| ABSTRACT | 第7-8页 |
| 摘要 | 第9-11页 |
| 1. CHAPTER ONE GENERAL INTRODUCTION AND LITERATURE REVIEW | 第11-27页 |
| 1.1 BACKGROUND OF ORIENTAL FRUIT FLY | 第11-12页 |
| 1.2 CONTROL MEASURES | 第12-15页 |
| 1.2.1 Insecticide cover sprays | 第12页 |
| 1.2.2 Protein bait sprays | 第12页 |
| 1.2.3 Male annihilation technique | 第12-13页 |
| 1.2.4 Cultural controls | 第13页 |
| 1.2.5 Release of natural enemies | 第13页 |
| 1.2.6 Sterile insect technique | 第13-15页 |
| 1.3 MICRORNAS | 第15-19页 |
| 1.3.1 Biogenesis of micro RNAs | 第16-17页 |
| 1.3.2 Mechanisms of mi RNA mediated gene silencing | 第17-19页 |
| 1.4 SPERMATOGENESIS – AN INTRODUCTION | 第19-22页 |
| 1.4.1 Spermatogenesis in the model insect Drosophila melanogaster | 第19-20页 |
| 1.4.2 Genetic controls of spermatogenesis | 第20-22页 |
| 1.5 MICRORNAS AS REGULATORS OF SPERMATOGENESIS | 第22-23页 |
| 1.6 ROLE OF MITOFERRIN IN SPERMATOGENESIS | 第23-25页 |
| 1.7 THESIS OBJECTIVES | 第25-27页 |
| 2. CHAPTER TWO IDENTIFICATION AND CHARACTERIZATION OF MICRORNAS IN THETESTES OF BACTROCERA DORSALIS | 第27-53页 |
| 2.1 INTRODUCTION | 第27-29页 |
| 2.2 MATERIALS AND METHODS | 第29-37页 |
| 2.2.1 Insect rearing and RNA extraction | 第29-30页 |
| 2.2.2 Small RNA library preparation and sequencing | 第30页 |
| 2.2.3 Analysis of sequencing data | 第30-31页 |
| 2.2.4 RNA library preparation and comparison | 第31页 |
| 2.2.5 Target gene prediction | 第31-32页 |
| 2.2.6 Quantitative real-time PCR | 第32-33页 |
| 2.2.7 Stem-loop RT-PCR assay | 第33-34页 |
| 2.2.8 Target gene prediction | 第34-35页 |
| 2.2.9 Statistical analysis | 第35-37页 |
| 2.3 RESULTS | 第37-50页 |
| 2.3.1 Deep sequencing of small RNA libraries | 第37-38页 |
| 2.3.2 Identification of known and novel mi RNAs in the testis of B. dorsalis | 第38-40页 |
| 2.3.3 Differentially expressed known mi RNAs across different developmental stages ofthe testis | 第40-44页 |
| 2.3.4 The analysis of selected mi RNA-m RNA expression patterns | 第44-50页 |
| 2.4 DISCUSSION | 第50-53页 |
| 2.4.1 Identification of mi RNAs in the testes of B. dorsalis | 第50页 |
| 2.4.2 Characterization of testicular mi RNAs | 第50-53页 |
| 3. CHAPTER THREE THE ROLE OF MIR83P IN THE TESTES OF BACTROCERA DORSALIS | 第53-77页 |
| 3.1 INTRODUCTION | 第53-55页 |
| 3.2 MATERIALS AND METHODS | 第55-65页 |
| 3.2.1 Bioinformatics Analysis | 第55页 |
| 3.2.2 Mitoferrin 3′UTR cloning and ligation into expression vector | 第55-56页 |
| 3.2.3 Examining DNA by gel electrophoresis | 第56-57页 |
| 3.2.4 Isolation of DNA from agarose gel | 第57页 |
| 3.2.5 Bacterial transformation | 第57-58页 |
| 3.2.6 Plasmid Extraction and Purification | 第58-59页 |
| 3.2.7 mi RNA mimics, antagomi Rs and ds RNAs synthesis | 第59-60页 |
| 3.2.8 Quantitative real-time PCR | 第60-61页 |
| 3.2.9 Cell culture | 第61页 |
| 3.2.10 Freezing cells | 第61页 |
| 3.2.11 Dual-luciferase assay in He La cells | 第61-63页 |
| 3.2.12 Reproductive capacity of male flies | 第63页 |
| 3.2.13 Sperm viability assays and spermatozoa counts | 第63-64页 |
| 3.2.14 Statistical analysis | 第64-65页 |
| 3.3 RESULTS | 第65-74页 |
| 3.3.1 mi R83p interacts with the mitoferrin 3′UTR and represses mitoferrin expression | 第65-69页 |
| 3.3.2 Dietary delivery of ds RNA, mi R83p mimics/antagomi R successfully alters bmfrnexpression in testes and reduces reproductive capacity of male flies | 第69-74页 |
| 3.4 DISCUSSION | 第74-77页 |
| 3.4.1 Functional role of mi R83p in spermatogenesis of B. dorsalis | 第74-75页 |
| 3.4.2 Role of mi R83p in sperm viability | 第75-77页 |
| SUMMARY | 第77-78页 |
| REFERENCES | 第78-102页 |
| ACKNOWLEDGEMENTS | 第102-103页 |
| ARTICLE PUBLISHED | 第103-104页 |
