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Table of contents
CHAPTER 1
GENERAL INTRODUCTION
1.1. The arbuscular mycorrhiza symbiosis
1.2. Molecular mechanisms regulating the AM symbiosis
1.3. Calcium-regulated signaling events in cells
1.4. Thesis objectives
CHAPTER 2
MATERIAL AND METHODS
2.1. Biological materials and growth conditions
2.2. Estimation of mycorrhizal root colonization
2.3. Laser Capture Microdissection (LCM) of arbuscule-containing cortical root cells
2.4. Nucleic acid preparation from spores and roots
2.5. Fungal gene selection and primer design
2.6. Polymerase chain reaction (PCR)
2.7. PCR amplification on genomic DNA
2.8. Yeast complementation
2.9. Statistical analysis
CHAPTER 3
STUDIES OF G. MOSSEAE GENES EXPRESSED BEFORE ROOT CONTACT WITH A. SINICUM
3.1. Fungal gene expression monitored by reverse-transcription (RT)-PCR
3.2. Full length gene sequences and analyses
3.3. Localization of G. mosseae Gm152 gene activity in mycorrhizal roots of A. sinicus
3.4. Discussion and conclusions
CHAPTER 4
GROWTH AND MYCORRHIZA DEVELOPMENT IN WILD-TYPE AND MYCORRHIZADEFECTIVE MUTANT PLANTS
4.1. Medicago truncatula wild-type J5 and the mutant TRV25
4.2. Wild type Pisum sativum L. cv Finale and the mutant Pssym36
4.3. Discussion and conclusions
CHAPTER 5
SELECTION OF G. INTRARADICES GENES RELATED TO CALCIUM HOMEOSTASIS AND SIGNALLING IN ARBUSCULAR MYCORRHIZA INTERACTIONS
5.1. Homology searches and primer design for fungal genes based on selected ESTs in G. intraradices DAOM 197198 (syn. R. irregularis)
5.2. Fungal gene expression monitored by reverse-transcription (RT)-PCR
5.3. Discussion and conclusions
CHAPTER 6
EXPRESSION OF G. INTRARADICES GENES ENCODING CA2+-RELATED PROTEINS IN INTERACTIONS WITH WILD-TYPE OR MYC- MUTANT ROOTS OF MEDICAGO TRUNCATULA
6.1. Relative quantitative Real-time RT-PCR
6.2. Absolute quantitative Real-time RT-PCR
6.3. Discussion and conclusions
CHAPTER 7
LOCALIZATION OF GENE ACTIVITY IN G. INTRARADICES DURING PRESYMBIOTIC STAGES AND DEVELOPMENT WITH ROOTS
7.1. In situ RT-PCR
7.2. Laser cryo-microdissection
7.3. G. intraradices gene expression in P. sativum L. wild-type and mutant genotypes
7.4. Discussion and conclusions
CHAPTER 8
FUNCTIONAL CHARACTERIZATION OF THREE G. INTRARADICES BEG141 GENES
8.1. Full length gene sequencing and phylogenetic analyses
8.2. Yeast complementation assays
8.3. Discussion and conclusions
