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Characterization of Chromosomal and Repetitive Elements in the Genome of “Rana nigrovittata” (Anura, Ranidae): Revealed by Classical and Molecular Techniques

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Abstract

Karyotype study and microsatellites pattern in the genome of Rana nigrovittata were studied, with the aim to provide a standard karyotype, chromosome marker and the distribution of repetitive DNA elements, informative knowledge of cytogenetics and evolutionary events. Here, we analyzed the karyotype structure and the distribution of repetitive DNA sequence in this species using conventional banding and Fluorescence in situ hybridization techniques. The ten specimens (five males and five females) were collected from Phitsanulok province, Thailand. Mitotic metaphases were prepared from the bone marrows by the standard protocol. The result showed that R. nigrovittata had the diploid chromosome number of 2n = 26 and the fundamental number (NF) were 52 in both males and females. The karyotypes compose of six large metacentric, four large submetacentric, two medium metacentric, two medium submetacentric and 12 small submetacentric chromosomes. No sex related chromosome heteromorphism was observed in male (XY) or female (ZW) of this species. The NOR was observed in subcentromeric region on chromosome no. 11. The C-positive heterochromatin blocks are mainly distributed in the centromere of most chromosomes, while some additionally in paracentromeric and telomeric regions. The large heterochromatic blocks were found on chromosome no 6. Some of repetitive elements were scattered while some were specific in the karyotype. The combine of conventional banding and molecular cytogenetics provide information for a cytogenetic determination of the examined species.

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REFERENCES

  1. Beccari, N., Le Nombre des chromosomes dans les cellules genetales de Bufo viridis, CR. Assoc. Anat., 1926, vol. 21, pp. 29–31.

    Google Scholar 

  2. Birstein, V.J., Localization of NORs in karyotypes of four Rana species, Genetica, 1984, vol. 64, pp. 149–154. https://doi.org/10.1007/BF00115338

    Article  Google Scholar 

  3. Blommers-Schlösser, R.M.A., Cytotaxonomy of the Ranidae, Rhacophoridae, Hyperoliidae (Anura) from Madagascar with a note on the karyotype of two amphibians of the Seychelles, Genetica, 1978, vol. 48, pp. 23–40. https://doi.org/10.1007/BF00125283

    Article  Google Scholar 

  4. Chan-Ard, T., Grossmann, W., Gumprecht, A., et al., Amphibians and Reptiles of Peninsular Malaysia and Thailand (An Illustrated Checklist), Wuerselen, Germany: Bushmaster Publications, 1999.

    Google Scholar 

  5. Chaiyasut, K., Cytogenetics and Cytotaxonomy of the Family Zephyranthes, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 1989.

    Google Scholar 

  6. Cioffi, M.B.P. and Bertollo, L.A.C., Chromosomal distribution and evolution of repetitive DNAs in fish, in Repetitive DNAs Genome Dynamics, Garrido, R., Ed., Basel: Karger, 2012, vol. 7, pp. 197–221. https://doi.org/10.1159/000337950

  7. Frost, D.R., Ed., Amphibian Species of the World: A Taxonomic and Geographical Reference, Lawrence, Kansas: Allen Press, 1985.

    Google Scholar 

  8. Guillemin, C., Karyotypes de Rana temporaria (L.) et de Rana dalmatina (Bonaparte), Chromosoma, 1967, vol. 21, pp. 189–197. https://doi.org/10.1007/BF00343644

    Article  CAS  PubMed  Google Scholar 

  9. Heppich, S., Hybridogenesis in Rana esculenta: C-band karyotypes of Rana ridibunda, Rana lessonae and Rana esculenta, Z. Zool. Syst. Evol. Forsch., 1978, vol. 16, pp. 27–39. https://doi.org/10.1111/j.1439-0469.1978.tb00918.x

    Article  Google Scholar 

  10. Heppich, S. and Tunner, H.G., Chromosomal constitution and C-banding in homotypic Rana esculenta crosses, Mitt. Zool. Mus. Berlin., 1979, vol. 55, pp. 111–114. https://doi.org/10.1007/BF00273874

    Article  Google Scholar 

  11. Hills, D.M. and Green, D.M., Evolutionary changes of heterogametic sex in the phylogenetics history of amphibians, J. Evol. Biol., 1990, vol. 3, pp. 49–64. https://doi.org/10.1046/j.1420-9101.1990.3010049.x

    Article  Google Scholar 

  12. Howell, W.M. and Black, D.A., Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method, Experientia, 1980, vol. 36, pp. 1014–1015. https://doi.org/10.1007/BF01953855

    Article  CAS  PubMed  Google Scholar 

  13. Iizuka, K., Constitutive heterochromatin and nucleolus organizer regions in Japanese brown frogs, Rana japonica and Rana ornativentris, Jpn. J. Herpetol., 1989, vol. 13, pp. 15–20.

    Article  Google Scholar 

  14. Joshy, H.S.S., Kuramoto, M., Sreepada, K.S., et al., Karyotypic Variations in Three Indian Species of the Genus Rana (Anura: Ranidae) from the Western Ghats, India, Cytologia, 2006, vol. 71, no. 1, pp. 63–68. https://doi.org/10.1508/cytologia.71.63

    Article  Google Scholar 

  15. Khonsue, W., and Thirakhupt, K., A checklist of the amphibians in Thailand, Nat. Hist. J. Chulalongkorn Univ., 2001, vol. 1, pp. 69–82.

    Google Scholar 

  16. Kuramoto, M., Karyotypes of the six species of frogs native to the Ryukyu islands, Caryologia, 1972, vol. 25, no. 4, pp. 547–559. https://doi.org/10.1080/00087114.1972.10796509

    Article  Google Scholar 

  17. Kuramoto, M., Karyotypes of several frogs from Korea, Taiwan and the Philippines, Experientia, 1979, vol. 39, pp. 826–828. https://doi.org/10.1007/BF01978594

    Article  Google Scholar 

  18. Kuramoto, M., Karyotypes of several frogs from Korea, Taiwan and the Philippines, Experientia, 1980, vol. 36, pp. 826–828. https://doi.org/10.1007/BF01978594

    Article  CAS  PubMed  Google Scholar 

  19. Kuramoto, M., Karyological studies on some Philippine frogs, in Current Herpetology in East Asia, Matsui, M., Hidika, T., and Goris, R.C., Eds., Kyoto: Herpetology Society of Japan, 1989, pp. 115–121.

    Google Scholar 

  20. Kuramoto, M., A list of chromosome numbers of anuran amphibians, Bull. Fukuoka Univ. Educ., 1990, vol. 39, pp. 83–127.

    Google Scholar 

  21. Kuramoto, M. and Yong, H.-S., Karyotypes from several frog species from peninsular Malaya, Herpetologica, 1992, vol. 28, no. 4, pp. 434–438.

    Google Scholar 

  22. Liu, W. and Zan, R., A special karyotype in the genus Rana—an investigation of the karyotype, C-banding and Ag-stained NORs of Rana phrynodes Boulenger, Acta Genet. Sin., 1984, vol. 11, pp. 52–60. https://doi.org/10.3897/CompCytogen.v8i4.7623

    Article  Google Scholar 

  23. Matsui, M., Nishikawa, K., Khonsue, W., et al., Allozymic variation in Rana nigrovittata (Amphibia: Anura) within Thailand with special reference to the taxonomic status of R. mortenseni, Nat. Hist. J. Chulalong. Univ., 2001, vol. 1, no. 1, pp. 15–22.

    Google Scholar 

  24. Matsui, M., Hidetoshi, O., Michael, W.L., et al., Cytotaxonomic Studies of Three Ranid Species (Amphibia: Anura) from Hong Kong, Jpn. J. Herpetol., 1995, vol. 16, no. 1, pp. 12–18.https://doi.org/10.5358/hsj1972.16.1_12

  25. Miura, I., Sex chromosome differentiation in the Japanese brown frog, Rana japonica. Sex-related heteromorphism of the distribution pattern of constitutive heterochromatin in chromosome no. 4 of the Wakuya population, Zool. Sci., 1994, vol. 11, pp. 797–806.

    Google Scholar 

  26. Odierna, G., Vences, M., Aprea, G., et al., Chromosome data for Malagasy poison frogs (Amphibia: Ranidae: Mantella) and their bearing on the taxonomy and phylogeny, Zool. Sci., 2001, vol. 18, no. 4, pp. 505–514. https://doi.org/10.2108/zsj.18.505

    Article  Google Scholar 

  27. Pinkel, D., Straume, T., and Gray, J., Cytogenetic analysis using quantitative, high sensitivity, fluorescence hybridization, Proc. Natl. Acad. Sci. U. S. A., 1986, vol. 83, no. 9, pp. 2934–2938. https://doi.org/10.1073/pnas.83.9.2934

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Popov, P. and Dimitrov, B., Karyotype study of Rana camerani and comparisons with the other 26-chromosome European brown frog species (Amphibia, Anura), Cytobios., 1999, vol. 97, pp. 13–22.

    CAS  PubMed  Google Scholar 

  29. Sangpakdee, W., Phimphan, S., Tengjaroenkul, B., et al, Cytogenetic study of three microhylid species (Anura, Microhylidae) from Thailand, Cytologia, 2017, vol. 82, no. 1, spec. iss., pp. 67–74. https://doi.org/10.1508/cytologia.82.67

  30. Seto, T., Cytogenetic studies in lower vertebrates II. Karyological studies of several species of frogs (Ranidae), Cytologia, 1965, vol. 30, pp. 437–446. https://doi.org/10.1508/cytologia.30.437

    Article  Google Scholar 

  31. Schmid, M., Chromosome banding in Amphibia II. Constitutive heterochromatin and nucleolus organizer regions in Ranidae, Microhylidae and Rhacophoridae, Chromosoma, 1978, vol. 68, pp. 131–148. https://doi.org/10.1007/BF00287145

    Article  Google Scholar 

  32. Schmid, M., Chromosome banding in Amphibia IV. Differentiation GC- and AT-rich chromosome regions in Anura, Chromosoma, 1980, vol. 77, no. 1, pp. 215–234. https://doi.org/10.1007/BF00292043

    Article  Google Scholar 

  33. Schmid, M., Steinlein, C., Freiedl, R., et al., Chromosome banding in Amphibia. XV. Two types of Y chromosome heterochromatin hypervariability in Gastrotheca pseustes (Anura, Hylidae), Chromosoma, 1990, vol. 99, pp. 413–423.

    Article  Google Scholar 

  34. Shi, H.L., Zhang, C., Wu, M., et al., A study on the karyotype, C-banding and Ag-NORs in Rana nigromaculata, Hereditas, 2006, vol. 28, pp. 533–539.

    PubMed  Google Scholar 

  35. Singh, A.K. and Banerjee, R., Chromosomal diversity of Indian mammals, amphibians and reptiles, Rec. Zool. Surv. India, 2004, vol. 102 (parts 3–4), pp. 127–138.

  36. Spasić-Bošković, O., Tanić, N., Blagojević, J., et al., Comparative cytogenetic analysis of European brown frogs: Rana temporaria, R. dalmatina and R. graeca, Cytologia, 1997, vol. 50, no. 2, pp. 139–149. https://doi.org/10.1080/00087114.1997.10797393

    Article  Google Scholar 

  37. Stohler, R., Cytologische Untersuchungen an den Keimdrusen der mittel europaischen Kroten (Bufo viridis Laur., B. calamita Laur. and B. vulgaris Laur.), Z. Zellforsch., 1927, vol, 7, pp. 400–475. https://doi.org/10.1007/BF00372490

    Article  Google Scholar 

  38. Supaprom, T., Cytogenetics of Amphibians in Thailand, Ph.D. Dissertation, Mahidol University, 2003.

  39. Supaprom, T., Chantree, P., and Palasarn, W., Cytogenetics and cytotaxonomy of localized Amphibians in Northeastern Thailand, Research Project, Ubonratchathani University, 1999.

  40. Supaprom, T., Baimai, V., Karyotypes of ten species of Ranid frogs (Anura: Ranidae) from Thailand, Amphibia–Reptilia, 2004, vol. 25, no. 1, pp. 104–111.

    Google Scholar 

  41. Sumner, A.T., A simple technique for demonstrating centromeric heterochromatin, Exp. Cell. Res., 1972, vol. 75, no. 1, pp. 304–306. https://doi.org/10.1016/0014-4827(72)90558-7

    Article  CAS  PubMed  Google Scholar 

  42. Tautz, D. and Renz, M., Simple sequences are ubiquitous repetitive components of eukaryotic genomes, Nucleic Acids Res., 1984, vol. 12, no. 10, pp. 4127–4138. https://doi.org/10.1093/nar/12.10.4127

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This work was supported by Phetchabun Rajabhat University and Toxic Substances in Livestock and Aquatic Animals Research Group, Department of Biology, Faculty of Science, Khon Kaen University, Thailand.

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Authors and Affiliations

  1. Biology program, Faculty of Science and Technology, Phetchabun Rajabhat University, 67000, Phetchabun, Thailand

    Sumalee Phimphan & Surachest Aiumsumang

  2. Department of Biology, Faculty of Science, Khon Kaen University, 40002, Muang, Khon Kaen, Thailand

    Alongklod Tanomtong

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  1. Sumalee Phimphan
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Correspondence to Sumalee Phimphan.

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Phimphan, S., Aiumsumang, S. & Tanomtong, A. Characterization of Chromosomal and Repetitive Elements in the Genome of “Rana nigrovittata” (Anura, Ranidae): Revealed by Classical and Molecular Techniques. Cytol. Genet. 55, 583–589 (2021). https://doi.org/10.3103/S0095452721060104

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