| 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 |
