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桃蚜对新烟碱类杀虫剂抗性机制及R81T突变的LAMP检测

中国农业科学院博士学位论文评阅人、答辩委员会签名表第5-6页
中文摘要第6-8页
Abstract第8-9页
Abbreviations第16-19页
CHAPTER 1 Introduction第19-37页
    1.1 Myzus persicae biology第19-20页
        1.1.1 Life cycle第19-20页
        1.1.2 Significance第20页
    1.2 Hormesis第20-22页
    1.3 Insecticide and Myzus persicae第22-29页
        1.3.1 Insecticide resistance第23页
        1.3.2 Resistance mechanisms第23-29页
    1.4 Synergists第29-30页
    1.5 Resistance monitoring based on loop-mediated isothermal amplification(LAMP)第30-35页
        1.5.1 Working principle第32-35页
        1.5.2 Detection of LAMP amplicons第35页
    1.6 Objectives and hypothesis第35-37页
CHAPTER 2 Evaluation of Insecticides induced hormesis based on the demographic parameters and expression changes of CYP6CY3 and E4/FE4 in Myzus persicae第37-54页
    2.1 Introduction第37-38页
    2.2 Material and Methods第38-41页
        2.2.1 Insect rearing第38页
        2.2.2 Insecticides solution第38-39页
        2.2.3 Bioassay第39页
        2.2.4 Sublethal dose exposure第39页
        2.2.5 Analysis of demographic parameters第39-40页
        2.2.6 Total RNA extraction and cDNA synthesis第40页
        2.2.7 Gene expression第40-41页
        2.2.8 Statistical analysis第41页
    2.3 Results第41-51页
        2.3.1 Bioassays第41-43页
        2.3.2 Sublethal effects of neonicotinoids第43-46页
        2.3.3 Sublethal effects of pyrethroids第46-49页
        2.3.4 Insecticides resistance linked genes expression第49-51页
    2.4 Discussion第51-54页
CHAPTER 3 Resistance to imidacloprid and acetamiprid in a field population of Myzus Persicae in China第54-66页
    3.1 Introduction第54-55页
    3.2 Materials and Methods第55-60页
        3.2.1 M.persicae populations and rearing第55-56页
        3.2.2 Chemicals and Reagents第56页
        3.2.3 Detection of insecticide resistance第56页
        3.2.4 Synergist response assays第56-57页
        3.2.5 Total RNA isolation and cDNA synthesis第57页
        3.2.6 Quantitative real-time PCR第57页
        3.2.7 Sequence analysis of nAChR第57-60页
        3.2.8 Statistical analysis第60页
    3.3 Results第60-64页
        3.3.1 M.persicae collected from Lang fang(FP)resistance to two neonicotinoids第60页
        3.3.2 Synergism of PBO in FP population compared to susceptible clone(SC)第60-62页
        3.3.3 Expression levels of cytochrome P450 CYP6CY3 and esterase E4/EF4 in SC clone and FP population第62页
        3.3.4 Sequence analysis of M.persicae n AChRβ1 subunit第62-64页
    3.4 Discussion第64-66页
CHAPTER 4 Loop-mediated isothermal amplification for the detection of R81T mutation in nAChR with crude genomic DNA extracted from individual Myzus persicae第66-83页
    4.1 Introduction第66-67页
    4.2 Materials and Methods第67-72页
        4.2.1 M.persicae clones or populations第67-68页
        4.2.2 Genomic DNA extraction,and quantification and quality detection第68页
        4.2.3 Amplification of the part of intronic sequence of the nAChR第68-69页
        4.2.4 Design of LAMP primers第69-70页
        4.2.5 Optimization of LAMP reaction components第70-71页
        4.2.6 Optimization of LAMP reaction conditions第71页
        4.2.7 Assessment of LAMP on detection R81T mutant genotype with different populations第71-72页
    4.3 Results第72-81页
        4.3.1 Quantity and quality of DNA第72-74页
        4.3.2 Amplification of R81T point mutation region of nAChR第74页
        4.3.3 LAMP development第74-79页
        4.3.4 Genotyping of field populations detected with the developed LAMP第79-81页
    4.4 Discussion第81-83页
CHAPTER 5 General Discussion第83-87页
Future prospects第87-88页
ACKNOWLEDGEMENTS第88-90页
REFERENCES第90-108页
Author Biography第108-115页

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