红棕象甲肠道菌群介导调控宿主营养代谢促进幼虫的生长发育

发布时间：2021-02-01 01:37

　　红棕象甲（Red palm weevil,Rynchophorus ferrugineus Olivier）是一种危害全球棕榈植物的毁灭性蛀干害虫。最近的研究表明,红棕象甲的肠道内定殖着大量的细菌,而且肠道菌群结构的改变可以显著地影响该害虫的营养代谢。然而,关于肠道菌群对红棕象甲表型影响的具体机理目前仍不清楚。本研究发现,与对照组相比,肠道无菌的红棕象甲从卵到预蛹的发育时间明显延长,体重也明显降低。此外,检测发现无菌饲养的红棕象甲幼虫的血淋巴中蛋白质、葡萄糖和甘油三酯的含量显著地降低。有趣的是,将肠道菌群重新植入无菌幼虫的肠道可以显著地提高三种营养指数的水平,其中重新植入乳酸乳球菌Lactococcus lactis的无菌红棕象甲幼虫的血淋巴中蛋白质的含量恢复到正常水平,而植入阴沟肠杆菌Enterobacter cloacae可以显著地增加红棕象甲幼虫的血淋巴中甘油三酯和葡萄糖的含量。因此,我们的证据表明,肠道菌群可以通过调节红棕象甲幼虫营养代谢影响该害虫的发育,而且不同的肠道细菌对宿主生理水平有不同的影响。为进一步探明肠道菌群调节红棕象甲营养代谢的相关分子机理,我们克隆获得了RfIG... 

【文章来源】：福建农林大学福建省

【文章页数】：64 页

【学位级别】：硕士

【文章目录】：
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


【参考文献】：
期刊论文
[1]椰子大害虫—锈色棕榈象及其近缘种的鉴别（鞘翅目:象虫科）[J]. 张润志,任立,孙江华,吴坚,曾睿.  中国森林病虫. 2003(02)



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