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紅棕象甲腸道菌群介導調控宿主營養(yǎng)代謝促進幼蟲的生長發(fā)育

發(fā)布時間:2021-02-01 01:37
  紅棕象甲(Red palm weevil,Rynchophorus ferrugineus Olivier)是一種危害全球棕櫚植物的毀滅性蛀干害蟲。最近的研究表明,紅棕象甲的腸道內定殖著大量的細菌,而且腸道菌群結構的改變可以顯著地影響該害蟲的營養(yǎng)代謝。然而,關于腸道菌群對紅棕象甲表型影響的具體機理目前仍不清楚。本研究發(fā)現(xiàn),與對照組相比,腸道無菌的紅棕象甲從卵到預蛹的發(fā)育時間明顯延長,體重也明顯降低。此外,檢測發(fā)現(xiàn)無菌飼養(yǎng)的紅棕象甲幼蟲的血淋巴中蛋白質、葡萄糖和甘油三酯的含量顯著地降低。有趣的是,將腸道菌群重新植入無菌幼蟲的腸道可以顯著地提高三種營養(yǎng)指數(shù)的水平,其中重新植入乳酸乳球菌Lactococcus lactis的無菌紅棕象甲幼蟲的血淋巴中蛋白質的含量恢復到正常水平,而植入陰溝腸桿菌Enterobacter cloacae可以顯著地增加紅棕象甲幼蟲的血淋巴中甘油三酯和葡萄糖的含量。因此,我們的證據(jù)表明,腸道菌群可以通過調節(jié)紅棕象甲幼蟲營養(yǎng)代謝影響該害蟲的發(fā)育,而且不同的腸道細菌對宿主生理水平有不同的影響。為進一步探明腸道菌群調節(jié)紅棕象甲營養(yǎng)代謝的相關分子機理,我們克隆獲得了RfIG... 

【文章來源】:福建農(nóng)林大學福建省

【文章頁數(shù)】:64 頁

【學位級別】:碩士

【文章目錄】:
摘要
Abstract
1 Introduction
    1.1 Gut microbiota on digestion and degradation of polymers
        1.1.1 Gut microbiota on the degradation of lignin
        1.1.2 Gut microbiota on the degradation of cellulose and pectin
    1.2 Gut microbiota on detoxifying food
    1.3 Gut microbiota on fermentation
    1.4 Gut microbiota-mediated nutrient provisioning
    1.5 Gut microbiota on growth and development
    1.6 Background of Red Palm Weevil
    1.7 RPW and its gut microbiota
    1.8 Gut microbiota on expression of insulin growth factor binding protein Acid-labile subunit (IGFALS)
        1.8.1 Expression of IGFs across different tissues
        1.8.2 Effect of gut microbiota on the expression of IGFs in other animals
2. Materials and Methods
    2.1 Insect collection and maintenance
    2.2 Generating the axenic R. ferrugineus larvae
    2.3. Generating the gnotobiotic RPW larvae
    2.4 Effect of eliminating gut microbiota on physiological traits
        2.4.1 Egg hatching success and prepupal developmental duration
        2.4.2 RPW body mass
        2.4.3. Assessing the effect of gut microbiota on larva survival
    2.5. Effect of eliminating gut microbiota on RPW nutrition metabolism
        2.5.1 Collection of hemolymph
        2.5.2 Effect of eliminating gut microbiota on the concentration of protein
        2.5.3 Effect of eliminating gut microbiota on the concentration of glucose
        2.5.4 Effect of eliminating gut microbiota on the concentration of TAG
        2.5.5 Effect of eliminating gut microbiota on the concentration of trehalose
    2.6 Effect of eliminating gut microbiota on the expression of growth anddevelopment related Rf IGFALS gene
        2.6.1 Extracting total RNA and gene cloning
        2.6.2 Analysis on the expression level of Rf IGFALS by Real Time –q PCR
        2.6.3 Synthesis of ds RNA
        2.6.4 Injecting ds RNA into RPW larvae and verifying RNAi efficiency
        2.6.5 Hemolymph collection from Rf IGFALS knockdown insects
        2.6.6 Collection of fat body from Rf IGFALS knockdown insects
        2.6.7 Measurements of TAG level from hemolymph and fat body
        2.6.8 Measurements of glucose from hemolymph and fat body
    2.7 Statistical analysis
3. Results
    3.1 Effect of gut microbiota on an insect on hatching success, development,survival and body mass
    3.2 Effect of gut microbiota on the concentration of hemolymph Protein, Glucose,TG and trehalose in RPW larvae
    3.3 Impact of Lactococcus lactis and Enterobacter cloacaemonoassciation with RPW larvae on the concentration of hemolymphProtein, Glucose, TAG and Trehalose
        3.4.1 c DNA cloning and sequence analysis
        3.4.2 Rf IGFALS constitutively expressed in different tissues of RPW larvae
        3.4.3 The presence of gut microbiota induced the expression of Rf IGFALS
        3.4.4 The effect of Rf IGFALS ds RNA in the gut and fat body
        3.4.5 Effect of Silencing Rf IGFALS on TAG and Glucose content from in hemolymphand fat body
4 Discussion and Conclusion
References
Publication list
Appendix list of Abbreviation
Acknowledgement


【參考文獻】:
期刊論文
[1]椰子大害蟲—銹色棕櫚象及其近緣種的鑒別(鞘翅目:象蟲科)[J]. 張潤志,任立,孫江華,吳堅,曾睿.  中國森林病蟲. 2003(02)



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