| 摘要 | 第4-6页 |
| Abstract | 第6-7页 |
| Nomenclature | 第14-17页 |
| Chapter 1 Introduction | 第17-34页 |
| 1.1 Background, objective and significance of the subject | 第17-18页 |
| 1.2 Literature review | 第18-29页 |
| 1.2.1 Research progress on tomato gray mold | 第18-21页 |
| 1.2.2 Research progress on Gliocladium biological control | 第21-22页 |
| 1.2.3 Transgenic plants and their Application in Agriculture | 第22-25页 |
| 1.2.4 Transgenic strategies against diseases | 第25-27页 |
| 1.2.5 Strategies against bacterial and fungal disease | 第27-29页 |
| 1.3 Objectives of this research | 第29页 |
| 1.4 Main research content of this thesis | 第29-34页 |
| 1.4.1 Defense-related enzymes response | 第29-30页 |
| 1.4.2 Hypersensitive response | 第30-31页 |
| 1.4.3 Over-expression of plant defense related genes | 第31-32页 |
| 1.4.4 Function and transformation of atpA synthase | 第32-34页 |
| Chapter 2 Materials and Methods | 第34-68页 |
| 2.1 Materials | 第34-36页 |
| 2.1.1 Microbial strains | 第34页 |
| 2.1.2 Bacterial strains and plasmid | 第34页 |
| 2.1.3 Microbial culture Media and culture condition | 第34页 |
| 2.1.4 Bacterial culture media and culture condition | 第34-35页 |
| 2.1.5 Plants material and tissue culture Media | 第35-36页 |
| 2.1.6 Plant tissue culture conditions | 第36页 |
| 2.1.7 Other chemicals and materials | 第36页 |
| 2.2 Methods | 第36-68页 |
| 2.2.1 Sterilization | 第36-37页 |
| 2.2.2 Treatment and infection of tomato plant | 第37-38页 |
| 2.2.3 Disease Assessment of tomato leaves | 第38页 |
| 2.2.4 Enzyme Activity Assay PAL, PPO, POD, SOD, POX and LOX | 第38-40页 |
| 2.2.5 Assays of Nitric Oxide (NO) and Hydrogen Peroxide (H2O2) | 第40-41页 |
| 2.2.6 Analysis of two proteins in infected tomato plants: atpA and Lexyl2 | 第41-44页 |
| 2.2.7 Cloning of atpA gene | 第44-52页 |
| 2.2.8 Expression Analysis of atpA gene in tobacco plant | 第52-59页 |
| 2.2.9 Resistance of tobacco transgenic plants | 第59-67页 |
| 2.2.10 Statistic analysis | 第67-68页 |
| Chapter 3 Induced resistance in tomato leaves infected by gray mould | 第68-82页 |
| 3.1 Introduction | 第68-69页 |
| 3.2 Effects of C. rosea on tomato gray mould | 第69-70页 |
| 3.3 Effects of C. rosea on activities of protective enzymes in tomato infected with gray mould | 第70-75页 |
| 3.3.1 Effects of C. rosea on activities of PAL, POD and PPO | 第70-72页 |
| 3.3.2 Effects of C.rosea on activities of SOD, POX and LOX | 第72-75页 |
| 3.4 Effects of C.rosea treatment on activities of NO and H2O2 | 第75-76页 |
| 3.5 C. rosea promoted and increased the expression level of atpA and Lexyl2 gene in tomato leaves infected by gray mold | 第76-80页 |
| 3.6 Summary | 第80-82页 |
| Chapter 4 Expression and Analysis of atpA gene in tobacco transgenic plants | 第82-94页 |
| 4.1 Introduction | 第82页 |
| 4.2 Verification of atpA gene within the tomato leaves (PCR) | 第82-83页 |
| 4.3 Digestion, ligation and transformation of the recombinant clone | 第83-84页 |
| 4.4 PCR detection and restriction identification of positive clones | 第84页 |
| 4.5 Development of transgenic tobacco plants | 第84-88页 |
| 4.5.1 Callus induction | 第85页 |
| 4.5.2 Shoot and Root development | 第85-86页 |
| 4.5.3 Development in soil | 第86-88页 |
| 4.6 Expression and analysis of Tobacco transgenic plants | 第88-91页 |
| 4.6.1 PCR analysis of tobacco transgenic plant | 第88-89页 |
| 4.6.2 Real time- PCR analysis | 第89-91页 |
| 4.7 Transgenic expression analysis | 第91-93页 |
| 4.7.1 Southern blot | 第91-92页 |
| 4.7.2 Northern blot | 第92-93页 |
| 4.8 Summary | 第93-94页 |
| Chapter 5 Function of atpA gene in tobacco transgenic plants infected | 第94-112页 |
| 5.1 Introduction | 第94页 |
| 5.2 Changes of protective enzyme activities | 第94-98页 |
| 5.2.1 Changes of PAL and PPO | 第95-97页 |
| 5.2.2 Changes of POD and SOD | 第97-98页 |
| 5.3 Changes of second messenger activites | 第98-102页 |
| 5.4 Changes of proteins activites | 第102-105页 |
| 5.4.1 Changes of PR1 | 第102-103页 |
| 5.4.2 Changes of Ca2+Mg2+-ATPase and Ca2+-ATPase | 第103-105页 |
| 5.5 Changes of chlorophyll content | 第105-107页 |
| 5.6 Resistance to B. cinerea in tobacco plants expressing tomato atpA gene | 第107-111页 |
| 5.7 Summary | 第111-112页 |
| Conclusion | 第112-115页 |
| References | 第115-130页 |
| Papers published in the period of Ph.D. education | 第130-132页 |
| Acknowledgements | 第132-134页 |
| Curriculum vitea | 第134-135页 |
| APPENDIX | 第135-144页 |
| A. Plasmid Map | 第135页 |
| B. Microbial and Bacterial culture Media and PDA For 1 liter | 第135-136页 |
| C. Plant Tissue Culture Media | 第136-137页 |
| D. Extraction buffer of defense relative enzyme and Boric acid Buffer | 第137-138页 |
| E. Primers of atpA, Lexyl2 and β-actin genes | 第138页 |
| F. Plasmid DNA extraction solutions and TE Buffer | 第138-139页 |
| G. TAE and TBE buffers | 第139-140页 |
| H. CTAB Buffer | 第140页 |
| I. Southern blotting | 第140-142页 |
| J. Northern blotting | 第142-143页 |
| K. Phosphate and normal saline buffers | 第143页 |
| L. Sequence of PR1 gene | 第143-144页 |
| M. Anatomical solution | 第144页 |
