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Lecocq Génétique Et Phéromo UNIVERSITE DE MONS - HAINAUT - FACULTE DES SCIENCES - LABORATOIRE DE ZOOLOGIE Génétique et phéromones sexuelles des bourdons de la Corse Directeur de mémoire Mémoire de fin d’Etudes Prof. Pierre Rasmont Présenté par Thomas Lecocq Rapporteurs : Dr. Michael Terzo, En vue de l’obtention du grade Dr. Igor Eeckhaut et Dr. Denis Michez de Maitre en Sciences Biologiques Année académique 2008-2009 T. Lecocq – Génétique et phéromones sexuelles des bourdons de la Corse - 2009 - page - 2 T. Lecocq, 2009. Génétique et phéromones sexuelles des bourdons de la Corse . Mémoire de Master en Biologie des organismes et écologies, Université de Mons-Hainaut, 111 pp. Résumé : Huit taxons de bourdons vivent en Corse. Parmi ceux-ci, six sont endémiques et morphologiquement bien différenciés de leur plus proche parent continental (homologue), à tel point que deux d’entre eux sont considérés actuellement comme des espèces à part entière (B. perezi et B. pereziellus ). Les quatre autres bourdons endémiques sont communément classés comme de simples sous-espèces de taxons continentaux ( B. hortorum jonghei , B. lucorum renardi , B. ruderatus corsicola et B. terrestris xanthopus ). Ces espèces et sous-espèces endémiques se seraient retrouvées isolées en Corse à la fin de la dernière période glaciaire (- 13 000 ans). Enfin, les deux derniers bourdons corses sont en tous points identiques aux spécimens continentaux ( B. maxillosus italicus et B. pascuorum melleofacies ). Cependant, ces statuts taxonomiques ne reposent que sur des comparaisons morphologiques par rapport aux homologues avec le lot d’incertitudes que cela comporte. L’intérêt de vérifier ces statuts à l’aide de nouvelles techniques (étude génétique et étude du Species Mating Recognition System , SMRS) se fait dès lors sentir. En effet, l’étude des phéromones sexuelles et les études génétiques constituent des méthodes très puissantes pour statuer sur la taxonomie et les affinités entre taxons. C’est sur cet objectif que se penche ce travail. Pour ce faire, la quasi totalité des spécimens corses et quelques continentaux ont été récoltés sur le terrain. Le reste est issu de collection ou d’élevage ou encore de GenBank. Ces analyses ont nécessité de décrire la composition des phéromones sexuelles des mâles corses et de séquencer deux gènes d’intérêt (COI et EF-1α), jusqu’alors inconnus. Les mêmes analyses ont été effectuées sur les homologues continentaux, utilisés comme point de comparaison. Néanmoins, la difficulté que représente l’échantillonnage limite la certitude des résultats obtenus. Au final, deux taxons ( B. corsicola et B. xanthopus ) sont considérés non comme de simples sous-espèces mais comme des espèces à part entière. Pour les autres taxons, rien de certain ne peut malheureusement être avancé étant donné le manque d’échantillonnage. Mots clés : Bombus , Corse, Phéromones, Génétique, Taxonomie T. Lecocq – Génétique et phéromones sexuelles des bourdons de la Corse - 2009 - page - 3 Remerciements Je tiens à remercier par ces quelques lignes, toutes les personnes qui ont contribué de près ou de loin à l’élaboration du présent travail. Je remercie tout particulièrement le Prof. Pierre Rasmont de m’avoir accueilli dans son service, pour ses conseils judicieux et sa constante attention. Je remercie l’Office de l’Environnement de la Corse, la Direction Régionale de l’Environnement de Corse et Madame Marie-Cécile Andrei-Ruiz de l’Observatoire Conservatoire des Insectes de Corse pour leur contribution financière à ce travail. Je tiens à exprimer ma gratitude à Mademoiselle Audrey Coppée et au Dr Denis Michez de m’avoir guidé tout au long de cette année, pour leurs conseils avisés et leur disponibilité. Je tiens à remercier le Prof. Irena Valterova (Institut of Organic Chemistry and Biochemistry, Prague, République Tchèque) de m’avoir accueilli dans son service pour l’identification des molécules. Je remercie le Dr Déborah Lanterbecq, Madame Stéphanie Iserbyt, le Dr Murat Aytekin, le Dr Sébastien Patiny et Monsieur Thibaut De Meulemeester pour leur aide, leur conseil et leur soutien. Je tiens à exprimer ma reconnaissance à A. Roelandts pour son aide lors de la récolte de matériel en Corse. Je n’oublie pas mes proches qui m’ont été indispensable dans les moments les plus difficiles. Que toutes les personnes qui ont contribué à la réalisation de ce travail et que je n’ai pas citées ici sachent que je ne les oublie pas et que je les remercie. T. Lecocq – Génétique et phéromones sexuelles des bourdons de la Corse - 2009 - page - 4 Table des matières 1. Introduction ................................................................................................................................... 7 1.1. Les bourdons de la Corse ........................................................................................................ 7 1.2. Les homologues continentaux ................................................................................................. 8 1.3. Insularité et dérive génique ................................................................................................... 10 1.4. Peuplement de la Corse ......................................................................................................... 18 1.5. Concept d’espèce et de sous-espèce ...................................................................................... 23 1.6. Objectifs ................................................................................................................................ 27 2. Matériel et méthode ..................................................................................................................... 28 2.1. Matériel biologique ............................................................................................................... 28 2.1.1. Récolte des spécimens ................................................................................................... 28 2.1.2. Systématique et détermination des spécimens ............................................................... 30 2.1.3. Nombre de spécimens corses collectés .......................................................................... 30 2.2. Analyse des sécrétions labiales céphaliques .......................................................................... 30 2.2.1. Origine des spécimens utilisés pour les analyses phéromonales ................................... 30 2.2.2. Dissection des glandes labiales céphaliques .................................................................. 35 2.2.3. Analyse chimique par GC-MS ...................................................................................... 36 2.2.4. Analyses statistiques ...................................................................................................... 36 2.2.4.1. Elaboration de la matrice de données .................................................................... 36 2.2.4.2. Analyse en composante principale (ACP) ............................................................. 39 2.2.4.3. Analyse en coordonnée principale (PCoordA) ...................................................... 39 2.2.4.4. Dendrogramme ...................................................................................................... 40 2.3. Analyses génétiques .............................................................................................................. 40 2.3.1. Echantillonnage des taxons ........................................................................................... 40 2.3.2. Analyses moléculaires ................................................................................................... 45 2.3.2.1. Extraction de l’ADN .............................................................................................. 46 2.3.2.2. PCR ....................................................................................................................... 46 2.3.2.3. Séquençage ............................................................................................................ 47 T. Lecocq – Génétique et phéromones sexuelles des bourdons de la Corse - 2009 - page - 5 2.3.3. Alignement et analyse de phylogénie moléculaire ........................................................ 47 3. Résultats ....................................................................................................................................... 49 3.1. Analyse des sécrétions labiales céphaliques .......................................................................... 49 3.1.1. Chromatogrammes et identification des composés des sécrétions labiales céphaliques des différentes espèces .................................................................................................................. 49 3.1.1.1. B. ruderatus corsicola et ses homologues ............................................................. 49 3.1.1.2. B. maxillosus italicus et ses homologues ............................................................... 53 3.1.1.3. B. perezi et ses homologues ................................................................................... 56 3.1.1.4. B. lucorum renardi et son homologue ................................................................... 59 3.1.1.5. B. terrestris xanthopus et son homologue ............................................................. 62 3.2. Analyses statistiques .............................................................................................................. 66 3.3. Analyses génétiques .............................................................................................................
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