Saccharothrix Algeriensis NRRL B-24137
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Acknowledgements THÈSE En vue de l'obtention du DOCTORAT DE L’UNIVERSITÉ DE TOULOUSE Délivré par Institut National Polytechnique De Toulouse Discipline ou spécialité : Ingénierie microbienne et enzymatique Présentée et soutenue par Saima MUZAMMIL Le 13/07/2012 Saccharothrix algeriensis NRRL B-24137: biocontrol properties, colonization and induced systemic resistance towards Botrytis cinerea on grapevine and Arabidopsis thaliana JURY M. AZIZ Aziz Maître de Conférences, Université de Reims Champagne-Ardenne M. REY Patrice Professeur, Université de Bordeaux Mme. MATHIEU Florence Professeur, Université de Toulouse Mme STRUB Caroline Maître de Conférences, Université de Montpellier II M. LEBRIHI Ahmed Professeur, Université de Toulouse M. COMPANT Stéphane Maître de Conférences, Université de Toulouse Ecole doctorale : École doctorale: Sciences Ecologiques, Vétérinaires, Agronomiques et Bioingénieries Unité de recherche : LGC UMR 5503 (CNRS/UPS/INPT) Directeur de Thèse : Pr. LEBRIHI Ahmed (INP-ENSAT) Co-Directeur de Thèse : Dr. COMPANT Stéphane (INP-ENSAT) 1 Acknowledgements Acknowledgements First of all I am thankful to the almighty ALLAH, whose blessings are always with me. My sincere appreciation goes to my supervisor Professor Ahmed LEBRIHI and co- supervisor Doctor Stéphane COMPANT, whose scientific approach, careful reading and constructive comments were valuable. Their timely and efficient contributions helped me to shape my research work into its final form and I express my sincerest appreciation for their assistance in any way that I may have asked. I deem it utmost pleasure to avail this opportunity to express the heartiest gratitude and deep sense of obligation to Stéphane COMPANT for his dexterous guidance, critical judgment, sympathetic attitude and inspiring efforts to inculcate in me the spirit of motivation during the course of my research work. He was always available whenever I need his assistance and guidance, especially, during thesis writing. Sincere thanks are due to Professor Florence MATHIEU for her kindness and helping in thesis work. I also wish to thank the “Higher Education Commission (HEC)” of Pakistan, its leadership and the staff who in their limited resources supported me financially for my studies in France. I must also mention the services provided by SFERE (Société Française d' Exportation des Ressources Educatives) to facilitate my living in France. I fervently extend my zealous thanks to the members of my thesis jury Dr. AZIZ Aziz, Pr. REY Patrice, Pr. MATHIEU Florence, Dr. STRUB Caroline and my two directors. I would like to reflect my gratitude to Professor Nasserdine SABAOU, Professor Abdelghani ZITOUNI and Atika MEKLAT who always helped me in every aspect of my work and gave me strong support. Special thanks to Amine YEK and Omrane TOMATIA who always supported me in every difficult situation. A bundle of thanks to Rayenne SARIA, Zhuoran.YU and Clotilde GRAILLON for their great help and nice cooperation and unforgettable company. 2 Acknowledgements I am also indebted to my colleagues at Laboratoire de Genie Chimique (LGC) and I would like to reflect my gratitude particularly Carol VERHEECKE, Safwan SAKER, Philippe ANSON and Marion DARRIET who always welcomed, helped and appreciated me. Finally, I am forever indebted to my family to my mother and father. A special thanks to my husband Muhammad Hussnain Siddique who always shares my problems with an ultimate solution, and cooperated at each and every crucial step of my life. My humble thanks to my sisters and brother. I dedicate this thesis in honor of my family. 3 Summary Summary Résumé………………………..........................................................................................7 Abstract……………………….........................................................................................8 List of Abbrevations………….........................................................................................9 Foreward……………………….....................................................................................10 Chapter I: Introduction………………………………………………………….........11 I. Generality on rhizospheric and/or endophytic bacteria………………. ............... 12 1. Definition and History ........................................................................................ 12 2. Sources and Niches of colonization of PGPR and endophytes ........................... 14 3. Functions of rhizo- and endophytic bacteria ....................................................... 19 3.1 Direct plant growth promotion by PGPR and endophytic bacteria ............... 19 (a) Biological nitrogen fixation ................................................................... 19 (b) Solubilisation of phosphorus .................................................................. 20 (c) Phytohormone production ....................................................................... 21 (d) Production of sulphur ............................................................................. 21 3.2 Indirect effect by PGPR and/or Endophytic bacteria .................................... 22 II. Use of rhizosphere and endophytic bacteria for biocontrol of phytopathogens .22 1. Competition for Space and Nutrients .................................................................. 24 2. Antibiosis ............................................................................................................. 25 3. Lytic enzymes secretions ..................................................................................... 27 4. Induced Systemic Resistance .............................................................................. 27 III. Use of rhizospheric and endophytic bacteria and their secondary metabolites to protect grapevine towards diseases ......................................................................... 34 Compant S., Brader G., Muzammil S., Sessitsch A., Lebrihi A. and Mathieu F. Use of beneficial bacteria and their secondary metabolites to control grapevine pathogen diseases. Accepted with minor changes in Biocontrol ............................................................... 36 4 Summary IV. The case of rhizo- and endophytic actinobacteria and rare species for plant health: an interesting area for biocontrol studies ...................................................... 71 1. Interesting genera and species for biocontrol and secretion of bioactive products ……………………………………………………………………….………..….71 2. Actinosynnemataceae and Saccharothrix spp., uncommon bacteria ............ …74 3. Saccharothrix algeriensis NRRL B-24137: a model used for characterization of new bioactive metabolites ..................................................................................... 76 4. Does Saccharothrix algeriensis NRRL B-24137 can be used to protect plants to- wards phytopathogens? But it came from desert soil and can secrete some antibio- tics!…………………………………………………………………………….…78 5. Saccharothrix algeriensis NRRL B-24137/grapevine-Arabidopsis thaliana/Botrytis cinerea as a model?....................................................................79 Objectives of the thesis .................................................................................................. 80 Chapter II ....................................................................................................................... 81 Muzammil S., Mathieu F., Lebrihi A., and Compant S. Direct biocontrol activity of Saccharothrix algeriensis NRRL B-24137 towards Botrytis cinerea Pers. Unpublished....................................................................................................................82 Chapter III ..................................................................................................................... 97 Compant S., Muzammil S., Lebrihi A., and Mathieu F. Visualization of root colonization of grapevine by the Saharan soil isolate Saccharothrix algeriensis NRRL B-24137 by using DOPE-FISH microscopy and a probe specific to Saccharothrix spp. Plant and Soil submitted...............................................................................................98 Chapter IV ................................................................................................................... 116 Muzammil S., Graillon C., Mathieu F., Lebrihi A., and Compant S. Systemic resistance of grapevine plants towards Botrytis cinerea induced by the desert soil bacterium Saccharothrix algeriensis NRRL B-24137. Unpublished ................................................................................................................. 117 5 Summary Chapter V ..................................................................................................................... 127 Muzammil S., Graillon C., Saria R., Mathieu F., Lebrihi A., and Compant S. The desert isolate Saccharothrix algeriensis NRRL B-24137 can be endophytic in Arabidopsis, induces ISR towards Botrytis cinerea as well as allows to determine new mechanisms involved. Molecular Plant-Microbe Interaction, submitted .................................................... 128 Chapter VI Conclusions and Future Prospects............................................................................. 153 References .................................................................................................................... 166 Curriculum Vitae of the PhD candidate ..................................................................... 195 6 Résumé Résumé Au cours de cette thèse, un isolat de sol de désert, Saccharothrix algeriensis NRRL B-24137,