| Abstract (Chinese) | 第6-8页 |
| Abstract (English) | 第8-9页 |
| Abbreviation | 第16-18页 |
| Chapter Ⅰ. Introduction | 第18-39页 |
| 1.1 Initiation of leaf senescence | 第18-31页 |
| 1.1.1 Age dependent developmental processes | 第19页 |
| 1.1.2 Phytohormones regulating leaf senescence | 第19-28页 |
| 1.1.2.1 Gibberellic acid | 第19-20页 |
| 1.1.2.2 Auxin | 第20-21页 |
| 1.1.2.3 Cytokinins | 第21-22页 |
| 1.1.2.4 Abscisic acid | 第22-23页 |
| 1.1.2.5 Brassinosteroids | 第23-24页 |
| 1.1.2.6 Ethylene | 第24-25页 |
| 1.1.2.7 Jasmonic acid | 第25-26页 |
| 1.1.2.8 Salicylic acid | 第26-27页 |
| 1.1.2.9 Strigolactone | 第27-28页 |
| 1.1.3 Genetical regulation of leaf senescence | 第28-31页 |
| 1.2 Progression of leaf senescence | 第31-33页 |
| 1.2.1 Loss of chlorophyll | 第31页 |
| 1.2.2 Lipids degradation | 第31-32页 |
| 1.2.3 Nitrogen re-mobilization | 第32页 |
| 1.2.4 Protein degradation | 第32-33页 |
| 1.2.5 Nitrogen translocation from source to sink | 第33页 |
| 1.2.6 Autophagy | 第33页 |
| 1.3 Leaf senescence in agriculture setting | 第33-35页 |
| 1.3.1 Early senescence | 第34页 |
| 1.3.2 Delay senescence | 第34-35页 |
| 1.4 Leaf senescence manipulation strategies for crop improvement | 第35-39页 |
| 1.4.1 Plant hormone biology-based technology | 第35-36页 |
| 1.4.2 Senescence-specific transcription factor biology-based technology | 第36-37页 |
| 1.4.3 Translation initiation factor biology-based technology | 第37-39页 |
| Chapter Ⅱ. Screening Arabidopsis T-DNA mutants for identification of leucine-rich repeat receptor-like kinase (LRR RLKs) genes involved in leaf senescence | 第39-60页 |
| 2.1 Introduction | 第39-40页 |
| 2.2 Materials and methods | 第40-46页 |
| 2.2.1 Plant materials and growth conditions | 第40页 |
| 2.2.2 Arabidopsis T-DNA insertion mutants | 第40页 |
| 2.2.3 Plant genomic DNA extraction | 第40-41页 |
| 2.2.4 Identification of homozygous T-DNA insertion mutants | 第41-42页 |
| 2.2.4.1 Verification of T-DNA insertions and isolation of homozygous lines | 第41页 |
| 2.2.4.2 Plant RNA extraction | 第41-42页 |
| 2.2.4.3 cDNA synthesis | 第42页 |
| 2.2.5 Phenotypic screening of homozygous T-DNA mutants for leaf senescence | 第42页 |
| 2.2.5.1 Measurements of chlorophyll content and fluorescence | 第42页 |
| 2.2.6 Gain of function analysis of Senescence-associated Receptor-like Kinasel | 第42-44页 |
| 2.2.6.1 Isolation and amplification of SRKI | 第42-43页 |
| 2.2.6.2 Construction of over-expression vector and Arabidopsis transformation | 第43-44页 |
| 2.2.7 Gain of function analysis of Senescence-associated Receptor-like | 第44页 |
| 2.2.7.1. Isolation and amplification of SRK2 | 第44页 |
| 2.2.7.2 Construction of overexpression vector | 第44页 |
| 2.2.8 Antibiotic stocks | 第44页 |
| 2.2.9 Sequencing | 第44-45页 |
| 2.2.10 Preparation of Agrobacterium tumefaciens cells | 第45页 |
| 2.2.11 Transformation of Agrobacteria | 第45页 |
| 2.2.12 Bacterial stock | 第45页 |
| 2.2.13 Arabidopsis transformation | 第45-46页 |
| 2.2.14 Planting of TO transgenic Arabidopsis plants on selection media | 第46页 |
| 2.3 Results | 第46-56页 |
| 2.3.1 Available insertion mutants | 第46页 |
| 2.3.2 PCR based genotyping of T-DNA lines | 第46-49页 |
| 2.3.3 Phenotypic screening of RLKs mutants | 第49-53页 |
| 2.3.3.1 Expression pattern of srks in rosette leaves during leaf | 第50-53页 |
| 2.3.4 Characterization and functional analysis of SRK1 | 第53-54页 |
| 2.3.4.1 Gain of function analysis of SRK1 | 第53-54页 |
| 2.3.5 Characterization and functional analysis of SRK2 | 第54-56页 |
| 2.3.5.1 Gain of function analysis of SRK2 | 第55-56页 |
| 2.4 Discussion | 第56-60页 |
| 2.4.1 T-DNA mutagenesis | 第56页 |
| 2.4.2 PCR based identification of T | 第56-57页 |
| 2.4.3 Loss of function analysis of LRR RLK mutants regulating leaf senescence | 第57-58页 |
| 2.4.4 Receptor Receptor-like kinases (SRK1 and SRK2) involved in leaf senescence | 第58-60页 |
| Chapter Ⅲ. Inducible gene expression system for leaf senescence regulation in tobacco | 第60-78页 |
| 3.1 Introduction | 第60-61页 |
| 3.2 Material and methods | 第61-67页 |
| 3.2.1 Primers designing for amplification of NAP genes | 第61-62页 |
| 3.2.2 AtNAP gene cloning in pEasyBlunt vector | 第62页 |
| 3.2.3 Plasmids for ethanol inducible system | 第62-63页 |
| 3.2.4 Plasmid Construction for inducible gene expression system | 第63-64页 |
| 3.2.4.1 Restriction enzyme digestion of pEasyBlunt:AtNAP and | 第63页 |
| 3.2.4.2 Restriction enzyme digestion of pBJ36_AlcA:AtNAP and | 第63-64页 |
| 3.2.4.3 Media preparation | 第64页 |
| 3.2.5 Transformation of Plasmid into Bacteria | 第64-65页 |
| 3.2.6 Agrobacterium mediated Tobacco transformation with Ethanol Inducible | 第65-66页 |
| 3.2.6.1 Preparation of Agrobacterium Culture | 第65页 |
| 3.2.6.2 Preparation of Plant material | 第65页 |
| 3.2.6.3 Co-cultivation with Agrobacteria | 第65页 |
| 3.2.6.4 Preparation of media | 第65页 |
| 3.2.6.5 Phase selection of leaf explants | 第65页 |
| 3.2.6.6 Acclimatization | 第65-66页 |
| 3.2.6.7 Planting of T1 generation and Transgene selection | 第66页 |
| 3.2.7 Treatment with ethanol | 第66页 |
| 3.2.8 Cloning of NtNAP2 (Ntab0758860.1) in pMLBART-based constructs | 第66页 |
| 3.2.9 Agrobacterium mediated tobacco transformation | 第66-67页 |
| 3.3 Results | 第67-75页 |
| 3.3.1 Construction of pMLBART-based constructs for ethanol inducible system | 第67-70页 |
| 3.3.1.1 Amplification of AtNAP from cDNA/cloning | 第67页 |
| 3.3.1.2 AtNAP insertion in pBJ36_AlcA vector | 第67-68页 |
| 3.3.1.3 Construction of pMLBART_AlcR based ethanol inducible system | 第68-70页 |
| 3.3.2 Agrobacterium-mediated tobacco transformation with | 第70-71页 |
| 3.3.2.1 Agrobacterium transformation with pMLBART_alcR/AtNAP construct | 第70页 |
| 3.3.2.2 Co-cultivation of tobacco leaf discs with | 第70页 |
| 3.3.2.3 Shoot regeneration | 第70页 |
| 3.3.2.4 Selection of Transgenic Plants | 第70-71页 |
| 3.3.2.5 Acclimatization of transgenic plantlets | 第71页 |
| 3.3.2.6 Selection of homozygous transgenic plants | 第71页 |
| 3.3.3 Preliminary characterization of alc-mediated AtNAP gene expression | 第71-75页 |
| 3.3.3.1 Ethanol induction | 第71页 |
| 3.3.3.2 Leaf senescence induction in soil grown transgenic plants | 第71-72页 |
| 3.3.3.3 Leaf senescence induction in young plantlets under controlled | 第72-75页 |
| 3.3.3.3.1 Vapors response | 第72页 |
| 3.3.3.3.2 Foliar spray | 第72-73页 |
| 3.3.3.3.3 Detached leaves | 第73-75页 |
| 3.4 Discussion | 第75-78页 |
| Chapter Ⅳ. Conclusion | 第78-79页 |
| References | 第79-106页 |
| Acknowledgement | 第106-108页 |
| Author Biography | 第108页 |
