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A Complementary Method for Production of Tetraploid Crassostrea gigas Using Crosses Between Diploids and Tetraploids with Cytochalasin B Treatments

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Abstract

We present a new method to produce tetraploid Crassostrea gigas by cytochalasin B inhibition of polar body 2 expulsion in diploid females crossed with tetraploid males. This offers a means of direct introgression of genetic characters from selected diploid to tetraploid lines, avoiding a triploid step. Offspring larval ploidy shifted over time and depended on size, with tetraploids more frequent among the smaller larvae and triploids among the large. Viable tetraploids were found at 4 and 6 months, indicating the technique was successful. The possibility that gynogenesis occurred was tested by microsatellite analysis to confirm the presence of paternally inherited alleles. These were present in all animals of the 2n × 4n + CB (female first) cross. However, a 4n × 2n + CB cross produced triploids, including some gynogens. Our method illustrates for the first time that diploid C. gigas eggs, if selected for large size, can give viable tetraploid offspring.

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Acknowledgments

We thank the technical staff at Ifremer La Tremblade for the care and preparation of the parental animals used in this study and Florence Cornette for practical assistance during the larval rearing period. Some preliminary experiments leading to this study were conducted and tested by Raphaël Brizard. Delphine Lallias and Nicolas Taris gave invaluable advice on the application of microsatellites.

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Author notes

  1. Christophe Ledu & André Gérard

    Present address: IFREMER, Centre de Nantes, Nantes Cedex 03, 44311, France

Authors and Affiliations

  1. Institut Français pour la Recherche et Exploitation de la Mer (IFREMER), Laboratoire d’ Génétique et Pathologie, La Tremblade, 17390, France

    Helen McCombie, Christophe Ledu, Pascal Phelipot, Sylvie Lapègue, Pierre Boudry & André Gérard

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  1. Helen McCombie
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  2. Christophe Ledu
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  3. Pascal Phelipot
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Correspondence to Helen McCombie.

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McCombie, H., Ledu, C., Phelipot, P. et al. A Complementary Method for Production of Tetraploid Crassostrea gigas Using Crosses Between Diploids and Tetraploids with Cytochalasin B Treatments. Mar Biotechnol 7, 318–330 (2005). https://doi.org/10.1007/s10126-004-0440-2

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