母代效应影响飞蝗胚胎滞育调控的分子机制
发布时间：2021-08-11 03:06
飞蝗是全球范围的、最具破坏性的农业害虫。前人对它的生态学、生物学和防控方法有较全面的研究。如:飞蝗子代滞育受母代光周期影响。研究表明,FOXO在调控昆虫滞育中发挥重要作用。但光周期诱导母代飞蝗调控子代滞育的分子机制仍不清楚。本研究分别对飞蝗脂肪体、卵巢和中枢神经系统进行了转录组测序。其中长日照和短日照处理组对比,脂肪体和卵巢中有71个共有差异表达基因,中枢神经系统有84个差异表达基因涉及滞育诱导并且与FOXO信号途径密切相关,包括三羧酸循环、糖酵解,氧化磷酸化和PI3K-Akt途径等。随后,在所有处理组中随机选取36个差异基因进行qRT-PCR验证。结果显示差异基因的相对表达量与转录组数据一致。进一步选取rai1,shkC和takeout三个基因研究它们的滞育调控功能。我们分别克隆了这三个基因,合成了它们的双链RNA并注射羽化72小时内的飞蝗雌成虫。非滞育诱导条件的光周期为16 h:8h(L:D,温度28°C,相对湿度为60%。非滞育诱导条件的光周期为10 h:14 h（L:D）,温度28°C,相对湿度为60%。结果表明,在长日照或短日照下与对照相比,干扰候选基因后,候选基因的相对表达...
【文章来源】：中国农业科学院北京市
【文章页数】：81 页
【学位级别】：博士
【文章目录】：
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
【参考文献】：
期刊论文
[1]有效积温与东亚飞蝗越冬卵滞育关系研究[J]. 王洁,涂雄兵,郝昆,范要丽,张泽华. 中国生物防治学报. 2014(06)
[2]应用绿僵菌防治东亚飞蝗田间试验[J]. 陆庆光,邓春光,陈长风. 昆虫天敌. 1996(04)
[3]东亚飞蝗（Locusta migratoria manilensis Meyen）在中国的发生动态[J]. 马世骏. 昆虫学报. 1958(01)
本文编号：3335319
【文章来源】：中国农业科学院北京市
【文章页数】：81 页
【学位级别】：博士
【文章目录】：
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
【参考文献】：
期刊论文
[1]有效积温与东亚飞蝗越冬卵滞育关系研究[J]. 王洁,涂雄兵,郝昆,范要丽,张泽华. 中国生物防治学报. 2014(06)
[2]应用绿僵菌防治东亚飞蝗田间试验[J]. 陆庆光,邓春光,陈长风. 昆虫天敌. 1996(04)
[3]东亚飞蝗（Locusta migratoria manilensis Meyen）在中国的发生动态[J]. 马世骏. 昆虫学报. 1958(01)
本文编号：3335319
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