10种真菌对三种昆虫的毒力差异与调控宿主基因表达模式研究
发布时间：2021-05-09 12:54
近年来,微生物杀虫剂的应用越来越受到人们的重视。在昆虫的病原微生物中,昆虫病原真菌（EPF）占据着非常重要的地位,与其他微生物相比,EPF可以很容易地通过表皮渗透进入昆虫体内,其他微生物需要通过口器摄入才能引起感染。在真菌穿透昆虫表皮的过程中,昆虫和病原真菌之间发生了一系列复杂的相互作用,最终,病原真菌导致昆虫死亡。整个过程中涉及多种生物化学和生理机制,虽然以前的研究揭示了其中一些机制,但很多机制仍然未被揭示。本研究利用三个属（Metarhizium,Isaria和Beauveria）的病原真菌对三种昆虫（米象Sitophilus20oryzae.桃蚜Myzus20persicae,豌豆芽Acyrthosiphon20pisum）的致病效率和病理学进行了研究。对米象毒力最强的菌株进行了筛选。结果表明,在分生孢子剂量为1×107/ml时,各种病原菌的LT50值不同,ME-33（ILT-01）Isaria20cateniannulatus为3.63±0.11天、Qin-2120Beauveria20bassiana为4.17±0.18天Qin-1...
【文章来源】：西北农林科技大学陕西省 211工程院校 985工程院校 教育部直属院校
【文章页数】：113 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
CHAPTER1 Introduction
1.1 Literature Review,Potential role of Entomopathogenic Fungi in Biological Control of Insect Pests
1.2 History of biological control and the EPF background
1.3 EPF Usage,Benefits and Limitations
1.3.1 Efficacy over the Chemical Pesticides usage
1.3.2 EPF infection mode advantages over other Entomopathogens
1.4 Habitats and Uses of Entomopathogenic fungi
1.5 Characterization of Entomopathogenic Fungi and Biology
1.6 An overview on EPF Classification
1.7 Dispersal and host specificity of EPF
1.8 Myco-bio control of insect pests,progress and development
1.9 Indigenous EPF,a well option for biological control
1.10 Current status and economic importance Entomopathogenic Fungi
1.10.1 Beauveria Species
1.10.2 Metarhizium Species
1.10.3 Isaria Species
1.11 Tested insect models in study
1.11.1 Rice Weevil(Sitophilus oryzae.)
1.11.2 Pest Status
1.11.3 An overview on the Biology and Ecology of Pest
1.12 Green Peach Aphid(Myzus persicae)
1.12.1 Pest Status
1.12.2 Biology and Ecology of Green Peach Aphid
1.13 Pea Aphid(Acyrthosiphon pisum)
1.13.1 Pest Status
1.13.2 Biology and Ecology of Pea Aphid
CHAPTER2 Pathogenicity of the Indigenous isolates Metarhizium,Isaria and Beauveria against the Sitophilus oryzae(Coleoptera:Curculionidae)Under the Lab conditions
2.1 Introduction
2.2 Materials and Methods
2.2.1 Insect culture
2.2.2 Source of fungal isolates
2.2.3 Preparation of fungal conidia
2.2.4 Initial pathogenicity assay for EPF isolates against S.oryzae
2.2.5 Virulence assay of EPF against S.oryzae
2.2.6 Statistical analysis
2.3 Results
2.3.1 Fungal Isolates tested by Immersion Method at Single Concentration Bioassay
2.3.2 Fungal Isolates tested via Food Mix Method at Single Concentration Bioassay
2.3.3 Comparison of Immersion and Food Mix Fungus Isolates
2.3.4 Multiple conidial concentration bioassays
2.3.5 LT50 at various conidial concentrations
2.4 Discussion
2.5 Conclusion
CHAPTER3 Potential of Four Entomopathogenic Fungi Isolates as Biological Control Agents against two Aphid Species under Laboratory Conditions
3.1 Introduction
3.2 Materials and Methods
3.2.1 Plants and Insects stock
3.2.2 Preparation of fungus culture,conidial suspension and Gemination test
3.2.3 Pathogenicity of EPF Isolates
3.2.4 Concentration–mortality response
3.2.5 Efficacy of fungal isolates at different temperatures
3.2.6 Statistical analysis
3.3 Results
3.4 Discussion
3.5 Conclusions
CHAPTER4 Biochemical Study,Post infection effect of Entomopathogenic fungi on the profile activity of some Anti-Oxidative and Detoxifying Enzymes in Acyrthosiphon pisum
4.1 Introduction
4.2 Materials and Methods
4.2.1 Plants and Insects Rearing
4.2.2 Fungal culture,conidial suspension and viability test
4.2.3 Bioassay and sample collection
4.2.4 Samples for Enzymes preparation
4.2.5 Protein concentration test
4.3 Detoxification enzymes activity,post exposure to fungal infection
4.3.1 GS-Glutathione S-Transferase enzyme activity
4.3.2 CarE-Carboxylesterase enzyme activity
4.3.3 AKP-Alkaline phosphate enzyme activity
4.4 Anti-oxidative enzymes activity assay
4.4.1 SOD-Superoxide dismutase enzyme activity
4.4.2 POD-Peroxidase enzyme activity
4.4.3 CAT-Catalase enzyme activity
4.4.4 Statistical analysis
4.5 Results and Discussion
4.5.1 Effect of I.fumosorosea and M.anisopliae isolate on the actitivity of anti-oxidative enzymes on the post infection
4.5.2 Effect of I.fumosorosea and M.anisopliae isolate on the Detoxification enzymes Post infection
4.6 Conclusion
CHAPTER5 Evaluation of transcriptome analysis indicates the differential gene expression for Acyrthosiphon pisum as induced by Isaria fumosorosea
5.1 Introduction
5.2 Materials and Methods
5.2.1 Insect and plant stock
5.2.2 Fungal culture,conidial suspension and sample collection
5.2.3 Biological assay of A.pisum adults infected with Isaria fumosorosea SP-535 isolate and sample prepration for RNA-Seq
5.2.4 RNA Isolation cDNA Library and Illumina sequencing
5.2.5 Alignment and annotation
5.2.6 Quality control
5.2.7 Comparative analysis
5.2.8 Gene functional annotation
5.2.9 Differential expression analysis,For the samples with biological replicates
5.2.10 GO enrichment analysis
5.2.11 KEGG pathway enrichment analysis
5.3 Results
5.3.1 Features and Statistics of sequenced cDNA libraries
5.3.2 Novel genes detection
5.3.3 Functional annotation
5.3.4 Overall distribution of gene expression levels in all samples
5.3.5 Differential expression analysis
5.3.6 Correlation assessment of biological replicates
5.3.7 Identification of differentially expressed genes
5.3.8 Clustering analysis of differentially expressed genes
5.3.9 GO classification of differentially expressed genes
5.3.10 KEGG pathway enrichment analysis of differentially expressed genes
5.4 Discussion
5.5 Conclusion
CHAPTER6 Discussion and Conclusion
6.1 General discussion
6.2 Conclusion
REFERENCES
Acknowledgements
Author’s Introduction
【参考文献】：
期刊论文
[1]我国古代农业技术的优良传统之一——生物防治[J]. 彭世奖. 中国农业科学. 1983(01)
本文编号：3177338
【文章来源】：西北农林科技大学陕西省 211工程院校 985工程院校 教育部直属院校
【文章页数】：113 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
CHAPTER1 Introduction
1.1 Literature Review,Potential role of Entomopathogenic Fungi in Biological Control of Insect Pests
1.2 History of biological control and the EPF background
1.3 EPF Usage,Benefits and Limitations
1.3.1 Efficacy over the Chemical Pesticides usage
1.3.2 EPF infection mode advantages over other Entomopathogens
1.4 Habitats and Uses of Entomopathogenic fungi
1.5 Characterization of Entomopathogenic Fungi and Biology
1.6 An overview on EPF Classification
1.7 Dispersal and host specificity of EPF
1.8 Myco-bio control of insect pests,progress and development
1.9 Indigenous EPF,a well option for biological control
1.10 Current status and economic importance Entomopathogenic Fungi
1.10.1 Beauveria Species
1.10.2 Metarhizium Species
1.10.3 Isaria Species
1.11 Tested insect models in study
1.11.1 Rice Weevil(Sitophilus oryzae.)
1.11.2 Pest Status
1.11.3 An overview on the Biology and Ecology of Pest
1.12 Green Peach Aphid(Myzus persicae)
1.12.1 Pest Status
1.12.2 Biology and Ecology of Green Peach Aphid
1.13 Pea Aphid(Acyrthosiphon pisum)
1.13.1 Pest Status
1.13.2 Biology and Ecology of Pea Aphid
CHAPTER2 Pathogenicity of the Indigenous isolates Metarhizium,Isaria and Beauveria against the Sitophilus oryzae(Coleoptera:Curculionidae)Under the Lab conditions
2.1 Introduction
2.2 Materials and Methods
2.2.1 Insect culture
2.2.2 Source of fungal isolates
2.2.3 Preparation of fungal conidia
2.2.4 Initial pathogenicity assay for EPF isolates against S.oryzae
2.2.5 Virulence assay of EPF against S.oryzae
2.2.6 Statistical analysis
2.3 Results
2.3.1 Fungal Isolates tested by Immersion Method at Single Concentration Bioassay
2.3.2 Fungal Isolates tested via Food Mix Method at Single Concentration Bioassay
2.3.3 Comparison of Immersion and Food Mix Fungus Isolates
2.3.4 Multiple conidial concentration bioassays
2.3.5 LT50 at various conidial concentrations
2.4 Discussion
2.5 Conclusion
CHAPTER3 Potential of Four Entomopathogenic Fungi Isolates as Biological Control Agents against two Aphid Species under Laboratory Conditions
3.1 Introduction
3.2 Materials and Methods
3.2.1 Plants and Insects stock
3.2.2 Preparation of fungus culture,conidial suspension and Gemination test
3.2.3 Pathogenicity of EPF Isolates
3.2.4 Concentration–mortality response
3.2.5 Efficacy of fungal isolates at different temperatures
3.2.6 Statistical analysis
3.3 Results
3.4 Discussion
3.5 Conclusions
CHAPTER4 Biochemical Study,Post infection effect of Entomopathogenic fungi on the profile activity of some Anti-Oxidative and Detoxifying Enzymes in Acyrthosiphon pisum
4.1 Introduction
4.2 Materials and Methods
4.2.1 Plants and Insects Rearing
4.2.2 Fungal culture,conidial suspension and viability test
4.2.3 Bioassay and sample collection
4.2.4 Samples for Enzymes preparation
4.2.5 Protein concentration test
4.3 Detoxification enzymes activity,post exposure to fungal infection
4.3.1 GS-Glutathione S-Transferase enzyme activity
4.3.2 CarE-Carboxylesterase enzyme activity
4.3.3 AKP-Alkaline phosphate enzyme activity
4.4 Anti-oxidative enzymes activity assay
4.4.1 SOD-Superoxide dismutase enzyme activity
4.4.2 POD-Peroxidase enzyme activity
4.4.3 CAT-Catalase enzyme activity
4.4.4 Statistical analysis
4.5 Results and Discussion
4.5.1 Effect of I.fumosorosea and M.anisopliae isolate on the actitivity of anti-oxidative enzymes on the post infection
4.5.2 Effect of I.fumosorosea and M.anisopliae isolate on the Detoxification enzymes Post infection
4.6 Conclusion
CHAPTER5 Evaluation of transcriptome analysis indicates the differential gene expression for Acyrthosiphon pisum as induced by Isaria fumosorosea
5.1 Introduction
5.2 Materials and Methods
5.2.1 Insect and plant stock
5.2.2 Fungal culture,conidial suspension and sample collection
5.2.3 Biological assay of A.pisum adults infected with Isaria fumosorosea SP-535 isolate and sample prepration for RNA-Seq
5.2.4 RNA Isolation cDNA Library and Illumina sequencing
5.2.5 Alignment and annotation
5.2.6 Quality control
5.2.7 Comparative analysis
5.2.8 Gene functional annotation
5.2.9 Differential expression analysis,For the samples with biological replicates
5.2.10 GO enrichment analysis
5.2.11 KEGG pathway enrichment analysis
5.3 Results
5.3.1 Features and Statistics of sequenced cDNA libraries
5.3.2 Novel genes detection
5.3.3 Functional annotation
5.3.4 Overall distribution of gene expression levels in all samples
5.3.5 Differential expression analysis
5.3.6 Correlation assessment of biological replicates
5.3.7 Identification of differentially expressed genes
5.3.8 Clustering analysis of differentially expressed genes
5.3.9 GO classification of differentially expressed genes
5.3.10 KEGG pathway enrichment analysis of differentially expressed genes
5.4 Discussion
5.5 Conclusion
CHAPTER6 Discussion and Conclusion
6.1 General discussion
6.2 Conclusion
REFERENCES
Acknowledgements
Author’s Introduction
【参考文献】：
期刊论文
[1]我国古代农业技术的优良传统之一——生物防治[J]. 彭世奖. 中国农业科学. 1983(01)
本文编号：3177338
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