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Extensive polymorphism and chromosomal characteristics of ribosomal DNA in a loach fish, Cobitis vardarensis (Ostariophysi, Cobitidae) detected by different banding techniques and fluorescence in situ hybridization (FISH)

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Abstract

When surveying the karyotype diversity of European loaches of the genus Cobitis to identify species involved in hybrid polyploid complexes, an extensive polymorphism in number and location of NORs was discovered in C. vardarensis using Ag-staining, C-banding, CMA3-fluorescence and fluorescence in situ hybridization (FISH). This species had 2n = 50, the karyotype contained 13 pairs of metacentric, 10 pairs of submetacentric and two pairs of subtelocentric chromosomes. The NOR-bearing chromosomes included one medium-sized metacentric pair with a large CMA3-positive heterochromatic pericentromeric block, one small metacentric as well as one large submetacentric pairs. Ribosomal sites were always located in telomeres of these chromosomes. Each of the pair of NOR-bearing chromosomes occurred in three variants – (1) presence and/or (2) absence of NORs on both homologues and (3) heterozygous combination where only one of the homologues bears NORs. Altogether, 10 different NOR cytotypes from 27 theoretically possible ones were discovered among 20 indviduals examined. The number of NORs ranged from two to five per specimen. The results regarding the number and locations of NORs as revealed by banding techniques were confirmed using FISH with rDNA probe. NOR sites were of CMA3-positive, suggesting that ribosomal sites are associated with GC-rich DNA. Very similar structural polymorphism with multiple NORs is expressed in the Danubian loach C. elongatoides indicating a close relationship between both species.

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References

  • Boroń, A., 1999. Banded karyotype of spined loach Cobitis taenia and triploid Cobitis from Poland. Genetica 105: 293–300.

    Google Scholar 

  • Boroń, A. & J. Kotusz, 1999. A preliminary report and karyotype of a new Cobitis species in the ichthyofauna of Poland. Cytobios 98: 59–64.

    Google Scholar 

  • Collares-Pereira, M.J. & P. Ráb, 1999. NOR polymorphism in the Iberian species Chondrostoma lusitanicum (Pisces:Cyprinidae) - re-examination by FISH. Genetica 105: 301–303.

    Google Scholar 

  • De Lucchini, S., I. Nardi, G. Barsacchi, R. Bastistoni & F. Andronico, 1993. Molecular cytogenetics of the ribosomal (18S+28S and 5S) DNA loci in primitive and advanced urodele amphibians. Genome 36: 762–773.

    Google Scholar 

  • Economidis, P.S. & T.T. Nalbant, 1996. A study of the loaches of the genera Cobitis and Sabanejewia (Pisces, Cobitidae) of Greece, with description of six new taxa. Trav. Mus. Natl. Hist. Nat. ‘Grigore Antipa' 36: 295–347.

    Google Scholar 

  • Erkanan, F., F.G. Atalay-Ekmekçi & T.T. Nalbant, 1998. Four new species and one new subspecies of the genus Cobitis (Pisces: Ostariophysi: Cobitidae) from Turkey. Tr. J. Zool. 22: 9–15.

    Google Scholar 

  • Erkanan, F., F.G. Atalay-Ekmekçi & T.T. Nalbant, 1999. A review of the genus Cobitis in Turkey (Pisces: Ostariophysi: Cobitidae). Hydrobiologia 403: 13–26.

    Google Scholar 

  • Fujiwara A., S. Abe, E. Yamaha, F. Yamazaki & M.C. Yoshida, 1998. Chromosomal localization and heterochromatin association of ribosomal regions in salmonid fishes. Chrom. Res. 6: 463–471.

    Google Scholar 

  • Galetti P.M. & E.M. Rasch, 1993. Chromosome studies in Poecilia latipunctata with NOR polymorphism as shown by silver nitrate and chromomycin A3 (Teleostei: Poeciliidae). Ichthyol. Explor. Freshwaters 4: 269–277.

    Google Scholar 

  • Galetti P.M., F. Foresti, L.A.C. Bertollo & O. Moreira-Filho,1984. Characterization of eight species of Anostomidae (Cyprini formes) fish on the basis of the nucleolar organizing region. Caryologia 37: 401–406.

    Google Scholar 

  • Gollmann G, Y. Bouvet, Y. Karakousis & C. Triantaphyllidis, 1997. Genetic variability in Chondrostoma from Austrian, French and Greek rivers (Teleostei, Cyprinidae). J. Zool. Syst. Evol. Res. 35: 165–169.

    Google Scholar 

  • Haaf, T. & M. Schmid, 1984. An early stage of ZW/ZZ sex chromosome differentiation in Poecilia sphenops var. melanistica (Poeciliidae, Cyprinodontiformes). Chromosoma 89: 37–41.

    Google Scholar 

  • Howell, W.M. & D.A. Black, 1980. Controlled silver staining of nucleolus organizer regions with a protective colloidal developer: a 1–step method. Experientia 36: 1014–1015.

    Google Scholar 

  • Jankun, M., P. Martínez, B.G. Padro, P. Ráb, M. Rábová & L. Sanchez, 2000. rRNA genes map to chromosomes 10, 11 and 12 in European whitefish (Coregonus lavaretus) and to chromosomes 1, 5, 9 and 10 in vendace (Coregonus albula). Chrom. Res. 8: 455.

    Google Scholar 

  • Levan, A., K. Fredga & A.A. Sandberg, 1964. Nomenclature for centromeric position on chromosomes. Hereditas 52: 201–220.

    Google Scholar 

  • Long, E.O. & I.D. David, 1980. Repeated genes in eucaryotes. Ann. Rev. Biochem. 49: 727–764.

    Google Scholar 

  • Martínez P., A. Vinas, C. Bouza, J. Arias, R. Amaro & L. Sanchez, 1991. Cytogenetical characterization of hatchery stocks and natural populations of sea and brown trout from northwestern Spain. Heredity 66: 9–17.

    Google Scholar 

  • Mellink C.H.M., A.A. Bosma, N.A. De Haan & A.A. Macdonald, 1992. Numerical variation of nucleolar organizer regions after silver staining in domestic and wild Suidae (Mammalia). Anim. Genet. 23: 231–239.

    Google Scholar 

  • Moss, T. & V.Y. Stefanovsky, 1995. Promotion and regulation of ribosomal transcription by RNA polymerase I. Prog. Nucl. Acid Res. 50: 207–250.

    Google Scholar 

  • Padilla, J.A., J.L. Fernández-García, A. Rabasco, M. Martinez-Trancon, I. Rodriguez de Ledesma & J.J. Perez-Regardera, 1993. Characterization of the karyotype of the tench (Tinca tinca L.) and analysis of its chromosomal heterochromatic regions by C-banding, Ag-staining, and restriction endonuclease banding. Cytogenet. Cell Genet. 62: 220–223.

    Google Scholar 

  • Pendás, A.M., P. Morán & E. García-Vazquez, 1993. Multichromosomal location of ribosomal RNA genes and heterochromatin associations in brown trout. Chrom. Res. 1: 63–67.

    Google Scholar 

  • Phillips, R.B., K.D. Zajicek & F.M. Utter, 1986. Chromosome banding in salmonid fishes: nucleolar organizer regions in Oncorhynchus. Can. J. Genet. Cytol. 28: 502–510.

    Google Scholar 

  • Ozouf-Costaz, C., E. Pisano, C. Bonillo & R. Williams, R. 1996: Ribosomal RNA location in the Antarctic fish Champsocephalus gunnari (Notothenioidei, Channichthyidae) using banding and fluorescence in situ hybridization. Chrom. Res. 4: 557–561.

    Google Scholar 

  • Ráb, P. & P. Roth, 1988. Cold-blooded vertebrates, pp. 115–124 in Methods of Chromosome Analysis, edited by P. Balíček, J.Forejt, J. & J. Rubeš., Czechoslovak Biol. Soc. Publishers, Brno.

    Google Scholar 

  • Ráb, P. & O. Slavík, 1996. Diploid-triploid-tetraploid complex of the spined loach, genus Cobitis in Pšovka Creek: the first evidence of new species of Cobitis in the ichthyofauna of the Czech Republic. Acta Univ. Carol - Biol. 39: 201–214.

    Google Scholar 

  • Ráb, P., K.M. Reed, F.A. Ponce de Leon & R.B. Phillips, 1996. A new method for detection nucleolus organizer regions in fish chromosomes using denaturation and propidium iodide staining. Biotech. Histochem. 71: 157–162.

    Google Scholar 

  • Ráb, P., M. Rábová, K.M. Reed & R.B. Phillips, 1999. Chromosomal characteristics of ribosomal DNA in the primitive semionotiform fish, longnose gar Lepisosteus osseus. Chrom. Res. 7: 475–480.

    Google Scholar 

  • Ráb, P., M. Rábová, J. Bohlen & S. Lusk, 2000a. Genetic differentiation of the two hybrid diploid-polyploid complexes of loaches, genus Cobitis (Cobitidae) involving C. taenia, C. elongatoides and C. sp. in the Czech Republic: karyotypes and cytogenetic diversity. Folia Zool. 49(Suppl. 1): 55–66.

    Google Scholar 

  • Ráb, P., M. Rábová, P.S. Economidis & C. Triantaphyllidis, 2000b. Banded karyotype of Greek endemic cyprinid fish Pachychilon macedonicum. Ichthyol. Res. 47: 107–110.

    Google Scholar 

  • Reed, K.M. & R.B. Phillips, 1995. Molecular cytogenetic analysis of the double-CMA3 chromosome of lake trout, Salvelinus namaycush. Cytogenet. Cell Genet. 70: 104–107.

    Google Scholar 

  • Rodrigues, E. & M.J. Collares-Pererira, 1996. NOR polymorphism in the Iberian species Chondrostoma lusitanicum (Pisces:Cyprinidae). Genetica 98: 59–63.

    Google Scholar 

  • Schmid M., W. Feichtinger, R. Weimer, C. Mais, F. Bolaños & P. León, 1995. Chromosome banding in Amphibia. XXI. Inversion polymorphism and multiple nucleolus organizer regions in Agalychnis callidryas (Anura, Hylidae). Cytogenet. Cell Genet. 69: 18–26.

    Google Scholar 

  • Sola L., A.R. Rossi, V. Iaselli, E.M. Rasch & P.J. Monaco, 1992. Cytogenetics of bisexual/unisexual species of Poecilia. II. Analysis of heterochromatin and nucleolar organizer regions in Poecilia mexicana maxicana by C-banding and DAPI, quinacrine, chromomycin A3, and silver staining. Cytogenet. Cell Genet. 60: 229–235.

    Google Scholar 

  • Souza I.L. & O. Moreira-Filho, 1995. Cytogenetic diversity in the Astyanax scabripinnis species complex (Pisces, Characidae). I. Allopatric distribution in a small stream. Cytologia 60: 1–11.

    Google Scholar 

  • Takai, A. & Y. Ojima, 1986. Some features on nucleolus organizer regions on fish chromosomes, pp: 899–909 in Indo-Pacific Fish Biology: Proc. 2nd Int. Conf. Indo-Pacific Fishes, edited by T. Uyeno, R. Arai, T. Taniuchi, T. & K. Matsura. Ichthyological Society of Japan, Tokyo.

    Google Scholar 

  • Volleth, M., 1987. Differences in location of nucleolus organizer regions in European vespertilinid bats. Cytogenet. Cell Genet. 44: 186–197.

    Google Scholar 

  • Vasil'ev, V.P., 1995. Karyotype diversity and taxonomic nonhomogeneity of Cobitis ‘taenia’ (Pisces, Cobitidae). Dokl. Akad. Nauk. 342: 839–842.

    Google Scholar 

  • Zhuo L., K.M. Reed & R.B. Phillips, 1995. Hypervariability of ribosomal DNA at multiple chromosomal sites in lake trout (Salvelinus namaycush). Genome 38: 487–496.

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Fish Genetics, Institute of Animal Physiology and Genetics, Czech Academy of Sciences, 277 21, Liběchov, Czech Republic

    M. Rábová

  2. Laboratory of Fish Genetics, Institute of Animal Physiology and Genetics, Czech Academy of Sciences, 277 21, Liběchov, Czech Republic

    P. Ráb

  3. Service commun de Systématique moléculaire (CNRS FR 1541) & Laboratoire d’Ichtyologie generale et appliqueé, Museum national d’histoire naturelle, 43 rue Cuvier, 75231, Paris Cedex 05, France

    C. Ozouf-Costaz

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  1. M. Rábová
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  2. P. Ráb
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  3. C. Ozouf-Costaz
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Rábová, M., Ráb, P. & Ozouf-Costaz, C. Extensive polymorphism and chromosomal characteristics of ribosomal DNA in a loach fish, Cobitis vardarensis (Ostariophysi, Cobitidae) detected by different banding techniques and fluorescence in situ hybridization (FISH). Genetica 111, 413–422 (2001). https://doi.org/10.1023/A:1013763903513

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