogenetic. Gebrüder Borntraeger, Berlin. John, B. 1990. Meiosis. Cambridge University Press, Cambridge. p. 396. John, B. and Hewitt, G. M. 1968. Patterns and pathways of chromosome evolution within the Orthoptera. Chromosoma 25: 40–74. Jones, G. H. 1974. Corralated complated of chiasma variation and control of chiasma distribution in rye. Heredity 22: 333– 347. Karabağ, T. 1983. Ankara vilayeti dahilinde mevcut çekirgelerin ekolojik, coğrafik ve sistematik durumları üzerine araştırmalar. Ankara Üniversitesi yayınları, Ankara. p. 121. Karaca, İ., Aslan, B., Demirözer, O. and Karsavuran, Y. 2006. Isparta ili Orthoptera faunası üzerine ön bir değerlendirme. Süleyman Demirel Üniversitesi Ziraat Fakültesi Dergisi 1: 49–52. Koca, S. 1993. Chorthippus dorsatus, Ch. loratus ve Ch. brunneus (Acrididae: Orthoptera) erkeklerinde bazı fiziksel ve kimyasal etmenlerin kiazma frekansı ve meiotik bölünmeye etkileri. Doktora Tezi, Cumhuriyet Üniversitesi, Fen Bilimleri Enstitüsü, Sivas. Koca, S. and Tunçbaş, O. 2006. Poecilimon sanctipauli (Brunner von Wattenwyl, 1878) ve Chorthippus loratus (Fishcher- Waldheim, 1846)’un karyotip analizleri. 18. Ulusal Biyoloji Kongresi, Kuşadası, Aydın. Levan, A., Fredga K. and Sandberg, A. A. 1964. Nomenclature for centromeric position on chromosomes. Hereditas 52: 201–220. Li, N., Wei W. and Ren, B. 2008. C-banding karyotypes of two species of Chorthippus (Orthoptera: Acrypteridae) from China. Entomol. News 119: 1–10. Lopez-Fernandes, C., Rufas, J. S., de la Vega, C. G. and Gosalvez, J. 1984. Cytogenetic studies on Chorthippus jucundus (Fisch.) (Orthoptera) III. The meiotic consequences of a spontaneous centric fusion. Genetica 63: 3–7. Messina, A., Ippolito, S. and Lombardo, F. 1975. Cariologia di alcume specie europee di Phaneropterinae (Insecta, Orthoptera). Animalia 2: 215–224. Oraler Temizkan, G. 1994. Genetik. I. Temel Genetik. İstanbul Üniversitesi Fen Fakültesi Basımevi, İstanbul. p. 281. Rocha, M. F., Melo, N. F. and Souza, M. J. 2011. Comparative cytogenetic analysis of two grasshopper species of the tribe Abracrini (Ommatolampinae, Acrididae). Genet. Mol. Biol. 34: 214–219. Sevgili, H., Demirsoy, A. and Durmuş, Y. 2011. Orthoptera and Mantodea fauna of Kazdağı (Ida) National Park with data on the calling songs of some bush-crickets. Turk. Zool. Derg. 35: 631–652. Sevgili, H. and Heller, K. G. 2003. A new species of the genus Isophya Brunner von Wattenwyl from Turkey (Orthoptera, Tettigoniidae, Phaneropterinae). Tijdschr. Entomol. 146: 39–44. Sybenga, J. 1975. Meiotic Configurations. Monograhps on Theoretical and Applied Genetics I. Springer-Verlag, Berlin, New York. Teoh, S. B. and Yong, H. S. 1983. A spontaneous centric fusion heterozygote in the tropical grasshopper Valanga nicrocornis (Burmeister). Caryologia 36: 165–173. 516 F. Çakmak and S. Koca Cytologia 79(4)

Turkoglu, S. 2001. Türkiye’de yayılış gösteren bazı çekirge (Insecta: Orthoptera) türlerinde karyolojik incelemeler. Doktora Tezi, Cumhuriyet Üniversitesi, Fen Bilimleri Enstitüsü, Sivas. Turkoglu, S. and Koca, S. 2002a. Karyotype, C- and G- band patterns and DNA content of Callimenus (Bradyporus) macrogaster macrogaster. J. Insect Sci. 2: 1–4. Turkoglu, S. and Koca, S. 2002b. Chromosomes of Oedipoda schochi schochi and Acrotylus insbricus (Orthoptera, Acrididae, Oedipodinae). Karyotype and C- and G-band patterns. Turk. Zool. Derg. 26: 327–332. Turkoglu, S., Koca, S. and Akpınar, N. 2003. Karyological observations on the ﬁeld cricket, Gryllus campestris L. (Gryllidae: Orthoptera). Zool. Middle East 28: 113–117. Ünal, M. 2000. First data on Orthoptera of Mount Köroğlu, NW Anatolia, with description of three new taxa. Entomol. News 113: 275–288. Ünal, M. 2008. Bolu ve Düzce illeri Caelifera (Orthoptera) faunası. Bitki Koruma Bülteni 48: 1–31. Ünal, M. 2012. Türkiye Orthoptera Faunası. Erişim Tarihi: 19.05.2012. Viseras, C. E. and Camacho, J. P. M. 1984. The B-choromosomes of Locusta migratoria detection of negative correlation between mean chiasma frequency and rate of accumulation of the B’s; A reanalysis of the available data about the transmission of these B-chromosomes. Genetica 64: 155–164. Warchalowska-Sliwa, E. 1984. Karyological studies on Polish Orthoptera species of the Tettigoniidea superfamily. I. Karyotypes of families: Ephippigeridae, Phaneropteridae, Meconemidae, Conocephalidae. Folia Biol. (Krakow) 32: 253–269. Warchalowska-Sliwa, E., Maryanska-Nadachowska A. and Kostia, D. 1993. Chromosomes of Diesrtammena unicolor unicolor Brunnes von Wattenwvyl, 1888 and Paratachycines (Hemitachycines) boldyrevi (Uvarov, 1926) (Orthoptera, Rhaphidophoridae, Aemodogryllinae) Karyotypes, C-bands and NORs. Folia Biol. (Krakow) 41: 77– 82. White, M. J. D. 1968. Karyotypes and nuclear size in the spermatogenesis of gasshoppers belonging to the subfamilies Gomphomastacinae, Chininae and Biroellinae (Orthoptera, Eumastacidae). Caryologia 21: 167–179. White, M. J. D. 1973. Animal Cytology and Evolution. 3rd Ed. Cambridge University Press, Cambridge.