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Development of Novel Microsatellite DNA Markers from the Pacific Oyster Crassostrea gigas

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Abstract

We document the potential of novel microsatellites as a genetic tool in furthering our understanding of the Crassostrea gigas genetic structure. From the microsatellite-enriched libraries we constructed, 123 repeat regions that had sufficient sequence information to design polymerase chain reaction primer sets were isolated. From these, 9 primer pairs were screened in a C. gigas population of 67 individuals to evaluate the genetic variability. All but 1 of the 9 loci showed allelic variation (number of alleles, 2–20; observed heterozygosity, 0.119–0.925; unbiased expected heterozygosity, 0.139–0.914). Considerable discrepancy of genotypic proportions from the Hardy-Weinberg equilibrium was observed at 1 locus with an apparent heterozygote deficiency. Several loci were successfully amplified in 3 other related species with the appropriate allele size: 6 loci in C. sikamea, 4 loci in C. ariakensis, and 5 loci in C. nippona.

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References

  1. M.A. Banks C. Waters D. Hedgecock (1993) ArticleTitleDiscrimination between closely related Pacific oyster species (Crassostrea) via mitochondrial DNA sequences coding for large subunit rRNA. Mol Mar Biol Biotechnol 2 129–136 Occurrence Handle1:CAS:528:DyaK2cXktVaitr0%3D Occurrence Handle8103411

    CAS  PubMed  Google Scholar 

  2. M.A. Banks D.J. McGoldrick W. Borgeson (1994) ArticleTitleGametic incompatibility and genetic divergence of Pacific and Kumamoto oysters Crassostrea gigas and C. sikamea. Mar Biol 121 127–135

    Google Scholar 

  3. P. Bentzen C.T. Taggart D.E. Ruzzante D. Cook (1996) ArticleTitleMicrosatellite polymorphism and the population structure of Atlantic cod (Gadus morpha) in the Northwest Atlantic. Can J Fish Aquat Sci 53 2106–2721

    Google Scholar 

  4. D. Botstein R.L. White M. Skolnick R.W. Davis (1980) ArticleTitleConstruction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet 32 314–331 Occurrence Handle1:CAS:528:DyaL3cXkvFamtbc%3D Occurrence Handle6247908

    CAS  PubMed  Google Scholar 

  5. Buroker, N.E., Hershberger, W.K., Chew, K.K. (1979) Population genetics of the family Ostreidae, I: intraspecific studies of Crassostrea gigas and Saccostrea commercialis.

  6. D.F. Callen A.D. Thompson Y. Shen H.A. Phillips R.I. Richards J.C. Mulley G.R. Sutherland (1993) ArticleTitleIncidence of null alleles in the (AC)n microsatellite markers. Am J Hum Genet 52 922–927

    Google Scholar 

  7. G.R. Carvalho L. Hauser (1994) ArticleTitleMolecular genetics and the stock concept in fisheries. Rev Fish Biol Fisheries 4 326–350

    Google Scholar 

  8. A. Dempster N. Laird D. Rubin (1977) ArticleTitleMaximum likelihood estimation from incomplete data via the EM algorithm. J R Statist Soc 39 1–38

    Google Scholar 

  9. J.A. DeWoody J.C. Avise (2000) ArticleTitleMicrosatellite variation in marine, freshwater, and anadromous fishes compared with other animals. J Fish Biol 56 461–473 Occurrence Handle10.1006/jfbi.1999.1210 Occurrence Handle1:CAS:528:DC%2BD3cXht1Khu7s%3D

    Article  CAS  Google Scholar 

  10. M.L. Egglestontott A. Delvalle S. Dileanis E. Wictum A.T. Bowling (1997) ArticleTitleA single base transversion in the flanking region of an equine microsatellite locus affects amplification of one allele. Anim Genet 28 438–440 Occurrence Handle9589585

    PubMed  Google Scholar 

  11. Y. Fujio (1979) ArticleTitleEnzyme polymorphism and population structure of the Pacific oyster, Crassostrea gigas. Tohoku J Agr Res 30 32–42 Occurrence Handle1:CAS:528:DyaE1MXlvFSjurY%3D

    CAS  Google Scholar 

  12. P.M. Gaffney T.M. Scott (1984) ArticleTitleGenetic heterozygosity and production traits in natural and hatchery populations of bivalves. Aquaculture 42 289–302

    Google Scholar 

  13. D. Hedgecock G. Li M.A. Banks Z. Kain (1999) ArticleTitleOccurrence of the Kumamoto oyster Crassostrea sikamea in the Ariake Sea, Japan. Mar Biol 133 65–68 Occurrence Handle10.1007/s002270050443

    Article  Google Scholar 

  14. A. Huvet P. Boudry M. Ohresser C. Delsert F. Bonhomme (2000) ArticleTitleVariable microsatellites in the Pacific oyster Crassostrea gigas and other cupped oyster species. Anim Genet 31 68–79 Occurrence Handle10.1046/j.1365-2052.2000.00579.x

    Article  Google Scholar 

  15. T. Imai S. Sakai (1961) ArticleTitleStudy of breeding of Japanese oyster, Crassostrea gigas. Tohoku J Agric Res 12 125–171

    Google Scholar 

  16. A.G. Jones C.A. Stockwell D. Walker J.C. Avise (1998) ArticleTitleThe molecular basis of a microsatellite null allele from the White Sands pupfish. J Hered 89 339–342 Occurrence Handle1:CAS:528:DyaK1cXls1aqtLc%3D

    CAS  Google Scholar 

  17. H.A. Lessios (1992) ArticleTitleTesting electrophoretic data for agreement with Harding-Weinberg expectations. Mar Biol 112 517–523

    Google Scholar 

  18. C.J. Lundy P. Moran C. Rico R.S. Milner G.M. Hewitt (1999) ArticleTitleMacrogeographical population differentiation in oceanic environments: a case study of European hake (Merluccius merluccius), a commercially important fish. Mol Ecol 8 1889–1898 Occurrence Handle1:STN:280:DC%2BD3c%2FoslKjtA%3D%3D Occurrence Handle10620232

    CAS  PubMed  Google Scholar 

  19. A. Magoulas B. Gjetvaj V. Terzoglou E. Zouros (1998) ArticleTitleThree polymorphic microsatellites in the Japanese oyster, Crassostrea gigas (Thunberg). Anim Genet 29 69–70 Occurrence Handle1:CAS:528:DyaK1cXksFKlurk%3D Occurrence Handle9682462

    CAS  PubMed  Google Scholar 

  20. T.C. Marshall J. Slate L.E.B. Kruuk J.M. Pemberton (1998) ArticleTitleStatistical confidence for likelihood-based paternity inference in natural populations. Mol Ecol 7 639–655 Occurrence Handle1:STN:280:DyaK1c3ps1Olsw%3D%3D Occurrence Handle9633105

    CAS  PubMed  Google Scholar 

  21. D.J. McGoldrick D. Hedgecock L.J. English P. Baoprasertkul R.D. Ward (2000) ArticleTitleThe transmission of microsatellite alleles in Australian and North American stocks of the Pacific oyster (Crassostrea gigas): selection and null alleles. J Shellfish Res 19 779–788

    Google Scholar 

  22. M. O'Connell J.M. Wright (1997) ArticleTitleMicrosatellite DNA in fishes. Rev Fish Biol Fisheries 7 331–363 Occurrence Handle10.1023/A:1018443912945

    Article  Google Scholar 

  23. M. O'Connell M.C. Dillon J.M. Wright P. Bentzen S. Merkouris J. Seeb (1998) ArticleTitleGenetic structuring among Alaskan Pacific herring populations identified using microsatellite variation. J Fish Biol 53 150–163 Occurrence Handle10.1006/jfbi.1998.0697

    Article  Google Scholar 

  24. K. Okoshi K. Mori T. Nomura (1987) ArticleTitleCharacteristics of shell chamber formation between the two local races in the Japanese oyster, Crassostrea gigas. Aquaculture 67 313–320

    Google Scholar 

  25. H. Ozaki Y. Fujio (1985) ArticleTitleGenetic differentiation in geographical populations of the Pacific oyster (Crassostrea gigas) around Japan. Tohoku J Agr Res 36 49–61

    Google Scholar 

  26. J.M. Pemberton J. Slate D.R. Bancroft J.A. Barrett (1995) ArticleTitleNonamplifying alleles at microsatellite loci: a caution for parentage and population studies. Mol Ecol 4 249–252 Occurrence Handle1:STN:280:ByqB2Mzkslw%3D Occurrence Handle7735527

    CAS  PubMed  Google Scholar 

  27. W.R. Rice (1989) ArticleTitleAnalyzing tables of statistical tests. Evolution 43 223–249

    Google Scholar 

  28. S. Schneider J.M. Kueffer D. Roessli L. Excoffier (1997) ARLEQUIN Ver.1.1: A Software for Population Genetic Data Analysis. Genetics and Biometry Laboratory University of Geneva, Switzerland

    Google Scholar 

  29. M. Sekino N. Takagi M. Hara H. Takahashi (2000) ArticleTitleMicrosatellites in rockfish Sebastes thompsoni (Scorpaenidae). Mol Ecol 9 634–636 Occurrence Handle10.1046/j.1365-294x.2000.00882-3.x Occurrence Handle1:CAS:528:DC%2BD3cXjsFGjsr0%3D Occurrence Handle10792708

    Article  CAS  PubMed  Google Scholar 

  30. P.W. Shaw G.J. Pierce P.R. Boyle (1999) ArticleTitleSubtle population structure within a highly vagile marine invertebrate, the veined squid Loligo forbesi, demonstrated with microsatellite DNA markers. Mol Ecol 8 407–418 Occurrence Handle1:CAS:528:DyaK1MXjtVKht7s%3D

    CAS  Google Scholar 

  31. K. Summers W. Amos (1997) ArticleTitleBehavioral, ecological and molec glaruenetic analyses of reproductive strategies in the Amazonian dart-poison frog, Dendrobates ventrimaculatus. Behav Ecol 8 260–267

    Google Scholar 

  32. H. Takahashi K. Nirasawa T. Furukawa (1996) ArticleTitleAn efficient method to clone chicken microsatellite repeat sequences. Jpn Poultry Sci 33 292–299 Occurrence Handle1:CAS:528:DyaK28XmtFyhur8%3D

    CAS  Google Scholar 

  33. A.E. Wilbur P.M. Gaffney (1997) ArticleTitleA nuclear marker for the molecular identification of Kumamoto oyster (Crassostrea sikamea) broodstock. J Shellfish Res 16 358

    Google Scholar 

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Acknowledgements

We acknowledge T. Bunya, Miyagi Prefectural Fisheries Research Center, for providing the oyster samples. This study was supported by a grant-in-aid (Research Project for Utilizing Advanced Technologies in Agriculture, Forestry and Fisheries) from the Ministry of Agriculture, Forestry and Fisheries, Japan.

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Authors and Affiliations

  1. Tohoku National Fisheries Research Institute, Fisheries Research Agency, Shinhama, Shiogama, Miyagi 985-0001, Japan

    Masashi Sekino

  2. Research Institute of Seto Inland Sea, Fisheries Research Agency, Maruishi, Ohno, Saeki, Hiroshima 739-0452, Japan

    Masami Hamaguchi

  3. National Institute of Fisheries Science, Fisheries Research Agency, Fukuura, Kanazawa, Yokohama 236-8648, Japan

    Futoshi Aranishi

  4. Department of Biotechnology, Shinmito, Ishinomaki Senshu University, Minamisakai, Ishinomaki, Miyagi 986-8580, Japan

    Kenji Okoshi

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  1. Masashi Sekino
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  2. Masami Hamaguchi
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  3. Futoshi Aranishi
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  4. Kenji Okoshi
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Correspondence to Masashi Sekino.

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Sekino, M., Hamaguchi, M., Aranishi, F. et al. Development of Novel Microsatellite DNA Markers from the Pacific Oyster Crassostrea gigas . Mar. Biotechnol. 5, 227–233 (2003). https://doi.org/10.1007/s10126-002-0104-z

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  • DOI: https://doi.org/10.1007/s10126-002-0104-z

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