| 摘要 | 第6-7页 |
| Abstract | 第7-8页 |
| Abbreviations | 第11-17页 |
| CHAPTER-Ⅰ Literature review | 第17-34页 |
| 1.1 Introduction to Geminiviruses | 第17页 |
| 1.2 Begomoviruses,their genomes and associated satellites | 第17-21页 |
| 1.2.1 Betasatellites and their genomic organization | 第20页 |
| 1.2.2 Tomato yellow leaf curl China virus(TYLCCNV) | 第20-21页 |
| 1.2.3 βC1 functions in the development of viral symptoms | 第21页 |
| 1.3 Effect of viral infection on chloroplast and photosynthesis | 第21-30页 |
| 1.3.1 Key modifications in the plant’s photosynthesis apparatus induced by plant viruses | 第22-27页 |
| 1.3.1.1 Ultrastructure of the chloroplast is modified | 第22-25页 |
| 1.3.1.2 Viral replication complexes/proteins are targeted to the chloroplasts | 第25-26页 |
| 1.3.1.3 Metabolomic,transcriptomic and proteomic alterations in the photosynthetic machinery are induced | 第26页 |
| 1.3.1.4 Viral proteins interact with photosynthesis and chloroplast-related proteins | 第26-27页 |
| 1.3.2 PSII extrinsic proteins and their role in plant-virus interactions | 第27-30页 |
| 1.3.2.1 Manganese-stabilizing protein/PsbO | 第28页 |
| 1.3.2.2 PSII-related protein of oxygen-evolving complex/PsbP | 第28页 |
| 1.3.2.3 Oxygen-evolving enhancer protein-3/PsbQ | 第28-29页 |
| 1.3.2.4 PSII-related10 kDa protein/PsbR | 第29-30页 |
| 1.3.2.5 Interaction of PsbP with pathogenicity proteins of plant viruses | 第30页 |
| 1.4 Chlorophyll fluorescence techniques | 第30-33页 |
| 1.4.1 Imaging chlorophyll fluorescence transients | 第31-32页 |
| 1.4.2 Imaging chlorophyll fluorescence kinetics to monitor plant virus infections | 第32-33页 |
| 1.5 Objectives | 第33-34页 |
| CHAPTER-Ⅱ Materials and Methods | 第34-49页 |
| 2.1 Plant materials and growth conditions | 第34页 |
| 2.2 Agroinoculation of plants | 第34页 |
| 2.3 Nondestructive measurement of chlorophyll contents | 第34-35页 |
| 2.4 Chlorophyll fluorescence imaging | 第35-36页 |
| 2.5 Measurement of fluorescene transients in N.benthamiana with transgenic expression of βC1 | 第36页 |
| 2.6 Total RNA extraction,synthesis of cDNA and RT-qPCR analysis | 第36-38页 |
| 2.6.1 Extraction of total RNA | 第36-37页 |
| 2.6.2 cDNA synthesis | 第37页 |
| 2.6.3 RT-PCR | 第37-38页 |
| 2.6.4 RT-qPCR | 第38页 |
| 2.7 Gene amplification,cloning and construction of expression vectors | 第38-43页 |
| 2.7.1 Nucleotide sequence accession numbers and primer design | 第38-39页 |
| 2.7.2 Amplification of PsbO,PsbP,PsbQ and PsbR | 第39页 |
| 2.7.3 Gel purification of PCR products | 第39-40页 |
| 2.7.4 Ligation of PCR products into pEASY?-T5 Zero vector | 第40-41页 |
| 2.7.5 Transformation of ligated products into E.coli | 第41页 |
| 2.7.6 Detection of positive constructs by colony PCR | 第41-42页 |
| 2.7.7 Plasmid extraction from positive colonies | 第42页 |
| 2.7.8 Verification of positive constructs by restriction enzyme analysis | 第42-43页 |
| 2.7.9 Ligation of genes into bait(pGBKT7)and prey(pGADT7)vectors | 第43页 |
| 2.8 Yeast two-hybrid test | 第43-47页 |
| 2.8.1 Preparation of yeast(Y-187)competent cells | 第43-44页 |
| 2.8.2 Small-scale transformation of yeast competent cells | 第44-45页 |
| 2.8.3 Selection of interacting proteins using high stringency media | 第45-46页 |
| 2.8.4 Extraction of plasmid from co-transformed yeast | 第46-47页 |
| 2.8.5 Verification of co-transformed yeast by PCR | 第47页 |
| 2.9 Sequence homology and phylogenetic analysis of OEC proteins | 第47-49页 |
| CHAPTER-Ⅲ Results | 第49-64页 |
| 3.1 TYLCCNV A+β infecion and transgene of βC1 induced abnormal phenotype in N.benthamiana | 第49-50页 |
| 3.2 Relative chlorophyll contents were decreased in TYLCCNV A,TYLCCNV A+β infected and βC1 transgenic N.benthamiana | 第50-52页 |
| 3.3 Photosynthetic efficiency was hindered in TYLCCNV A,TYLCCNV A+β infected and βC1 transgenic N.benthamiana | 第52-56页 |
| 3.4 Expressions of PSII-related genes were significantly reduced under prolonged influence of βC1 | 第56-58页 |
| 3.5 Expression of NPQ-related PsbS was significantly upregulated by betasatellite infection | 第58-59页 |
| 3.6 Yeast two hybrid assay revealed interaction of PsbP with βC1 | 第59-60页 |
| 3.7 Multiple sequence alignment and phylogenetic analysis of OEC proteins | 第60-61页 |
| 3.8 Multiple sequence alignment and phylogenetic analysis of PSII-related PsbS protein | 第61-64页 |
| CHAPTER-Ⅳ Discussion | 第64-68页 |
| CHAPTER-Ⅴ Conclusion | 第68-70页 |
| References | 第70-85页 |
| Acknowledgements | 第85-86页 |
| Author biography | 第86-87页 |
| Appendix | 第87-100页 |
