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Impacts of Transgenic Bt Rice on Non-Target Brown Planthopper

ACKNOWLEDGEMENTS第6-11页
ABSTRACT第11-12页
LIST OF ABBREVIATIONS第13-15页
LIST OF FIGURES第15-17页
LIST OF TABLES第17-19页
AIM OF THE STUDY第19-27页
CHAPTER 1: LITERATURE OVERVIEW第27-47页
    1:1 Brown Planthopper - Threat to Rice Production第27-31页
        1:1:1 Brown Planthopper problem in China第29-30页
        1:1:2 Brown Planthopper problem in Sri Lanka第30-31页
    1:2 Genetically Modified Rice第31-34页
        1:2:1 Insect-Resistant Genetically Modified Rice第31-34页
            1:2:1:1 Crystal delta endotoxins from Bacillus thuringiensis第32-34页
            1:2:1:2 Protease inhibitors第34页
            1:2:1:3 Plant lectins第34页
    1.3 Effect of transgenic Bt rice on BPH第34-38页
    1.4 Safety consideration of Bt rice第38-41页
        1.4.1 Food safety of Bt Rice第39-40页
        1.4.2 Environmental concerns第40页
        1.4.3 Socio-economic concerns第40-41页
    1.5 The immune system in insects第41-47页
        1.5.1 Cellular immune responses第42-44页
            1.5.1.1 Types of hemocytes第42-43页
            1.5.1.2 Immune functions of hemocytes第43-44页
        1.5.2 Humoral immune responses第44-47页
            1.5.2.1 Antimicrobial peptides (AMPs)第44-45页
            1.5.2.2 Prophenoloxidase activating system第45-47页
CHAPTER 2: CHARACTERIZATION OF HEMOCYTES OF THE BROWNPLANTHOPPER; Nilaparvata lugens Stal(Hemiptera:Delphacidae)第47-63页
    2.1 Introduction第47-49页
    2.2 Materials and methods第49-52页
        2.2.1 Experimental insects第49页
        2.2.2 Hemocyte morphotypes:Observation of hemocytes on stained第49-50页
        2.2.3 Hemocyte morphotypes: Observation of the hemocytesimmediately after bleeding第50页
        2.2.4 Differential hemocyte count第50-51页
        2.2.5 Total hemocyte count第51-52页
        2.2.6 Statistical Analysis第52页
    2.3 Results第52-60页
        2.3.1 Morphological observations第52-56页
            2.3.1.1 Prohaemocytes第52-53页
            2.3.1.2 Plasmatocytes第53页
            2.3.1.3 Granulocytes第53-54页
            2.3.1.4 Oenocytoids第54页
            2.3.1.5 Adipohaemocyte第54页
            2.3.1.6 Spherulocytes第54-56页
        2.3.2 Differential hemocyte count第56-59页
        2.3.3 Total hemocyte population第59-60页
    2.4 Discussion第60-63页
CHAPTER 3: IMPACTS OF TRANSGENIC BT RICE ON HEMOCYTES OF BROWNPLANTHOPPER; Nilaparvata lugens Stal (Homoptera:Delphacidae)第63-83页
    3.1 Introduction第63-65页
    3.2 Materials and methods第65-69页
        3.2.1 The experimental population of N. lugens第65-66页
        3.2.2 Test plants第66页
        3.2.3 Rearing the experimental populations of BPH第66-67页
        3.2.4 Total hemocyte count第67-68页
        3.2.5 Differential hemocyte count第68页
        3.2.6 Histopathology of Hemocytes第68-69页
        3.2.7 Statistical Analysis第69页
    3.3 Results第69-79页
        3.3.1 Alterations onTHC of BPH after espouse to Bt toxin第69-70页
        3.3.2 Alterations on DHC of BPH after espouse to Bt toxin第70-78页
        3.3.3 Hemocyte abnormalities第78-79页
    3.4 Discussion第79-83页
CHAPTER 4: PHENOLOXIDASE ACTIVITY IN BROWN PLANTHOPPER;Nilaparvata lugens Stal (Homoptera:Delphacidae) FEEDING ON BT RICE第83-93页
    4.1 Introduction第83-85页
    4.2 Materials and methods第85-88页
        4.2.1 The experimental population of N.lugens第85页
        4.2.2 Experimental plant materials第85页
        4.2.3 Rearing of N. lugens on experimental plants第85页
        4.2.4 Hemocyte Lysate Supernatant(HLS)Preparation第85-86页
        4.2.5 Assay of PO Activity第86-87页
        4.2.6 Kinetic parameters(Vmax and Km)of PO第87页
        4.2.7 Statistical Analysis第87-88页
    4.3 Results第88-91页
        4.3.1 Phenoloxidase activity(PO)第88页
        4.3.2 Determination of kinetic parameters of PO第88-91页
    4.4 Discussion第91-93页
CHAPTER 5: DYNAMICS OF YEAST-LIKE SYMBIOTES IN BROWNPLANTHOPPER Nilaparvata lugens Stal(Hemiptera:Delphacidae)TO BT RICE第93-106页
    5.1 Introduction第93-94页
    5.2 Materials and methods第94-96页
        5.2.1 Insects第94页
        5.2.2 Plants第94-95页
        5.2.3 Feeding of insects on experimental plants第95页
        5.2.4 Quantification of YLS in BPH第95-96页
        5.2.5 Statistical Analysis第96页
    5.3 Results第96-104页
    5.4 Discussion第104-106页
CHAPTER 6: DIFFERENTIAL GENE EXPRESSION IN RESPONSE TO BT RICE INBROWN PLANTHOPPER Nilaparvata lugens Stak(Hemiptera:Delphacidae)第106-123页
    6.1 Introduction第106-108页
    6.2 Materials and methods第108-113页
        6.2.1 Test plant materials第108页
        6.2.2 Test insects第108-109页
        6.2.3 Total RNA Isolation and cDNA Synthesis第109-111页
        6.2.4 Quantitative real-time PCR(qRT-PCR)analysis第111-113页
        6.2.5 Statistical Analysis第113页
    6.3 Results第113-119页
        6.3.1 Immune-related genes第113-116页
        6.3.2 Detoxification related genes第116页
        6.3.3 Digestion-related gene第116-119页
    6.4 Discussion第119-123页
CHAPTER 7: BT RICE ON THE PHYSIOLOGICAL DEVELOPMENT OF BROWNPLANTHOPPER Nilaparvata lugens Stal(Hemiptera:Delphacidae)第123-134页
    7.1 Introduction第123-124页
    7.2 Materials and methods第124-126页
        7.2.1 Stock colonies of BPH第124页
        7.2.2 Plant materials第124-125页
        7.2.3 Oviposition preference of BPH第125页
        7.2 4 Survival,growth and development of BPH第125-126页
        7.2.5 Statistical analysis第126页
    7.3 Results第126-132页
        7.3.1 Oviposition preference opf BPH第126-127页
        7.3.2 Survical,growth and development of BPH第127-132页
    7.4 Discussion第132-134页
CHAPTER 8: PROTEOMIC ANALYSIS OF BROWN PLANTHOPPER;Nilaparvatalugens Stal(Homoptera:Delphacidae)FEEDING ON BT RICE:IDENTIFICATION OFBacillus thuringiens BINDING PROTEINS第134-154页
    8.1 Introduction第134-136页
    8.2 Materials and methods第136-146页
        8.2.1 Insects rearing第136-137页
        8.2.2 Purification of Cry toxin-binding proteins and Cry-binding assays第137-146页
            8.2.2.1 Crude Protein extraction from BPH第137页
            8.2.2.2 One dimensional-polyacrylamide gel electrophoresis(1DE-PAGE)第137-139页
            8.2.2.3 Two dimensional-polyacrylamide gel electrophoresis(2DE-PAGE)第139-143页
            8.2.2.4 Detection of Cry toxin by Western blot analysis第143-144页
            8.2.2.5 Cry binding assays第144-145页
            8.2.2.6 Silver staining for visualizes the protein spots第145-146页
    8.3 Results第146-152页
        8.3.1 Detection of Cry toxin by Western blot analysis第146页
        8.3.2 BPH Binding of Cry toxin to BHP第146-152页
    8.4 Discussion第152-154页
CHAPTER 9: GENERAL CONCLUSIONS AND FUTURE PERSPECTIVES第154-157页
REFERENCES第157-188页

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