Mitochondrial and nuclear DNA phylogeography of two cupped oysters Crassostrea gigas and Crassostrea angulata

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1 BIOLOGY & EVOLUTION OF THE BIVALVIA Cambridge September 1999 Mitochondrial and nuclear DNA phylogeography of two cupped oysters Crassostrea gigas and Crassostrea angulata P. Boudry & A. Huvet Laboratoire de Génétique & Pathologie BP La Tremblade France -

2 Introduction 1793 Crassostrea gigas (Thunberg) 1819 Crassostrea angulata (Lamarck) 1868 Accidental introduction of C. angulata from Portugal into France 1950's Up to 100,000 tons per year of C. angulata produced in France 1966 First case of introduction of C. gigas into France 1970 Major mortality leading to disappearance of C. angulata by Massive introduction of C. gigas from Canada and Japan into Europe 1974 "Portuguese and Japanese oysters are the same species" (Menzel, 1974): they are inter-fertile and morphologically identical. 1980's Allozyme-based studies showed no genetic differentiation between C. angulata and C. gigas (Buroker et al. 1979, Mattuicci & Villani, 1983).

3 How do we explain the presence of these two conspecific taxa on opposite sides of the world? 2 hypotheses: (1) Common ancestry: C. gryphoides (Stenzel, 1971; Durve, 1986). (2) Introduction: From Japan to Portugal or reverse (Buroker et al., 1971). This study: Genetic differentiation among C. angulata and C. gigas populations using mitochondrial and nuclear DNA markers.

4 Sampling of C. gigas & C. angulata Seudre Arcachon Biarritz Rio Mira Faro Cadiz + Taiwan (2) and Japan 9 populations 383 individuals Samples were first classified as C. angulata or C. gigas according to the information provided by the suppliers.

5 Mitochondrial markers (Boudry et al. 1998) PCR amplification of a 710nt mitochondrial fragment (Cytochrome Oxydase C subunit I) 9 restriction enzymes tested 4 restriction enzymes showed polymorphism 6 haplotypes

6 Mitochondrial DNA Neighbor-joining tree based on Reynolds' genetic distance (Reynolds, 1983) Taiwan 1 Faro Taiwan 2 Cadiz C. angulata haplotype A 92 % 0.1 Rio Mira Biarritz Japan Arcachon Seudre C. gigas haplotype C 95 %

7 Microsatellite markers (Huvet, 1998) PCR amplification of 3 polymorphic loci (Magoulas et al., 1998) CG44 CG49 CG108 Total number of alleles Alleles/population Observed heterozygosity (Ho) Gene diversity (He)

8 Microsatellite markers Neighbor-joining tree based on Reynolds' genetic distance (Reynolds, 1983) Japan (35) Biarritz (32) Arcachon (31) C. gigas Seudre (32) Rio Mira (18) Taiwan 2 (27) C. angulata Faro (24) Cadiz (27) 0.01 Taiwan 1 (26) Population (mean number of alleles)

9 Comparison between mitochondrial and microsatellite data Global Fst Mitochondrial DNA 0.81 Microsatellites Microsatellite pairwise Fst values R 2 = 0.49*** Significant microsatellite pairwise Fst values 0.05 Mitochondrial pairwise Fst values R 2 = 0.54*** Significant mitochondrial pairwise Fst values

10 Comparison between mitochondrial and microsatellite data Neighbor-joining tree based on Reynolds' genetic distance (Reynolds, 1983) 0.01 Japan Biarritz Arcachon Seudre Rio Mira Taiwan 2 Faro Cadiz Taiwan 1 Biarritz Japan Arcachon Seudre Faro Taiwan 2 Rio Mira Cadiz Taiwan 1 0.1

11 Conclusion Portuguese oysters originated from Asia, probably from Taiwan. The Asian origin of C. angulata was supported by mtdna sequence data and the divergence date between C. angulata and C. gigas was estimated at 1 to 2 million years (O'Foighil et al. 1998). Portuguese oysters were probably transported by ships from "Formosa", reached by European seafarers during the XVII th century. These naturalised in Southern Europe and then were described by Lamarck. Microsatellite markers show no evidence of drift following this introduction. Mitochondrial and markers give qualitatively but not quantitatively congruent results, probably due to different evolutionary forces on these markers (drift, mutation).

12 Distribution of allele frequencies in terms of allele size for the locus CG allele frequency Haplotype A Haplotype C allele size (bp)

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