飛蝗是全球范圍的、最具破壞性的農(nóng)業(yè)害蟲。前人對它的生態(tài)學、生物學和防控方法有較全面的研究。如:飛蝗子代滯育受母代光周期影響。研究表明,FOXO在調(diào)控昆蟲滯育中發(fā)揮重要作用。但光周期誘導母代飛蝗調(diào)控子代滯育的分子機制仍不清楚。本研究分別對飛蝗脂肪體、卵巢和中樞神經(jīng)系統(tǒng)進行了轉(zhuǎn)錄組測序。其中長日照和短日照處理組對比,脂肪體和卵巢中有71個共有差異表達基因,中樞神經(jīng)系統(tǒng)有84個差異表達基因涉及滯育誘導并且與FOXO信號途徑密切相關,包括三羧酸循環(huán)、糖酵解,氧化磷酸化和PI3K-Akt途徑等。隨后,在所有處理組中隨機選取36個差異基因進行qRT-PCR驗證。結(jié)果顯示差異基因的相對表達量與轉(zhuǎn)錄組數(shù)據(jù)一致。進一步選取rai1,shkC和takeout三個基因研究它們的滯育調(diào)控功能。我們分別克隆了這三個基因,合成了它們的雙鏈RNA并注射羽化72小時內(nèi)的飛蝗雌成蟲。非滯育誘導條件的光周期為16 h:8h(L:D,溫度28°C,相對濕度為60%。非滯育誘導條件的光周期為10 h:14 h（L:D）,溫度28°C,相對濕度為60%。結(jié)果表明,在長日照或短日照下與對照相比,干擾候選基因后,候選基因的相對表達... 

【文章來源】：中國農(nóng)業(yè)科學院北京市

【文章頁數(shù)】：81 頁

【學位級別】：博士

【文章目錄】：
摘要
Abstract
Abbreviations
CHAPTER1 INTRODUCTION
    1.1 Background of migratory locust
    1.2 Damage behavior of migratory locusts
    1.3 Chemical control and their disadvantages
    1.4 Biological control
    1.5 Genetic and hormonal control
        1.5.1 Insect diapause
        1.5.2 Maternal photoperiod effect on diapause
        1.5.3 Role of retinoids in insect diapause
        1.5.4 Role of circadian clock genes during insect diapause
        1.5.5 Role of Insulin signaling/FOXO pathways in diapause induction
        1.5.6 Role of insulin receptors
        1.5.7 ROS activate the insulin signaling/FOXO
    1.6 Molecular technique’s used in the present study
    1.7 Objectives
    1.8 Techanical route of the present study
Chapter2 Transcriptomic analysis from fat body and ovary of L.migratoria
    2.1 Summary
    2.2 Materials and Methods
        2.2.1 Ethics Statement
        2.2.2 Insect rearing
        2.2.3 Dissection and sample collection method
        2.2.4 RNA extraction
        2.2.5 c DNA and RNA sequencing
        2.2.6 Sequence assembly,annotation and DEGs analysis
        2.2.7 Gene expressions by qRT-PCR
        2.2.8 Statistical analysis
    2.3 Results
        2.3.1 Transcriptomic analysis
        2.3.2 Differentially expressed genes(DEGs)between long photoperiod and short photoperiods in L.migratoria
        2.3.3 Correlation analysis of Differentally Expressed Genes(DEGs)amongst long and short photoperiods
        2.3.4 Different Expressed Genes(DEGs)functions in Cluster wise series
        2.3.5 KEGG pathways classification
        2.3.6 Validations of DEGs by quantitative real-time PCR(qRT-PCR)
    2.4 Discussion
Chapter3 Molecular cloning of rai1,shkC and their effect on maternal offspring diapause of L.migratoria
    3.1 Summary
    3.2 Material and Methods
        3.2.1 Insect rearing and sample collection method
        3.2.2 RNA Extraction
        3.2.3 cDNA Synthesis
        3.2.4 Molecular cloning and sequencing of rai1 and shkC
        3.2.5 dsRNA synthesis
        3.2.6 RNA interference by qRT-PCR
        3.2.7 Egg collection and diapause detection
        3.2.8 SOD and ROS activity detection
        3.2.9 Statistical analysis
    3.3 Results
        3.3.1 Cloning and Sequencing of rai1 and shkC
        3.3.2 RNAi efficiency and maternal diapause
        3.3.3 SOD and ROS activity changes after RNAi rai
    3.4 Discussion
Chapter4 Transcriptomic analysis from CNS of L.migratoria
    4.1 Summary
    4.2 Material and Methods
        4.2.1 Insect rearing
        4.2.2 Dissection and sample collection method
        4.2.3 RNA extraction and RNA-Seq
        4.2.4 Sequence assembly,annotation and DEGs analysis
        4.2.5 cDNA synthesis and qRT-PCR
    4.3 Results
        4.3.1 Transcriptomes sequencing and annotation assembly
        4.3.2 Different expressed genes(DEGs)from CNS of L.migratoria
        4.3.3 Functional classification by GO enrichment of DEGs
        4.3.4 KEGG pathway classification
        4.3.5 Gene expressions by using qRT-PCR
    4.4 Discussion
Chapter5 Molecular cloning of takeout gene and their effect on maternal offspring diapause of L.migratoria
    5.1 Summary
    5.2 Material and Methods
        5.2.1 Insect colony rearing and tissues collection
        5.2.2 RNA extraction
        5.2.3 cDNA synthesis
        5.2.4 Molecular cloning and sequencing of takeout gene
        5.2.5 dsRNA synthesis
        5.2.6 RNA interference
        5.2.7 Egg collection and diapause detection
    5.3 Results
        5.3.1 Cloning and sequencing of takeout gene
        5.3.2 RNA interference of dstakeout and maternal diapause
    5.4 Discussion
Chapter6 Conclusion
References
Appendix
Acknowledgement
Author Biography


【參考文獻】：
期刊論文
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