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Table of contents
CHAPTER I: INTRODUCTION
I. EARLY CELLULAR EVENTS TRIGGERED BY WOUNDING AND HERBIVORY
1. Sensing cell wall integrity and cell death
2. Ion fluxes and other second messengers
3. Production of Reactive Oxygen Species (ROS)
4. Damage-Associated Molecular Patterns (DAMPs) are intercellular signaling molecules produced upon wounding
5. JA is a central actor of plant responses to wounding and insects
6. Long-distance signaling
7. How to differentiate herbivory from mechanical wounding?
1. A great transcriptional reprogramming largely dependent on JA
2. Various defense responses
III. MITOGEN-ACTIVATED PROTEIN KINASE (MAPK) MODULES ARE KEY SIGNALING ACTORS OF ENVIRONMENTAL PERCEPTION
1. MAPKs are important stress responsive modules in plants
2. Spare data suggest a complex MAPK role in wounding and herbivory interaction
IV. OBJECTIVES OF THE PHD WORK: UNVEILING THE FUNCTION OF MKK3 MODULES IN WOUNDING SIGNALING
1. MAPKs and signal transduction before this work, a state of knowledge
2. Objectives: characterizing the activation of a MAPK module upon wounding stress
CHAPTER II: RESULTS & DISCUSSIONS
I. TOWARD A GENERAL MODEL FOR THE ACTIVATION OF MKK3-RELATED MODULES
1. Identification of MAP3Ks able to interact with MKK3 using the yeast 2-hybrid system
2. Functional validation of MAP3Ks using a transient expression system
3. Sub-clade III MAP3Ks are strongly transcriptionally regulated by stresses
4. Discussion
II. IDENTIFICATION AND CHARACTERIZATION OF NEW MAPK MODULES ACTIVATED BY WOUNDING
1. Wounding activates an MKK3 module in Arabidopsis thaliana
2. The wounding-induced activation of the iconic MPK3 and 6 is not dependent on MKK3
3. The hunt of upstream MAP3K(s)
4. Discussion
Characterization of a MAPK module involved in Arabidopsis response to wounding
III. IDENTIFICATION OF SIGNALING ELEMENTS ACTING UPSTREAM MKK3 UPON WOUNDING
1. Exogenous application of some candidate second messengers can activate an MKK3-dependent module
2. Wounding-induced activation of the MKK3-dependent module is disrupted in mutants of the JA signaling pathway
3. MKK3-MPK2 is not activated in systemic leaves
4. Discussion
IV. ACTIVATION OF AN MKK3-DEPENDENT MODULE BY CELL WALL-DAMAGING PATHOGENS
1. Botrytis cinerea activates an MKK3-dependent module in Arabidopsis thaliana 68
2. Spodoptera littoralis activates an MKK3-dependent module in Arabidopsis thaliana
3. Discussion
V. STUDY OF THE ROLE OF AN MKK3-DEPENDENT MODULE IN WOUNDING-RELATED RESPONSES
1. Plants impaired in MKK3 signaling do not show clear detectable phenotype upon Botrytis infection
2. Plants impaired in MKK3 signaling do not properly respond to herbivorous insects
3. Wounding-induced trichome formation
4. Transcriptomic analysis of mkk3-1 plants submitted to wounding did not highlight any MKK3-regulated genes
5. Discussion
CHAPTER III: CONCLUSIONS & PERSPECTIVES
I. A MAPK SIGNALING NETWORK ACTIVATED IN RESPONSE TO WOUNDING
1. Toward a sequential and interdependent activation of two MAPK modules
2. Spatial organization of MAPK activation upon wounding
II. A NOVEL MKK3-DEPENDENT MODULE ACTIVATED BY ENVIRONMENTAL CONSTRAINTS THROUGH TRANSCRIPTIONAL REGULATION OF MAP3K GENES
1. An emerging general working model…
2. …coexisting with other models?
3. Open questions about the functioning of sub-clade III MAP3Ks-MKK3-C-group MAPKs modules
III. A ROLE FOR MKK3 IN STRESS RESPONSES?
1. A well-conserved MAPK through evolution
2. A role in herbivory signaling?
3. Characterized phenotypes in other stress and developmental contexts
4. The hunt of MPK1/2/7/14 substrates
CHAPTER IV: MATERIALS & METHODS
I. MATERIALS
1. Plant material
2. Culture media and conditions
3. Antibodies
4. Vector backbones
5. Buffers and solutions
II.METHODS
1. Plant methods
2. Molecular biology methods
3. Biochemistry methods
CHAPTER V: REFERENCES
ANNEX: Review – Convergence of Multiple MAP3Ks on MKK3 Identifies a Set of Novel Stress MAPK Modules
