| 中文摘要 | 第13-16页 |
| ABSTRACT | 第16-20页 |
| CHAPTER ONE GENERAL INTRODUCTION AND LITERATURE OF REVIEW | 第21-75页 |
| 1. INTRODUCTION | 第21-26页 |
| 1.1 Background of the present study | 第21-23页 |
| 1.2 Rationale/justification of the present study | 第23-25页 |
| 1.3 Scope of the present study | 第25-26页 |
| 1.4 Objectives of the present study | 第26页 |
| 2. LITERATURE OF REVIEW | 第26-75页 |
| 2.1 Small brown planthopper | 第27-31页 |
| 2.2 Insecticides | 第31-45页 |
| 2.3 Cytochrome P450 monooxygenases (P450s) | 第45-52页 |
| 2.4 Cytochrome P450 monooxygenases (P450s) in insects | 第52-64页 |
| 2.5 Cytochrome P450s and redox partners | 第64-65页 |
| 2.6 Insecticides degradation mechanism by insect P450s | 第65-75页 |
| CHAPTER TWO MATERIALS AND METHODS | 第75-83页 |
| 2.1 Chemicals and Reagents | 第75-76页 |
| 2.2 Biological Materials | 第76页 |
| 2.2.1 Experimental insects | 第76页 |
| 2.2.2 Spodoptera frugiperda (Sf9) cell lines | 第76页 |
| 2.3 Tested P450 genes (Family 4) of L. striatellus | 第76页 |
| 2.4 Functional expression of P450 genes in Sf9 cell and microsomal protein isolation | 第76-79页 |
| 2.4.1 Gene Cloning | 第76-77页 |
| 2.4.2 Construction of expression vector (recombinant plasmid) | 第77页 |
| 2.4.3 Cell culture and stable transfer | 第77-78页 |
| 2.4.4 Preparation of cell lysate/microsome | 第78-79页 |
| 2.5 Total RNA Isolation, cDNA Synthesis and RT-PCR detection of expression in Sf9cell | 第79页 |
| 2.6 SDS-PAGE analysis | 第79页 |
| 2.7 Carbon monoxide (CO) difference spectroscopy | 第79-80页 |
| 2.8 Determination of Enzyme activities | 第80-81页 |
| 2.8.1 Oxidative metabolism against model substrate | 第80页 |
| 2.8.2 Metabolism of probe substrates | 第80-81页 |
| 2.9 HPLC analysis of insecticide degradation | 第81页 |
| 2.10 UPLC-MS and MS/MS analysis and identification of metabolites | 第81-82页 |
| 2.11 P450 homology modeling and substrate docking | 第82-83页 |
| CHAPTER THREE DELTAMETHRIN IS DEGRADED BY CYP439A1v3; A CYTOCHROME P450OVER-EXPRESSED IN RESISTANT STRAIN OF Laodelphax striatellus | 第83-99页 |
| 3.1 Abstract | 第83-84页 |
| 3.2 Introduction | 第84-85页 |
| 3.3 Materials and Methodology | 第85-87页 |
| 3.3.1 Chemical and biological materials | 第85页 |
| 3.3.2 P450 gene used in this study | 第85-86页 |
| 3.3.3 Vector construction for functional Expression in Sf9 cell | 第86-87页 |
| 3.3.4 CYP439A1v3 transfection in Sf9 Cells | 第87页 |
| 3.3.5 Methodology for biochemical analysis of P450 gene | 第87页 |
| 3.4 Results | 第87-95页 |
| 3.4.1 Functional expression of L. striatellus CYP439Alv3 | 第87-89页 |
| 3.4.2 CYP439A1v3 catalytic activity against standard P450 model substrates | 第89-90页 |
| 3.4.3 CYP439A1v3 catalytic activity for insecticide metabolism | 第90-92页 |
| 3.4.4 Identification of deltamethrin metabolites | 第92-93页 |
| 3.4.5 CYP439A1v3 homology modeling and substrate docking | 第93-95页 |
| 3.5 Discussion | 第95-99页 |
| CHAPTER FOUR RESISTANCE IRRELEVANT CYP417A2v2 WAS FOUND DEGRADINGINSECTICIDE IN Laodelphax striatellus | 第99-113页 |
| 4.1 Abstract | 第99-100页 |
| 4.2 Introduction | 第100-102页 |
| 4.3 Materials and Methodology | 第102-103页 |
| 4.3.1 Chemical and biological materials | 第102页 |
| 4.3.2 P450 genes used in this study | 第102页 |
| 4.3.3 Vector construction for functional Expression in Sf9 cell | 第102-103页 |
| 4.3.4 P450 genes transfection in Sf9 Cells | 第103页 |
| 4.3.5 Methodology for biochemical analysis of P450 gene | 第103页 |
| 4.4 Results | 第103-110页 |
| 4.4.1 Functional expression of L. striatellus P450s | 第103-104页 |
| 4.4.2 Enzymatic activity of CYP417A2v2 with CO-difference spectrum | 第104-106页 |
| 4.4.3 CYP417A2v2 catalytic activity against standard P450 model substrates | 第106页 |
| 4.4.4 CYP417A2v2 capability to metabolize insecticides | 第106-108页 |
| 4.4.5 Identification of imidacloprid metabolite | 第108-110页 |
| 4.5 Discussion | 第110-113页 |
| CHAPTER FIVE FIPRONIL WAS FOUND TO BE DEGRADED BY A CYTOCHROME P450MONOOXYGENASE CYP426A1 IN Laodelphax striatellus | 第113-125页 |
| 5.1 Abstract | 第113-114页 |
| 5.2 Introduction | 第114-116页 |
| 5.3 Materials and Methodology | 第116-117页 |
| 5.3.1 Chemical and biological materials | 第116页 |
| 5.3.2 P450 gene used in this study | 第116页 |
| 5.3.3 Vector construction for functional Expression in Sf9 cell | 第116-117页 |
| 5.3.4 CYP426A1 transfection in Sf9 Cells | 第117页 |
| 5.3.5 Methodology for biochemical analysis of P450 gene | 第117页 |
| 5.4 Results | 第117-121页 |
| 5.4.1 Functional expression of L. striatellus CYP426A1 | 第117-119页 |
| 5.4.2 Catalytic activity of CYP426Alagainst standard P450 model substrates | 第119页 |
| 5.4.3 CYP426A1 catalytic activity for insecticide metabolism | 第119-121页 |
| 5.5 Discussion | 第121-125页 |
| SUMMARY | 第125-127页 |
| REFERENCES | 第127-155页 |
| PUBLICATIONS | 第155-157页 |
| ACKNOWLEDGEMENTS | 第157页 |
