| Acknowledgements | 第1-18页 |
| List of Tables | 第18-19页 |
| List of Figures | 第19-22页 |
| Abstract | 第22-26页 |
| 摘要 | 第26-28页 |
| Chapter 1 General Introduction | 第28-32页 |
| Chapter 2 Literature Review | 第32-44页 |
| ·Biology of M. oryzae | 第32-37页 |
| ·Blast disease cycle | 第32-34页 |
| ·Appressorium formation | 第34-36页 |
| ·Blast symptoms | 第36-37页 |
| ·Plant penetration and colonization | 第37-43页 |
| ·Signaling pathways | 第37-42页 |
| ·G protein signaling | 第37-38页 |
| ·Autophagy | 第38-39页 |
| ·Mitogen-activated protein (MAP) kinase signaling | 第39-40页 |
| ·Cyclic adenosyl monophosphate (CAMP) signaling | 第40-41页 |
| ·Ca~(2+) -mediated signaling | 第41-42页 |
| ·Mucilage | 第42页 |
| ·Hydrophobin | 第42页 |
| ·Melanin | 第42-43页 |
| ·Concluding remarks | 第43-44页 |
| Chapter 3 Materials and Methods | 第44-84页 |
| ·Growth conditions of fungal strains | 第44-45页 |
| ·Media composition | 第45-51页 |
| ·Preparation of competent E. coli cells (DH5α) | 第51页 |
| ·Molecular cloning and disruption of targeted genes | 第51-62页 |
| ·Bioinformatics analysis | 第51页 |
| ·Primer designing (for targeted genes of interest) | 第51-52页 |
| ·PCR amplification (up/dn flanking sequences of targeted gene) | 第52-53页 |
| ·Nucleic acids | 第53-54页 |
| ·Ligation (up/dn fragment in pUCm-T vector) | 第54页 |
| ·Transformation (E. coli DH5α competent cells with pUCm-T vector) | 第54-55页 |
| ·Plasmid pUCm-T isolation and confirmation | 第55-57页 |
| ·Purification (up/dn fragment) | 第57页 |
| ·Ligation (up/dn purified fragment into pBS vector) | 第57-59页 |
| ·Transformation (pBS into E. coli DH5α competent cel | 第59-60页 |
| ·Plasmid pBS isolation and confirmation | 第60页 |
| ·Purification and ligation | 第60页 |
| ·Transformation (pCAMBIA-1300 into AGL-1 cells) | 第60-61页 |
| ·Confirmation of pCAMBIA-1300 (PCR and restriction digestion) | 第61页 |
| ·ATMT transformation (using Guy-11 strain conidia) | 第61-62页 |
| ·Confirmation of the gene knock-out transformants (Check PCR) | 第62-64页 |
| ·Genomic DNA isolation (Quick and safe DNA extraction method) | 第63页 |
| ·Check PCR confirmation (Guy-11 and mutants gDNA) | 第63-64页 |
| ·Mono-conidial isolation and storage of deleted transformants | 第64页 |
| ·Southern blot analysis | 第64-73页 |
| ·High quality genomic DNA isolation (2% CTAB method) | 第65-66页 |
| ·Southern blotting (DIG High Prime DNA Labeling) | 第66-73页 |
| ·Reverse Transcription PCR (Guy-11, mutants and rescued strains RNA) | 第73-75页 |
| ·RNA isolation (Guy-11, mutants and rescued strains mycelium) | 第73-74页 |
| ·RT (Reverse transcription) PCR | 第74-75页 |
| ·Real Time PCR (Guy-11, mutants and rescued strains cDNA) | 第75-76页 |
| ·Phenotypic studies (Guy-11, mutants and rescued strains) | 第76-81页 |
| ·Assays for mycelium radial growth and conidiation | 第76-77页 |
| ·Assays for mycelium radial growth on MM, MM-N and MM-C medium | 第77-78页 |
| ·Analysis for conidial germination and appressorium formation | 第78页 |
| ·Cell wall integrity test (Congo red) | 第78-79页 |
| ·Stress response under chemical pressure (ZnSO_4, CuSO_4 and CdCl_2) | 第79页 |
| ·Wettable phenotype (H_2O and 0.2% gelatin) | 第79页 |
| ·Osmotic and oxidative stress response (NaCl and H_2O_2) | 第79页 |
| ·Plant cultivars and growth conditions | 第79-80页 |
| ·Infection assays (Rice and barley) | 第80-81页 |
| ·Mating with 2539 strain | 第81页 |
| ·Complementation of the mutant with the wild-type allele | 第81-82页 |
| ·Generation of GFP fusion constructs | 第82-84页 |
| Chapter 4 An Mitochondrial Fission Protein(MoFIS1) Is Essential for the Blast FungusMagnaporthe oryzae to cause Full Pathogenicity | 第84-104页 |
| ·Abstract | 第84-85页 |
| ·Introduction | 第85-87页 |
| ·Results | 第87-100页 |
| ·Identification and isolation of MoFIS1 | 第87-88页 |
| ·Disruption of MoFIS1 | 第88-89页 |
| ·MoFIS1 affects mycelium radial growth and conidiation | 第89-91页 |
| ·MoFIS1 mutants showed reduced growth on MM, MM-N and MM-C medium | 第91-92页 |
| ·MoFIS1 role in conidial germination and appressorium formatio | 第92-93页 |
| ·MoFIS1 does not affect cell wall integrity | 第93-94页 |
| ·MoFIS1 role in surface hydrophobicity | 第94-95页 |
| ·MoFIS1 is involved in stress response under chemicals pressure | 第95页 |
| ·MoFIS1 role in hypersensitivity to osmotic and oxidative stress | 第95-97页 |
| ·MoFIS1 is critical for full virulence of M. oryzae | 第97-98页 |
| ·MoFIS1 is not required for sexual reproduction | 第98-99页 |
| ·Complementation of the mutant MoFISl with the wild-type MoFIS1 allele | 第99页 |
| ·Subcellular localization of MoFIS1-GFP fusion protein | 第99-100页 |
| ·Discussion | 第100-102页 |
| ·Conclusions | 第102-104页 |
| Chapter 5 Molecular Characterization of a Carbon Catabolite De-repressing (MoYPL1) Genein Magnaporthe oryzae | 第104-117页 |
| ·Abstract | 第104页 |
| ·Introduction | 第104-106页 |
| ·Results | 第106-115页 |
| ·Isolation of Mo YPL1 | 第106-108页 |
| ·Disruption of Mo YPL1 | 第108-109页 |
| ·Assay for mycelium radial growth and conidiation | 第109-110页 |
| ·Assay for conidial germination and appressorium formation | 第110页 |
| ·Mutants of MoYPL1 showed normal growth on MM, MM-C and MM-N medium | 第110-111页 |
| ·MoYPL1 plays no role in cell wall integrity | 第111-112页 |
| ·MoYPL1 is not involved in stress response under chemicals pressure | 第112-113页 |
| ·MoYPL1 plays no role in hypersensitivity to osmotic and oxidative stress | 第113页 |
| ·Pathogenicity test (barley) | 第113-114页 |
| ·MoYPL1 is not required for sexual reproduction | 第114-115页 |
| ·Discussion | 第115-116页 |
| ·Conclusions | 第116-117页 |
| Chapter 6 PCR Cloning and Molecular Characterization of an Endonuclease (MoNUC1) Genein Magnaporthe oryzae | 第117-125页 |
| ·Abstract | 第117页 |
| ·Introduction | 第117-119页 |
| ·Results | 第119-123页 |
| ·Isolation of MoNUC1 | 第119-120页 |
| ·Disruption of MoNUC1 | 第120-121页 |
| ·Assay for mycelium radial growth and conidiation | 第121页 |
| ·Assay for conidial germination and appressorium formation | 第121页 |
| ·Pathogenicity test (barley) | 第121-122页 |
| ·Mating of MoNUC1 mutants with 2539 strain | 第122-123页 |
| ·Discussion | 第123-124页 |
| ·Conclusions | 第124-125页 |
| Chapter 7 Molecular Characterization of an Apoptosis-Inducing Factor (MoAIF1) Gene inMagnaporthe oryzae | 第125-133页 |
| ·Abstract | 第125页 |
| ·Introduction | 第125-126页 |
| ·Results | 第126-131页 |
| ·Isolation of MoAIF1 | 第126-127页 |
| ·Disruption of MoAIF1 | 第127-129页 |
| ·Assay for mycelium radial growth and conidiation | 第129页 |
| ·Assay for conidial germination and appressorium formation | 第129页 |
| ·Pathogenicity test (barley) | 第129-131页 |
| ·Mating of MoAIF1 mutants with 2539 strain | 第131页 |
| ·Discussion | 第131-132页 |
| ·Conclusions | 第132-133页 |
| Chapter 8 Disruption of a Zinc-Regulated Transcription (MoZAP1) Gene does not affectPathogenicity in Magnaporthe oryzae | 第133-142页 |
| ·Abstract | 第133页 |
| ·Introduction | 第133-135页 |
| ·Results | 第135-140页 |
| ·Isolation of MoZAP1 | 第135-136页 |
| ·Disruption of MoZAP1 | 第136-138页 |
| ·Assay for mycelium radial growth and conidiation | 第138页 |
| ·Assay for conidial germination and appressorium formation | 第138页 |
| ·Pathogenicity test (barley) | 第138-140页 |
| ·Mating of MoZAP1 mutants with 2539 strain | 第140页 |
| ·Discussion | 第140-141页 |
| ·Conclusions | 第141-142页 |
| Chapter 9 Molecular Characterization of a Nitrogen Sensor (MoSTM1) Gene in Magnaportheoryzae | 第142-151页 |
| ·Abstract | 第142页 |
| ·Introduction | 第142-144页 |
| ·Results | 第144-149页 |
| ·Isolation of MoSTM1 | 第144-145页 |
| ·Disruption of MoSTM1 | 第145-147页 |
| ·Assay for mycelium radial growth and conidiation | 第147页 |
| ·Assay for conidial germination and appressorium formation | 第147页 |
| ·Pathogenicity test (barley) | 第147-149页 |
| ·MoSTM1 plays no role in sexual reproduction | 第149页 |
| ·Discussion | 第149-150页 |
| ·Conclusions | 第150-151页 |
| Chapter 10 Calpains-Related Genes (MoCAPN1, MoCAPN3 and MoCAPN4) are not Requiredfor Pathogenicity in Magnaporthe oryzae | 第151-170页 |
| ·Abstract | 第151页 |
| ·Introduction | 第151-153页 |
| ·Results | 第153-168页 |
| ·Isolation of MoCAPN1, MoCAPN3 and MoCAPN4 | 第153-157页 |
| ·Disruption of MoCAPN1, MoCAPN3 and MoCAPN4 | 第157-160页 |
| ·Assay for mycelium radial growth and conidiation | 第160-161页 |
| ·Assay for conidial germination and appressorium formation | 第161页 |
| ·Mutants of MoYPL1 showed normal growth on MM, MM-C and MM medium | 第161-162页 |
| ·MoCAPN1, MoCAPN3 and MoCAPN4 play no role in cell wall integrity | 第162页 |
| ·MoCAPN1, MoCAPN3 and MoCAPN4 are not involved in stress response under chemicals pressure | 第162-163页 |
| ·MoCAPN1, MoCAPN3 and MoCAPN4 play no role in hypersensitivity to osmotic and oxidative stress | 第163-164页 |
| ·Pathogenicity test (rice and barley) | 第164-167页 |
| ·MoCAPN1, MoCAPN3 and MoCAPN4 play no role in sexual reproduction | 第167-168页 |
| ·Discussion | 第168-169页 |
| ·Conclusions | 第169-170页 |
| Conclusions and Future Perspectives | 第170-172页 |
| References | 第172-195页 |
| List of publications | 第195页 |
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