植物毒素诱导甜菜夜蛾对杀虫剂产生耐受性差异的机制研究
发布时间：2024-05-25 05:32
在植物-昆虫互作中,棉酚等植物毒素通过干扰食草昆虫的基本代谢、生化和生理途径在植物防御中发挥重要作用。甜菜夜蛾是许多重要作物的主要害虫,其寄主包括蔬菜、棉花和观赏植物。在本论文中,我们研究了植物毒素对甜菜夜蛾对溴氰菊酯、氯虫苯甲酰胺等农药的耐药性的影响。采用棉酚对甜菜夜蛾幼虫进行饲喂预处理,再经杀虫剂溴氰菊酯连续筛选10代,Gos-SEL(棉酚筛选)种群产生了113.29倍的抗药性。在相同条件下,Delta-SEL(溴氰菊酯筛选)种群显示出69.76倍的抗性增加以及相关解毒酶活性的提高。同时,与Gos-SEL种群和实验室敏感种群(SS种群)比较,发现Delta-SEL种群的繁殖力以及雄性和雌性寿命显着降低。此外,与单独使用溴氰菊酯相比,当用溴氰菊酯和棉酚预处理的饲料喂养昆虫时,甜菜夜蛾中细胞色素P450的活性显着增强。总之,与对照组(SS-种群)相比,在Delta-SEL和Gos-SEL种群中甜菜夜蛾的繁殖能力显着降低。主要解毒酶细胞色素P450单加氧酶和酯酶的升高可能在棉酚诱导甜菜夜蛾种群提高对溴氰菊酯的耐受性中起重要作用。单宁是棉田中最重要的植物毒素之一。将单宁酸添加到人工饲料中饲...
【文章页数】：156 页
【学位级别】：博士
【文章目录】：
Abstract
摘要
CHAPTER1 Literature Review
1.1 Introduction of beet armyworm(Spodoptera exigua)
1.1.1 Biology and distribution
1.1.2 Geographical distribution
1.1.3 Host plants of beet armyworm
1.2 Insect-plant interaction and phytotoxins
1.3 Insect pests and pesticides
1.3.1 Overview of insect pests and insecticides
1.3.1.1 Pyrethroid insecticides
1.3.1.2 Diamide insecticides
1.3.2 Insecticides resistance in beet armyworm S.exigua
1.3.2.1 Overview of insecticides resistance mechanism in insect pests
1.3.2.2 Behavioral Resistance
1.3.2.3 Reduced penetration resistance(Delayed penetration)
1.3.3 Molecular mechanisms of metabolic base resistance to insecticides
1.3.3.1 Cytochrome P450-mediated insecticides resistance in beet armyworm
1.3.3.2 Esterase-mediated insecticides resistance in beet armyworm
1.3.3.3 Glutathione S-transferase-mediated insecticides resistance in beet armyworm
1.4 Eco-Friendly integrated pest Management approach for controlling beet armyworm S.exigua
1.4.1 Biotechnological Approaches for the management of beet armyworm
1.4.1.1 Use of RNA interference(RNAi)technique to control beet armyworm
1.4.1.2 Use of CRISPR/Cas9 technique to control beet armyworm
1.4.1.3 Use of sterilize insect technique(SIT)to control beet armyworm
1.4.2 Use of host plant resistance
1.5 Biological control
1.5.1 Cultural control practices
1.5.2 Objectives of the study
CHAPTER2 Gossypol‐induced fitness gain and increased resistance to deltamethrin in beet armyworm,Spodoptera exigua(Hübner)
2.1 Introduction
2.2 Materials and methods
2.2.1 Experimental insects rearing technique
2.2.2 Chemicals
2.2.3 Preparation of insecticide and gossypol-supplemented diets
2.2.4 Toxicity bioassays
2.2.5 Selection of gossypol& deltamethrin-resistant strains
2.2.6 Life table construction
2.2.7 Enzyme assays
2.2.7.1 Assays of P450 PNOD activities
2.2.7.2 Assays of Glutathione S-transferase(GST)activity
2.2.7.3 Esterase activity towards a-naphthyl acetate(a-NA)
2.2.8 Statistical analysis
2.2.9 Age-stage,Two-sex Life Table Analysis
2.3 Results
2.3.1 Toxicity of Gos-SEL and Delta-SEL population
2.3.2 Pre-adult developmental time for Gos-SEL and Delta-SEL strains
2.3.3 Adult longevity and growth metrics of S.exigua Gos-SEL and Delta-SEL
2.3.4 Pre-adults Pupal,Adult’s survival rate and Hatchability of the Gos-SEL and Delta-SEL population of Spodoptera exigua
2.3.5 Fitness comparison
2.3.6 Detoxification enzymes activity for Gos-SEL and Delta-SEL strains
2.3.7 Survival rate,life expectancy,reproductive value and fecundity of the Gos-SEL and Delta-SEL populations of Spodoptera exigua
2.4 Discussion
CHAPTER3 Enhanced effects of dietary tannic acid with chlorantraniliprole on life table parameters and nutritional physiology of Spodoptera exigua(Hübner)
3.1 Introduction
3.2 Materials and Methods
3.2.1 Insects breeding technique
3.2.2 Chemicals
3.2.3 Preparation of insecticide plus tannic acid-supplemented diets
3.2.4 Enhanced effects of tannic acid with of chlorantraniliprole against S.exigua
3.2.5 Effect of dietary tannic acid with chlorantraniliprole on feeding index of third-stage larvae of S.exigua
3.2.6 Measurement of nutrient contents
3.2.6.1 Total protein content
3.2.6.2 Measurement of lipid content
3.2.6.3 Carbohydrate content
3.2.7 Effect of dietary tannic acid alone and combination with chlorantraniliprole on the developmental growth and population parameters of S.exigua
3.2.8 Statistical analysis
3.2.9 Life Table Analysis
3.3 Results
3.3.1 Toxicity of tannic acid with chlorantraniliprole and tannic acid or chlorantraniliprole alone to third-stage larvae of S.exigua
3.3.2 Toxic effects of tannic acid alone and combined with chlorantraniliprole on the development and growth of S.exigua
3.3.3 Toxic effects of tannic acid alone and combined with chlorantraniliprole on the population growth parameters of S.exigua
3.3.4 Enhancement effects of tannic acid combined with chlorantraniliprole on feeding indices of S.exigua
3.3.5 Effects of tannic acid alone and combined with chlorantraniliprole on the total nutrient contents of S.exigua
3.4 Discussion
CHAPTER4 Knock-down of gossypol-inducing cytochrome P450 genes reduced deltamethrin sensitivity in Spodoptera exigua(Hübner)
4.1 Introduction
4.2 Materials and methods
4.2.1 Insect culture
4.2.2 Chemicals
4.2.3 Preparation of treatment diets
4.2.4 Toxicological analysis of deltamethrin tolerance in larvae
4.2.5 Effect of PBO on toxicity of insecticides
4.2.6 The effect of0.1%gossypol diet on bodyweight
4.2.7 Samples Preparation for P450 Enzyme Activity
4.2.8 Measurement of P450 Enzyme activity
4.2.9 Samples preparation
4.2.9.1 RNA extraction and cDNA synthesis
4.2.9.2 Quantitative real-time PCR
4.2.9.3 dsRNA Synthesis
4.2.9.4 Administration of dsRNA by Droplet-Feeding
4.2.9.5 Combined Effects of dsRNA on Mortality
4.2.9.6 Analysis of the Silencing Effect
4.2.10 Statistical Analysis
4.3 Results
4.3.1 Induced Effect of Gossypol to Deltamethrin Tolerance and Synergism Assessment
4.3.2 Effect of gossypol diet on larval body weight
4.3.3 Effect of gossypol on midgut P450 activity
4.3.4 Effect of gossypol,flavone and deltamethrin on expression response of P450 genes
4.3.5 Silencing effect of dsCYP6AB14 and dsCYP9A98 on larval mortality
4.3.6 Combined effect of target dsCYP6AB14+dsCYP9A98 genes on larval mortality
4.3.7 Effect of silencing by dsRNA
4.4 Discussion
CHAPTER5 Conclusion and Prospects
5.1 Conclusion
5.2 Prospects
References
Supplementary Appendix 1
List of Publications
Acknowledgement
本文编号：3981774
【文章页数】：156 页
【学位级别】：博士
【文章目录】：
Abstract
摘要
CHAPTER1 Literature Review
1.1 Introduction of beet armyworm(Spodoptera exigua)
1.1.1 Biology and distribution
1.1.2 Geographical distribution
1.1.3 Host plants of beet armyworm
1.2 Insect-plant interaction and phytotoxins
1.3 Insect pests and pesticides
1.3.1 Overview of insect pests and insecticides
1.3.1.1 Pyrethroid insecticides
1.3.1.2 Diamide insecticides
1.3.2 Insecticides resistance in beet armyworm S.exigua
1.3.2.1 Overview of insecticides resistance mechanism in insect pests
1.3.2.2 Behavioral Resistance
1.3.2.3 Reduced penetration resistance(Delayed penetration)
1.3.3 Molecular mechanisms of metabolic base resistance to insecticides
1.3.3.1 Cytochrome P450-mediated insecticides resistance in beet armyworm
1.3.3.2 Esterase-mediated insecticides resistance in beet armyworm
1.3.3.3 Glutathione S-transferase-mediated insecticides resistance in beet armyworm
1.4 Eco-Friendly integrated pest Management approach for controlling beet armyworm S.exigua
1.4.1 Biotechnological Approaches for the management of beet armyworm
1.4.1.1 Use of RNA interference(RNAi)technique to control beet armyworm
1.4.1.2 Use of CRISPR/Cas9 technique to control beet armyworm
1.4.1.3 Use of sterilize insect technique(SIT)to control beet armyworm
1.4.2 Use of host plant resistance
1.5 Biological control
1.5.1 Cultural control practices
1.5.2 Objectives of the study
CHAPTER2 Gossypol‐induced fitness gain and increased resistance to deltamethrin in beet armyworm,Spodoptera exigua(Hübner)
2.1 Introduction
2.2 Materials and methods
2.2.1 Experimental insects rearing technique
2.2.2 Chemicals
2.2.3 Preparation of insecticide and gossypol-supplemented diets
2.2.4 Toxicity bioassays
2.2.5 Selection of gossypol& deltamethrin-resistant strains
2.2.6 Life table construction
2.2.7 Enzyme assays
2.2.7.1 Assays of P450 PNOD activities
2.2.7.2 Assays of Glutathione S-transferase(GST)activity
2.2.7.3 Esterase activity towards a-naphthyl acetate(a-NA)
2.2.8 Statistical analysis
2.2.9 Age-stage,Two-sex Life Table Analysis
2.3 Results
2.3.1 Toxicity of Gos-SEL and Delta-SEL population
2.3.2 Pre-adult developmental time for Gos-SEL and Delta-SEL strains
2.3.3 Adult longevity and growth metrics of S.exigua Gos-SEL and Delta-SEL
2.3.4 Pre-adults Pupal,Adult’s survival rate and Hatchability of the Gos-SEL and Delta-SEL population of Spodoptera exigua
2.3.5 Fitness comparison
2.3.6 Detoxification enzymes activity for Gos-SEL and Delta-SEL strains
2.3.7 Survival rate,life expectancy,reproductive value and fecundity of the Gos-SEL and Delta-SEL populations of Spodoptera exigua
2.4 Discussion
CHAPTER3 Enhanced effects of dietary tannic acid with chlorantraniliprole on life table parameters and nutritional physiology of Spodoptera exigua(Hübner)
3.1 Introduction
3.2 Materials and Methods
3.2.1 Insects breeding technique
3.2.2 Chemicals
3.2.3 Preparation of insecticide plus tannic acid-supplemented diets
3.2.4 Enhanced effects of tannic acid with of chlorantraniliprole against S.exigua
3.2.5 Effect of dietary tannic acid with chlorantraniliprole on feeding index of third-stage larvae of S.exigua
3.2.6 Measurement of nutrient contents
3.2.6.1 Total protein content
3.2.6.2 Measurement of lipid content
3.2.6.3 Carbohydrate content
3.2.7 Effect of dietary tannic acid alone and combination with chlorantraniliprole on the developmental growth and population parameters of S.exigua
3.2.8 Statistical analysis
3.2.9 Life Table Analysis
3.3 Results
3.3.1 Toxicity of tannic acid with chlorantraniliprole and tannic acid or chlorantraniliprole alone to third-stage larvae of S.exigua
3.3.2 Toxic effects of tannic acid alone and combined with chlorantraniliprole on the development and growth of S.exigua
3.3.3 Toxic effects of tannic acid alone and combined with chlorantraniliprole on the population growth parameters of S.exigua
3.3.4 Enhancement effects of tannic acid combined with chlorantraniliprole on feeding indices of S.exigua
3.3.5 Effects of tannic acid alone and combined with chlorantraniliprole on the total nutrient contents of S.exigua
3.4 Discussion
CHAPTER4 Knock-down of gossypol-inducing cytochrome P450 genes reduced deltamethrin sensitivity in Spodoptera exigua(Hübner)
4.1 Introduction
4.2 Materials and methods
4.2.1 Insect culture
4.2.2 Chemicals
4.2.3 Preparation of treatment diets
4.2.4 Toxicological analysis of deltamethrin tolerance in larvae
4.2.5 Effect of PBO on toxicity of insecticides
4.2.6 The effect of0.1%gossypol diet on bodyweight
4.2.7 Samples Preparation for P450 Enzyme Activity
4.2.8 Measurement of P450 Enzyme activity
4.2.9 Samples preparation
4.2.9.1 RNA extraction and cDNA synthesis
4.2.9.2 Quantitative real-time PCR
4.2.9.3 dsRNA Synthesis
4.2.9.4 Administration of dsRNA by Droplet-Feeding
4.2.9.5 Combined Effects of dsRNA on Mortality
4.2.9.6 Analysis of the Silencing Effect
4.2.10 Statistical Analysis
4.3 Results
4.3.1 Induced Effect of Gossypol to Deltamethrin Tolerance and Synergism Assessment
4.3.2 Effect of gossypol diet on larval body weight
4.3.3 Effect of gossypol on midgut P450 activity
4.3.4 Effect of gossypol,flavone and deltamethrin on expression response of P450 genes
4.3.5 Silencing effect of dsCYP6AB14 and dsCYP9A98 on larval mortality
4.3.6 Combined effect of target dsCYP6AB14+dsCYP9A98 genes on larval mortality
4.3.7 Effect of silencing by dsRNA
4.4 Discussion
CHAPTER5 Conclusion and Prospects
5.1 Conclusion
5.2 Prospects
References
Supplementary Appendix 1
List of Publications
Acknowledgement
本文编号：3981774
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