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These Doctorat. SADRATI Nouari .Pdf الجمهورية الجزائرية الديمقراطية الشعبية وزارة التعليم العالي و البحث العلمي جامعة فرحات عباس، سطيف Université Ferhat Abbas Sétif 1 1 كلية علوم الطبيعة و الحياة Faculté des Sciences de la Nature et de la Vie DEPARTEMENT DE MICROBIOLOGIE N°………………………………………….…………..…….……/SNV/2021 Thèse Présentée par SADRATI Nouari Pour l’obtention du diplôme de Doctorat en Sciences Filière : Biologie Spécialité : MICROBIOLOGIE Thème Isolement, identification et culture des champignons endophytes isolés à partir des plantes médicinales algériennes pour la production des métabolites secondaires biologiquement actifs Soutenue publiquement le 07/04/2021 Devant le Jury Président ROUAG Noureddine Pr. Université Ferhat Abbas - Sétif 1 Directeur de thèse HARZALLAH Daoud Pr. Université Ferhat Abbas - Sétif 1 Examinateurs DEHIMAT Laid Pr. Université Frères Mentouri - Constantine 1 GHADBANE Mouloud Pr. Université Mohamed Boudiaf - M’sila MEDJEKAL Samir MCA. Université Mohamed Boudiaf - M’sila Laboratoire de Microbiologie Appliquée Remerciements Avant toute chose, je remercie DIEU, le tout puissant, pour m’avoir donné le courage et la patience d’achever ce travail. Je remercie vivement mon encadrant le professeur HARZALLAH Daoud pour les orientations et les conseils qu’il n’a pas manqués de me prodiguer durant la réalisation de ce travail, pour sa patience et sa compréhension. Qu’il trouve ici l’expression de ma gratitude et de mon respect. Ma vive reconnaissance va au Pr ROUAG Noureddine, professeur à l’Université Ferhat Abbas Sétif 1 d’avoir accepté de présider le jury de soutenance. J’adresse mes remerciements au Pr DEHIMAT Laid, professeur à l’Université des frères mentouri Constantine 1, Pr GHADBANE Mouloud, et Pr MEDJEKAL Samir, de l’Université Mohamed BOUDIAF - M'Sila, d’avoir accepté de juger ce travail. Je présente ici le témoignage de ma sincère gratitude pour leur aide. Je ne remercierai jamais assez Pr DEMIREL Rasime, pour son amitié et son accueil, votre aide m’a été très précieuse, Merci. Je voudrais également adresser mes remerciements au Pr. ROLAIN Jean Marc et son équipe, surtout pour leur accueil chaleureux, ils m’ont beaucoup aidé pour compléter mon travail de recherche. Un grand merci à mon beau père, BENDAOUD El-Amine de m’avoir corrigé la thèse. A toutes les personnes qui, de près comme de loin, m'ont encouragée aux moments opportuns. Dédicaces A mes très chers parents, sans eux je n’aurai pu être ce que je suis, en reconnaissance de leurs efforts, leurs amours et leurs encouragements durant toutes mes études et mes recherches. A ma petite famille surtout, ma femme Amina et ma petite fille Nour El-Imène Sakina. Sommaire I ....................................................................................................................................................... الملخص Résumé ................................................................................................................................................... II Abstract ................................................................................................................................................ III Liste des figures ................................................................................................................................... IV Liste des tableaux ................................................................................................................................ VI Liste des abréviations ......................................................................................................................... VII Introduction ........................................................................................................................................... 1 I. Revue bibliographique I. Revue bibliographique ......................................................................................................................... 4 I.1. Généralités sur les champignons endophytes ................................................................................ 4 I.1.1. Définition des endophytes ...................................................................................................... 4 I.1.2. Diversité et classification des champignons endophytes ....................................................... 4 I.1.3. Critères de sélection des plantes et d’isolement efficace des endophytes .............................. 5 I.1.3.1. Choix des plantes hôtes et du site d’échantillonnage ..................................................... 5 I.1.3.2. La taille et l’âge des échantillons ................................................................................... 6 I.1.3.3. Influence des saisons ...................................................................................................... 6 I.1.4. Mode de transmission des champignons endophytes ............................................................ 7 I.1.5. Interaction endophyte-hôte .................................................................................................... 7 I.2. Molécules bioactives des champignons endophytes ..................................................................... 8 I.2.1. Molécules antibactériennes .................................................................................................... 9 I.2.2. Molécules antifongiques ...................................................................................................... 10 I.2.2.1. Molécules contre les dermatophytes ............................................................................ 10 I.2.2.1. Molécules contre les phytopathogènes ........................................................................ 11 I.2.3. Molécules anticancéreuses ................................................................................................... 13 I.2.4. Molécules antioxydantes ...................................................................................................... 14 I.2.5. Molécules antivirales ........................................................................................................... 16 I.2.6. Molécules antiparasitaires .................................................................................................... 18 I.2.7. Molécules antidiabétiques .................................................................................................... 19 I.2.8. Molécules antiinflammatoires .............................................................................................. 21 I. 3. Plantes médicinales étudiées ...................................................................................................... 22 I. 3.1. Juniperus oxycedrus L. ....................................................................................................... 22 I. 3.2. Pistacia lentiscus L. ............................................................................................................ 24 II. Matériel et Méthodes II. Matériel et Méthodes ........................................................................................................................ 25 II.1. Matériel biologique.................................................................................................................... 25 II.1.1. Matériel microbien ............................................................................................................. 25 II.1.2. Matériel végétal .................................................................................................................. 26 II.1.3. Lignées cellulaires .............................................................................................................. 26 II.1.4. Produits chimiques ............................................................................................................. 26 II.2. Échantillonnage ......................................................................................................................... 26 II.3. Isolement et purification des champignons endophytes ............................................................ 27 II.4. Dépistage préliminaire de l’activité antimicrobienne ................................................................ 28 II.4.1. Criblage de l'activité antibactérienne et antilevurienne ...................................................... 28 II.4.2. Criblage de l'activité antifongique ...................................................................................... 28 II.5. Identification des champignons endophytes sélectionnés ......................................................... 29 II.5.1. Identification morphologique ............................................................................................. 29 II.5.2. Identification moléculaire des champignons ...................................................................... 30 II.5.2.1. Extraction de l’AND total et purification ................................................................... 30 II.5.2.2. Amplification par PCR, visualisation et purification des amplicons .......................... 30 II.5.2.3. Réaction de séquence (BigDye) et séquençage .......................................................... 31 II.5.2.4. Analyse phylogénétique ............................................................................................. 31 II.6. Sélection du milieu de culture optimum .................................................................................... 32 II.7. Choix du meilleur solvant d'extraction et du type de fermentation ........................................... 32 II.7.1. Fermentation
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