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ZERIBI Rihana.Pdf Université Mohamed Khider de Biskra Faculté des Sciences Exactes et des Sciences de la Nature Département Sciences de la Nature et la Vie Filière Sciences biologiques Référence …..…... / 2020 MÉMOIRE DE MASTER Spécialité : Microbiologie Appliquée Présenté et soutenu par : ZERIBI Rihana Le: 30 /09/2020 Caractérisation biochimique et symbiotique des BNL isolés à partir de la plante Genista saharae Jury : President Mme H. GHITTI MCB Université de Biskra Examinateur M. AGLI ABDENACER Pr Université de Biskra Rapporteur Mme M.DJOUAMAA MAA Université de Biskra Année universitaire : 2019/2020 Remerciements Remerciements Avant tout je remercier DIEU (Allah) tout puissant de m’avoir donné le courage, la volonté et la patience pour terminer ce travail. Ensuite j’adresse mes remerciements à mon enseignante Mme DJOUAMAA Manel d’avoir bien voulu diriger ce travail et pour tous ses conseils fructueux et ses encouragement. Mes remerciements vont également à l’ensemble des membres de jury Je tenais à exprimer nos sincères remerciements à l’enseignante Mme MOKRANI Djamila Au terme de ce travail, il est agréable de remercier toutes les personnes qui ont contribué de près ou de loin à la réalisation de ce travail : les Ingénieures de laboratoire. Ce mémoire fondamentalement est un travail individuel, elle n’aurait pu être menée à bien sans une équipe de collègues qui contribuent au bon fonctionnement du laboratoire, avec lesquels il est possible d’échanger des conseils et des suggestions, et qui assurent une atmosphère de travail donnant. Je remercie pour cela tous mes collègues. Dédicace Dédicace Je dédie ce travail à : Mes très chers parents qui ont fait beaucoup de sacrifiées et continuent d’en faire pour me voir réussir, qui ont veillé à mon instruction et qui m’ont soutenu durant toutes ces années. Mes très chers frères : Hatim, Tarek, Alladin. Ma tendre sœur : Salsabil. Mon beau-frère : Abderrahmane Haddar. Mes belles sœurs : Karima et Sarrah. A mes collègues de Microbiologie Appliquée promotion 2020. A mon ex binôme et amie intime : Barkat Yamina. Grand merci à tous ceux qui m’ont soutenu de près ou de loin dans l’élaboration de ce travail. Rihana.Zéribi Table de matières Table de matières Contents Remerciements ........................................................................................................................... 3 Dédicace ..................................................................................................................................... 4 Table de matières ....................................................................................................................... 5 Liste des Tableaux ..................................................................................................................... 7 Liste des Figures ........................................................................................................................ 8 Liste des abréviations ................................................................................................................. 9 Introduction générale ................................................................................................................. 1 Chapitre 1 : Synthèse Bibliographique ...................................................................................... 5 1.1. Les légumineuses : .......................................................................................................... 5 1.1.1. Généralités : .............................................................................................................. 5 1.1.2. Classification des légumineuses : ............................................................................. 5 1.1.3. Légumineuses spontanée sahariennes : .................................................................... 8 1.1.4. Taxonomie et caractéristiques du genre Genista : .................................................... 8 1.1.5. Espèce Genista saharae : ......................................................................................... 9 1.2. Les bactéries nodulants les légumineuses (BNL): ........................................................ 10 1.2.1. Le rhizobia : ............................................................................................................ 10 1.2.2. Les différentes espèces nodulant la légumineuse Genista saharae: ....................... 13 1.3. L’Association rhizobia-légumineuse fixatrice d'azote : ................................................ 13 1.3.1. La symbiose légumineuse rhizobium-: .................................................................. 13 1.3.2. Formation des nodules : .......................................................................................... 15 1.3.3. Processus de nodulation : ....................................................................................... 15 1.3.4. Fixation d’azote par la symbiose rhizobienne : ...................................................... 16 Chapitre 2 : Matériels et Méthodes .......................................................................................... 21 2.1. Présentation de la région d’étude : ............................................................................... 21 2.2. Mode opératoire : .......................................................................................................... 22 2.2.1. Collecte des nodules : ............................................................................................. 22 2.2.2. Conservation des nodules : ..................................................................................... 22 2.2.3. Isolement, purification et conservation des souches à partir des nodules : ............ 23 2.2.4. Caractères morphologiques : .................................................................................. 27 2.2.5. Caractérisation symbiotique : ................................................................................. 28 2.2.6. Caractérisation biochimique des isolats : ............................................................... 29 Chapitre 3 : Résultats et Discussions ....................................................................................... 31 Table de matières 3.1. Caractérisation morphologique : ................................................................................... 31 3.1.1. Caractérisation macroscopique : ............................................................................. 31 3.1.2. Caractérisation microscopique : ............................................................................. 31 3.2. Résultats de test de nodulation : .................................................................................... 32 3.2.1. Résultats de Mahdhi (2007) : .................................................................................. 32 3.2.2. Résultats de Chaich (2017) : ................................................................................... 34 3.2.3. Résultats de Ahnia (2019) : .................................................................................... 35 3.3. Caractérisation biochimique des BNL associés à G. saharae :..................................... 39 3.3.1. Résultats de Mahdhi : ............................................................................................. 39 3.3.2. Résultats de Dekak : ............................................................................................... 41 Conclusion ............................................................................................................................... 46 Bibliographie............................................................................................................................ 49 Annexes.................................................................................................................................... 58 Résumé ..................................................................................................................................... 66 Liste des Tableaux Liste des Tableaux Tableau 1: Taxonomie des légumineuses (Baba arbi,2016). ................................................................. 6 Tableau 2: Taxonomie de Genista (Tourene, 2018). ............................................................................. 9 Tableau 3: Exemples d'associations entre Rhizobia et Légumineuses (Selami, 2017). ....................... 12 Tableau 4: Les différentes espèces nodulant la légumineuse Genista saharae. ................................... 13 Tableau 5: La nodulation de Genista saharae par différentes souches (Mahdhi, 2007). ..................... 32 Tableau 6: La nodulation de Genista saharae inoculée par les souches représentative et la souche type de référence E.garamanticus (Chaich, 2018). ....................................................................................... 34 Tableau 7: Gamme d'hôtes des souches de Retama sphaerocarpa (Ahnia, 2019). .............................. 37 Tableau 8: Croissance des souches dans différents sucres et acides aminés (Mahdhi, 2007). ............ 40 Liste des Figures Liste des Figures Figure 1: Phylogénie de léguminosae (Chaich, 2018). .......................................................................... 7 Figure 2: Photo de Genista saharae Coss & Dur (Kherraze et al., 2010) ............................................. 10 Figure 3: Déférentes étapes de l'établissement de la symbiose rhizobia-légumineuse (Aouadj et Saidi Sief 2015). ............................................................................................................................................
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