Anne-Leïla Meistertzheim, Christine Paillard , Alain Van Wormhoudt, Dario Moraga and Sylvain Huchette

1. Genetic variability and selection of abalone (Haliotis tuberculata) strains for a better growth rate and resistance to their specific pathogen (WP5) Grant Agreement no 222 156 Anne-Leïla Meistertzheim, Christine Paillard, Alain Van Wormhoudt, Dario Moraga and Sylvain Huchette

2. 15.5 Cherbourg 16 Guernsey 16 Jersey 16.5 15.5 17 St-Malo Morlaix Brest France 16 17 Concarneau 16 Lorient 16.5 20 17 18 Nantes 20 19 20 19 Huchette & Clavier, 2004 Historyof abalone mortalities Up to 60 - 80% of estimated stock Always in september Temperature high Spawning period  V. harveyi was isolated from moribund abalone in 1998 (VCR) Nicolas et al. 2002

4. Objectives Haliotis tuberculata Vibrio harveyi 1 µm Scea Scea France Haliotis Haliotis Travers et Paillard (2008) Identification of genetic markers associated with phenotype ”best growing” and “resistance to vibriosis” Methodology : Identification of genes potentially regulated by two successive bacterial challenges (Suppression Subtractive Hybridization) Polymorphism analysis of candidate genes Biological traits (sex, maturation, growth) Phenotype/genotype coupling

5. 100 80 60 Cumulative mortality (%) 1st Infection 2nd Infection 40 20 0 0 10 20 30 40 50 60 Material and methods Experimental design and results Conditioning fast for several days at 15°C then 19°C of individuals of 2,5 years old from different families Two successive infections using GFP ORM4 V. harveyi at 19°C Moribunds Survivors Time (days) • Number of died ormers per day • Checking presence of pathogen on moribunds (ORM4 - GFP V. harveyi) • Sex and size identification

6. Moribunds Results • High number of small males • Moribunds : Big and small / male and female

7. 60 50 40 ratio( Nb big/Nb total) 30 20 female male 10 0 0 10 20 30 40 50 60 70 Time (days) • Growth of males << Growth of females • Better resistance to infections of big males Results • Number of ratio big/small male increase significantly throughout infections • Sampling of muscle • RNA extraction and DNA extraction • Genetic study

8. Angiogenesis Cell cycle regulation and proliferation Cellular respiratory Cytoskeleton organization Endocytosis General and energetic metabolisms Homeostasis Ferritine NADH deshydrogenase sub. 2 Malate dehydrogenase CoxI and III Muscular activity Neurotransmission Protein synthesis Stress response Unknown function Species identification (Barcoding) Glutamine synthetase Clathrin ESTs • Several physiological processes • High number of ESTs Results 221 sequences potentially regulated by two bacterial challenges in H. tuberculata with among them 67% unknown (EST) Up-regulated genes in survivors Up-regulated genes in moribunds Cellular adhesion Down-regulated genes in survivors Down- regulated genes in moribunds Meistertzheim et al., in prep

9. Genetic structure approach using COI Frequency of the 5 haplotypes in the four different group of abalones moribunds

10. Genetic differences between size class associated with sex Genetic structure using COI Test significance on haplotypic frequencies distribution

11. Genetic study with others candidate genes on size groups on sex groups on moribunds and resistant individuals to bacterial challenge • Genetic markers developed for diagnostic tools in situ • Difference between sexes on growth • Genetic differentiation of groups (sex and size) Conclusion and perspectives • Moribund : females + males • Growth of females > growth of males • Big resistant males more resistant than small ones • Genetic study difference between small and big males Genetic similarities of COI between small males and big females big females and small males