| Table of Contents | 第1-8页 |
| 摘要 | 第8-9页 |
| Abstract | 第9-10页 |
| Abbreviations | 第10-11页 |
| Chapter 1 General Introduction | 第11-26页 |
| ·The arbuscular mycorrhiza symbiosis | 第12-15页 |
| ·AM developmental stages | 第12-15页 |
| ·Molecular mechanisms regulating the AM symbiosis | 第15-19页 |
| ·Recognition and exchange of molecular signals for symbiosis establishment | 第15-16页 |
| ·Symbiosis-related transcriptional changes during AM formation | 第16-19页 |
| ·Calcium-regulated signalling events in cells | 第19-25页 |
| ·Ca~(2+) flux in organelles | 第20-23页 |
| ·Ca~(2+) related gene expression related to AM establishment | 第23-24页 |
| ·Ca~(2+) involvement in plant-biotrophic pathogen interactions | 第24-25页 |
| ·Thesis objectives | 第25-26页 |
| Chapter 2 Material and Methods | 第26-50页 |
| ·Biological materials and growth conditions | 第27-31页 |
| ·Astragalus sinicus L.+Glomus mosseae(EG225) | 第27-28页 |
| ·Medicago truncatula+Glomus intraradices DAOM197198(syn.Rhizophagus irregularis Blaszk.,Wubet,Renker & Buscot) | 第28-29页 |
| ·Medicago truncatula+Glomus intraradices BEG141 | 第29-30页 |
| ·Pisum sativum L.+Glomus intraradices BEG141 | 第30-31页 |
| ·Estimation of mycorrhizal root colonization | 第31-32页 |
| ·Laser Capture Microdissection(LCM) of arbuscule-containing cortical root cells | 第32-33页 |
| ·Tissue preparation | 第32页 |
| ·Dissection of arbuscule-containing cells | 第32页 |
| ·RNA extraction from laser-dissected cells | 第32-33页 |
| ·One-step RT-PCR | 第33页 |
| ·Nucleic acid preparation from spores and roots | 第33-35页 |
| ·RNA extraction | 第33-34页 |
| ·cDNA synthesis | 第34-35页 |
| ·Genomic DNA extraction from fungal spores | 第35页 |
| ·Plasmid DNA preparation | 第35页 |
| ·Fungal gene selection and primer design | 第35-39页 |
| ·Gene selection | 第35-37页 |
| ·Primer design | 第37-39页 |
| ·Polymerase chain reaction (PCR) | 第39-44页 |
| ·Reverse-transcription (RT-PCR) | 第39页 |
| ·Relative quantitative real-time PCR | 第39-40页 |
| ·Absolute quantitative real-time PCR | 第40-41页 |
| ·In situ RT-PCR | 第41-42页 |
| ·Rapid Amplification of cDNA Ends (RACE) | 第42-44页 |
| ·PCR amplification on genomic DNA | 第44-47页 |
| ·Inverse PCR | 第45-46页 |
| ·TAIL-PCR | 第46-47页 |
| ·Yeast complementation | 第47-49页 |
| ·Yeast strains | 第47-48页 |
| ·Plasmid construction and yeast transformation | 第48-49页 |
| ·Growth media and tests | 第49页 |
| ·Statistical analysis | 第49-50页 |
| Chapter 3 Studies of G. mosseae genes expressed before root contact with A. sinicum | 第50-59页 |
| ·Fungal gene expression monitored by reverse-transcription (RT)-PCR | 第51-52页 |
| ·Full length gene sequences and analyses | 第52-55页 |
| ·Localization of G. mosseae Gm152 gene activity in mycorrhizal roots of A. sinicus | 第55-57页 |
| ·Discussion and conclusions | 第57-59页 |
| Chapter 4 Growth and mycorrhiza development in wild-type and mycorrhiza-defective mutantplants | 第59-65页 |
| ·Medicago truncatula wild-type J5 and the mutant TRV25 | 第60-62页 |
| ·Wild type Pisum sativum L. cv Finale and the mutant Pssym36 | 第62-63页 |
| ·Discussion and conclusions | 第63-65页 |
| Chapter 5 Selection of G. intraradices genes related to calcium homeostasis and signalling inarbuscular mycorrhiza interactions | 第65-75页 |
| ·Homology searches and primer design for fungal genes based on selected ESTs in G. intraradices DAOM 197198 (syn. R. irregularis) | 第66-70页 |
| ·Fungal gene expression monitored by reverse-transcription (RT)-PCR | 第70-74页 |
| ·Gene expression in G. intraradices DAOM197198 spores and mycorrhizal roots of M. truncatula | 第70-74页 |
| ·Discussion and conclusions | 第74-75页 |
| Chapter 6 Expression of G. intraradices genes encoding Ca~(2+))-related proteins in interactions withwild-type or Myc-mutant roots of Medicago truncatula | 第75-87页 |
| ·Relative quantitative Real-time RT-PCR | 第76-81页 |
| ·Gene expression in G. intraradices DAOM197198 spores and mycorrhizal M. truncatula roots | 第76页 |
| ·Gene expression in G. intraradices BEG141 spores | 第76-77页 |
| ·Gene expression in G. intraradices BEG141 associated with wild-type and mutant M. truncatula roots | 第77-81页 |
| ·Absolute quantitative Real-time RT-PCR | 第81-83页 |
| ·Discussion and conclusions | 第83-87页 |
| Chapter 7 Localization of gene activity in G. intraradices during presymbiotic stages anddevelopment with roots | 第87-96页 |
| ·In situ RT-PCR | 第88-89页 |
| ·Laser cryo-microdissection | 第89-91页 |
| ·G. intraradices gene expression in P. sativum L. wild-type and mutant genotypes | 第91-94页 |
| ·Discussion and conclusions | 第94-96页 |
| Chapter 8 Functional characterization of three G. intraradices BEG141 genes | 第96-110页 |
| ·Full length gene sequencing and phylogenetic analyses | 第97-101页 |
| ·Yeast complementation assays | 第101-108页 |
| ·Growth tests | 第101-102页 |
| ·Complementation of vcx1 mutants | 第102-103页 |
| ·Complementation of a pmr1 mutant | 第103-108页 |
| ·Discussion and conclusions | 第108-110页 |
| Chapter 9 General discussion and conclusions | 第110-119页 |
| References | 第119-129页 |
| Appendix | 第129-131页 |
| Acknowledgements | 第131页 |
