綠僵菌菌株(IMI330189)是一種能有效防治害蟲的昆蟲病原真菌。MAD1蛋白是由綠僵菌產(chǎn)生、協(xié)助綠僵菌附著在昆蟲體表對昆蟲產(chǎn)生毒力的一種蛋白。本實(shí)驗(yàn)通過餌劑飼喂的方法研究了綠僵菌菌株(IMI330189)和MAD1蛋白單獨(dú)或混合取食對飛蝗的免疫相關(guān)酶和Toll通路基因的影響。研究表明當(dāng)綠僵菌菌株(IMI330189)和MAD1蛋白混合處理飛蝗時(shí),其累計(jì)死亡率最高為55%。MAD1+IMI330189混合處理使飛蝗PO的活性增加,單獨(dú)使用MAD1蛋白處理時(shí)致使飛蝗體內(nèi)的PO、POD和SOD活性降低。在處理72小時(shí)后,檢測了飛蝗的中腸和體內(nèi)Toll通路4個(gè)基因(MyD88、Cactus、Pelle和CaN)的表達(dá)量,發(fā)現(xiàn)當(dāng)MAD1蛋白和MAD1+IMI330189混合處理后,MyD88在中腸和蟲體的表達(dá)明顯下降,但在中腸與蟲體的下調(diào)程度不一致。IMI330189顯著提高了Cactus在中腸和體內(nèi)的表達(dá)。而MAD1+IMI330189混合處理顯著降低了Cactus在中腸和體內(nèi)的表達(dá)。單獨(dú)使用MAD1蛋白和IMI330189處理可顯著增加Pelle基因在飛蝗中腸中的表達(dá),而混合處理MAD1...

【文章頁數(shù)】：80 頁

【學(xué)位級別】：博士

【文章目錄】：
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



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