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Publis013-Cbgp-063 Dumas Thès UNIVERSITE MONTPELLIER II SCIENCES ET TECHNIQUES DU LANGUEDOC THESE Pour obtenir le diplôme de doctorat Formation doctorale : Biologie de l'évolution et écologie Ecole Doctorale : Systèmes Intégrés en Biologie, Agronomie, Géosciences, Hydrosciences, Environnement, SIBAGHE Étude intégrative du statut des deux variants adaptatifs à la plante hôte de Spodoptera frugiperda (Lepidoptera : Noctuidae) Présentée et soutenue publiquement par Pascaline DUMAS le 29 octobre 2013 Devant le jury composé de : Emmanuelle d’ALENÇON, Chargée de recherche, INRA Montpellier (Directrice de Thèse) Denis BOURGUET, Directeur de recherche, INRA Montpellier (Examinateur) Michel CUSSON, Chercheur scientifique, RNCan Québec (Rapporteur) Gaël KERGOAT, Chargé de recherche, INRA Montpellier (Invité, co-Directeur de Thèse) Emmanuelle JOUSSELIN, Chargée de recherche, INRA Montpellier (Représentante Ecole Doctorale) Jean-François SILVAIN, Directeur de recherche, CNRS Gif-sur-Yvette (Examinateur) Jean-Christophe SIMON, Directeur de recherche, INRA Rennes (Rapporteur) 1 2 3 4 UNIVERSITE MONTPELLIER II SCIENCES ET TECHNIQUES DU LANGUEDOC THESE Pour obtenir le diplôme de doctorat Formation doctorale : Biologie de l'évolution et écologie Ecole Doctorale : Systèmes Intégrés en Biologie, Agronomie, Géosciences, Hydrosciences, Environnement, SIBAGHE Étude intégrative du statut des deux variants adaptatifs à la plante hôte de Spodoptera frugiperda (Lepidoptera : Noctuidae) Présentée et soutenue publiquement par Pascaline DUMAS le 29 octobre 2013 Devant le jury composé de : Emmanuelle d’ALENÇON, Chargée de recherche, INRA Montpellier (Directrice de Thèse) Denis BOURGUET, Directeur de recherche, INRA Montpellier (Examinateur) Michel CUSSON, Chercheur scientifique, RNCan Québec (Rapporteur) Gaël KERGOAT, Chargé de recherche, INRA Montpellier (Invité, co-Directeur de Thèse) Emmanuelle JOUSSELIN, Chargée de recherche, INRA Montpellier (Représentante Ecole Doctorale) Jean-François SILVAIN, Directeur de recherche, CNRS Gif-sur-Yvette (Examinateur) Jean-Christophe SIMON, Directeur de recherche, INRA Rennes (Rapporteur) 5 Financements : Cette thèse a été financée pour moitié par l’INRA département SPE et par l’Université Montpellier II Laboratoires de recherche : - UMR 1333 Diversité, Génome et Interactions Microorganismes-Insectes (DGIMI) Université Montpellier II Bâtiment 24, 4ème étage Place Eugène Bataillon 34 095 Montpellier Cedex 5 - UMR 1062 Centre de Biologie pour la Gestion des Population (CBGP) Campus International de Baillarguet CS 30016 34 988 Montferrier-sur-Lez 6 Remerciements Je tiens à remercier chacun des membres de ce jury, Denis Bourguet, Michel Cusson, Emmanuelle Jousselin, Jean-François Silvain et Jean-Christophe Simon pour avoir accepté de participer à la soutenance de cette thèse. Je tiens également à remercier M. Philippe Fournier et son successeur M. Patrick Tailliez pour leur accueil au laboratoire DGIMI. Je remercie également leurs homologues du CBGP, M. Denis Bourguet puis Mme Flavie Vanlerberghe-Masutti pour m’avoir permis d’effectuer une partie de ma thèse dans leur laboratoire. Emma, Gaël, merci à tous les deux pour votre encadrement pendant ces trois ans. Cela a été un vrai honneur d’être sous votre tutelle. Jusqu’au bout vous avez été présents et nos discussions m’ont beaucoup apportées, spécialement lors de ces derniers mois. Emma, je souhaite vous remercier du fond du cœur pour le soutien que vous m’avez apporté tout au long de cette thèse. Vous m’avez toujours fait confiance et je me suis toujours sentie à l’aise sous votre direction. Votre disponibilité, votre engagement et votre ouverture d’esprit sont un vrai modèle pour la future scientifique que je souhaiterais devenir. Gaël, toi aussi je te remercie sincèrement. Avec toi, c’était la découverte permanente : de nouveaux concepts, de nouvelles techniques, de nouveaux continents… Merci pour tes conseils toujours pertinents, pour ta rigueur et ton soutien. Merci de m’avoir poussée vers le haut. Je souhaite également remercier toute l’équipe EHA pour leurs conseils et leur aide pendant cette thèse. Un merci tout spécial à Sylvie pour son encadrement à la paillasse et pour ses conseils techniques mais également pour tout le reste! Merci à Philippe, François et Nicolas ainsi qu’à ma collègue de bureau Imène, la seule et unique ! Et bien sûr, merci à Marion. Un très grand merci à tous les membres de l’insectarium et particulièrement à Clotilde et Gaëtan. Merci pour m’avoir formée aux élevages et pour avoir été toujours disponibles quand j’avais des soucis de boîtes ou de Poitout ! Sans votre travail, je n’aurais rien pu faire. Même s’il n’y a pas d’équipe au CBGP, je tiens à remercier Anne-Laure pour son soutien technique et pas seulement ! Merci également pour m’avoir supportée en mission 7 poulette. Un merci très chaleureux à Anne et Bruno pour ne pas m’avoir laissée dans la forêt le jour où on s’est perdu…lors de la mission en Guyane. Ces trois années de thèses se sont partagées entre deux laboratoires différents ; un pied au laboratoire DGIMI et l’autre au CBGP. Grâce à cela j’ai eu la chance de rencontrer deux fois plus de gens intéressants, qui à un moment ou l’autre de ma thèse m’ont aidée, conseillée, soutenue et beaucoup apporté. Je tiens donc à remercier toutes les personnes du laboratoire DGIMI et du CBGP. Un grand merci aux « glougloux » de DGIMI : Sylvie, Amélie, Cécilia, Aurélie, Imène, Laure, Doriane, Anne-Sophie, Fanny, Véro, Marie-Elsie, Apolline, Marion. Nous avons beaucoup ri ensemble et vous m’avez apporté un soutien permanent tout au long de cette thèse. Je suis vraiment heureuse de vous avoir rencontrées, merci les filles ! Bien-sûr je ne vous oublie pas les garçons. Un autre immense merci aux poulettes du CBGP : Anne-Laure, Joséphine, Aléna, Laure, Caro, Marie et Morgane (poulette de SupAgo !). Je vous remercie pour tout : votre accueil, le soutien moral et scientifique, les encouragements de ces derniers mois, les soirées filles, les conseils… Vous avez rendu ma vie à Montpellier encore plus agréable ! Merci à tous les autres du CBGP avec qui j’ai partagé des moments formidables autour d’une bière ou lors d’une partie de pétanque : Fabien, Jeff, Max et Virginie, Manu, Ashraf, Guillaume, Laurent, Julie, Réjane, Manue, Armelle, Alex … Il y a aussi les copains du Limousin et de la Dordogne, Poppy, Léa, Sakout, Clément, Mathieu, Villou merci pour votre amitié qui dure depuis si longtemps et merci à Skype de nous avoir donné la possibilité de nous parler tous les jours (ou presque) pendant ces trois ans qui ne nous ont pas permis de nous voir autant que nous l’aurions souhaité. Et puis merci à maman, « ma papa » et Manie, merci pour votre soutien inconditionnel et pour vos encouragements. Vous m’avez manqué pendant ces 9 derniers mois, j’ai hâte de vous revoir enfin. Pour finir, merci à Paco, pendant cette thèse tu as été ma boussole, ma Grande Ourse. Avec toi, la vie est plus belle mais bon, « suffit la parlotte !» Tout ce que je n’écris pas ici, tu le sais déjà. 8 Liste des Figures Figure 1 : Représentation schématique de la notion d’espèce. Figure 2 : Différentes terminologies des états de caractères en phylogénie. Figure 3 : Exemples de cas d’espèces cryptiques, de convergence évolutive et de dimorphisme sexuel. Figure 4 : Figure du Coalescent standard. Figure 5 : Histogrammes du nombre de publication par année pour une recherche sur Web of science comprenant les mots integrative taxonomy. Figure 6 : Représentation schématique des étapes théoriques de la spéciation allopatrique vicariante et de la spéciation péripatrique. Figure 7 : Représentation schématique des étapes théoriques de la spéciation sympatrique et de la spéciation parapatrique. Figure 8 : Classification des mécanismes d’isolement reproducteur. Figure 9 : Représentation schématique d’incompatibilité cytoplasmique. Figure 10 : Représentation schématique de l’incompatibilité de type Dobzhansky–Muller. Figure 11 : Evolution des incompatibilités génétiques qui peuvent induire la non-viabilité et la stérilité des hybrides. Figure 12 : Mécanismes génétiques qui lient la sélection divergente à l’isolement reproducteur. Figure 13 : Représentation schématique de la vision en île ou en continent de la divergence génomique. Figure 14 : Aires de répartition des six plus grandes espèces de ravageurs du genre Spodoptera. Figure 15 : Cycle de vie de Spodoptera frugiperda. Figure 16 : Diagramme de la composition des différents composés présents dans le cocktail de phéromones sexuelles du variant maïs et riz de S. frugiperda. Figure 17 : Photographies de dommages engendrés par les larves de S. frugiperda. Figure 18 : Organigramme présentant chronologiquement les différentes étapes de la thèse. Figure 19 : Photographie du protocole expérimental utilisé pour réaliser des croisements intra et inter variants. Figure 20 : Comparaison entre variant maïs et variant riz des séquences d’un fragment du gène COI. Figure 21 : Représentation schématique du processus de crossing-over lors de la formation des gamètes. Figure 22 : Carte du continent américain présentant l’échantillonnage de Spodoptera frugiperda. Figure 23 : Photographies prises lors de l’échantillonnage sur le terrain de FAW adultes. Figure 24 : Analyse de barcode pour une région du gène COI. Figure 25 : Schéma de la méthode utilisée pour assembler les contigs et les scaffolds. Figure 26 : Pipeline d’analyses bio-informatiques réalisées pour annoter les régions flanquantes des microsatellites étudiés et comparer les séquences du variant riz et du variant maïs de S.
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