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亚洲玉米螟Cry1Ah抗性遗传规律与分子基础研究

摘要第6-8页
Abstract第8-9页
Abbreviations第14-16页
1.Introduction第16-30页
    1.1 General overview第16页
    1.2 Significance of Ostrinia furnacalis第16-17页
    1.3 ACB Integrated Control Technology第17页
    1.4 Introduction of Bacillus thuringiensis第17-22页
        1.4.1 Cry toxins:a diverse and large family of insecticidal protein第20-22页
        1.4.2 Transgenic crops:an environmentally friendly alternative for insect control第22页
        1.4.3 Pyramiding or cry gene stacking第22页
    1.5 Mechanism of Cry toxins第22-24页
        1.5.1 Dual mode of action of cry toxins第24页
    1.6 A threat to the technology:resistance to Cry toxins第24-26页
    1.7 Inheritance of the resistance第26-27页
    1.8 Significance of Cry1Ah toxin第27-29页
    1.9 Objectives第29-30页
2.Materials and methods第30-43页
    2.1 Insect strains第30页
    2.2 Bt toxins第30-31页
    2.3 Quantification of Cry1Ah toxin expressed in Bt maize tissues第31页
    2.4 Plant tissue bioassay第31-32页
    2.5 Diet Bioassay第32页
    2.6 Inheritance experiment第32-33页
    2.7 Preparation of BBMV from ACB larvae第33-34页
    2.8 Purification of Cry1Ah toxin第34-35页
    2.9 Pull down assay第35-36页
        2.9.1 Cry1Ah binding with medium第35页
        2.9.2 NHS-activated-Cry1Ah sepharose binding with BBMV proteins第35-36页
        2.9.3 SDS-PAGE analysis第36页
    2.10 Identification of binding proteins第36页
    2.11 RNA extraction and cDNA synthesis第36-37页
    2.12 Validation of gene expression第37-38页
    2.13 Library preparation for Transcriptome sequencing第38页
    2.14 Assembly and Functional Gene Annotation第38-39页
    2.15 Screening of differentially expressed genes between ACB-AhR and ACB-BtS第39页
    2.16 Protein quantification and database search using iTRAQ labeling第39-40页
    2.17 GO classification of differentially expressed genes and proteins pathway enrichment analysis第40-41页
    2.18 Correlation analysis of proteome and transcription groups第41页
    2.19 RT-qPCR for expression analysis第41页
    2.20 Statistical analysis第41-43页
3.Results第43-69页
    3.1 Resistance evolution to Cry1Ah in ACB第43页
    3.2 Cross-resistance to different Bt toxins第43-44页
    3.3 Survival on plant tissues第44-45页
    3.4 Quantification of Cry1Ah toxin第45页
    3.5 Maternal effects and sex linkage第45-46页
    3.6 Estimation of the degree of dominance第46-47页
    3.7 Number of loci influencing resistance第47-51页
    3.8 APN and ALP activities in ACB larval midgut第51页
    3.9 Cry1Ah-binding profiles in BBMV of ACB第51-53页
    3.10 Cry1Ah-binding proteins identified by LC-MS/MS第53页
    3.11 Primer evaluation and amplification efficiency第53-56页
    3.12 Expression profiles of Cry1Ah receptor genes第56-57页
    3.13 RNA-Seq and sequence assembly第57-59页
    3.14 Differentially expressed genes between Cry1Ah-resistant and susceptible strains of ACB第59-63页
    3.15 Cry1Ah-induced differentially expressed proteins between ACB-AhR and ACB-BtS strains第63-64页
    3.16 Gene Ontology and pathway enrichment第64-68页
    3.17 Validation of differentially expressed genes by RT-qPCR第68-69页
4.Discussion第69-80页
    4.1 Evolution and the inheritance of Cry1Ah resistance strait in ACB第69-72页
    4.2 Cry1Ah-binding proteins identified in ACB midgut through pull-down assay第72-75页
    4.3 molecular understanding of the resistance in ACB at transcriptome(RNA-seq)and proteome(i TRAQ)level第75-80页
5.General Conclusions第80-81页
References第81-99页
Acknowledgements第99-101页
Brief introduction of the author第101-103页
Appendix第103-115页

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