Karyomorphological delineation, and the NOR loci on the sex chromosome in three species of Chrysopeleinid (Chrysopeleinae: Colubridae) from Thailand




Abstract. Donbundit N, Bausriyod P, Tanomtong A, Srisamoot N, Sumontha M, Thongnetr W, Patawang I, Supiwong W, Ditcharoen S, Muanglen N, Kaewmad P. 2022. Karyomorphological delineation, and the NOR loci on the sex chromosome in three species of Chrysopeleinid (Chrysopeleinae: Colubridae) from ThailandBiodiversitas 23: 3813-3819The description of chromosomal characteristics of three snakes from Thailand namely, Ahaetulla fusca, A. prasina, and Chrysopelea ornata was by using conventional staining and Ag-NOR banding techniques. The results showed that the same diploid chromosome number (2n) = 36 (16 macrochromosomes + 20 microchromosomes) and the fundamental number (NF) was 54 while the karyotype formulae in these three species are as follows: A. fusca; 2n(36) = Lm6 + Sm2+ Ssm4 + Sa2+ ZZ/ZW + 20 microchromosomes, A. prasina; 2n(36) = Lm6+ Sm6+ Sa2+ ZZ/ZW + 20 microchromosomes and C. ornata; 2n(36) = Lm4+ Lsm2+ Sm2 + Ssm4+ Sa2+ ZZ/ZW + 20 microchromosomes. Besides, the ZZ/ZW sex chromosome was observed in all species, whereas C. ornata was discovered the heteromorphic sex chromosome for the first time. Nucleolar organizer regions (NORs) are located on telomeric region of one pair in all species, while located on microchromosomes in A. fusca and A. prasina, whereas C. ornata was detected on sex chromosomes. Therefore, the Z chromosome revealed the NOR on the telomeric region of the long arm, while NOR of the W chromosome was located on the telomeric region of the short arm.


Chan-ard T, Parr JWK, Nabhitabhata J. 2015. A Field Guide to the Reptiles of Thailand, Oxford University Press, New York. pp. 187-192.
Chooseangjaew S, Tanyaros S, Maneechot N, Buasriyot P, Getlekha N, Tanomtong A. 2017. Chromosomal characteristics of the tropical oyster, Crassostrea belcheri Sowerby, 1871 (Ostreoida, Ostreidae) by conventional and Ag-NOR banding techniques. Cytologia 82: 3-8. DOI: 10.1508/cytologia.82.3.
David P, Nadolski B, Ganesh SR, Adhikari OD, Srikanthan AN. 2020. A nomenclatural review of Ahaetulla nasuta (LACÉPÈDE, 1789): resurrection of Ahaetulla fusca (DUMÉRIL, BIBRON ET DUMÉRIL, 1854) for the populations of the Indochinese region (Reptilia, Squamata, Colubridae). Russ J Herpetol 29 (1): 19-34. DOI: 10.30906/1026-2296-2022-29-1-19-34.
Donbundit N, Thongnetr W, Patawang I, Tanomtong A. 2020. First Study on Chromosomal Feature and NORs Localization of the Yellow-spotted Keelback Snake, Fowlea flavipunctatus (Squamata, Natricinae) in Thailand. STEJ 6 (1): 37-45.
Falcione C, Hernando A, Barrasso DA, Pietro DOD. 2016. Karyotypes of four species of Xenodontini snakes (Serpentes: Dipsadidae) and implications for taxonomy. Contrib to Zool 85 (3): 265-273. DOI: 10.1163/18759866-08503001.
Falcione C, Hernando A, Bressa MJ. 2018. Comparative cytogenetic analysis in Erythrolamprus snakes (Serpentes: Dipsadidae) from Argentina. An Acad Bras Cienc 90 (2): 1417-1429. DOI: 10.1590/0001-3765201820170374.
Figueroa A. 2022. Chrysopeleinae Cope, 1893 (Squamata: Colubridae) as the valid name for the family-group currently known as Ahaetuliinae Figueroa, McKelvy, Grismer, Bell & Lailvaux, 2016 (Squamata: Colubridae). Zootaxa 5120 (4): 595-597. DOI: 10.11646/zootaxa.5120.4.9.
Figueroa A, McKelvy AD, Grismer LL, Bell CD, Lailvaux SP. 2016. A species-level phylogeny of extant snakes with description of a new colubrid subfamily and genus. Plos One. DOI: 10.1371/journal.pone.0161070.
Gamble T, Zarkower D. 2012. Sex determination. Curr. Biol. 22: 258-262. DOI: 10.1016/j.cub.2012.02.054.
Holden J, Poyarkov NA. 2021. A range extension for Dryophiops rubescens (Gray, 1835) with the first record of the species from Vietnam. Herpetology Notes 14: 795-798. DOI: 10.6084/m9.figshare.14744310.
Jiang K, Ren J, Guo J, Wang Z, Ding L, Li J. 2020. A new species of the genus Dendrelaphis (Squamata: Colubridae) from Yunnan Province, China, with discussion of the occurrence of D. cyanochloris (Wall, 1921) in China. Zootaxa 4743 (1): 001-020. DOI: 10.11646/zootaxa.4743.1.1.
Mallik AK, Achyuthan NS, Ganesh SR, Pal SP, Vijayakumar SP, Shanker K. 2019. Discovery of a deeply divergent new lineage of vine snake (Colubridae: Ahaetuliinae: Proahaetulla gen. nov.) from the southern Western Ghats of Peninsular India with a revised key for Ahaetuliinae. Plos One 14 (7): e0218851.DOI: 10.1371/journal.pone.0218851.
Matsubara K, O’Meally D, Sarre SD, Georges A, Srikulnath K, Ezaz T. 2019. ZW Sex chromosomes in Australian Dragon Lizards (Agamidae) Originated from a Combination of Duplication and Translocation in the Nucleolar Organising Region. Genes 10: 861. DOI: 10.3390/genes10110861.
Matsubara K, Tarui H, Toriba M, Yamada K, Nishida-Umehara C, Agata K, Matsuda Y. 2006. Evidence for different origin of sex chromosomes in snakes, birds, and mammals and step-wise differentiation of snake sex chromosomes. Proc Natl Acad Sci USA 103: 18190-18195. DOI: 10.1073/pnas.0605274103.
Mengden GA. 1982. Chromosomal evolution and the phylogeny of elapid snakes. [Ph. D. Thesis]. Australian National University, Canberra, pp. 55-74. DOI: 10.25911/5d6c3d988ec88.
Mengden GA, Stock AD. 1980. Chromosomal evolution in Serpentes; a comparison of G and C chromosome banding patterns of some colubrid and boid genera. Chromosoma 79: 53-64. DOI: 0009-5915/80/0079/0053/$02.40.
Mezzasalma M, Andreone F, Branch WR, Glaw F, Guarino FM, Nagy ZT, Odierna G, Aprea G. 2014. Chromosome evolution in pseudoxyrhophiine snakes from Madagascar: a wide range of karyotypic variability. Biol J Linn Soc 112: 450-460. DOI: 10.1111/BIJ.12280.
Mezzasalma M, Andreoneb F, Glawc F, Petracciolia A, Odiernaa G, Guarino FM. 2016. A karyological study of three typhlopid species with some inferenceson chromosome evolution in blindsnakes (Scolecophidia). Zoologischer Anzeiger 264: 34-40. DOI: 10.1016/j.jcz.2016.07.001.
Mohapatra PP, Dutta SK, Kar NB, Das A, Murthy BHCK, Deepak V. 2017. Ahaetulla fusca anomala (Annandale, 1906) (Squamata: Colubridae), resurrected as a valid species with marked sexual dichromatism. Zootaxa 4263: 318-332. DOI: 10.11646/zootaxa.4263.2.6.
Oguiura N, Ferrarezzi H, Batistic RF. 2009. Cytogenetics and molecular data in snakes: a phylogenetic approach. Cytogenet Genome Res 127: 128-142. DOI: 10.1159/000295789.
Paim FG, Nirchio M, Oliveira C, Rossi AR. 2020. Sex Chromosomes and Internal Telomeric Sequences in Dormitator latifrons (Richardson 1844) (Eleotridae: Eleotrinae): An Insight into Their Origin in the Genus. Genes 11: 659. DOI: 10.3390/genes11060659.
Patawang I, Tanomtong A, Getlekha N, Phimphan S, Pinthong K, Neeratanaphan L. 2017. Standardized Karyotype and Idiogram of Bengal Monitor Lizard, Varanus bengalensis (Squamata, Varanidae). Cytologia 82 (1): 75-82. DOI: 10.1508/cytologia.82.75.
Patawang I, Tanomtong A, Jumrusthanasan S, Kakampuy W, Neeratanaphan L, Pinthong K. 2014 Chromosomal characteristics of NORs and karyological analysis of tokey gecko, Gekko gecko (Gekkonidae, Squamata) from mitotic and meiotic cell division. Cytologia 79 (3): 315-324. DOI: 10.1508/cytologia.79.315.
Patawang I, Tanomtong A, Kaewmad P, Chuaynkern Y, Duengkae P. 2016. New record on karyological analysis and first study of NOR localization of parthenogenetic brahminy blind snake, Ramphotyphlops braminus (Squamata, Typhlopidae) in Thailand. Nucleus 59 (1): 61-66. DOI: 10.1007/s13237-015-0154-z.
Phimphan S, Chaiyasan P, Suwannapoom C, Reungsing M, Juntaree S, Tanomtong A, Supiwong W. 2020. Comparative karyotype study of three Cyprinids (Cyprinidae, Cyprininae) in Thailand by classical cytogenetic and FISH techniques. Comp Cytogen 14 (4): 597-612. DOI: 10.3897/CompCytogen.v14i4.54428.
Pinthong K, Tanomtong A, Getlekha N, Sangpadee W, Sangpakdee K, Sanoamuang L. 2013. First Cytogenetic Study of Puff-Faced Water Snake, Homalopsis buccata (Squamata, Colubridae) by Conventional Staining, Ag-NOR Banding and GTG-Banding Techniques. Cytologia 78 (2): 141-150. DOI: DOI: 10.1508/cytologia.78.141.
Proskuryakova AA, Kulemzina AI, Perelman PL, Serdukova NA, Ryder OA, Graphodatsky AS. 2018. The Case of X and Y Localization of Nucleolus Organizer Regions (NORs) in Tragulus javanicus (Cetartiodactyla, Mammalia). Genes 9: 312. DOI: 10.3390/genes9060312.
Pyron RA, Bunbrink FT, Wiens JJ. 2013. A phylogeny and revised classification of Squamata, including 4161 species of lizards and snakes. BMC Evol Biol 13: 93. DOI: DOI: 10.1186/1471-2148-13-93.
Pyron RA, Hendry CR, Chou VM, Lemmon EM, Lemmon AR, Burbrink FT. 2014. Effectiveness of phylogenomic data and coalescent species-tree methods for resolving difficult nodes in the phylogeny of advanced snakes (Serpentes: Caenophidia). Mol Phylogenet Evol 81: 221-231. DOI: 10.1016/j.ympev.2014.08.023.
Rao L, Turlapati R, Patel M, Panda B, Tosh D, Mangalipalli, S, Singh L. 2009. Cytogenetic characterization and fluorescence in situ hybridization of (GATA)10 repeats on established primary cell cultures from Indian water snake (Natrix piscator) and Indian mugger (Crocodylus palustris) embryos. Cytogenet Genome Res 127: 287-296. DOI: 10.1159/000304046
Rooijen JH, Vogel G. 2013. A revision of the taxonomy of Dendrelaphis caudolineatus (Gray, 1834) (Serpentes: Colubridae). Zootaxa 3272: 1-25. DOI: 10.11646/zootaxa.3271.1.
Rovatsos M, Johnson Pokorná M, Kratochvíl L. 2015. Differentiation of sex chromosomes and karyotype characterisation in the dragonsnake Xenodermus javanicus (Squamata: Xenodermatidae). Cytogenet Genome Res 147: 48-54. DOI: 10.1159/000441646.
Rovatsos M, Vuki? J, Lymberakis P, Kratochv?l L. 2015. Evolutionary stability of sex chromosomes in snakes. Proc Biol Sci 282. DOI: 10.1098/rspb.2015.1992.
Sangpakdee W, Phimphan S, Tengjaroenkul B, Pinthong K, Neeratanaphan L, Tanomtong A. 2017. Cytogenetic study of three microhylid species (Anura, Microhylidae) from Thailand. Cytologia 82: 67-74. DOI: 10.1508/cytologia.82.67.
Scacchetti PC, Utsunomia R, Pansonato-Alves JC, Vicari MR, Artoni RF, Oliveira C, Foresti F. 2015. Chromosomal Mapping of Repetitive DNAs in Characidium (Teleostei, Characiformes): Genomic Organization and Diversification of ZW Sex Chromosomes. Cytogenet Genome Res 146: 136-143. DOI: 10.1159/000437165.
Sharma GP, Nakhasi U. 1980. Karyological studies on six species of Indian snakes (Colubridae: Reptilia). Cytobios 27 (107-108): 177-92.
Silva A, Weerawansa P, Pilapitiya S, Maduwage T, Siribaddana S. 2013. First Authenticated Case of Sri Lankan Flying Snake (Chrysopelea taprobanica) Bite. Wild Environ Med 24 (3), 273-276. DOI: 10.1016/j.wem.2013.01.008.
Singchat W, O’Connor RE, Tawichasri P, Suntronpong A, Sillapaprayoon S, Suntrarachun S, Muangmai N, Baicharoen S, Peyachoknagul S, Chanhome L, Griffin D, Srikulnath K. 2018. Chromosome map of the Siamese cobra: did partial synteny of sex chromosomes in the amniote represent “a hypothetical ancestral super-sex chromosome” or random distribution?. BMC Genomics 19: 939. DOI: 10.1186/s12864-018-5293-6.
Singh L. 1974. Chromosomes of Six Species of Indian Snakes. Herpetologica 30 (4): 419-429. DOI: jstor.org/pss/3891442.
Supanuam P, Kaewsri S, Patawang I, Juntarat S, Jumrusthanasan S, Tanomtong A. 2020. First Cytogenetic Report of Painted Bronzeback (Dendrelaphis pictus, Colubrinae) by Giemsa’s Staining. KKU Sci J 48 (3): 326-332.
Supiwong W, Jiwyam, Sreeputhorn K, Maneechot N, Bertollo LAC, Cioffi MB, Getlekha N, Tanomtong A. 2017. First report on classical and molecular cytogenetics of archerfish, Toxotes chatareus (Perciformes: Toxotidae). Nucleus 60 (3): 349-359. DOI: 10.1007/s13237-017-0216-5.
Tanomtong A, Supiwong W, Jearranaiprepame P, Khakhong S, Kongpironchuen C, Getlekha N. 2014. A new natural autotetraploid and chromosomal characteristics of dwarf snakehead fish, Channa gachua (Perciformes, Channidae) in Thailand. Cytologia 79 (1): 15-27. DOI: 10.1508/cytologia.79.15.
Thongnetr W, Aiumsumang S, Kongkaew R, Tanomtong A, Suwannapoom C, Phimphan S. 2021. Cytogenetic characterisation and chromosomal mapping of microsatellite and telomeric repeats in two gecko species (Reptilia, Gekkonidae) from Thailand. Comp Cytogen 15 (1): 41-52. DOI: 10.3897/compcytogen.v15.i1.58208.
Uetz P, Ho?ek J. 2013. The Reptile Database. www.reptile-database.org/.
Uno Y, Nishida C, Tarui H, Ishishita S, Takagi C, Nishimura O, Ishijima J, Ota H, Kosaka A, Matsubara K, Murakami Y, Kuratani S, Ueno N, Agata K, Matsuda Y. 2012. Inference of the protokaryotypes of amniotes and tetrapods and the evolutionary processes of microchromosomes from comparative gene mapping. Plos One 7, e53027. DOI: 10.1371/journal.pone.0053027.
Wallach V, Williams KL, Boundy J. 2014. Snakes of the World: A Catalogue of Living and Extinct Species. CRC Press, New York.
Waters PD, Patel HR, Ruiz-Herrera A, Álvarez-González L, Lister NC, Simakov O, Ezaz T, Kaur P, Frere C, Grützner F, Georges A. 2021. Microchromosomes are building blocks of bird,reptile, and mammal chromosomes. Proc Natl Acad Sci USA 118 (45): 1-11. DOI: 10.1073/pnas.2112494118.
Zaher H, Murphy RW, Arredondo JC, Graboski R, Machado-Filho PR, Mahlow K, Montingelli GG, Quadros AB, Orlov NL, Wilkinson M, Zhang YP, Grazziotin FG. 2019. Large-scale molecular phylogeny, morphology, divergence-time estimation, and the fossil record of advanced caenophidian snakes (Squamata: Serpentes). Plos One 14 (5): e0216148. DOI: 10.1371/journal.pone.0216148.
Zheng Y, Wiens JJ. 2016. Combining phylogenomic and supermatrix approaches, and a time-calibrated phylogeny for squamate reptiles (lizards and snakes) based on 52 genes and 4162 species. Mol Phylogenet Evol 94: 537-547 DOI: 10.1016/j.ympev.2015.10.009.

Most read articles by the same author(s)