| 中文摘要 | 第4-6页 |
| Abstract | 第6-8页 |
| Abbreviations | 第9-14页 |
| CHAPTER 1 REVIEW | 第14-36页 |
| 1.0 Introduction | 第14-15页 |
| 1.1 Botrytis cinerea | 第15-32页 |
| 1.1.1 Classification | 第16-18页 |
| 1.1.2 Host Range and Symptoms | 第18-20页 |
| 1.1.3 Life cycle and Epidemiology | 第20-21页 |
| 1.1.4 Disease control | 第21-25页 |
| 1.1.5 B.cinerea virulence under Molecular Approaches | 第25-32页 |
| 1.2 Alcohol Dehydrogenases | 第32-35页 |
| 1.2.1 Molecular Characterization of ADHs | 第33-34页 |
| 1.2.2 The Roles of Alcohol Dehydrogenase in fungal pathogen | 第34-35页 |
| 1.3 Objectives | 第35-36页 |
| CHAPTER 2 MATERIALS AND METHODS | 第36-62页 |
| 2.1 Materials and Equipment | 第36-39页 |
| 2.2 Experimental strains and Growth conditions | 第39-44页 |
| 2.2.1 Experimental strains | 第39-40页 |
| 2.2.2 Growth conditions | 第40-43页 |
| 2.2.3 Reagents and solutions | 第43-44页 |
| 2.3 Molecular Methods | 第44-58页 |
| 2.3.1 Fungal strain | 第44页 |
| 2.3.2 Sequence analysis and design primers | 第44-46页 |
| 2.3.3 qRT-PCR Analysis of Gene Expression | 第46-47页 |
| 2.3.4 Extractiongenomic DNA of B.cinerea | 第47-48页 |
| 2.3.5 PCR reaction system and conditions: | 第48-49页 |
| 2.3.6 Transformation method | 第49-50页 |
| 2.3.7 Plasmid purification | 第50页 |
| 2.3.8 Detection of the concentration of the plasmid | 第50页 |
| 2.3.9 Construction of vector | 第50-56页 |
| 2.3.10 Electric Transformation of the plasmid into Agrobacterium | 第56-57页 |
| 2.3.11 Transformation to B. cinerea | 第57页 |
| 2.3.12 Complementation of mutant | 第57-58页 |
| 2.4 Morphology observation and pathogenicity assessment | 第58-60页 |
| 2.4.1 Microscopic observation | 第58页 |
| 2.4.2 Features of Growth rate, Conidial and Sclerotium | 第58-59页 |
| 2.4.3 Stress responses | 第59页 |
| 2.4.4 Pathogenicity assay | 第59-60页 |
| 2.5 Statistical analyses | 第60-62页 |
| CHAPTER 3 RESULTS | 第62-90页 |
| 3.1 Identification of Alcohol Dehydrogenases (BcADHs) Genes in B.cinerea | 第62页 |
| 3.2 Phylogenetic analysis of BcADHs from different organisms | 第62-63页 |
| 3.3 Real-time PCR demonstration of the up-regulation of the threegenes during infection | 第63-65页 |
| 3.4 Generation of BcADH1 gene knockout mutant △bcadh1 and itscomplementary strains △bcadh1/C | 第65-69页 |
| 3.5 Generation of BcADH2 gene knockout mutant △bcadh2 and itscomplementary strain Abcadh2/C | 第69-70页 |
| 3.6 Generation of BcADH3 gene knockout mutant △bcadh3 and itscomplementary strain △bcadh3/C | 第70-72页 |
| 3.7 Morphological observations of conidiospore shape and size | 第72-75页 |
| 3.8 Characteristics analysis of conidial germination | 第75-77页 |
| 3.9 Features of Radial growth, Sporulation and Sclerotia formation | 第77-80页 |
| 3.10 BcADH genes mediate formation of fungal infection cushion | 第80-81页 |
| 3.11 Stress Responseof BcADHs genes KO results in B. cinerea | 第81-88页 |
| 3.12 Pathogenicity assay | 第88-90页 |
| CHAPTER 4 DISCUSSION | 第90-96页 |
| CHAPTER 5 CONCLUSIONS | 第96-97页 |
| References | 第97-122页 |
| Acknowledgement | 第122-124页 |
| Curriculum vitae | 第124页 |
