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Marine Biology

, Volume 150, Issue 6, pp 1183–1189 | Cite as

Chromosomal mapping of major ribosomal rRNA genes in the hard clam (Mercenaria mercenaria) using fluorescence in situ hybridization

  • Yongping Wang
  • Ximing GuoEmail author
Research Article

Abstract

Karyotype and chromosomal location of the major ribosomal RNA genes were studied in the hard clam (Mercenaria mercenaria Linnaeus) using fluorescence in situ hybridization (FISH). Metaphase chromosomes were obtained from early embryos. Internal transcribed spacers (ITS) between major RNA genes were amplified and used as FISH probes. The probes were labeled with digoxigenin-11-dUTP by polymerase chain reaction and detected with fluorescein-labeled anti-digoxigenin antibodies. FISH with the ITS probes produced two to four signals per nucleus or metaphase. M. mercenaria had a haploid number of 19 chromosomes with a karyotype of seven metacentric, four metacentric or submetacentric, seven submetacentric, and one submetacentric or subtelocentric chromosomes (7M + 4M/SM + 7SM + 1SM/ST). Two ITS loci were observed: one located near the centromere on the long arm of Chromosome 10 and the other at the telomere of the short arm of Chromosome 12. FISH signals on Chromosome 10 are strong and consistent, while signals on Chromosome 12 are variable. This study provides the first karyotype and chromosomal assignment of the major RNA genes in M. mercenaria. Similar studies in a wide range of species are needed to understand the role of chromosomal changes in bivalve evolution.

Keywords

Internal Transcribe Spacer Bivalve Hard Clam Haploid Number Mercenaria Mercenaria 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This study was supported by grants from the U.S. Sea Grant (B/T-9801; R/OD-2003-1), U.S. Department of Agriculture (96-35205-3854) and New Jersey Commission on Science and Technology (02-2042-007-11), and from China’s Natural Science Foundation (39825121) and China’s High-tech 863 program (2001AA628150). This is publication IMCS-2006-13 and NJSG-06-641.

References

  1. Cornet M (1993) A short-term culture method for chromosome preparation from somatic tissues of adult mussel (Mytilus edulis). Experientia 49:87–90CrossRefGoogle Scholar
  2. DeWit JMJ (1980) Origins of polyploids. In: Lewis WH (ed) Polyploidy biological relevance. Plenum, New York, pp 3–15CrossRefGoogle Scholar
  3. Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15Google Scholar
  4. Fernández-Tajes J, González-Tizón A, Martínez-Lage A, Méndez J (2003) Cytogenetics of the razor clam Solen marginatus (Mollusca : Bivalvia : Solenidae). Cytogenet Genome Res 101:43–46CrossRefGoogle Scholar
  5. Fontana F, Lanfredi M, Chicca M, Congiu L, Tagliavini J, Rossi R (1999) Fluorescence in situ hybridization with rDNA probes on chromosomes of Acipenser ruthenus and Acipenser naccarii ( Osteichthyes Acipenseriforms). Genome 42:1008–1012CrossRefGoogle Scholar
  6. González-Tizón A, Martínez-Lage A, Mariñas L, Feire R, Cornudella L, Méndez J (1998) Cytogenetic characterization of Donax trunculus (Mollusca, Bivalvia). In: Proceedings of the 13th international chromosome conference: abstract 109Google Scholar
  7. González-Tizón AM, Martínez-Lage A, Rego I, Ausió J, Méndez J (2000) DNA content, karyotypes, and chromosomal location of 18S-5.8S-28S ribosomal loci in some species of bivalve molluscs from the Pacific Canadian coast. Genome 43:1065–1072CrossRefGoogle Scholar
  8. Guo X, Allen SK Jr (1997) Fluorescence in situ hybridization of the vertebrate telomere sequence to chromosome ends of the Pacific oyster, Crassostrea gigas Thunberg. J Shellfish Res 16:87–89Google Scholar
  9. Insua A, Lopez-Pinon MJ, Mendez J (1998) Characterization of Aequipecten opercularis (Bivalvia: Pectinidae) chromosomes by different staining techniques and fluorescent in situ hybridization. Genes Genet Syst 73:193–200CrossRefGoogle Scholar
  10. Insua A, Freire R, Mendez J (1999) The 5S rDNA of the bivalve Cerastoderma edule: nucleotide sequence of the repeat unit and chromosomal location relative to 18S-28S rDNA. Genet Sel Evol (Paris) 31:509–518CrossRefGoogle Scholar
  11. Ito M, Ohmido N, Akiyama Y, Fukui K, Koba T (2000) Characterization of Spinach chromosomes by condensation patterns and physical mapping of 5S and 45S rDNA by FISH. J Am Soc Hort Sci 125:59–62Google Scholar
  12. King M (1993) Species evolution: the role of chromosome change. Cambridge University Press, CambridgeGoogle Scholar
  13. Leitão A, Thiriot-Quiévreux C, Boudry P, Malheiro I (1999) A ‘G’ chromosome banding study of three cupped oyster species: Crassostrea gigas, Crassostrea angulata and Crassostrea virginica (Mollusca: Bivavia). Genet Sel Evol 31:519–527CrossRefGoogle Scholar
  14. Levan A, Fredga D, Sandberg AA (1964) Nomenclature for centromeric position on chromosomes. Hereditas 52:201–220CrossRefGoogle Scholar
  15. Li X, Havenhand JN (1997) Karyotype, nucleolus organiser regions and constitutive heterochromatin in Ostrea angasi (Mollusca: Bivalvia): evidence of taxonomic relationships within the Ostreidae. Mar Biol 127:443–448CrossRefGoogle Scholar
  16. Martínez JL, Morán P, García-Vázquez E, Pendás AM (1996) Chromosomal localization of the major and 5S rDNA gene in the European eel ( Anguilla anguilla). Cytogenet Cell Gene 73:149–152CrossRefGoogle Scholar
  17. Martínez A, Mariñas L, González-Tizón A, Méndez J (2002) Cytogenetic characterization of Donax trunculus (Bivalvia : Donacidae) by means of karyotyping, fluorochrome banding and fluorescent in situ hybridization. J Molluscan Stud 68:393–396CrossRefGoogle Scholar
  18. Mendez J, Pasantes JJ, Martinez-Exposito MJ (1990) Banding pattern of mussel (Mytilus galloprovincialis) chromosomes induced by 2X SSC/Giemsa-stain treatment. Mar Biol 106:375–377CrossRefGoogle Scholar
  19. Menzel RW, Menzel MY (1965) Studies on chromosomes to two species of quahog clams and their hybrids. Biol Bull 129:181–188CrossRefGoogle Scholar
  20. Nakamura HK (1985) A review of molluscan cytogenetic information based on the CISMOCH-computerized index system for molluscan chromosomes. Bivalvia, Polyplacophora and Cephalopoda. Venus Jpn J Malacol 44:193–226Google Scholar
  21. Pauls E, Affonso PR (2000) The karyotype of Nodipecten nodosus (Bivalvia: Pectinidae). Hydrobiologia 420:99–102CrossRefGoogle Scholar
  22. Plohl M, Prats E, Martínez-Lage A, González-Tizón A, Méndez J, Cornudella L (2002) Telomeric localization of the vertebrate-type hexamer repeat, (TTAGGG)n, in the wedgeshell clam Donax trunculus and other marine invertebrate genomes. J Biol Chem 277:19839–19846CrossRefGoogle Scholar
  23. Rodriguez-Romero F, Laguarda-Figueras A, Uribe-Alcocer M, Rojas-Lara ML (1979) Distribution of “G” bands in the karyotype of Crassostrea virginica. Venus Jpn J Malacol 38:180–184Google Scholar
  24. Shishido R, Sano Y, Fukui K (2000) Ribosomal DNAs: an exception to the conservation of gene order in rice genomes. Mol Gen Genet 263:586–591CrossRefGoogle Scholar
  25. Tagliavini J, Williot P, Congiu L, Chicca M, Lanfredi M, Rossi R, Fontana F (1999) Molecualr cytogenetic analysis of the karyotype of the European Atlantic sturgeon, Acipenser sturio. Heredity 83:520–525CrossRefGoogle Scholar
  26. Thiriot-Quiévreux C (1994) Advances in cytogenetics of aquatic organisms. In: Beaumont AR (ed) Genetics and evolution of aquatic organisms. Chapman and Hall, London, pp 369–388Google Scholar
  27. Thiriot-Quiévreux C (2002) Review of the literature on bivalve cytogenetics in the last ten years. Cah Biol Mar 43:17–26Google Scholar
  28. Wada K, Scarpa J, Allen SK Jr (1990) Karyotype of the dwarf-surf clam Mulinia lateralis (Say 1822) (Mactridae, Bivalvia). J Shellfish Res 9:279–281Google Scholar
  29. Wang Y, Guo X (2001) Chromosomal mapping of the vertebrate telomeric sequence (TTAGGG)n in four bivalve molluscs by fluorescence in situ hybridization. J Shellfish Res 20:1187–1190Google Scholar
  30. Wang Y, Guo X (2004) Chromosomal rearrangement in pectinidae revealed by rRNA Loci and implications in bivalve evolution. Biol Bull 207:247–256CrossRefGoogle Scholar
  31. Wang Y, Xu Z, Guo X (2004) Differences in the rDNA-bearing chromosome divide the Asian-Pacific and Atlantic species of Crassostrea (Bivalvia, Mollusca). Biol Bull 206:46–54CrossRefGoogle Scholar
  32. Wang Y, Xu Z, Pierce JC, Guo X (2005) Characterization of eastern oyster (Crassostrea virginica Gmelin) chromosomes by fluorescence in situ hybridization with bacteriophage P1 clones. Mar Biotechnol 7:207–214CrossRefGoogle Scholar
  33. White MJD (1978) Modes of speciation. W.H. Freeman, San FranciscoGoogle Scholar
  34. Xu Z, Guo X, Gaffney PM, Pierce JC (2001) Chromosomal location of the major ribosomal RNA genes in Crassostrea virginica and Crassostrea gigas. Veliger 44:79–83Google Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Haskin Shellfish Research Laboratory, Institute of Marine and Coastal SciencesRutgers UniversityPort NorrisUSA
  2. 2.Experimental Marine Biology Laboratory, Institute of OceanologyChinese Academy of SciencesShandongPeople’s Republic of China

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