| 中文摘要 | 第6-7页 |
| Abstract | 第7-8页 |
| List of abbreviations | 第9-18页 |
| Chapter 1 Introduction | 第18-33页 |
| 1.1 Background | 第18页 |
| 1.2 Genome and characteristics of Xoo | 第18-19页 |
| 1.3 Cyclic di-GMP,a novel bacterial second messenger | 第19-28页 |
| 1.3.1 Synthesis and degradation of c-di-GMP | 第19-20页 |
| 1.3.2 Receptors or effector proteins of c-di-GMP | 第20-25页 |
| 1.3.3 Regulation of downstream phenotypes by c-di-GMP | 第25-28页 |
| 1.4 C-di-GMP signaling pathways | 第28-30页 |
| 1.4.1 C-di-GMP regulation of signaling pathways in Xoo | 第29-30页 |
| 1.5 Objectives | 第30-33页 |
| 1.5.1 Technical route | 第32-33页 |
| Chapter 2 Functional analysis of downstream components of Filp/PXO_02715 | 第33-53页 |
| 2.1 Materials and Methods | 第34-42页 |
| 2.1.1 Bacterial strains,plasmids and primers | 第34页 |
| 2.1.2 Regents and equipments | 第34页 |
| 2.1.3 Strains culture media and culture conditions | 第34-35页 |
| 2.1.4 Antibiotic concentrations | 第35页 |
| 2.1.5 Escherichia coli strain | 第35页 |
| 2.1.6 PXO99A and other mutant strains | 第35页 |
| 2.1.7 Rice variety IR 24 | 第35页 |
| 2.1.8 Genomic DNA extraction | 第35-36页 |
| 2.1.9 Construction of complementary strains | 第36-39页 |
| 2.1.10 Pathogenicity test | 第39-40页 |
| 2.1.11 Sliding and swimming motility | 第40页 |
| 2.1.12 Extracellular polysaccharide(EPS)assay | 第40-41页 |
| 2.1.13 Biofilm formation | 第41页 |
| 2.1.14 Statistical analyses | 第41-42页 |
| 2.2 Results | 第42-51页 |
| 2.2.1 Bioinformatics analysis of proteins downstream of Filp and PXO_02715 | 第42-43页 |
| 2.2.2 Construction of complementary strains | 第43-45页 |
| 2.2.3 Phenotype detection of mutant and complementary strains | 第45-51页 |
| 2.3 Discussion | 第51-53页 |
| Chapter 3 Phosphodiesterase activity of PXO_00476 and PXO_04753 | 第53-63页 |
| 3.1 Material and methods | 第54-56页 |
| 3.1.1 Site-directed mutagenesis | 第54-55页 |
| 3.1.2 Protein expression and purification | 第55页 |
| 3.1.3 Inclusion body Protein Purification | 第55页 |
| 3.1.4 Purification of soluble recombinant proteins | 第55-56页 |
| 3.1.5 Phosphodiesterase colorimetric assay | 第56页 |
| 3.1.6 In vitro proteins phosphodiesterase assay and UPLC analysis | 第56页 |
| 3.2 Results | 第56-61页 |
| 3.2.1 Alignment and bioinformatics analysis of EAL and HD-GYP domains | 第56-57页 |
| 3.2.2 Construction of site directed mutant strains | 第57-59页 |
| 3.2.3 HD-GYP and EAL domain proteins exhibit PDE activity in vitro | 第59-61页 |
| 3.3 Discussions | 第61-63页 |
| Chapter 4 Detection the protein-protein interactions by yeast two-hybrid system | 第63-72页 |
| 4.1 Overview | 第63页 |
| 4.2 Yeast two hybrid system | 第63-64页 |
| 4.3 Material and methods | 第64-68页 |
| 4.3.1 Amplification of target genes | 第64页 |
| 4.3.2 Cloning of genes with middle vector | 第64页 |
| 4.3.3 Construction of target prey vector plasmid | 第64-65页 |
| 4.3.4 Transformation of prey fusion vector into yeast | 第65页 |
| 4.3.5 Y2H assay | 第65-66页 |
| 4.3.6 Strains culture media and buffer preparations | 第66-68页 |
| 4.4 Results | 第68-71页 |
| 4.4.1 Vector constructions | 第68-69页 |
| 4.4.2 Interaction between DBD-Filp/PXO_02715 with their downstream components | 第69-71页 |
| 4.5 Discussion | 第71-72页 |
| Chapter 5 Subcellular localization patterns of Filp/PXO_02715 and their downstream components in Xoo | 第72-79页 |
| 5.1 Material and methods | 第72-74页 |
| 5.1.1 Construction of plasmids and Xoo strains carrying GFP-fusion proteins | 第72-73页 |
| 5.1.2 Western blotting analysis | 第73页 |
| 5.1.3 Preparations of Xoo strains for total protein expression | 第73页 |
| 5.1.4 Western blotting | 第73-74页 |
| 5.1.5 Fluoresce microscopy | 第74页 |
| 5.2 Results | 第74-77页 |
| 5.2.1 Vector constructions and different bacterial strains | 第74-75页 |
| 5.2.2 Confirmation of GFP-fusion proteins expression | 第75页 |
| 5.2.3 Subcellular localization of different GFP-fusion proteins in Xoo strains | 第75-77页 |
| 5.3 Discussion | 第77-79页 |
| Chapter 6 Summary | 第79-81页 |
| Literature cited | 第81-100页 |
| Appendices | 第100-105页 |
| Acknowledgements | 第105-107页 |
| Author biography | 第107-110页 |
