| 摘要 | 第6-7页 |
| Abstract | 第7-8页 |
| LIST OF ABBREVIATIONS | 第12-15页 |
| CHAPTER1 INTRODUCTION | 第15-32页 |
| 1.1 General introduction | 第15-18页 |
| 1.2 Review of literature | 第18-32页 |
| 1.2.1 Heterosis and hybrid rice | 第18页 |
| 1.2.2 Three-line hybrid system | 第18-19页 |
| 1.2.3 Two-line hybrid system | 第19-20页 |
| 1.2.4 Developed EGMS lines for two-line hybrid system | 第20-22页 |
| 1.2.5 Important loci that control EGMS | 第22页 |
| 1.2.6 Advantages of two-line hybrid system | 第22-23页 |
| 1.2.7 Lipid transfer protein | 第23-24页 |
| 1.2.8 Role of lipid transfer protein in plants | 第24-26页 |
| 1.2.9 Clustered regularly interspaced short palindromic repeat/Cas(CRISPR/Cas)system | 第26-27页 |
| 1.2.10 CRISPR/Cas9 system | 第27-28页 |
| 1.2.11 The application of Cas9/sgRNA system for plant genome editing | 第28-29页 |
| 1.2.12 Success of CRISPR/Cas9 system in plants | 第29-30页 |
| 1.2.13 The future application of CRISPR/Cas9 in agriculture | 第30-32页 |
| CHAPTER2 Generation of New Thermo-sensitive Genic Male Sterile Rice Line by Knocking out TMS5 Gene based on CRISPR/Cas9 System | 第32-61页 |
| 2.1 Introduction | 第32-33页 |
| 2.2 Materials and methods | 第33-45页 |
| 2.2.1 Plant materials and growing conditions | 第33页 |
| 2.2.2 Vector construction and rice transformation | 第33-35页 |
| 2.2.3 Extraction of total DNA from leaves(SDS method) | 第35-37页 |
| 2.2.4 PCR amplification system(KOD-FX method) | 第37页 |
| 2.2.5 Mutation detection and analysis of transgenic plants | 第37-38页 |
| 2.2.6 Phenotypic analysis of T-DNA-free segregants | 第38页 |
| 2.2.7 Procedure of RNA extraction | 第38-40页 |
| 2.2.8 Synthesis of cDNA | 第40页 |
| 2.2.9 Procedure of Real-time PCR(RT-PCR) | 第40-42页 |
| 2.2.10 Extraction of protein and Western blot analysis | 第42-44页 |
| 2.2.11 The performance of F_1 hybrids made with YK17S | 第44页 |
| 2.2.12 Fertilizer management | 第44页 |
| 2.2.13 Data collection of yield traits | 第44-45页 |
| 2.2.14 Statistical analysis | 第45页 |
| 2.3 Results | 第45-58页 |
| 2.3.1 The mutation of TMS5 via CRISPR/Cas | 第45-47页 |
| 2.3.2 Morphological features of the tms5 mutants | 第47-49页 |
| 2.3.3 The effect of temperature on the pollen production in the tms5 mutants | 第49-52页 |
| 2.3.4 The suitability of YK17S1 for two-line hybrid breeding | 第52-58页 |
| 2.4 Discussion | 第58-60页 |
| 2.5 Conclusion | 第60-61页 |
| CHAPTER3 The Function Analysis of a Putative Lipid Transfer Protein OsLTPL164 in Grain Development and Quality in Rice | 第61-88页 |
| 3.1 Introduction | 第61-62页 |
| 3.2 Materials and methods | 第62-73页 |
| 3.2.1 Phylogenetic analysis | 第62页 |
| 3.2.2 Plant materials and growing conditions | 第62-63页 |
| 3.2.3 Vector construction and rice transformation | 第63页 |
| 3.2.4 Mutation detection and analysis of transgenic plants | 第63-64页 |
| 3.2.5 Transcription profiling | 第64页 |
| 3.2.6 Determination of dry matter accumulation | 第64页 |
| 3.2.7 Data collection for yield traits | 第64页 |
| 3.2.8 Determination of total starch content | 第64-66页 |
| 3.2.9 Determination of amylose content | 第66页 |
| 3.2.10 Determination of total protein content | 第66-67页 |
| 3.2.11 Determination of total lipid and fatty acid content | 第67页 |
| 3.2.12 GUS staining | 第67-68页 |
| 3.2.13 Vector construction and plasmid extraction for subcellular localization | 第68-69页 |
| 3.2.14 Isolation and transformation of rice protoplasts | 第69-73页 |
| 3.2.15 Sample preparation and microscopy | 第73页 |
| 3.2.16 Seed germination assays | 第73页 |
| 3.2.17 Statistical analysis | 第73页 |
| 3.3 Results | 第73-84页 |
| 3.3.1 Structural and sequence analysis of OsLTPL | 第73-74页 |
| 3.3.2 The Mutation of OsLTPL164 via CRISPR/Cas | 第74-76页 |
| 3.3.3 Expression pattern,GUS assay and subcellular localization of OsLTPL | 第76-78页 |
| 3.3.4 Morphological features of cr-ltp164 mutant | 第78-81页 |
| 3.3.5 Effects of knockout OsLTPL164 on quality traits | 第81页 |
| 3.3.6 Effects of knockout OsLTPL164 on seed germination | 第81页 |
| 3.3.7 The expression of fatty acid biosynthesis related genes | 第81-84页 |
| 3.4 Discussion | 第84-87页 |
| 3.5 Conclusion | 第87-88页 |
| Chapter4 Major findings and future perspectives | 第88-89页 |
| 4.1 Major findings | 第88页 |
| 4.2 Future perspectives | 第88-89页 |
| REFERENCES | 第89-100页 |
| LIST OF PUBLICATION DURING PH.D STUDY | 第100-101页 |
| ACKNOWLEDGEMENTS | 第101-103页 |
| AUTHOR’S RESUME | 第103页 |
