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Manuscrit Nathe Ok UNIVERSITE CHEIKH ANTA DIOP DE DAKAR ECOLE DOCTORALE: SCIENCES DE LA VIE, DE LA SANTE ET DE L’ENVIRONNEMENT FACULTE : SCIENCES ET TECHNIQUES Année : 2011 N° d’ordre : 003 THESE DE DOCTORAT Spécialité : Biotechnologies Végétales et Microbiennes et Amélioration des Plantes Présentée par : Nathalie DIAGNE Titre : Diversité, fonctionnement et incidence sur la symbiose actinorhizienne des champignons mycorhiziens associés aux Casuarina equisetifolia dans leur zone endémique et dans leur zone d’introduction Soutenue le 16 Juin 2011 devant le jury composé de : Président : M. Diaga DIOUF Maître de conférences - UCAD Rapporteurs : M. Kandioura NOBA Professeur titulaire, Directeur de thèse - UCAD M. Laurent LAPLAZE Directeur de Recherches - IRD Examinateurs : M. Djibril SANE Maître de conférences - UCAD M. Robin DUPONNOIS Directeur de Recherches, Co-Directeur - IRD ECOLE DOCTORALE: SCIENCES DE LA VIE, DE LA SANTE ET DE L’ENVIRONNEMENT FACULTE : SCIENCES ET TECHNIQUES THESE DE DOCTORAT Spécialité : Biotechnologies Végétales et Microbiennes et Amélioration des Plantes Résumé Nom et prénom de la Candidate : DIAGNE Nathalie Titre de la thèse: Diversité, fonctionnement et incidence sur la symbiose actinorhizienne des champignons mycorhiziens associés aux Casuarinaceae dans leur zone endémique et dans leur zone d’introduction Date et lieu de soutenance : 16 Juin 2011 UCAD Dakar Jury : Président: M. Diaga DIOUF Maître de conférences - UCAD Rapporteurs : M. Kandioura NOBA Professeur titulaire, Directeur de thèse -UCAD M. Laurent LAPLAZE Directeur de Recherches - IRD Examinateurs : M. Djibril SANE Maître de conférences - UCAD M. Robin DUPONNOIS Directeur de Recherches, Co-Directeur - IRD Résumé : Depuis plusieurs décennies, des essences forestières à croissance rapide ont été introduites en Afrique de l’Ouest afin de réhabiliter des milieux dégradés, c'est le cas de Casuarina equisetifolia, un arbre australien introduit au Sénégal. Une technique utilisée pour améliorer la croissance des arbres plantés est l'inoculation avec des champignons mycorhiziens, généralement faite avec des souches présentes dans la zone d'introduction ou connues pour leur agressivité vis à vis de la plante hôte. Cependant, l'effet de l'inoculation est très variable selon la souche utilisée, et il n'existe aucune étude sur l'efficacité de l'inoculation avec des souches issues de la zone d'origine de l'arbre introduit. Or ces souches peuvent être potentiellement intéressantes car dans ce cas une co-évolution longue entre les deux partenaires symbiotiques peut aboutir à une optimisation de la symbiose. Dans ce travail nous nous sommes intéressés à l'impact sur la croissance et sur la communauté bactérienne du sol des complexes microbiens (champignons arbusculaires et Frankia ) associés à C. equisetifolia dans sa zone d’origine (Australie) et dans sa zone d’introduction (Sénégal) selon trois approches: (1) en étudiant la diversité des champignons endomycorhiziens et de la communauté microbienne associée aux C. equisetifolia dans leur zone d’origine et d’introduction, (2) en explorant l’impact de ces complexes symbiotiques sur la croissance de C. equisetifolia et la diversité fonctionnelle des communautés microbiennes du sol (3) en caractérisant la fonctionnalité de la symbiose mycorhizienne et actinorhizienne chez C. equisetifolia inoculé avec un complexe microbien d’origine australienne ou sénégalaise. Nos résultats montrent que la composition de la communauté fongique et son impact sur la communauté bactérienne du sol est variable suivant les origines des sols, mais qu'une diversité accrue est observée pour les échantillons australiens. Nous avons aussi montré que l’inoculation a un effet positif sur la croissance de C. equisetifolia et modifie la diversité structurale et fonctionnelle de la communauté microbienne tellurique ; cet effet positif sur la croissance est plus marqué chez les plantes inoculées avec l'inoculum sénégalais, où le métabolisme est plus actif et le transport symbiotique de phosphate plus accru. Mots clés : Casuarina equisetifolia , Mycorhizes à arbuscules, Frankia , Diversité, Communauté bactérienne Dédicaces Dédicaces A MES PARENTS A MES FRERES ET SOEURS A MES BELLES SOEURS A MES NEVEUX ET NIECES A TOUTES LES FAMILLES DIAGNE, DIOUF, NDOUR, SENE, FAYE, NDIAYE et MBOW A tous ceux qui me sont chers Remerciements Remerciements Je tiens à remercier Pr. Ibrahima NDOYE, responsable de la formation doctorale Sciences de la vie, de la Santé et de l’Environnement de m’avoir autorisé à s’inscrire à cette école. Mes sincères remerciements pour vos conseils récurrents. Je remercie Pr. Samba N. SYLLA chef de département de Biologie Végétale pour avoir autorisé cette inscription en thèse et merci pour sa sympathie. Je remercie Pr. Amadou BA responsable du Laboratoire Commun de Microbiologie (LCM, ISRA, UCAD) pour l’accueil sympathique au sein de ce laboratoire et pour ses précieuses remarques. Je remercie également Dr. Serge HAMON responsable de l’UMR DIADE et Dr. Didier BOGUSZ pour l’accueil chaleureux au sein de l’équipe Rhizogenèse et particulièrement pour son enthousiasme. Je tiens à remercier l’Institut de Recherche pour le Développement (IRD, DSF) pour le financement de ces travaux de thèse. Mes sincères remerciements à mes encadreurs Dr. Sergio SVISTOONOFF, Dr. Diegane DIOUF, Dr. Robin DUPONNOIS et Pr. Kandioura NOBA. Je témoigne de l’intérêt avec lequel, ils ont suivi cette thèse, de leur rigueur dans le travail bien fait, de leur disponibilité et particulièrement de leur sympathie. J’exprime ma profonde gratitude au Dr. Diaga DIOUF, pour l’honneur qu’il m’a fait d’avoir accepté de juger ce travail et d’en présider le jury et surtout je tiens à lui exprimer ma reconnaissance pour l’enthousiasme qu’il a toujours manifesté. Encore merci pour les discussions scientifiques. Je remercie vivement Dr. Djibril SANE qui a accepté de juger ce travail. Qu’il trouve ici mes sincères remerciements. Je remercie Dr. Laurent LAPLAZE pour avoir accepté d’être rapporteur de cette thèse. Merci pour les conseils très précieux qui m’ont aidé à perfectionner ce document. Mes sincères remerciements au Dr. Aboubakry KANE pour sa participation à ce projet de thèse. Un grand merci pour les remarques et les conseils pertinents qui m’ont aidé à améliorer ce manuscrit. Je tiens à remercier Dr. Claudine FRANCHE, pour ses conseils avisés et ses remarques très pertinentes qui m’ont aidé à perfectionner ce travail. Je remercie également Pr. Tahir DIOP et Pr. Mamadou GUEYE pour leurs conseils et leurs encouragements. Je remercie Florence AUGUY et Dr. Valérie HOCHER pour leur aide et conseils en biologie moléculaire. Remerciements Merci au Dr. Ezékiel BAUDOUIN pour son aide dans les expériences d’étude de la diversité génétique des microorganismes. Je remercie Dr Jacques ESCOUTE, Dr. Jean Luc VERDEIL et Dr Marc LARTAUD pour leur aide en imagerie. Un grand merci au Dr. Abdala G. DIEDHIOU pour sa disponibilité et pour les discussions scientifiques fructueuses. Un grand merci à Jocelyne BONNEAU pour avoir accepté d’apporter des corrections à la rédaction de ce document. Un grand merci à Christine OUATTARA qui a participé avec efficacité à ce projet de thèse. Je ne saurais oublier, dans mes remerciements Cheikh NDIAYE pour l’assistance qu’il m’a apportée durant ces trois années de thèse. Toute ma sympathie va également à tout le personnel du LCM : Jean BAKHOUM, Maimouna CISSOKHO, Mathieu FAYE, Paul TENDENG, Francis do REGO, Anna Marthe KONTE, Marie Claire DASYLVA, Tatiana WADE, Ramatoulaye MBAYE, Mansour THIAO, François KOUADJO, Saliou FALL, Omar TOURE, Ousseynou GUEYE. Je remercie tous les membres de l’équipe Rhizogenèse Hassen, Antony, Alexandre, Daniel Khalid, Soazig, Mikael. Une pensée particulière aux filles Virginie, Amandine, Meriem, Stella, Alice, Anaïs, Faten et Joane pour les agréables moments passés ensemble. Je remercie tous mes collègues du LCM Niokhor BAKHOUM, Amadou DIENG, Fatou NDOYE, Dioumacor FALL, Abdoulaye SOUMARE, Godar SENE, Fatou GUEYE, Arsène SANON, Mansour NGNING, Fatou DIOUF, Fatoumatou FALL, Marie BOURY, Mikaella EBENYE, Mamadou DIALLO, Ibou DIOP et Seynabou SENE pour leur aide et pour la bonne ambiance dans laquelle se sont déroulés ces travaux. J’ai une pensée particulièrement à tous mes amis : Boubacar DIEDHIOU, Papa Makhtar DRAME, Augustin DIOUF, Souleymane DOUCOURE, Massila SENGHOR, Jean Birame SARR, Cheikh Lô, Myriem DIATTA, Ambroise NGNING et Gaston DIATTA pour leur écoute, leur disponibilité et leurs conseils. Un grand merci à mes frères, mes sœurs, mes belles sœurs pour leur soutien constant durant toutes mes études. A tous ceux qui ont contribué à la réalisation de ce travail, je leur témoigne toute ma reconnaissance. Abréviations Liste des abréviations AA Acide Aminé ABA Acide abcissique ACP Analyse de composante principale ADN Acide Désoxyribonucléique ADNc ADN complémentaire AFTD Agroforestry tree database AIA Acide indole acétique AJ Acide jasmonique ALP Alcaline phosphatase AMT Transporteur d’ammonium « Amonium transporter » APP Appareil de prépénétration ARISA Automated Approach for Ribosomal Intergenic Spacer Analysis ARN Acide ribonucléique AS Acide salicylique ATP Adénine Triphosphate BAM Bactéries Auxiliaires de Mycorhization BAP « Buffered mineral medium Added of Phosphatidylcholine» BGA Between Group Analysis BLAST-N Local Alignment Search Tool CaCMK Calcium Calmoduline Kinase CAS Chrome Azurol S CBE chlorale black E CIRAD Centre de coopération Internationale en Recherche Agronomique pour le Développement CK Cytokinine CLSM Confocal Laser
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