| 摘要 | 第7-8页 |
| ABSTRACT | 第8页 |
| CHAPTER I INTRODUCTION AND REVIEW OF LITRATURE | 第18-30页 |
| 1.1. Plant defense responses | 第18-19页 |
| 1.2. Elicitor-induced resistance | 第19-21页 |
| 1.3. Rice defense response | 第21-23页 |
| 1.4. Abiotic and biotic stresses-related pathways | 第23-27页 |
| 1.4.1. ABA pathway | 第23-24页 |
| 1.4.2. SA signaling pathway | 第24-25页 |
| 1.4.3. JA signaling pathway | 第25页 |
| 1.4.4. NO signaling in plant defense | 第25-26页 |
| 1.4.5. Reactive oxygen species production and programmed cell death | 第26-27页 |
| 1.5. Molecular approaches identifying elicitor-induced disease resistance | 第27-28页 |
| 1.6. Fungal elicitors trigger disease resistance in rice | 第28-29页 |
| Objectives | 第29-30页 |
| CHAPTER II Fungal elicitor Mo Hrip2 triggers expression of pathogenesis-related genes and synthesis of SA and JA in rice leaves | 第30-45页 |
| 2.1. Introduction | 第30-31页 |
| 2.2. Materials and methods | 第31-36页 |
| 2.2.1. Protein elicitor purification | 第31-32页 |
| 2.2.2. Mo Hrip2 purification using column chromatography | 第32页 |
| 2.2.3. Confirmation of Mo Hrip2 by SDS-PAGE and tobacco leaf HR response | 第32-33页 |
| 2.2.4. Rice growth conditions and sample preparation | 第33页 |
| 2.2.5. Total RNA isolation | 第33-34页 |
| 2.2.6. c DNA synthesis for q RT-PCR | 第34页 |
| 2.2.7. q RT-PCR | 第34-35页 |
| 2.2.8. SA quantification by HPLC | 第35页 |
| 2.2.9. JA quantification by GC-MS | 第35-36页 |
| 2.2.10. Data analysis | 第36页 |
| 2.3 Results | 第36-42页 |
| 2.3.1 Expression profile of defense-related TFs and SA and JA pathway markergenes | 第36-38页 |
| 2.3.2 Expression profile of defense-related genes over time | 第38-40页 |
| 2.3.3 Endogenous levels of SA and JA in rice leaves | 第40-42页 |
| 2.4 Discussion | 第42-45页 |
| CHAPTER III Fungal elicitor Mo Hrip2 induces disease resistance in rice leaves, triggering stress-related pathways proteins | 第45-59页 |
| 3.1 Introduction | 第45-46页 |
| 3.2 Materials and methods | 第46-50页 |
| 3.2.1 Protein elicitor purification | 第46页 |
| 3.2.2 Mo Hrip2 purification using column chromatography | 第46页 |
| 3.2.3 Confirmation of Mo Hrip2 by SDS-PAGE and tobacco leaf HR response | 第46页 |
| 3.2.4 Rice growth conditions and sample preparation | 第46页 |
| 3.2.5 Total proteins extraction for 2D-PAGE | 第46-47页 |
| 3.2.6 2D-PAGE | 第47-48页 |
| 3.2.7 IEF using Strip Holder for gel rehydration and IPGphore II system | 第48页 |
| 3.2.8 Equilibrating Immobiline Dry Strip gels | 第48页 |
| 3.2.9 2D SDS-PAGE | 第48页 |
| 3.2.10 Gel staining with coomassie blue | 第48-49页 |
| 3.2.11 Gel scanning and differential display proteins analysis | 第49页 |
| 3.2.12 MS/MS detection and database searching | 第49页 |
| 3.2.13 Total RNA isolation | 第49-50页 |
| 3.2.14 c DNA synthesis | 第50页 |
| 3.2.15 q PCR | 第50页 |
| 3.3 Results | 第50-56页 |
| 3.3.1 Differential display proteins treated with Mo Hrip2 | 第50-53页 |
| 3.3.2 Identification of differentially display proteins by MS/MS analysis | 第53-54页 |
| 3.3.3 q RT–PCR analysis of differential display proteins | 第54-56页 |
| 3.4 Discussion | 第56-59页 |
| 3.4.1 ROS and PCD–related proteins | 第56-57页 |
| 3.4.2 Transcription and signal transduction–related proteins | 第57页 |
| 3.4.3 PR proteins | 第57-58页 |
| 3.4.4 Photosynthesis and energy–related proteins | 第58-59页 |
| CHAPTER IV Elucidating of Mo Hrip2 binding proteins in rice | 第59-75页 |
| 4.2 Introduction | 第59-65页 |
| 4.2.1 Protein elicitor purification | 第59页 |
| 4.2.2 Mo Hrip2 purification using column chromatography | 第59-60页 |
| 4.2.3 Confirmation of Mo Hrip2 by SDS-PAGE and tobacco leaf HR response | 第60页 |
| 4.2.4 Rice growth conditions and sample preparation | 第60页 |
| 4.2.5 Total protein extraction from rice leaves | 第60页 |
| 4.2.6 Protein-protein interaction using pull-down poly His-Tag protein interaction kit | 第60-61页 |
| 4.2.7 SDS-PAGE and staining with Coomassie blue | 第61页 |
| 4.2.8 MS/MS detection and database searching | 第61页 |
| 4.2.9 Total RNA isolation and c DNA synthesis | 第61-62页 |
| 4.2.10 Regular PCR | 第62-63页 |
| 4.2.11 Gel electrophoresis and gene purification | 第63页 |
| 4.2.12 Gene cloning and confirmation | 第63-64页 |
| 4.2.13 GST-pulls down | 第64-65页 |
| 4.2.13.1 Digestion of gene, p GEX-6p-2 vector with the restriction enzymes (RE) | 第64页 |
| 4.2.13.2 Ligation of gene into p GEX-6p-2 vector | 第64-65页 |
| 4.2.13.3 Transformation of p GEX-6p-2 vector into BL21 E. coli | 第65页 |
| 4.3 Results | 第65-73页 |
| 4.3.1 Identification of Mo Hrip2-interacting proteins by poly His-Tag Pulls-downmethod | 第65页 |
| 4.3.2 Mass spectrometry analysis of the proteins | 第65-66页 |
| 4.3.3 Role of binding proteins in hydrogen peroxide production and cell signaling | 第66-67页 |
| 4.3.4 Gene cloning and transformation | 第67-68页 |
| 4.3.5 Poly His Tag-Pulls down for single protein-protein interaction | 第68页 |
| 4.3.6 Bioinformatics analysis | 第68-73页 |
| 4.3.7 Predicted Mo Hrip2-induced disease resistance pathway in rice | 第73页 |
| 4.4 Discussion | 第73-75页 |
| CONCLUSION | 第75-76页 |
| REFERENCES | 第76-89页 |
| ACKNOWLEDGEMENTS | 第89-90页 |
| Resume | 第90-93页 |
