Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 75(4): 103-109, 2022 Firenze University Press www.fupress.com/caryologia ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.36253/caryologia-1875 Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Citation: Weera Thongnetr, Suphat Prasopsin, Surachest Aiumsumang, Sukhonthip Ditcharoen, Alongklod Tanomtong, Prayoon Wongchantra, Wutthisak Bunnaen, Sumalee Phim- phan (2022). First report of chromosome and karyological analysis of Gekko nutaphandi (Gekkonidae, Squamata) from Thailand: Neo-diploid chromo- some number in genus Gekko. Caryo- logia 75(4): 103-109. doi: 10.36253/cary- ologia-1875 Received: September 11, 2022 Accepted: December 13, 2022 Published: April 28, 2023 Copyright: © 2022 Weera Thongn- etr, Suphat Prasopsin, Surachest Aiumsumang, Sukhonthip Ditchar- oen, Alongklod Tanomtong, Prayoon Wongchantra, Wutthisak Bunnaen, Sumalee Phimphan. This is an open access, peer-reviewed article pub- lished by Firenze University Press (http://www.fupress.com/caryologia) and distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All rel- evant data are within the paper and its Supporting Information files. Competing Interests: The Author(s) declare(s) no conflict of interest. First report of chromosome and karyological analysis of Gekko nutaphandi (Gekkonidae, Squamata) from Thailand: Neo-diploid chromosome number in genus Gekko Weera Thongnetr1, Suphat Prasopsin2, Surachest Aiumsumang3,*, Sukhonthip Ditcharoen4, Alongklod Tanomtong5, Prayo on Wongchantra6, Wutthisak Bunnaen7, Sumalee Phimphan3 1 Division of Biology, Department of Science, Faculty of Science and Technology, Raja- mangala University of Technology Krungthep, Bangkok 10120, Thailand 2 Research Academic Supports Division, Mahidol University, Kanchanaburi Campus, Sai- yok, Kanchanaburi 71150, Thailand 3 Biology program, Faculty of Science and Technology, Phetchabun Rajabhat University, Phetchabun 67000, Thailand 4 Division of Biology, Faculty of Science and Technology, Rajamangala University of Tech- nology Thanyaburi, Khlong Luang, Pathum Thani 12120, Thailand 5 Program of Biology, Faculty of Science, Khon Kaen University, Muang, Khon Kaen 40002, Thailand 6 Center of Environmental Education Research and Training, Faculty of Environment and Resource Studies,Mahasarakham University 44150, Thailand 7 Mahasarakham University Demonstration School (Secondary) Kham Riang Sub-Dis- trict, Kantharawichai District, Maha Sarakham, 44150, Thailand *Corresponding author. E-mail: topthesun1978@gmail.com, surachest.aiu@pcru.ac.th Abstract. The karyotypes of red-eyed Gecko are not reported yet. Herein, we describe the karyotypes of red-eyed Gecko (Gekko nutaphandi Bauer, Sumontha & Pauwels, 2008) from Thailand. Gecko chromosome preparations were directly conducted from bone marrow and testis. Chromosomal characteristics were analyzed by Giemsa stain- ing, Ag-NOR banding as well as fluorescence in situ hybridization (FISH) using micro- satellites d(GC)15 probe. The results showed that the number of diploid chromosomes is 2n=34, while the fundamental number (NF) is 46 in both males and females. The types of chromosomes were 4 large metacentric, 6 large submetacentric, 2 medium telocentric, 2 small metacentric and 20 small telocentric chromosomes. The results of conventional Giemsa staining presented the diploid chromosome number differentia- tion even in the same genus. NORs are located at the secondary constriction to the tel- omere on the long arm of chromosome pair 5. There are no sex differences in karyo- types between males and females. FISH with d(GC)15 sequences were also displayed at the telomeres of most other chromosomes. We found that during metaphase I the homologous chromosomes showed synapsis, which can be defined as 19 ring bivalents and 17 haploid chromosomes (n=17) at metaphase II as a diploid species. The karyo- type formula is as follows: 2n (34) = L4m+L6sm+M2t+S2m+S20t. Keywords: Chromosome, Gekko nutaphandi, Karyotype, Red-eyed Gecko. 104 Thongnetr Weera et al. INTRODUCTION Geckos of the genus Gekko Laurenti, 1768 are mem- bers of a relatively large putatively monophyletic group of lizards that also includes Lepidodactylus Fitzinger, 1843, Luperosaurus Gray, 1845, Pseudogekko Taylor, 1922, and Ptychozoon Kuhl & van Hasselt, 1822 (Kluge 1968; Russell 1972; Bauer et al. 2008). The genus Gekko itself is species rich (84 species) and is generally charac- terized by regional endemism across its broad range in tropical Asia. Recently, G. nutaphandi, is described from Kanchanaburi Province in central western Thailand (Bauer et al. 2008). G. nutaphandi is most similar to G. gecko, G. smithii, G. siamensis, G. verreauxi, and G. albo- fasciolatus Günther, 1872, which have previously been considered to be closely related based on their shared possession of a suite of features. Information about karyotypes in Gekko is scarce and fragmented, and usually based on conventional staining technique; only a few studies have been pub- lished (19 species of 84 species), all of them recent in genus Gekko, namely, G. gecko: 2n=38 (Singh 1974; Sol- leder and Schmid 1984; Wu and Zhao 1984; Trifonov et al. 2011; Qin et al. 2012; Patawang et al. 2014), G. hokouensis: 2n=38 (Chen et al. 1986; Kawai et al. 2009; Shibaike et al. 2009), G. japonicus: 2n=38 (Yoshida and Itoh 1974; Shibaike et al. 2009; Trifonov et al. 2011), G. shibatai and G. vertebralis: 2n=38 (Shibaike et al. 2009), G. Vittatus and G. ulikovskii: 2n=38 (Trifonov et al. 2011), G. tawaensis: 2n=38 (Ota 1989a; Shibaike et al. 2009), G. taylori: 2n=42 (Ota and Nabhitabhata 1991), G. monarchus: 2n=44 (Ota et al. 1990), G. yakuensis, G. petricolus and G. smithii: 2n=38-42 (Ota 1989a), G. kikuchii: 2n=44 (Ota 1989a), G. chinensis: 2n=40 (Lau et al. 1997), G. subpalmatus: 2n=38 (Wu and Zhao 1984), G. swinhonis: 2n=38 (Chen et al. 1986), G. petricolus: 2n=34 (Thongnetr et al. 2022), Dixonius hangseesom: 2n=40, D. Siamensis: 2n=40, D. melanostictus: 2n=42 (Patawang et al. 2022) and Cyrtodactylus inthanon: 2n=40 (Prasopsin et al. 2022) and shown in Table 1. This fact is probably due to the difficulty in obtain- ing samples for cytogenetic analysis in some species or to problems in obtaining metaphase cells by cell culture induction. Moreover, cytogenetic studies using con- ventional staining techniques provide valuable infor- mation on the excellent karyotype diversity shown by these animals. Analyses of cytogenetic markers, includ- ing the number and karyotype formula, sex determina- tion, B chromosomes, number and location of nucleolar organizer regions (NORs), heterochromatin distribution, G-banding, and R-banding, treatments with base-specif- ic fluorochromes and fluorescence in situ hybridization techniques allowed the cytogenetic characterization of populations, species, and supra-specific groups (Affonso and Galetti Jr. 2005). The present study is the first report on the chromosomal characteristics of G. nutaphandi determined using conventional staining, Ag-NOR band- ing, and fluorescence in situ hybridization techniques. MATHERIAL AND METHODS Sample collection, chromosome preparation and chromo- some staining We obtained specimens of G. nutaphandi that were collected from Kanchanaburi Province, Western Thai- land. Chromosomes were directly prepared in vivo (Ota 1989a; Qin et al. 2012) using the following method. With 20% of Giemsa solution, the slides were convention- ally stained for 30 minutes (Patawang et al. 2014). After that, the slides were rinsed thoroughly with running tap water to remove excess stain and placed in air-dry at room temperature. Ag-NOR banding was analysed according to the method of Howell and Black (1980). Two drops each of 50% silver nitrate and 2% gelatine solutions were added to the slides, respectively. Then, they were sealed with cover glasses and incubated at 60°C for 5-10 minutes. They were also soaked in distilled water until the cover glasses were separated. Finally, the slides were placed in air-dry at room temperature. They were observed under the microscope. The use of micro- satellite investigations which were described by Kubat et al. (2008), was followed here with slight modifications. These sequences were directly labeled with Cy3 at the 5 -́terminal during synthesis by Sigma (St. Louis, MO, USA.) Fluorescence in situ hybridization (FISH) was performed under highly rigorous conditions on mitotic chromosome spreads (Pinkel et al. 1986). Chromosomal checks, karyotyping and idiograming Chromosome counting was carried out on mitotic metaphase cells under the light microscope for 30 cells per specimen to determine the diploid number (2n). Twenty clearly observable and well-spread metaphase cells were selected and photographed from each male and female. The short arm length (Ls) and the long arm length (Ll) of each chromosome were measured to cal- culate the total length of the chromosome for 20 well- spread metaphase cells. The chromosome types were classified from the method of Turpin and Lejeune (1965) as metacentric, submetacentric, acrocentric, and telocen- tric chromosomes. 105First report of chromosome and karyological analysis of Gekko nutaphandi (Gekkonidae, Squamata) from Thailand RESULTS AND DISCUSSION Diploid chromosome number, fundamental number and karyotype The Gecko is a large genus (84 species) of Gekkoni- dae family and until now, no study has investigated the karyotype of G. nutaphandi. Furthermore, this is the first report on cytogenetic characterization to use con- ventional Giemsa staining, NOR-banding, and FISH techniques for this species. For G. nutaphandi, the results indicated a diploid chromosome number (2n) of 34 in all studied samples (Figure 1). This result dif- Table 1. The karyotype reviews among the genes Gekko (Gekkonidae, Squamata). Species 2n NF Karyotype formulas NORs FISH Locations References Gekko gekko 38 50 6m+4sm+2a+26t P4 - Thailand Patawang et al. (2014) 38 44 6bi-arms+32uni-arms - - - Singh (1974) 38 - - - - - Wu and Zhao (1984) 38 46 8bi-arms+30uni-arms - - - Solleder and Schmid (1984) 38 - - - - - Trifonov et al. (2011) 38 50 8 m+2sm+2st(a)+26t Laos Qin et al. (2012) 38 48 8 m+2sm+28t - - China Qin et al. (2012) G. hokouensis 38 56 4 m+6sm+20t+8bi-arms* P19 - China, Chen et al. (1986) 38 58/59 2 m+8sm+10a+16t+Z(t)+W(a) - - Japan Kawai et al. (2009) 38 58 4 m+8sm+18t+8bi-arms* P19 - Taiwan Shibaike et al. (2009) 38 58/59 4 m+8sm+16t+8bi-arms*+Z(t)W(sm) P19 - Japan Shibaike et al. (2009) 38 42 2m+18sm+16a+ZW P19 FISH mapping Thailand Srikulnath (2015) G. japonicus 38 - - - - - Yoshida and Itoh (1974) 38 58 4m+8sm+8st+18t P17 - Japan Shibaike et al. (2010) 38 - - - - - Trifonov et al. (2011) G. shibatai 38 58 4m+8sm+18t+8bi-armed P19 - Japan Shibaike et al. (2009) G. vertebralis 38 62 4m+14sm+14t+6bi-arme* P19 - Japan Shibaike et al. (2009) G. vittatus 38 - - - Trifonov et al. (2011) G. ulikovskii 38 - - - Trifonov et al. (2011) G. tawaensis 38 58 4m+8sm+18t+8bi-armed P19 - Japan Shibaike et al. (2009) 38 56 - P19 - Japan Ota (1989a) G. taylori 42 - - - - Thailand Ota and Nabhitabhata (1991) G. monarchus 44 46 - - - Malaysia Ota et al. (1990) G. yakuensis 38 56 - P19 - Japan Ota (1989a) G. petricolus 38 54 - - - Thailand Ota (1989a) G. kikuchii 44 50 - - - Taiwan Ota (1989a) G. chinensis 40 46 6bi-armed+34uni-armed - - China Lau et al. (1997) G. smithii 38 48 - - - - Ota (1989a) G. subpalmatus 38 58 - - - - Wu and Zhao (1984) G. swinhonis 38 66 - - - Chen et al. (1986) G. petricolus 38 54 4m+2sm+10a+22t P17 (CA)15, (GAA)10 Thailand Thongnetr et al. (2022) G. nutaphandi 34 46 6m+6sm+22t P5 (GC)15 Thailand Present study Dixonius hangseesom 40 42 2m+38t P13 - Thailand Patawang et al. (2022) D. siamensis 40 42 2m+38t P13 - Thailand Patawang et al. (2022) D. melanostictus 42 44 2a+40t P8 - Thailand Patawang et al. (2022) Cyrtodactylus inthanon 40 58 12m+4sm+2a+22t P12 (CA)15 (GC) 15 (CAG)10 (GAA)10 Thailand Prasopsin et al. (2022) Note: 2n = diploid chromosome number, NORs = nucleolar organizer regions, NF = fundamental number (number of chromosome arms), bi-arm = bi-armed chromosome, m = metacentric, sm = submetacentric, a = acrocentric, t = telocentric chromosome, *=small bi-arms chromosome, and - = not available. 106 Thongnetr Weera et al. fers from most species of the genus Gecko. The 2n of this genus range from 38 to 44. (Singh 1974; Yoshida and Itoh 1974; Solleder and Schmid 1984; Wu and Zhao 1984; Chen et al. 1986; Ota 1989a; Ota et al. 1990; Ota and Nabhitabhata 1991; Lau et al. 1997; Kawai et al. 2009; Shibaike et al. 2009; Trifonov et al. 2011; Qin et al. 2012; Patawang et al. 2014; Patawang et al. 2022 and Pra- sopsin et al. 2022. The fundamental number (NF) of G. nutaphandi was 46 in both males and females. The karyotype con- sisted of 4 large metacentric, 6 large submetacentric, 2 medium telocentric, 2 small metacentric, and 20 small telocentric chromosomes (Table 1). These results of NF are agreeable with the previous reports of G. gecko (Sol- leder and Schmid 1984), G. monarchus (Ota et al. 1990), and G. chinensis (Lau et al. 1997). The NFs of the genus Gekko range from 44 to 62, and karyotypes are com- posed of both mono- and bi-arms chromosomes. Nir- chio et al. (2002) proposed that species with high NF is an advanced state or apomorphic character, whereas one with low NF is a primitive state or plesiomorphic char- acter. Thus, the G. nutaphandi seems to be a more prim- itive karyotype than other species. The karyotype for- mula for this species is 2n (34) = L4m+L6sm+M2t+S2m+S20t. There is no evidence of differentiated sex chromosomes in this species which accord with all species of this genus (Singh 1974; Yoshida and Itoh 1974; Solleder and Schmid 1984; Wu and Zhao 1984; Chen et al. 1986; Ota 1989a; Ota et al. 1990; Ota and Nabhitabhata 1991; Lau et al. 1997; Kawai et al. 2009; Shibaike et al. 2009; Tri- fonov et al. 2011; Qin et al. 2012; Patawang et al. 2014; Patawang et al. 2022 and Prasopsin et al. 2022. The present study on the meiotic cell division of G. nutaphandi found that during metaphase I (meiosis I), the homologous chromosomes showed synapsis, which can be defined as the 17 bivalent (Figure 2A), and 17 haploid chromosomes at metaphase II (Figure 2B) as diploid species. The largest metacentric chromosome pair 1 is the largest bivalent. No diakinesis and meta- phase I cell with partially paired bivalents are speculated to be heteromorphic sex chromosomes, and no meta- phase II cells with condensed chromosomes are specu- lated to be the sex chromosome. Chromosome markers from Ag-NOR banding The development of Ag-NOR staining technique (Howell and Black 1980) to detect metaphase chromo- some sites of NORs has greatly facilitated comparative studies of NORs variation. Silver staining of NORs is considered as one of the standard banding methods and has assumed considerable importance in characterizing a species’ karyotype. The present study was firstly accom- plished by using Ag-NOR staining in G. nutaphandi. The Ag-NOR positions were shown on the long arm near the centromere of the telocentric chromosome pair 5 (sub- telomeric NOR) (Figure 3). The single pair of NOR is the same as in Gecko species. Compared with other geckos, the NOR regions showed two NORs appearing near telo- meric region of small bi-armed or mono-armed chromo- some. An example of the previous reports of the genus Gekko had two NORs on one pair of small bi-arms chro- mosomes. G. gecko had two NORs on the near telomere of mono-arms chromosome pair 4 (Patawang et al. 2014), and G. petricolus had two NORs on the long arm near telomere of bi-arms chromosome pair 5 (Thongnetr et al. 2022), while there were G. shibatai, G. Yakuensis, Figure 1. Metaphase chromosome plates and karyotypes of G. nuta- phandi (A) male (B) female by conventional technique. Scale Bars = 10 µm. Figure 2. Metaphase chromosome plates and karyotypes of G. nuta- phandi (A) Metaphase I (B) Metaphase II by conventional tech- nique. Scale Bars = 10 µm. 107First report of chromosome and karyological analysis of Gekko nutaphandi (Gekkonidae, Squamata) from Thailand G. hokouensis, G. tawaensis, G. vertebralis and G. yakue- nsis which had two NORs on the long arm near the tel- omere of small bi-arms chromosome pair 19 (Ota 1989b; Chen et al. 1986; Shibaike et al. 2009). The use of NORs in explaining kinships depends to a considerable extent on the uniformity of this characteristic and the degree of variety within a taxon (Yüksel and Gaffaroğlu 2008). The idiogram shows a continuous length gradation of chromosomes. The size differences between the larg- est and smallest chromosomes exhibit approximately two-fold. The data of the chromosome measurement on mitotic metaphase cells (from all specimens) are shown in Table 2. Idiograms by conventional Giemsa staining and Ag-NOR banding are shown in Figure 4. Microsatellite pattern Microsatellites or simple sequence repeats (SSRs) are oligonucleotides of 1–6 base pairs in length, forming excessive tandem repeats of usually 4 to 40 units (Tautz and Renz 1984; Ellegren 2004; Chistiakov et al. 2006). They show ample distribution throughout eukaryotic genomes, scattered or clustered in euchromatin and het- erochromatin. They are highly polymorphic regarding Figure 3. Metaphase chromosome plates and karyotypes of G. nuta- phandi (A) male (B) female by Ag-NOR banding technique. Scale Bars = 10 µm. Table 2. Mean length of short arm chromosome (Ls), length of long arm chromosome (Ll), length of total chromosomes (LT), relative length (RL), centromeric index (CI), and standard deviation (SD) from 20 metaphases of male and female Red-eyed Gecko (Gekko nuta- phandi), 2n (diploid)=34. Chro.pairs Ls Ll LT CI±SD RL±SD Chro. size Chro. type 1 0.932 1.131 2.035 0.548±0.012 0.132±0.007 Large metacentric 2 0.835 1.160 1.961 0.602±0.015 0.120±0.005 Large submetacentric 3 0.769 1.303 2.001 0.626±0.023 0.115±0.007 Large submetacentric 4 0.784 1.113 1.833 0.586±0.020 0.113±0.007 Large metacentric 5* 0.410 1.003 1.372 0.673±0.019 0.084±0.004 Large submetacentric 6 0.000 1.216 1.216 1.000±0.000 0.066±0.003 Medium telocentric 7 0.000 1.016 1.016 1.000±0.000 0.060±0.003 Small telocentric 8 0.000 1.049 1.049 1.000±0.000 0.055±0.002 Small telocentric 9 0.000 0.720 0.720 1.000±0.000 0.046±0.003 Small telocentric 10 0.000 0.800 0.800 1.000±0.000 0.037±0.004 Small telocentric 11 0.000 0.629 0.629 1.000±0.000 0.032±0.003 Small telocentric 12 0.000 0.595 0.595 1.000±0.000 0.029±0.003 Small telocentric 13 0.000 0.615 0.615 1.000±0.000 0.026±0.003 Small telocentric 14 0.000 0.596 0.596 1.000±0.000 0.023±0.004 Small telocentric 15 0.000 0.464 0.464 1.000±0.000 0.021±0.003 Small telocentric 16 0.195 0.220 0.374 0.520±0.038 0.024±0.004 Small metacentric 17 0.000 0.326 0.326 1.000±0.000 0.016±0.003 Small telocentric Note: Chro.=chromosome, * NORs bearing chromosomes (satellite chromosome). Figure 4. Idiogram represents chromosome type, size, and Ag-NOR banding of G. nutaphandi. 108 Thongnetr Weera et al. copy number deviation (Ellegren 2004). Fluorescence hybridization indicates the (GC)15 repeat showing abun- dance at the telomeric ends of most chromosomes (Fig- ure 5), verifying the findings from other gekko groups investigated to date (Srikulnath 2015). The distribution of microsatellites was not only restricted to heterochro- matin but also dispersed in euchromatic regions of the chromosomes (Getlekha et al. 2016). Nonetheless, closely related fish species involved in recent speciation events could present a differential pattern in the distribution and quantity of microsatellite sequences on chromo- some. In conclusion, we present the first Karyotype, NOR phenotype, and microsatellite patterns (GC)15 on the chromosomes are specific to species in the G. nutaphan- di. More species and techniques should be further stud- ied for more information about the chromosomal diver- sity and chromosomal evolution in this genus. ACKNOWLEDGEMENTS This research project was financially supported by Thailand science research and innovation (TSRI). Khon Kaen University and Phetchabun Rajabhat University. REFERENCES Affonso PRAM, Galetti Jr PM. 2005. Chromosomal diversification of reef fishes from genus Centropyge (Perciformes, Pomacanthidae). Genetica 123: 227– 233. Bauer AM, Sumontha M, Pauwels SGO. 2008. A new red-eyed Gekko (Reptilia: Gekkonidae) from Kan- chanaburi Province, Thailand. Zootaxa 1750(1750): 32–42. Chen J, Peng X, Yu D. 1986. Studies on the karyotype of three species of the genus Gekko. Acta Herpetologica Sinica 5: 24–29. Chistiakov DA, Hellemans B, Volckaert FAM. 2006. Microsatellites and their genomic distribution, evolu- tion, function and applications: a review with special reference to fish genetics. Aquaculture 255: 1–29. htt- ps://doi.org/10.1016/j.aquaculture.2005.11.031 Ellegren H. 2004. Microsatellites: simple sequences with complex evolution. Nat Rev Genet 5: 435–445. htt- ps://doi.org/10.1038/nrg1348 Getlekha N, Molina WF, Cioffi MB, Yano CF, Maneechot N, Bertollo LAC, Supiwong W, Tanomtong A. 2016. Repetitive DNAs highlight the role of chromosomal fusions in the karyotype evolution of Dascyllus spe- cies (Pomacentridae, Perciformes). Genetica 144(2): 203–211. https://doi.org/10.1007/s10709-016-9890-5 Howell WM, Black DA. 1980. Controlled silver-staining of nucleolus organizer regions with a protective col- loidal developer: a 1-step method. Experientia 36: 1014–1015. Kawai A, Ishijima J, Nishida C, Kosaka A, Ota H, Kohno S, Matsuda Y. 2009. The ZW sex chromosomes of Gekko hokouensis (Gekkonidae, Squamata) represent highly conservedhomology with those of avian spe- cies. Chromosoma 118: 43–51. Kluge, A.G. 1968. Phylogenetic relationships of the gek- konid lizard genera Lepidodactylus Fitzinger, Hemi- phyllodactylus Bleeker, and Pseudogekko Taylor. Phil- ippine Journal of Science, 95: 331–352. Kubat Z, Hobza R, Vyskot B, Kejnovsky E. 2008. Micros- atellite accumulation in the Y chromosome of Silene latifolia. Genome 51: 350–356. Lau MW, Ota H, Bogadek A. 1997. Chromosomes poly- morphismand karyotype of Gekko chinensis (Tokay- nidae: Reptilia) from Hong Kong. J Herpetol 31: 137–139. Nirchio M, Turner BJ, Perez JE, Gaviria JI, Cequea H. 2002. Karyotypes of three species of toadfish (Batrachoididae: Teleostei) from Venezuela. Sci Mar 66(1): 1–4. Ota H. 1989a. Karyotypes of five species of Gekko (Gek- konidae: Lacertilia) from East and Southeast Asia. Herpetologica 45 (4): 438-443. Ota H. 1989b. A review of the geckos (Lacertilia: Reptil- ia) of the Ryukyu Archipelago and Taiwan. In: Mat- sui M, Hikida T, Goris RC. (eds.). Current Herpe- tology in East Asia. Herpetological Society of Japan, Kyoto. Ota H, Hikida T, Matsui M, Mori A. 1990. Karyotype of Gekko monarchus (Squamata: Gekkonidae) from Sarawak, Malaysia. Japanese Journal of Herpetology 13: 136–138. Ota H, Nabhitabhata J. 1991. A new species of Gekko (Gekkonidae: Squamata) from Thailand. Copeia 1991: 503–509. Figure 5. Metaphase chromosome plate of G. petricolus (A) by FISH technique of DAPI (B) microsatellite probe (GC)15. Scale Bars = 10 µm. 109First report of chromosome and karyological analysis of Gekko nutaphandi (Gekkonidae, Squamata) from Thailand Prasopsin S, Muanglen N, Ditcharoen S, Suwannapoom C, Tanomtong A, Thongnetr W. 2022. First Report on Classical and Molecular Cytogenetics of Doi Inthanon Bent-toed Gecko, Cyrtodactylus inthanon Kunya et al., 2015 (Squamata: Gekkonidae) in Thai- land. Caryologia, 75(2): 109–117. Patawang I, Tanomtong A, Jumrusthanasan S, Kakam- puy W, Neeratanaphan L, Pinthong K. 2014. Chro- mosomal characteristics of NORs and karyological analysis of tokay gecko, Gekko gecko )Gekkonidae, Squamata) from mitotic and meiotic cell division. Cytologia 79(3): 315–324. Patawang I, Prasopsin S, Suwannapoom C, Tanomtong A, Keawmad P, Thongnetr W. (2022). Chromosomal description of three Dixonius (Squamata, Gekkoni- dae) from Thailand. Caryologia, 75(2): 101–108. Pinkel D, Straume T, Gray J. 1986. Cytogenetic analy- sis using quantitative, high sensitivity, fluorescence hybridization. Proceedings of the National Academy of Sciences of the United States of America 83: 2934– 2938. Qin XM, Li HM, Zeng ZH, Zeng DL, Guan QX. 2012. Genetic variation and differentiation of Gekko gecko from different populations based on mitochondrial cytochrome b gene sequences and karyotypes. Zoo- logical Science 29: 384–389. Russell AP. 1972. The Foot of Gekkonid Lizards: A Study in Comparative and Functional Anatomy. [Disserta- tion]. University of London, London. Singh L. 1974. Study of mitotic and meiotic chromo- somes in seven species of lizards. Proceedings of the Zoological Society 27: 57–79. Solleder E, Schmid M. 1984. XX/XY sex chromosomes in Gekko gecko (Sauria, Reptilia). Amphibia-Reptilia 5: 339–345. Srikulnath K, Uno Y, Nishida C, Ota H, Matsuda Y. 2015. Karyotype reorganization in the Hokou Gecko (Gek- ko hokouensis, Gekkonidae): The Process of Micro- chromosome Disappearance in Gekkota. PLoS ONE. DOI:10.1371/journal.pone.0134829 Shibaike Y, Takahashi Y, Arikura I, Iiizumi R, Kitakawa S, Sakai M, Imaoka C, Shiro H, Tanaka H, Akakubo N, Nakano M, Watanabe M, Ohne K, Kubota S, Kohno S, Ota H. 2009. Chromosome evolution in the lizard genus Gekko (Gekkonidae, Squamata, Reptilia) in the East Asian islands. Cytogenet Genome Res 127: 182– 190. Solleder E, Schmid M. 1984. XX/XY sex chromosomes in Gekko gecko (Sauria, Reptilia) amphibia-Reptilia 5: 339–345. Tautz D, Renz M.1984. Simple sequences are ubiquitous repetitive components of eukaryotic genomes. Nucle- ic Acids Res 25: 4127–4138. https://doi. org/10.1093/ nar/12.10.4127 Thongnetr W, Aiumsumang S, Tanomtong A, Phimphan S. 2022. Classical chromosome features and micros- atellites repeat in Gekko petricolus (Reptilia, Gekkoni- dae) from Thailand. Caryologia 75(2): 81–88. Trifonov VA, Giovannotti M, O’Brien PCM, Wallduck M, Lovell F, Rens W, Parise-Maltempi PP, Caputo V, Ferguson-Smith MA. 2011. Chromosomal evolution in Gekkonidae. I. Chromosome painting between Gekko and Hemidactylus species reveals phylogenetic relationships within the group. Chromosome Res 19: 843–855. Turpin R, Lejeune J. 1965. Les Chromosomes Humains. Gauther Villars, Paris. Wu G, Zhao E. 1984. Studies on karyotypes of Gekko gecko and Gekko subpalmatus. Acta Herpetologica Sinica 3: 61–64. Yoshida M, Itoh M. 1974. Karyotype of the gecko, Gek- ko japonicus. Chromosome Information Service 17: 29–31. Yüksel E, Gaffaroğlu M. 2008. The analysis of nucleo- lar organizer regions in Chalcalburnus mossulensis (Pisces: Cyprinidae). Journal of Fisheriessciences.com 2: 587–591. Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Volume 75, Issue 3 - 2022 Firenze University Press