飞蝗对绿僵菌IMI330189和MAD1蛋白的免疫应答
发布时间：2023-02-18 21:32
绿僵菌菌株(IMI330189)是一种能有效防治害虫的昆虫病原真菌。MAD1蛋白是由绿僵菌产生、协助绿僵菌附着在昆虫体表对昆虫产生毒力的一种蛋白。本实验通过饵剂饲喂的方法研究了绿僵菌菌株(IMI330189)和MAD1蛋白单独或混合取食对飞蝗的免疫相关酶和Toll通路基因的影响。研究表明当绿僵菌菌株(IMI330189)和MAD1蛋白混合处理飞蝗时,其累计死亡率最高为55%。MAD1+IMI330189混合处理使飞蝗PO的活性增加,单独使用MAD1蛋白处理时致使飞蝗体内的PO、POD和SOD活性降低。在处理72小时后,检测了飞蝗的中肠和体内Toll通路4个基因(MyD88、Cactus、Pelle和CaN)的表达量,发现当MAD1蛋白和MAD1+IMI330189混合处理后,MyD88在中肠和虫体的表达明显下降,但在中肠与虫体的下调程度不一致。IMI330189显著提高了Cactus在中肠和体内的表达。而MAD1+IMI330189混合处理显著降低了Cactus在中肠和体内的表达。单独使用MAD1蛋白和IMI330189处理可显著增加Pelle基因在飞蝗中肠中的表达,而混合处理MAD1...
【文章页数】：80 页
【学位级别】：博士
【文章目录】：
摘要
Abstract
Abbreviations
CHAPTER 1 INTRODUCTION
1.1 Locust Background
1.2 Chemical control
1.3 Biological control
1.4 MAD1 Protein
1.5 Mode of infection of Metarhizium anisopliae
1.6 Host defense
1.7 The Phenoloxidase System
1.8 Antioxidant enzymes and immune related genes
1.9 Toll Pathway
1.10 Objectives
1.11 Technical route of study
Chapter 2 Influence of Metarhizium anisopliae(IMI330189)and MAD1 Protein on enzymatic activities and Toll-related genes of Locusta migratoria
2.1 Summary
2.2 Materials and Methods
2.2.1 Ethics Statement
2.2.2 Locusta migratoria rearing methods and experimental conditions
2.2.3 Fungal cultures
2.2.4 MAD1 protein culture
2.2.5 Bioassay
2.2.6 Preparation of enzyme solution
2.2.7 Enzyme Activity Assay
2.2.7.1 Phenoloxidase
2.2.7.2 Superoxide dismutase and Peroxidase
2.2.8.Real-Time qRT-PCR
2.2.8.1 Total RNA Isolation and cDNA synthesis
2.2.8.2 Quantitative real-time reverse transcription-polymerase reaction
2.3 Data analysis
2.4 Results
2.4.1 Virulence to Locusta migratoria
2.4.2 Enzymatic activities in Locusta migratoria
2.4.3 Relative quantitative expression of Myd88 m RNA in Locusta migratoria
2.4.4 Relative quantitative expression of Cactus m RNA in Locusta migratoria
2.4.5 Relative quantitative expression of Pelle m RNA in Locusta migratoria
2.4.6 Relative quantitative expression of Ca N m RNA in Locusta migratoria
2.5 Discussion
Chapter 3 Involvement of Multi-Function Oxidase,Chitinase and Attacin in the resistance of Locusta migratoria against Metarhizium anisopliae strain(IMI330189)and and MAD1 Protein
3.1 Summary
3.2 Material and Methods
3.2.1 Locusta migratoria rearing and Experimental conditions
3.2.2 Fungal Culture
3.2.3 MAD1 protein culture
3.2.4 Bioassay
3.2.5 Enzyme preparation
3.2.6 Enzyme activity assay
3.2.7 Data analysis
3.3 Results
3.3.1 Application of Metarhizium anisopliae and MAD1 causing virulence to Locusta migratoria
3.3.2 Influence of Metarhizium anisopliae and MAD1 protein on the activity of MFO in Locusta migratoria
3.3.3 Influence of Metarhizium anisopliae and MAD1 Protein on the activity of Attacin in the Locusta migratoria
3.3.4 Influence of Metarhizium anisopliae and MAD1 protein on the activity of Chitinase in Locusta migratoria
3.4 Discussion
Chapter 4 The immune response of Locusta migratoria to Metarhizium anisopliae and MAD1 Protein
4.1 Summary
4.2 Material and Methods
4.2.1 Locust rearing and Experimental conditions
4.2.2 Fungal culture
4.2.3 MAD1 protein culture
4.2.4 Bioassay
4.2.5 Enzyme assay and enzyme activity assay
4.3 Real-Time qRT PCR
4.3.1 Total RNA extraction and cDNA synthesis
4.3.2 Quantitative Real-Time reverse transcription-polymerase chain reaction
4.3.3 Data analysis
4.4 Results
4.4.1 Virulence to Locusta migratoria
4.4.2 Influence of Metarhizium.anisopliae strain(IMI330189)and MAD1 on the activity of Chitinase in Locusta migratoria
4.4.3 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of PO in the Locusta migratoria
4.4.4 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of SOD in the Locusta migratoria
4.4.5 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of POD in the Locusta migratoria
4.4.6 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of ROS in the Locusta migratoria
4.4.7 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of AMP in the Locusta migratoria
4.4.8 Relative quantitative expression of Chitinase m RNA in the Locusta migratoria
4.4.9 Relative quantitative expression of Diptericin m RNA in the Locusta migratoria
4.4.10 Relative quantitative expression of GNBP3 m RNA in the Locusta migratoria
4.5 Discussion
Chapter 5 Conclusion
References
Appendix
Acknowledgements
Author Biography
本文编号：3745573
【文章页数】：80 页
【学位级别】：博士
【文章目录】：
摘要
Abstract
Abbreviations
CHAPTER 1 INTRODUCTION
1.1 Locust Background
1.2 Chemical control
1.3 Biological control
1.4 MAD1 Protein
1.5 Mode of infection of Metarhizium anisopliae
1.6 Host defense
1.7 The Phenoloxidase System
1.8 Antioxidant enzymes and immune related genes
1.9 Toll Pathway
1.10 Objectives
1.11 Technical route of study
Chapter 2 Influence of Metarhizium anisopliae(IMI330189)and MAD1 Protein on enzymatic activities and Toll-related genes of Locusta migratoria
2.1 Summary
2.2 Materials and Methods
2.2.1 Ethics Statement
2.2.2 Locusta migratoria rearing methods and experimental conditions
2.2.3 Fungal cultures
2.2.4 MAD1 protein culture
2.2.5 Bioassay
2.2.6 Preparation of enzyme solution
2.2.7 Enzyme Activity Assay
2.2.7.1 Phenoloxidase
2.2.7.2 Superoxide dismutase and Peroxidase
2.2.8.Real-Time qRT-PCR
2.2.8.1 Total RNA Isolation and cDNA synthesis
2.2.8.2 Quantitative real-time reverse transcription-polymerase reaction
2.3 Data analysis
2.4 Results
2.4.1 Virulence to Locusta migratoria
2.4.2 Enzymatic activities in Locusta migratoria
2.4.3 Relative quantitative expression of Myd88 m RNA in Locusta migratoria
2.4.4 Relative quantitative expression of Cactus m RNA in Locusta migratoria
2.4.5 Relative quantitative expression of Pelle m RNA in Locusta migratoria
2.4.6 Relative quantitative expression of Ca N m RNA in Locusta migratoria
2.5 Discussion
Chapter 3 Involvement of Multi-Function Oxidase,Chitinase and Attacin in the resistance of Locusta migratoria against Metarhizium anisopliae strain(IMI330189)and and MAD1 Protein
3.1 Summary
3.2 Material and Methods
3.2.1 Locusta migratoria rearing and Experimental conditions
3.2.2 Fungal Culture
3.2.3 MAD1 protein culture
3.2.4 Bioassay
3.2.5 Enzyme preparation
3.2.6 Enzyme activity assay
3.2.7 Data analysis
3.3 Results
3.3.1 Application of Metarhizium anisopliae and MAD1 causing virulence to Locusta migratoria
3.3.2 Influence of Metarhizium anisopliae and MAD1 protein on the activity of MFO in Locusta migratoria
3.3.3 Influence of Metarhizium anisopliae and MAD1 Protein on the activity of Attacin in the Locusta migratoria
3.3.4 Influence of Metarhizium anisopliae and MAD1 protein on the activity of Chitinase in Locusta migratoria
3.4 Discussion
Chapter 4 The immune response of Locusta migratoria to Metarhizium anisopliae and MAD1 Protein
4.1 Summary
4.2 Material and Methods
4.2.1 Locust rearing and Experimental conditions
4.2.2 Fungal culture
4.2.3 MAD1 protein culture
4.2.4 Bioassay
4.2.5 Enzyme assay and enzyme activity assay
4.3 Real-Time qRT PCR
4.3.1 Total RNA extraction and cDNA synthesis
4.3.2 Quantitative Real-Time reverse transcription-polymerase chain reaction
4.3.3 Data analysis
4.4 Results
4.4.1 Virulence to Locusta migratoria
4.4.2 Influence of Metarhizium.anisopliae strain(IMI330189)and MAD1 on the activity of Chitinase in Locusta migratoria
4.4.3 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of PO in the Locusta migratoria
4.4.4 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of SOD in the Locusta migratoria
4.4.5 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of POD in the Locusta migratoria
4.4.6 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of ROS in the Locusta migratoria
4.4.7 Influence of Metarhizium anisopliae strain IMI330189 and MAD1 on the activity of AMP in the Locusta migratoria
4.4.8 Relative quantitative expression of Chitinase m RNA in the Locusta migratoria
4.4.9 Relative quantitative expression of Diptericin m RNA in the Locusta migratoria
4.4.10 Relative quantitative expression of GNBP3 m RNA in the Locusta migratoria
4.5 Discussion
Chapter 5 Conclusion
References
Appendix
Acknowledgements
Author Biography
本文编号：3745573
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