Iran. J. Ichthyol. (March 2021), 8(1): 14-18 Received: September 7, 2020 © 2021 Iranian Society of Ichthyology Accepted: February 4, 2021 P-ISSN: 2383-1561; E-ISSN: 2383-0964 doi: 10.22034/iji.v8i1.541 http://www.ijichthyol.org

Research Article

Karyology of an endemic simavicus (Balık & Bănărescu, 1978) (Teleostei: )

Muradiye KARASU AYATA*1, Sevgi ÜNAL KARAKUŞ2, Muhammet GAFFAROĞLU3

1Faculty of Health Sciences, Kırşehir Ahi Evran University, Kırşehir, . 2Department of Molecular Biology and Genetics, Faculty of Science, Bartın University, Bartın, Turkey. 3Department of Molecular Biology and Genetics, Faculty of Science and Art, Kırşehir Ahi Evran University, Kırşehir, Turkey. *Email: [email protected] Abstract: The diploid chromosome number and chromosome morphologies of endemic Oxynoemacheilus simavicus (Balık & Bănărescu, 1978) were revealed out from Gediz River, Turkey. Metaphase chromosomes were obtained from head kidney cells. The diploid chromosome number was found as 50 and karyotype was consisted 13 pairs of biarmed and 12 pairs of uniarmed chromosomes. Fundamental arm number was calculated as 76. The obtained results shall contribute to nemacheilid chromosomal studies.

Keywords: cytotaxonomy, Chromosome, Gediz River, Turkey.

Citation: Karasu Ayata, M.; Ünal Karakuş, S. & Gaffaroğlu, M. 2021. Karyology of an endemic Oxynoemacheilus simavicus (Balık & Bănărescu, 1978) (Teleostei: Nemacheilidae). Iranian Journal of Ichthyology 8(1): 14-18.

Introduction simavicus is an endemic loach of Turkish inland Besides molecular studies of , karyological water described from the Marmara Sea tributary studies are important in fish . Fish (Freyhof et al. 2011; Güçlü & Küçük 2015; Çiçek et karyology has been developed since 1960 (Chiarelli al. 2018, 2019). There is no karyological study of & Capanna 1973). In addition, karyological studies O. simavicus. Hence, this study aimed to study in fish have great importance in cytotaxonomy, karyological characteristics of O. simavicus. aquaculture, mutagenesis and genetic control (Esmaeili et al. 2015). Obtaining chromosomes and Materials and Methods karyotypes from fish species are often difficult. Thirteen specimens (six males and seven females) of However, improvements in methodological O. simavicus were collected using an electrofishing approaches have contributed to chromosome device from Tabakdere, Salihli, Manisa, Turkey analyses for evaluating fish karyotypic diversity (38°28′N, 28°03′E) in 2019. The individuals were (Bertollo et al. 2015). Chromosomal studies in fish transported alive to the laboratory. The fishes were are often limited to the determination of diploid treated with the guidelines of the local ethics chromosome number (2n), fundamental arm number committee. Metaphase chromosomes were obtained (FN) and chromosome morphology (Rábová et al. from head kidney cells according to Bertollo et al. 2015). (2015)’s air drying protocol. From each specimen, at From the Anatolian nemacheilid species, only least 10 chromosome slides were prepared. All nine of them have been studied cytogenetically analysed specimens are deposited in the Genetic (Tanrıkulu 2008; Değer 2011; Kılıç et al. 2011; Laboratory of Kırşehir Ahi Evran University, Gaffaroğlu et al. 2012; Gaffaroğlu et al. 2014; Ünal Turkey. All Giemsa stained slides were scanned with et al. 2016; Ayata et al. 2018). Oxynoemacheilus a Leica DM 3000 microscope (Leica Microsystems 14

Karasu Ayata et al.- Karyology of an endemic Oxynoemacheilus simavicus

GmbH, Germany) and metaphases were photographed with AKAS software (Argenit Mikrosistem, Turkey). At least 10 metaphases were examined per individual. Karyotype was arranged manually. Chromosomes in the karyotype were classified according to Levan et al. (1964). For calculating FN, meta-submetacentric (m-sm) chromosomes were taken as biarmed, whereas subtelo-acrocentric (st-a) chromosomes were considered as uniarmed.

Results A total of 130 metaphases were counted for chromosome number. The diploid chromosome number of O. simavicus was detected as 2n = 50 (Fig. 1A). The chromosome morphologies were as follows: six pairs of m, seven pairs of sm and 12 pairs of st-a chromosomes (Fig. 1B). The FN was calculated as 76. The largest chromosome pair of the karyotype was st-a chromosome. No heteromorphic sex chromosomes were observed in the karyotype.

Fig.1. (A) Metaphase of Oxynoemacheilus simavicus after Discussion Giemsa staining. (B) Arranged karyotype of O. simavicus. Sember et al. (2015) reported that despite the vast biodiversity within Nemacheilidae, the cytogenetics show some differences (Table 1). The number of of this group remain poorly explored. The fact that uniarmed chromosomes of O. simavicus is higher chromosomal studies have been reported in seven out than all other Anatolian Oxynoemacheilus species of 41 Anatolian Oxynoemacheilus species confirms (Table 1). Consequently, the FN of O. simavicus is this report (Table 1). From this limited data, the lowest from the other Anatolian karyotypes of all analysed Anatolian Oxynoemacheilus species (Table 1). Oxynoemacheilus species display the 2n = 50 (Table On the other hand, in contrast to 2n similarity, 1). However, chromosome morphologies and FNs there are some differences about the chromosome

Table 1. Karyological data for the representatives of the Oxynoemacheilus from Turkey.

Species 2n Chromosome morphology FN References O. panthera 50 14m+18sm+18a 82 Tanrıkulu (2008) O. argyrogramma 50 44m-sm+6a 94 Değer (2011) O. frenatus 50 32m-sm+18a 82 Değer (2011) O. sp. 50 30m-sm+20a 80 Değer (2011) O. tigris 50 18m+18sm+14a 86 Kılıç et al. (2011) O. angorae 50 8m+28sm+14st-a 86 Gaffaroğlu et al. (2014) O. atili 50 10m+18sm+22st-a 78 Ayata et al. (2018) O. simavicus 50 12m+14sm+24st-a 76 This study

15 Iran. J. Ichthyol. (March 2021), 8(1): 14-18

morphology of O. simavicus with other two AYATA for their help in the field. Anatolian nemacheilids i.e. Turcinoemacheilus kosswigi (Gaffaroğlu et al. 2012) and References Seminemacheilus lendlii (Ünal et al. 2016). Ayata, M.K.; Unal, S. & Gaffaroglu, M. 2018. Oxynoemacheilus simavicus possess a relatively high Chromosomal analysis of Oxynoemacheilus atili number of uniarmed chromosomes like T. kosswigi Erk’akan, 2012 (Teleostei, Nemacheilidae). Turkish (Gaffaroğlu et al. 2012). Therefore, the FN of Journal of Fisheries and Aquatic Sciences 18: 991-994. Bertollo, L.A.C.; Cioffi, M.B. & Moreira-Filho, O. 2015. O. simavicus is lower than S. lendlii (Ünal et al. Direct chromosome preparation from freshwater T kosswigi 2016) similar to those of . (Gaffaroğlu et teleost fishes. In: Ozouf-Costaz, C.; Pisano, E.; Foresti, al. 2012). F. & Toledo, L.F.A. (eds.), Fish cytogenetic Moreover, O. simavicus has the same 2n with techniques, ray-fin fishes and chondrichthyans, CRC O. persa and O. tongiorgii from Iran, however, Press, USA. pp. 21-26. chromosome morphologies have some differences Chiarelli, A.B. & Capanna, E. 1973. Checklist of fish (Esmaeili et al. 2015). The number of uniarmed chromosomes. In: Chiarelli, A.B. & Capanna, E. chromosomes of O. simavicus are much higher than (eds.), Cytotaxonomy and vertebrate evolution, O. persa and O. tongiorgii (only have four pairs) Academic Press, London. pp. 205-253. (Esmaeili et al. 2015). FN of O. simavicus is lower Çiçek, E.; Eagderi, S. & Sungur, S. 2019. Oxynoemacheilus phoxinoides than O. persa and O. tongiorgii (Esmaeili et al. (Erk'akan, Nalbant & Özeren, 2007): a junior synonym of Oxynoemacheilus 2015). Moreover, Sember et al. (2015) showed that angorae (Steindachner, 1897). FishTaxa 4(1): 13-17. 17 nemacheilid loach species have 2n=50 as Çiçek, E.; Fricke, R.; Sungur, S. & Eagderi, S. 2018. observed in this study. However, only two species Endemic freshwater fishes of Turkey. FishTaxa 3(4): have the same FN with O. simavicus. Sember et al. 1-39. (2015) reported that despite the conservatism of the Değer, D. 2011. The karyological investigations of some same 2n, nemacheilid loaches exhibited variability types from Cobitoidea from River System Tigris and on microstructural level. Chromosomal Euphrates. PhD thesis. University of Dicle, rearrangements like pericentric inversions, tandem Diyarbakır, Turkey. and centric fusions could be responsible for Esmaeili, H.R.; Pirvar, Z.; Ebrahimi, M. & Geiger, M.F. karyotype differentiation in the loaches (Sember et 2015. Karyological and molecular analysis of three al. 2015). This hypothesis should be true for the endemic loaches (: Cobitoidea) from Kor River Basin, Iran. Molecular Biology Research Anatolian Oxynoemacheilus species. Heteromorphic Communications 4(1): 1-13. sex chromosomes have not been detected in Freyhof, J.; Erk’akan, F.; Özeren, C. & Perdices, A. 2011. karyotype of O. simavicus as other Oxynoemacheilus An overview of the western palaearctic loach genus species (Değer 2011; Gaffaroğlu et al. 2014; Oxynoemacheilus (Teleostei: Nemacheilidae). Esmaeili et al. 2015; Ayata et al. 2018). Ichthyological Exploration of Freshwaters 22(4): 301- In conclusion, the chromosomal properties of the 312. Anatolian endemic nemacheilid loach, O. simavicus, Gaffaroğlu, M.; Karasu, M. & Unal, S. 2012. Karyotype is determined for the first time. It should be useful to of river loach Turcinoemacheilus kosswigi Bănărescu study chromosomal banding properties of and Nalbant, 1964 (, Balitoridae) from O. simavicus in the future studies. the Euphrates River, Turkey. Journal of Agricultural Science and Technology 14: 821-826. Acknowledgements Gaffaroğlu, M.; Ünal, S. & Ayata, M.K. 2014. Karyotype properties of Oxynoemacheilus angorae The authors thank to Yalçın KARASU and Ali 16

Karasu Ayata et al.- Karyology of an endemic Oxynoemacheilus simavicus

(Steindachner, 1897) (Teleostei, Nemacheilidae) from Anatolia. Journal of FisheriesSciences.com 8(4): 342- 345. Güçlü, S.S. & Küçük, F. 2015. The ichthyofauna of Gediz River (Turkey): taxonomic and zoogeographic features. Annual Research & Review in Biology 6(3): 202-214. Kılıç, B.; Gül, S.; Özkan, O.; Kaya, T.Ö.; Nur, G. & Aksu, P. 2011. Karyotype analysis in Orthrias tigris (Heckel, 1843), living in the Kura-Aras river basin. Afyon Kocatepe University Journal of Sciences 11: 43-49. Levan, A.; Fredga, K. & Sandberg, A.A. 1964. Nomenclature for centromeric position on chromosomes. Hereditas 52: 201-220. Rábová, M.; Völker, M.; Pelikánová, Š. & Ráb, P. 2015. Sequential chromosome banding in fishes. In: C.Ozouf-Costaz, E. Pisano, F. Foresti, L.F.A. Toledo, (Eds.). Fish cytogenetic techniques, ray-fin fishes and chondrichthyans, CRC Press, USA. pp. 92-102. Sember, A.; Bohlen, J.; Šlechtová, V.; Altmanová, M.; Symonová, R. & Ráb, P. 2015. Karyotype differentiation in 19 species of river loach fishes (Nemacheilidae, Teleostei): extensive variability associated with rDNA and heterochromatin distribution and ıts phylogenetic and ecological ınterpretation. BMC Evolutionary Biology 15: 251- 273. Tanrıkulu, D. 2008. Karyotype analysis of (Orthrias panthera, Heckel, 1843) from Kura-Aras river basin. MSc thesis. Kafkas University, Kars, Turkey. Ünal, S.; Ayata, M.K. & Gaffaroğlu, M. 2016. Cytogenetic analysis of Seminemacheilus lendlii (Hanko, 1925) (Teleostei: Nemacheilidae). Turkish Journal of Fisheries and Aquatic Sciences 16: 913-916.

17 Iran. J. Ichthyol. (March 2021), 8(1): 14-18 Received: September 7, 2020 © 2021 Iranian Society of Ichthyology Accepted: February 4, 2021 P-ISSN: 2383-1561; E-ISSN: 2383-0964 doi: 10.22034/iji.v8i1.541 http://www.ijichthyol.org

ﻣﻘﺎﻟﻪ ﭘﮋوﻫﺸﯽ ﮐﺎرﯾﻮﻟﻮژي ﮔﻮﻧﻪ ﺑﻮﻣﺰاد (Oxynoemacheilus simavicus(Balık & Bănărescu, 1978 (ﻣﺎﻫﯿﺎن اﺳﺘﺨﻮاﻧﯽ ﻋﺎﻟﯽ: ﺟﻮﯾﺒﺎرﻣﺎﻫﯿﺎن ﺑﺪون ﺧﺎردار)

ﻣﻮرادي ﮐﺎراﺳﻮ آﯾﺎﺗﺎ1*، ﺳﻮﮔﯽ اوﻧﺎل ﮐﺮﮐﻮش2، ﻣﺤﻤﺖ ﻏﻔﺎرﮔﻮﺷﻠﻮ3 1داﺷﮑﺪه ﻋﻠﻮم ﺑﻬﺪاﺷﺘﯽ، داﻧﺸﮕﺎه ﮐﺮﺷﻬﯿﺮ آﻫﯽ اوران، ﮐﺮﯾﺸﻬﯿﺮ، ﺗﺮﮐﯿﻪ. 2ﮔﺮوه زﯾﺴﺖ ﺷﻨﺎﺳﯽ ﺳﻠﻮﻟﯽ و ژﻧﺘﯿﮏ، داﻧﺸﮑﺪه ﻋﻠﻮم ﭘﺎﯾﻪ، داﻧﺸﮕﺎه ﺑﺎرﺗﯿﻦ ﺗﺮﮐﯿﻪ، ﺑﺎرﺗﯿﻦ، ﺗﺮﮐﯿﻪ. 3ﮔﺮوه زﯾﺴﺖ ﺷﻨﺎﺳﯽ ﺳﻠﻮﻟﯽ و ژﻧﺘﯿﮏ، داﺷﮑﺪه ﻫﻨﺮ و ﻋﻠﻮم، ﮐﺮﯾﺸﻬﯿﺮ، ﺗﺮﮐﯿﻪ.

ﭼﮑﯿﺪه: در اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺗﻌﺪاد ﮐﺮوﻣﻮزومﻫﺎي دﯾﭙﻠﻮﺋﯿﺪ و رﯾﺨﺖ ﮐﺮوﻣﻮزوﻣﯽ ﮔﻮﻧﻪ ﺑﻮﻣﺰاد (Oxynoemacheilus simavicus (Balık & Bănărescu 1978، از رودﺧﺎﻧﻪ ﮔﺪﯾﺰ، ﺗﺮﮐﯿﻪ ﻣﺸﺨﺺ ﺷﺪ. ﮐﺮوﻣﻮزومﻫﺎي ﻣﺘﺎﻓﺎز از ﺳﻠﻮلﻫﺎي ﻓﻮﻗﺎﻧﯽ ﮐﻠﯿﻪ ﮔﺮﻓﺘﻪ ﺷﺪ. ﺗﻌﺪاد ﮐﺮوﻣﻮزومﻫﺎي دﯾﭙﻠﻮﺋﯿﺪ، 50 ﻋﺪد و ﮐﺎرﯾﻮﺗﯿﭗ ﺷﺎﻣﻞ 13 ﺟﻔﺖ ﮐﺮوﻣﻮزوم ﺑﺎزودار و 12 ﺟﻔﺖ ﮐﺮوﻣﻮزوم ﺑﺪون ﺑﺎزو ﺑﻮد. ﺗﻌﺪاد ﺑﺎزوي اﺻﻠﯽ 76 ﻣﺤﺎﺳﺒﻪ ﺷﺪ. ﻧﺘﺎﯾﺞ ﻧﺘﺎﯾﺞ ﺣﺎﺻﻞ ﺑﻪ ﻣﻄﺎﻟﻌﺎت ﮐﺮوﻣﻮزوﻣﯽ ﺧﺎﻧﻮاده ﺟﻮﯾﺒﺎرﻣﺎﻫﯿﺎن ﺑﺪون ﺧﺎردار ﮐﻤﮏ ﺧﻮاﻫﺪ ﮐﺮد. ﮐﻠﻤﺎتﮐﻠﯿﺪي: ﺳﯿﺘﻮﺗﺎﮐﺴﻮﻧﻮﻣﯽ ﻣﺎﻫﯽ، ﮐﺮوﻣﻮزوم، رودﺧﺎﻧﻪ ﮔﺪﯾﺰ، ﺗﺮﮐﯿﻪ.

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