Advertisement

Genetica

, 137:245 | Cite as

Chromosomal localization of 18S and 5S rDNA using FISH in the genus Tor (Pisces, Cyprinidae)

  • Mamta Singh
  • Ravindra Kumar
  • N. S. Nagpure
  • B. Kushwaha
  • Indramani Gond
  • W. S. Lakra
Article

Abstract

Dual color fluorescence in situ hybridization (FISH) was performed to study the simultaneous chromosomal localization of 18S and 5S ribosomal genes in the genus Tor for the first time. The 18S and 5S rDNAs in four Tor species were amplified, sequenced and mapped on the metaphase chromosomes. The number and distribution of 18S and 5S rDNA clusters were examined on metaphase chromosome spreads using FISH. The specimens of T. chelynoides, T. putitora and T. progeneius showed six bright fluorescent signals of 18S rDNA and T. tor exhibited ten such signals. The 5S rDNA signals were present only on one pair of chromosomes in all the four Tor species. Ag-NORs were observed on two pairs of chromosomes in T. chelynoides, T. putitora, T. progeneius and four pairs in T. tor. Comparison of the observed 18S rDNA FISH signals and Ag-NORs strongly suggested a possible inactivation of NORs localized at the telomeres of a subtelocentric and telocentric chromosome pairs in all four species. The 5S rDNA contained an identical 120 bp long coding region and 81 bp long highly divergent non-transcribed spacers in all species examined. 18S and 5S rDNA sequencing and chromosomal localization can be a useful genetic marker in species identification as well as phylogenetic and evolutionary studies.

Keywords

18S 5S FISH rDNA Tor 

Notes

Acknowledgments

The authors are thankful to the Department of Biotechnology, the Ministry of Science and Technology, Government of India for funding.

References

  1. Almeida-Toledo LF, Ozouf-Costaz C, Foresti F, Bonillo C, Porto-Foresti F, Daniel-Silva MZF (2002) Conservation of the 5S-bearing chromosome pair and co-localization with major rDNA clusters in five species of Astyanax (Pisces, Characidae). Cytogenet Genome Res 97(3–4):229–233. doi: 10.1159/000066609 CrossRefPubMedGoogle Scholar
  2. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. doi: 10.1093/nar/25.17.3389 CrossRefPubMedGoogle Scholar
  3. Bertollo LAC, Takahashi CS, Moreira-Filho O (1978) Cytotaxonomic consideration on Hoplias lacrdae (Pisces, Erythrinidae). Braz J Genet 1:103–120Google Scholar
  4. Cabrero J, Camacho JP (2008) Location and expression of ribosomal RNA genes in grasshoppers: abundance of silent and cryptic loci. Chromosom Res 16(4):595–607. doi: 10.1007/s10577-008-1214-x CrossRefGoogle Scholar
  5. Ferro DAM, Neo DM, Moreira-Filho O, Bertollo LAC (2001) Nucleolar organizing regions, 18S and 5S rDNA in Astyanax scabripinnis (Pisces, Characidae): population distribution and functional diversity. Genetica 110:55–62. doi: 10.1023/A:1017963217795 CrossRefGoogle Scholar
  6. Fontana F, Lanfredi M, Congiu L, Leis M, Chicca M, Rossi R (2003) Chromosomal mapping of 18S–28S and 5S rRNA genes by two-colour fluorescent in situ hybridization in six sturgeon species. Genome 46:473–477. doi: 10.1139/g03-007 CrossRefPubMedGoogle Scholar
  7. Fujiwara A, Abe S, Yamaha E, Yamazaki F, Yoshida MC (1998) Chromosomal localization and heterochromatin association of ribosomal RNA genes loci and silver stained nucleolar organizer regions in salmonid fishes. Chromosom Res 6:463–471. doi: 10.1023/A:1009200428369 CrossRefGoogle Scholar
  8. Gromicho M, Coutanceau JP, Ozouf-Costaz C, Collares-Pereira MJ (2006) Contrast between extensive variation of 28S rDNA and stability of 5S rDNA and telomeric repeats in the diploid-polyploid Squalius alburnoides complex and in its maternal ancestor Squalius pyrenaicus (Teleostei, Cyprinidae). Chromosom Res 14:297–306. doi: 10.1007/s10577-006-1047-4 CrossRefGoogle Scholar
  9. Guillén AK, Hirai Y, Tanoue T, Hirai H (2004) Transcriptional repression mechanisms of nucleolus organizer regions (NORs) in humans and chimpanzees. Chromosom Res 12(3):225–237. doi: 10.1023/B:CHRO.0000021911.43225.eb CrossRefGoogle Scholar
  10. Hallenberg C, Nederby-Nielson J, Frederiksen S (1994) Characterization of 5S rRNA genes from mouse. Gene Amst 142:291–295. doi: 10.1016/0378-1119(94)90277-1 CrossRefGoogle Scholar
  11. Howell WM, Black DA (1980) Controlled silver staining of nucleolus organizer regions with a protective colloidal developer: a one step method. Experementia 36:1014–1015. doi: 10.1007/BF01953855 CrossRefGoogle Scholar
  12. Inafuku J, Nabeyama M, Kikuma Y, Saitoh J, Kubota S, Kohno S (2000) Chromosomal location and nucleotide sequences of 5S ribosomal DNA of two cyprinid species (Osteichthyes, Pisces). Chromosom Res 8:193–199. doi: 10.1023/A:1009292610618 CrossRefGoogle Scholar
  13. Khuda-Bukhsh AR, Nayak K (1982) Karyomorpological studies in two species of hill stream fishes from Kashmir, India: occurrence of a high number of chromosomes. Chromosom inf Sci (Japan) 33:12–14Google Scholar
  14. Kushwaha B, Srivastava SK, Nagpure NS, Ogale SN, Ponniah AG (2001) Cytogenetic studies in two species of mahseer, Tor khudree and Tor mussullah (Cyprinidae, Pisces) from India. Chromosom Sci 5:47–50Google Scholar
  15. Lakra WS (1996) Cytogenetic studies on endangered fish species: karyotypes of three species of mahseers, Tor putitora, T. tor and T. khudree (Cyprinidae Pisces). Cytobios 85:205–218Google Scholar
  16. Long EO, David ID (1980) Repeated genes in eukaryotes. Annu Rev Biochem 49:727–764. doi: 10.1146/annurev.bi.49.070180.003455 CrossRefPubMedGoogle Scholar
  17. Lucchini S, Nardi I, Barsacchi G, Batistoni R, Andronico F (1993) Molecular cytogenetics of the ribosomal (18S + 28S and 5S) DNA loci in primitive and advanced urodele amphibians. Genome 36:762–773. doi: 10.1139/g93-101 CrossRefPubMedGoogle Scholar
  18. Mantovani M, Abel LDS, Moreira-Filho O (2005) Conserved 5S and variable 45S rDNA chromosomal localization revealed by FISH in Astyanax scabripinnis (Pisces, Characidae). Genetica 123:211–216. doi: 10.1007/s10709-004-2281-3 CrossRefPubMedGoogle Scholar
  19. Martinez JL, Moran P, Garcia-Vazquez S, Pendas AM (1996) Chromosomal localization of major and 5S rRNA genes in the European eel (Anguilla anguilla). Cytogenet Cell Genet 73:149–152CrossRefPubMedGoogle Scholar
  20. Martins C, Galetti PM Jr (1999) Chromosomal localization of 5S rRNA genes in Leporinus fish (Anostomidae, Characiformes). Chromosom Res 7:363–367. doi: 10.1023/A:1009216030316 CrossRefGoogle Scholar
  21. Martins C, Galetti PM (2000) Conservative distribution of 5S rDNA loci in Schizodon (Pisces, Anastomidae) chromosomes. Chromosom Res 8:353–355. doi: 10.1023/A:1009243815280 CrossRefGoogle Scholar
  22. Martins C, Galetti PM Jr (2001) Two 5S rDNA array in neotropical fish species: is it a general rule for fishes? Genetica 111:439–446. doi: 10.1023/A:1013799516717 CrossRefPubMedGoogle Scholar
  23. Martins C, Wasko AP (2004) Organization and evolution of 5S ribosomal DNA in the fish genome. In: Williams CR (ed) Focus on Genome Research. Nova Science Publishers, Hauppauge, pp 289–319Google Scholar
  24. Moran P, Martinez JL, Garcia-Vazquez S, Pendas AM (1996) Sex linkage of 5S rDNA in rainbow trout (Oncorhynchus mykiss). Cytogenet Cell Genet 75:145–150. doi: 10.1159/000134466 CrossRefPubMedGoogle Scholar
  25. Nederby-Nielson J, Hallenberg C, Frederiksen S, Sorensen PD, Lomholt B (1993) Transcription of human 5S rRNA genes is influenced by an upstream DNA sequence. Nucleic Acids Res 26:3631–3636Google Scholar
  26. Pendas AM, Moran P, Garcia-Vazquez E (1993) Multi chromosomal location of ribosomal RNA genes and heterochromatin association in brown trout. Chromosom Res 1:63–67. doi: 10.1007/BF00710608 CrossRefGoogle Scholar
  27. Pendas AM, Moran P, Freije JP, Garcia-Vazquez S (1994) Chromosomal mapping and nucleotide sequences of two tandem repeats of Atlantic salmon 5S rDNA. Cytogenet Cell Genet 67:31–36. doi: 10.1159/000133792 CrossRefPubMedGoogle Scholar
  28. Sajdak SL, Reed KM, Phillips RB (1998) Intra individual and interspecies variation in the 5S rDNA of coregonid fish. J Mol Evol 46:680–688. doi: 10.1007/PL00006348 CrossRefPubMedGoogle Scholar
  29. Sambrook J, Russell I (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Plainsveiw New YorkGoogle Scholar
  30. Santos LVDR, Foresti F, Wasko AP, Oliveira C, Martins C (2006) Nucleotide sequence, genomic organization and chromosome localization of 5S rDNA in two species of Curimatidae (Teleostei, Characiformes). Genet Mol Biol 29(2):251–256. doi: 10.1590/S1415-47572006000200009 CrossRefGoogle Scholar
  31. Sarkar UK, Srivastava SM (2000) Endangered Mahseer in North East India. In: Ponniah AG, Sarkar UK (eds) Fish Biodiversity of North East India, NBFGR, NATP publ, Lucknow, India, pp 95–96Google Scholar
  32. Sola L, Rossi AR, Iaselli V, Rasch EM, Monaco PJ (1992) Cytogenetics of bisexual/unisexual species of Poecilia mexicana by C-banding and DAPI, quinacrine, chromomycin A3, and silver staining. Cytogenet Cell Genet 60:119–235CrossRefGoogle Scholar
  33. Suzuki H, Sakurai S, Matsuda Y (1996) Rat 5S rDNA spacer sequences and chromosomal assignment of the genes to the extreme terminal region of chromosome 19. Cytogenet Cell Genet 72:1–4. doi: 10.1159/000134149 CrossRefPubMedGoogle Scholar
  34. Tigano C, Rocco L, Ferrito V, Costagliola D, Pappalardo AM, Stinso V (2004) Chromosomal mapping and molecular characterization of ribosomal RNA genes in Lebias fasciata (Teleostei, Cyprinodontidae). Genetica 121:95–100. doi: 10.1023/B:GENE.0000019931.89458.dc CrossRefPubMedGoogle Scholar
  35. Wasko AP, Martins C, Wright JM, Galetti PM (2001) Molecular organization of 5S rDNA in fishes of genus Brycon. Genome 44:893–902. doi: 10.1139/gen-44-5-893 CrossRefPubMedGoogle Scholar
  36. Winterfeld G, Roser M (2007) Deposition of ribosomal DNAs in the chromosome of perennial oats (Poaceae: Aveneae). Bot J Linn Soc 155:193–210. doi: 10.1111/j.1095-8339.2007.00690.x CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Mamta Singh
    • 1
  • Ravindra Kumar
    • 1
  • N. S. Nagpure
    • 1
  • B. Kushwaha
    • 1
  • Indramani Gond
    • 1
  • W. S. Lakra
    • 1
  1. 1.National Bureau of Fish Genetic ResourcesLucknowIndia

Personalised recommendations