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Thèse Pour L’Obtention Du Grade De Docteur En Sciences

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Thèse Pour L’Obtention Du Grade De Docteur En Sciences الجمهورية الجزائرية الديمقراطية الشعبية REPUBLIQUE ALGERIENNE DEMOCRATIQUE ET POPULAIRE وزارة التعليم العالي والبحث العلمي MINISTERE DE L’ENSEIGNEMENT SUPERIEUR ET DE LA RECHERCHE SCIENTIFIQUE جامعة عبد الحميد ابن باديس. مستغانم. UNIVERSITE ABDELHAMID IBN BADIS DE MOSTAGANEM Faculté des Sciences de la Nature et de la Vie Département d’Agronomie THÈSE POUR L’OBTENTION DU GRADE DE DOCTEUR EN SCIENCES Option : Protection des végétaux Présentée par : Melle BADAOUI Mahdjouba (Ikram) Contribution à l'étude de la dynamique des populations de Tuta absoluta Meyrick (Lepidoptera ; Gelechiidae) et essais de contrôle biologique sur la culture de tomate » Devant le Jury : YOUCEF BENKADA Mokhtar Professeur Président Univ-Mostaganem BELABID Lakhdar Professeur Examinateur Univ-Mascara BELAHCEN Miloud Professeur Examinateur Centre univ -Ain Témouchent BENDAHMANE Boubekeur Seddik Professeur Examinateur Univ- Mostaganem BICHE Mohamed Professeur Examinateur I.N.A-Alger BERKANI Abdellah Professeur Directeur de thèse Univ-Mostaganem Année Universitaire : 2017/2018 REMERCIEMENTS Je remercie Dieu le Tout Puissant ALLAH de m’avoir donné les ressources morales, physiques, matérielles et intellectuelles pour accomplir cette thèse. Ce travail n'aurait certainement pas pu voir le jour sans l'assistance de plusieurs personnes. Je citerai à leur tête mon directeur de thèse M. BERKANI Abdallah., professeur à l’université de Mostaganem. Malgré la somme de ses responsabilités, sa disponibilité, et sa patience n’ont jamais manqué. Je lui suis reconnaissante de m’avoir appuyé à maintes reprises, pour toute la confiance qu’il m’a accordée, en acceptant de me diriger dans ce travail. Ses compétences en dynamique des populations ont été d'un précieux apport pour ce travail. Sa fermeté scientifique a permis de répondre à beaucoup d’interrogations concernant ce sujet de recherche. Je lui suis énormément reconnaissante pour les nombreuses lectures de ce manuscrit et pour le temps qu’il a consacré pour m’aider. Merci aussi d’être un père avant d’être un encadreur. Mes remerciements vont aussi à M. SIMONEAU Philippe, professeur à l’université d’Angers, à qui j'exprime toute ma gratitude pour m’avoir apporté ses connaissances en biologie moléculaire et m’avoir permis de réaliser des manipulations dans son laboratoire. Je le remercie chaleureusement pour son esprit généreux, sa gentillesse, sa compréhension, sa patience, son attention, ses conseils et ses regards critiques sur mes résultats durant les travaux de la caractérisation génétique des B. bassiana. Merci à tous les membres de l’équipe FungiSem (Laboratoire Phytopathologie et Semances) et l’équipe ALSA (Laboratoire Physiologie des Semances). Mes sincères remerciements s’adressent également à M. YOUCEF BENKADA Mokhtar, Professeur à l’Université de Mostaganem, qui m’a fait l'honneur d'accepter de présider le jury. Cette thèse n’aurait pas sa valeur sans le jury ; je tiens à remercier vivement ses membres, d’avoir eu l'amabilité d'accepter de faire partie du jury et de donner de leur temps précieux pour juger ce travail: M. BELABID Lakhdar, Professeur à l’Université de Mascara, M. BELAHCEN Miloud, Professeur à l’Université de Ain Témouchent, M. BENDAHMANE Boubekeur Seddik, Professeur à l’Université de Mostaganem et M. BICHE Mohamed, Professeur à l’Ecole Nationale Supérieure Agronomique d’Alger. Je tiens à exprimer mes vifs remerciements à tous les membres des équipes du laboratoire de protection des végétaux de l’Université de Mostaganem. Par ces quelques lignes je tiens aussi à remercier les personnes avec qui j’ai pu collaborer dans le cadre de mes travaux de recherche. Merci aux Professeurs KEDAD A. et SAHRAOUI L. de l’INA d’Alger, et aux Professeurs CHABASSE D. et BOUCHARA J.P de la CHU d’Angers. Merci à mes collègues Mme KOLAI N pour l’inventaire des auxilliaires de T. absoluta, Mme SAIAH F. et Mme BENNAMA R. maître de conférences à l’Université de Mostaganem pour leur aide précieuse et le partage de leur connaissance dans le monde des micro-organismes. Une grande part de mes remerciements s’adresse aux membres de ma famille pour leur soutien moral, particulièrement ma très chère mère NADJIBA, lumière de ma vie, à qui je dois ce que je suis. Je voudrais rendre hommage à la mémoire de mes chers amis (es) NAOUEL, WAFA et HALIM et mes chères enseignants M. MOKHBI A. et M. DIDI M. Que dieu vous accueil dans son vaste paradis. Je ne pourrais passer sous silence l'apport de mes amis (es) qui m’ont soutenu et ont partagé avec moi les meilleurs et les pires des moments, en particulier Farida qui a toujours sue m’aider, m’encourager et me motiver pour me faire avancer, je lui souhaite plein de courage et surtout beaucoup de bonheur avec sa famille. Merci également à Malika, Mostapha, Soulem, Zoula, Karima, Yasmine, Moufida, Soumia, Fouzia, Luiza, Fatima, Saida, Milouda et à ma voisine Malika. Un énorme merci à M. BOUZOUINA M., M. MKHARBECH A., M. CHEIKHI L., M. BENCHIKH M., Mme KETROUSSI N. et M. SOUANE A. pour leur gentillesse et leur aide. Je suis très reconnaissant à M. AZZEDINE et à tous les ouvriers de l’exploitation agricole de notre département. Je voudrais remercier et exprimer ma gratitude à tous les enseignants qui ont contribué à ma formation. BADAOUI Mahdjouba Ikram Résumé Résumé En Algérie, la culture de la tomate en plein champ et sous serre, a subi des dégâts considérables, provoqués par un phytophage redoutable : la mineuse de la tomate Tuta absoluta Meyrick (Lepidoptera: Gelechiidae). Le présent travail est une contribution à l’étude de ce déprédateur, il s’intéresse a plusieurs aspects et a été entamé par l’identification de l’espèce du ravageur par la méthode d’extraction des génitalia afin d’éviter toute confusion avec des espèces morphologiquement proches, ce qui nous a permis de déterminer avec exactitude qu’il s’agit bien de T. absoluta. Le deuxième volet de ce travail s’est axé sur l’étude de la dynamique des populations sous serre de ce déprédateur dans la région de Mostaganem sur deux années consécutives (2012 et 2013). Les résultats obtenus ont montré que le taux d’infestation de cette espèce est important et a atteint 100% la première année d’étude et 80% la seconde. L’étude de la densité et la distribution des différents stades évolutifs de T. absoluta, témoigne, d’un accroissement rapide des populations dans l’espace et dans le temps. L’inventaire de la faune auxiliaire de T. absoluta a révèlé la présence de deux punaises prédatrices (Nesidiocoris tenuis et Macrolophus sp.) et de deux espèces de parasitoïdes (Necremnus artynes et Diglyphus isaea), avec une prédominance de N. artynes. La troisième partie a été consacrée à la recherche des entompathogènes autochtones capables de réguler les populations de cet insecte nuisible, ce qui nous a permis d’isoler un certain nombre de bactéries et de champignons, où seuls les isolats du genre Beauveria étaient capables d’induire des infections vis-à-vis des larves de Tuta absoluta. Deux Beauveria sp. (BV1 et BV6) parmi les huit entomopathogènes isolés ont fait l’objet des tests approfondis in vitro pour évaluer leurs activités larvicides. Il s’est avéré que la souche BV6 était la plus efficace avec une DL50 de 10 2,97 spores/ml. La dernière étape de notre démarche avait comme objectif l’identification moléculaire et l’étude de la variabilité génétique des huit champignons entomopathogènes autochtones. L’analyse des régions ITS et du gène de facteur d’élongation (EF1-) montre l'appartenance des souches à Beauveria bassiana. En plus l’absence des introns dans le gène du grand ARN ribosomique (LSU rDNA) indique que nos souches sont des pseudobassiana décrite clade C. La diversité phylogénétique des huit Beauveria bassiana (pseudobassiana) a été examinée en analysant les inter-microsatellites (ISSR), les séquences nucléotidiques de deux locus des régions intergéniques mitochondriales (nad3- atp9 et atp6-rns) et les microsatellites (SSR). Les résultats des ISSR montrent une variation entre les champignons et le séquençage du locus ATP6 confirme ce polymorphisme et classe les huit souches en deux groupes distincts. Mots clés : Tuta absoluta, génitalia, dynamique des populations, champignons entomopathogènes, activité larvicide, caractérisation moléculaire. Abstract Abstract In Algeria, the cultivation of tomatoes in open fields and greenhouses has suffered from considerable damage, caused by harmful phytophagous: tomato leafminer Tuta absoluta Meyrick (Lepidoptera: Gelechiidae). The present work is a contribution to the study of this pest. It is interested in several aspects and has been initiated by the identification of the species of the pest through extraction of the genitalia in order to avoid any confusion with morphologically similar species, which enabled us to determine with precision that it is indeed T. absoluta. The second part of this work focuses on the study of the dynamics of the pest populations in Mostaganem region during two consecutive years (2012 and 2013). The results obtained show that the infestation rate of these species is high and reached 100% in the first year of study and 80% in the second year. The study of the density and distribution of the different evolutionary stages of T. absoluta shows a rapid increase of populations in space and time. The inventory of T. absoluta ancillary fauna revealed the presence of two predatory bugs (Nesidiocoris tenuis and Macrolophus sp.) and two species of parasitoids (Necremnus artynes and Diglyphus isaea), with a predominance of N. artynes. The third part of this work was devoted to the investigation of indigenous entomopathogens capable of regulating the populations of this insect pest, which enabled us to isolate a number of bacteria and fungi, where only isolates of the genus Beauveria were able to isolate, induce infections with Tuta absoluta larvae. Two Beauveria sp. (BV1 and BV6) of the eight isolated entomopathogens were subjected to extensive in vitro tests to evaluate their larvicidal activities.
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