Biologie De La Reproduction, Diversité Génétique Et Spatiale De Deux Espèces Du Genre Vanilla (Orchidaceae) Du Sud‐ Ouest De L’Océan Indien : V

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Biologie De La Reproduction, Diversité Génétique Et Spatiale De Deux Espèces Du Genre Vanilla (Orchidaceae) Du Sud‐ Ouest De L’Océan Indien : V UNIVERSITE DE LA REUNION Faculté des Sciences et Technologies UMR « Peuplements Végétaux et Bio‐agresseurs en Milieu Tropical » CIRAD – Université de la Réunion THESE Pour obtenir le diplôme de DOCTORAT Discipline : Sciences Formation Doctorale : Génétique Ecole Doctorale Sciences Technologies Santé Biologie de la reproduction, diversité génétique et spatiale de deux espèces du genre Vanilla (Orchidaceae) du sud‐ ouest de l’océan Indien : V. humblotii et V. roscheri. Implications pour leur conservation par Rodolphe GIGANT Soutenance publique du 02 Mars 2012, devant un jury composé de : Félix FOREST Chercheur, Royal Botanic Gardens, Kew, Rapporteur Tariq STEVART Chercheur, Missouri Botanical Garden, Rapporteur Michel DRON Professeur, Université Paris Sud, Examinateur Pascale BESSE Professeur, Université de la Réunion, Directrice de Thèse A papa... Remerciements REMERCIEMENTS Tant cette aventure a été riche de rencontres et d’enseignements sur les plans personnels, humains et professionnels, je tiens à remercier toutes les personnes avec qui j’ai pu échanger de façon formelle ou non et qui m’ont permis d’aboutir à ce travail de thèse. Tout d’abord, je suis reconnaissant envers Bernard Reynaud, Directeur du 3P, pour son accueil à l’UMR PVBMT. Je remercie Félix Forest, Tariq Stevart et Michel Dron pour avoir accepter de participer à mon jury de thèse. Mes remerciements infinis à LA Dream Team vanilla, mes encadrants Pascale Besse et Michel Grisoni pour leur investissement sans faille et les conditions de travail idéales qui m’ont été données, dans la confiance et la décontraction, sans oublier Jean‐Bernard Dijoux, Katia Jade, Séverine Bory, et Isabelle Fock‐Bastide. Mes sincères remerciements à mes collaborateurs à Mayotte Valérie Guiot, Guillaume Viscardi, Luc Gigord du CBNM, Danny Laybourne, Mao, Bako, Abdou de la DAF, et en Afrique Du Sud, Brigitte Church, Zack Diamini et ses rangers de l’EKZNW, sans qui ces recherches n’auraient pas vu le jour. Je tiens également à remercier Laurence Humeau, Thierry Pailler, Marie‐Hélène Chevallier, Hélène Delatte, Spencer Brown, André Charrier, Michel Noirot pour leurs conseils avisés tout au long de mes travaux ou leur participation aux réunions et comités de thèse. Je remercie Alexandre De Bruyn pour son travail de stage et pour les escapades de terrain partagées, mais aussi pour les moments de détente à Sodwana en compagnie de Chireen, Philippe et Eric qui nous ont accueilli chaleureusement et fait découvrir les plaisirs de ce coin, merci à eux aussi. Un clin d’œil également à l’ensemble des collègues et amis du 3P, notamment Vincent Lepitre, Cathy Jacquard, Magalie Thierry, Chloé Schmitt, Karine Vital, Anaïs Brugel, Frédérick Péréfares (et bien d’autres...) qui autour d’un café, d’un déjeuner ou entre deux génotypages m’ont apporté leur soutien au quotidien. Je remercie les membres de l’équipe d’entomologiste du 3P pour les identifications et les accès à leur matériel, Fredérick Chiroleu pour sa disponibilité dans les moments de crise statistique et Aude Chabirand pour sa sympathie et les accès au matériel de microcopie. Et bien sûr, je remercie ma famille, mes amours, Nelly, Lény et Mathys, pour leur soutien inconditionnel tout au long de ces trois années. Ce travail, les congrès et toutes ces rencontres n’auraient pas eu lieu sans le soutien financier de La Région Réunion et de l’école doctorale, alors je les remercie pour leur participation. TABLE DES MATIERES Table des matières TABLE DES MATIERES 1. CHAPITRE 1 : Introduction................................................................................................................1 1.1. Introduction générale...............................................................................................................1 1.2. Introduction bibliographique....................................................................................................6 Chapitre d’ouvrage Biodiversity and Evolution in the Vanilla Genus.......................................6 1.3. Présentation des vanilliers aphylles........................................................................................32 1.4. Objectifs de recherche de la thèse..........................................................................................49 2. CHAPITRE 2 Stratégie d’étude et méthodes...................................................................................51 2.1. Les collaborations...................................................................................................................51 2.2. Développement des marqueurs microsatellites ....................................................................53 Article 1 Nineteen polymorphic microsatellite markers from two African Vanilla species: across-species transferability and diversity in a wild population of V. humblotii from Mayotte. ................................................................................................................................54 3. CHAPITRE 3 Etude de V. humblotii à Mayotte...............................................................................59 3.1. Préambule.............................................................................................................................59 3.2. Article 2 Reproductive strategies and their consequences on fine-scale spatial genetic structure and diversity of the indigenous and endangered leafless Vanilla humblotii (Orchidaceae) from Mayotte (Comoros Archipelago, Indian Ocean)......................................61 3.2.1. Abstract.........................................................................................................................62 3.2.2. Introduction...................................................................................................................63 3.2.3. Materials and Methods.................................................................................................64 3.2.4. Results...........................................................................................................................73 3.2.5. Discussion......................................................................................................................80 3.2.6. Conclusion.....................................................................................................................87 3.3. Article 3 Impact of fragmentation on the genetic diversity of Vanilla humblotii Rchb. f. (Orchidaceae) in Mayotte (Comoros Archipelago, Indian Ocean) and implications for conservation..........................................................................................................................95 3.3.1. Abstract.........................................................................................................................96 3.3.2. Introduction..................................................................................................................97 3.3.3. Materials and Methods.................................................................................................99 3.3.4. Results.........................................................................................................................106 3.3.5. Discussion....................................................................................................................114 3.3.6. Implications for the conservation of V. humblotii in Mayotte.....................................123 4. CHAPITRE 4 Etude de V. roscheri en Afrique Du Sud...................................................................132 4.1. Préambule............................................................................................................................132 i Table des matières 4.2. Article 4 Active sexual reproduction but no genetic diversity in the range-edge distribution of Vanilla roscheri Rchb. f. (Orchidaceae) in South Africa....................................................134 4.2.1. Abstract........................................................................................................................135 4.2.2. Introduction.................................................................................................................136 4.2.3. Materials and Methods................................................................................................137 4.2.4. Results..........................................................................................................................144 4.2.5. Discussion.....................................................................................................................149 4.2.6. Implications for the conservation.................................................................................156 5. CHAPITRE 5 Discussion générale, conclusions et perspectives....................................................163 5.1. Apports fondamentaux à la biologie de la reproduction des vanilliers................................163 5.2. Importance de la reproduction végétative...........................................................................169 5.3. Les phénomènes de migration et d’isolement des populations chez les vanilliers aphylles de l’océan Indien.......................................................................................................................171 5.4. L’approche intégrative de la biologie de la reproduction et de la génétique des populations et la complémentarité des modèles.....................................................................................176 5.5. Implications pour la conservation
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