论文目录 | |
| DEDICATIONS | 第11-12页 |
| Acknowledgements | 第12-14页 |
| ABSTRACT | 第14-17页 |
| 摘要 | 第17-19页 |
| Abbreviations | 第19-26页 |
| Chapter 1. Introduction | 第26-29页 |
| Chapter 2. Review of Literature | 第29-51页 |
| 2.1 Sphingolipids | 第29-30页 |
| 2.2 Characters of Plant sphingolipid classes | 第30-36页 |
| 2.2.1 Glycosyl inositol phospho-ceramides (GIPCs) | 第30-31页 |
| 2.2.2 Glycosylceramides | 第31-33页 |
| 2.2.3 Ceramide | 第33-34页 |
| 2.2.4 Free LCBs | 第34-36页 |
| 2.3 Sphingolipid metabolism in plants | 第36-37页 |
| 2.4 Function of Sphingolipid | 第37-45页 |
| 2.4.1 Membrane structural componen | 第37-39页 |
| 2.4.2 Sphingolipids and Chloroplasts | 第39-40页 |
| 2.4.3 Cell signaling | 第40-41页 |
| 2.4.4 Programmed cell death | 第41-43页 |
| 2.4.5 Plant pathogen interactions | 第43-44页 |
| 2.4.6 Abiotic stress | 第44-45页 |
| 2.5 Serine palmitoyltransferase(SPT) | 第45-51页 |
| 2.5.1 Structure and mechanism of SPT reaction | 第45-47页 |
| 2.5.2 Functional Characterization of SPT Gene | 第47-51页 |
| Chapter 3. Cloning of Rice SPT genes and gene expression related studies under different aspects of brown planthopper infestation | 第51-68页 |
| 3.1 Materials and Methods | 第52页 |
| 3.1.1 Plant growth | 第52页 |
| 3.1.2 Insects | 第52页 |
| 3.1.3 Reagent | 第52页 |
| 3.2 | 第52-56页 |
| 3.2.1 Total RNA isolation and cDNA synthesis | 第52-53页 |
| 3.2.2 First-strand cDNA synthesis | 第53页 |
| 3.2.3 PCR amplification | 第53-54页 |
| 3.2.4 PCR product recovery | 第54页 |
| 3.2.5 PCR products were blunt plus A | 第54页 |
| 3.2.6 Connection transformation | 第54-55页 |
| 3.2.7 Positive clones were detected | 第55页 |
| 3.2.8 Plasmid extraction | 第55-56页 |
| 3.3 Multiple Alignments and Phylogenetic tree | 第56-57页 |
| 3.4 qRT-PCR (20μ1) | 第57页 |
| 3.5 Gene expression analysis | 第57-59页 |
| 3.6 Statistical analysis | 第59页 |
| 3.7 Results | 第59-64页 |
| 3.7.1 Gene Cloning | 第59-61页 |
| 3.7.2 Multiple alignments and Phylogenetic tree | 第61-64页 |
| 3.8 Gene expression study | 第64-68页 |
| 3.8.1 OsLCB2a1expression is up-regulated under BPH infestation | 第64-65页 |
| 3.8.2 OsLCB2a1 expression in different varieties, infestation time, plant parts and crop stages of rice | 第65-68页 |
| Chapter 4. Effect of exogenous phytosphingosine on sphingolipid profile,development and reproduction of the brown planthopper in rice | 第68-72页 |
| 4.1 Materials and Methods | 第69页 |
| 4.2 Results | 第69-72页 |
| Chapter 5. Transformation of OsLCB2a1 in Arabidopsis through floral dip method and determination of the T-DNA insertion position of OsLCB2a1 gene in Overexpressed arabidopsis plants | 第72-85页 |
| 5.1 Materials and methods | 第73-75页 |
| 5.1.1 Reagents | 第73页 |
| 5.1.2 Plant Material and Growth Conditions of Arabidopsis | 第73页 |
| 5.1.3 Confirmation of Homozygous in Mutant | 第73-74页 |
| 5.1.4 Construction of vector | 第74页 |
| 5.1.5 Generation of transgenic plants using floral dip method | 第74-75页 |
| 5.1.6 Transgenic Arabidopsis RT-PCR identification | 第75页 |
| 5.2 TAIL PCR | 第75-78页 |
| 5.3 Results and discussion | 第78-85页 |
| 5.3.1 Identification of homozygous mutant | 第78-79页 |
| 5.3.2 Identification of transgenic Arabidopsis thaliana | 第79页 |
| 5.3.3 Phenotype of transgenic Arabidopsis | 第79-80页 |
| 5.3.4 Transgenic Arabidopsis RT-PCR identification and qRT-PCR for gene expression | 第80-81页 |
| 5.3.5 TAIL PCR | 第81-85页 |
| Chapter 6. Measurement of sphingolipid in tested Arabidopsis plants and determination of OsLCB2a1 gene function as biotic stress response | 第85-95页 |
| 6.1 Materials and methods | 第86-88页 |
| 6.1.1 Chemicals | 第86页 |
| 6.1.2 Measurement of sphingolipid | 第86页 |
| 6.1.3 Fumonisin B 1-induced cell death | 第86-87页 |
| 6.1.4 No choice aphid test in wild type, over-expressed, mutant and mutant complementation plants | 第87页 |
| 6.1.5 Feeding behavior of aphid through EPG in wild type, over-expressed (OE),mutant and mutant complementation (MC) plants | 第87-88页 |
| 6.1.5.1 EPG technique | 第87页 |
| 6.1.5.2 Aphid rearing | 第87页 |
| 6.1.5.3 Aphid probing and feeding behaviour through EPG | 第87-88页 |
| 6.2 Results | 第88-95页 |
| 6.2.1 Measurement of sphingolipid | 第88-90页 |
| 6.2.2 Fumonisin B1 Induced cell death | 第90-91页 |
| 6.2.3 No choice Aphid test | 第91-92页 |
| 6.2.4 Aphid Probing and Feeding Behaviour through EPG | 第92-95页 |
| Chapter 7. Discussion | 第95-99页 |
| Chapter 8. Summary | 第99-102页 |
| Chapter 9. Overall conclusion and Future perspectives | 第102-103页 |
| References | 第103-115页 |
