Symbiotic Bacteria and the Fitness of Oriental Fruit Fly,Bac
發(fā)布時間：2021-03-13 02:21
橘小實蠅Bactrocera dorsalis（雙翅目,實蠅科）是一種世界范圍內(nèi)分布的重大果蔬害蟲,目前化學防治仍是控制橘小實蠅暴發(fā)危害的主要手段。隨著害蟲抗藥性的發(fā)展,以及人們對生態(tài)環(huán)境及健康問題的持續(xù)關(guān)注,迫切需要發(fā)展可持續(xù)的橘小實蠅綠色防控策略,其中包括昆蟲不育技術(shù)（SIT）及其相關(guān)生物防治技術(shù)。然而,經(jīng)過輻射后產(chǎn)生的不育雄蟲與野外雄蟲相比,面臨著交配競爭力低下的缺陷,并且大規(guī)模飼養(yǎng)的不育雄蟲與野外雄蟲相比,活力更弱。因此,本論文以橘小實蠅為研究對象,采用生化手段結(jié)合16S r RNA基因分析,分離鑒定其腸道及生殖器官的共生菌菌群種類及多樣性,通過在飼料中添加共生菌研究細菌對橘小實蠅適合度的影響;同時,通過向輻射不育的橘小實蠅雄蟲飼料中添加益生菌,探究益生菌對不育雄蟲交配競爭能力及雌蟲再交配行為的影響。此外,我們收集了中國和泰國的橘小實蠅（Bactrocera dorsalis）、南亞實蠅（B.tau）、木瓜實蠅（B.papayae）、楊桃實蠅（B.carambolae）四種實蠅害蟲的樣品,利用PCR技術(shù)檢測內(nèi)共生菌Wolbachia,Cardinium和Spiroplasma的...
【文章來源】：華中農(nóng)業(yè)大學湖北省 211工程院校 教育部直屬院校
【文章頁數(shù)】：115 頁
【學位級別】：博士
【文章目錄】：
Abstract
中文摘要
List of abbreviations
Chapter 1: General introduction
1.1 Introduction
1.2 Symbiotic bacteria in insects
1.2.1 Structure and community of bacteria associated with insect organ
1.2.1.1 Properties of bacteria in insect esophageal bulb
1.2.1.2 Properties of bacteria in insect gut
1.2.1.3 Properties of bacteria in reproductive organ
1.2.2 Bacteria isolation and identification in insects
1.2.2.1 Culture-dependent technique
1.2.2.2 Culture-independent technique
1.3 Function of symbiotic bacteria on insects
1.3.1 Bacteria provide host nutrients
1.3.2 Effects over host defense
1.3.3 Effects on development, longevity and other fitness parameters
1.4 Utilization of symbiotic bacteria towards on management of pests including sterile insect techniques （SIT） and incompatible insect technique （IIT）
1.4.1 The effects on mating performance in SIT
1.4.2 Development of wolbachia related IIT
1.5 Biology and management of Bactrocera dorsalis
1.5.1 General introduction and economic importance of B. dorsalis
1.5.2 The life history and distribution of B. dorsalis
1.5.3 The symbiotic bacteria in B. dorsalis and the application
1.5.4 Current management of B. dorsalis
1.5.4.1 Lure and kill approach
1.5.4.2 Male Annihilation Technique （MAT）
1.5.4.3 Area-wide integrated pest management
1.6 The objectives of this study
1.6.1 The flowchart of this study
Chapter 2: Isolation and identification of bacteria associated with gut and reproductive organ of laboratory oriental fruit fly, Bactrocera dorsalis
2.1 Introduction
2.2 Material and methods
2.2.1 Insect samples
2.2.2 Gut dissection and isolation of the intestinal culturable bacteria
2.2.3 Extracted DNA
2.2.4 PCR amplification and sequencing
2.3 Results and analyses
2.3.1 Identification of culturable bacteria in B. dorsalis
2.3.2 Taxonomic composition of bacteria at the phylum level
2.3.3 The influences tissue and sex on bacterial communities at genera level
2.4 Discussion
Chapter 3: Assessing the effect of gut bacteria manipulation on the development of the oriental fruit flies, Bactrocera dorsalis （Diptera: Tephritidiea）
3.1 Introduction
3.2 Material and Methods
3.2.1 The insect strain and rearing conditions
3.2.2 Growth rate of the bacteria
3.2.3 Preparation of experimental larval diet and feeding
3.2.4 The effect of bacteria on the developmental duration
3.2.5 The effect of bacteria on insect survival
3.2.6 The effect of gut bacterial isolates on pupal weight
3.2.7 Statistical analysis
3.3 Results and analyses
3.3.1 Gut bacterial community
3.3.2 The growth of gut symbiotic bacteria
3.3.3 Effect of bacteria on developmental duration
3.3.3.1 Larval developmental duration
3.3.3.2 Pupal developmental duration
3.3.3.3 Total developmental duration
3.3.4 The effect of bacteria on insect survival
3.3.4.1 Larval survival
3.3.4.2 Pupal survival
3.3.4.3 Total survival
3.3.5 Effect of bacteria on pupal weight
3.3.6 Association of weight, development time and survival
3.4 Discussion
Chapter 4: Effect of bacteria on mating competitiveness of the sterile male, Bactrocera dorsalis
4.1 Introduction
4.2 Materials and methods
4.2.1 Fly stocks and experiment design
4.2.2 Bacteria strains and the cultures
4.2.3 Effect of bacteria enriched diet on mating competitiveness of sterile males
4.2.4 Inhibition of female remating
4.2.5 Assessing probiotic effect on adult weight
4.2.6 Nutritional analyses
4.2.7 Statistical analysis
4.3 Results and analyses
4.3.1 Effects of Bacterial Inoculation on mating competitiveness of sterile males
4.3.2 Inhibition of female remating
4.3.3 Weight of adult flies
4.3.4 Biochemical analysis
4.3.4.1 Protein content
4.3.4.2 Triglyceride content
4.3.4.3 Glucose content
4.4 Discussion
Chapter 5: High-throughput next-generation of 16S r RNA gene reveals the core gut microbiome of the sterile oriental fruit fly, Bactrocera dorsalis
5.1 Introduction
5.2 Materials and methods
5.2.1 Flies
5.2.2 Gut dissection and DNA extraction
5.2.3 PCR amplification and illumina Mi Seq sequencing
5.2.4 Statistical and bioinformatics analysis
5.3 Results and analyses
5.3.1 The bacterial community
5.3.2 The bacterial composition at the phyla level
5.3.3 The bacterial composition at the family level
5.3.4 The bacterial composition at the genus level
5.4 Discussion
Chapter 6: Occurrence of the endosymbiotic bacteria across populations of Bactreocra fruit flies: Frequency and prevalence of Wolbachia, Cardinium, and Spiroplasma
6.1 Introduction
6.2 Material and methods
6.2.1 Insect sampling
6.2.2 DNA template preparation
6.2.3 PCR amplification and sequencing for reproductive symbionts
6.3 Results and analyses
6.3.1 Presence of Wolbachia
6.3.2 Presence of Cardiniun
6.3.3 Presence of Spiroplasma
6.4 Discussion
Chapter 7: General discussion and conclusion
References
Appendices
Curriculum Vitae
Acknowledgement
【參考文獻】：
期刊論文
[1]北京地區(qū)桔小實蠅的發(fā)生規(guī)律及防治措施研究[J]. 王曉梅,孫靜雙,李志強,段彥麗,張庭凱,高占月. 中國南方果樹. 2016(03)
[2]Bt殺蟲蛋白處理后二化螟幼蟲中腸細菌群落的變化[J]. 李振,韓蘭芝,劉玉娣,侯茂林. 昆蟲學報. 2016(03)
[3]不同抗藥性水平二化螟幼蟲中腸細菌群落多樣性分析[J]. 張玨鋒,何月平,陳建明. 昆蟲學報. 2013(09)
[4]Wolbachia與昆蟲精卵細胞質(zhì)不親和[J]. 袁林玲,劉晨,鄭雅,王玉鳳. 應用昆蟲學報. 2012(01)
[5]利用GARP生態(tài)位模型預測桔小實蠅（Bactrocera dorsalis）在中國的適生區(qū)域[J]. 周國梁,陳晨,葉軍,胡白石,劉鳳權(quán). 生態(tài)學報. 2007(08)
[6]桔小實蠅不育處理試驗研究初報[J]. 梁廣勤,梁帆,吳佳教,李惠河. 江西農(nóng)業(yè)大學學報. 2003(06)
本文編號：3079404
【文章來源】：華中農(nóng)業(yè)大學湖北省 211工程院校 教育部直屬院校
【文章頁數(shù)】：115 頁
【學位級別】：博士
【文章目錄】：
Abstract
中文摘要
List of abbreviations
Chapter 1: General introduction
1.1 Introduction
1.2 Symbiotic bacteria in insects
1.2.1 Structure and community of bacteria associated with insect organ
1.2.1.1 Properties of bacteria in insect esophageal bulb
1.2.1.2 Properties of bacteria in insect gut
1.2.1.3 Properties of bacteria in reproductive organ
1.2.2 Bacteria isolation and identification in insects
1.2.2.1 Culture-dependent technique
1.2.2.2 Culture-independent technique
1.3 Function of symbiotic bacteria on insects
1.3.1 Bacteria provide host nutrients
1.3.2 Effects over host defense
1.3.3 Effects on development, longevity and other fitness parameters
1.4 Utilization of symbiotic bacteria towards on management of pests including sterile insect techniques （SIT） and incompatible insect technique （IIT）
1.4.1 The effects on mating performance in SIT
1.4.2 Development of wolbachia related IIT
1.5 Biology and management of Bactrocera dorsalis
1.5.1 General introduction and economic importance of B. dorsalis
1.5.2 The life history and distribution of B. dorsalis
1.5.3 The symbiotic bacteria in B. dorsalis and the application
1.5.4 Current management of B. dorsalis
1.5.4.1 Lure and kill approach
1.5.4.2 Male Annihilation Technique （MAT）
1.5.4.3 Area-wide integrated pest management
1.6 The objectives of this study
1.6.1 The flowchart of this study
Chapter 2: Isolation and identification of bacteria associated with gut and reproductive organ of laboratory oriental fruit fly, Bactrocera dorsalis
2.1 Introduction
2.2 Material and methods
2.2.1 Insect samples
2.2.2 Gut dissection and isolation of the intestinal culturable bacteria
2.2.3 Extracted DNA
2.2.4 PCR amplification and sequencing
2.3 Results and analyses
2.3.1 Identification of culturable bacteria in B. dorsalis
2.3.2 Taxonomic composition of bacteria at the phylum level
2.3.3 The influences tissue and sex on bacterial communities at genera level
2.4 Discussion
Chapter 3: Assessing the effect of gut bacteria manipulation on the development of the oriental fruit flies, Bactrocera dorsalis （Diptera: Tephritidiea）
3.1 Introduction
3.2 Material and Methods
3.2.1 The insect strain and rearing conditions
3.2.2 Growth rate of the bacteria
3.2.3 Preparation of experimental larval diet and feeding
3.2.4 The effect of bacteria on the developmental duration
3.2.5 The effect of bacteria on insect survival
3.2.6 The effect of gut bacterial isolates on pupal weight
3.2.7 Statistical analysis
3.3 Results and analyses
3.3.1 Gut bacterial community
3.3.2 The growth of gut symbiotic bacteria
3.3.3 Effect of bacteria on developmental duration
3.3.3.1 Larval developmental duration
3.3.3.2 Pupal developmental duration
3.3.3.3 Total developmental duration
3.3.4 The effect of bacteria on insect survival
3.3.4.1 Larval survival
3.3.4.2 Pupal survival
3.3.4.3 Total survival
3.3.5 Effect of bacteria on pupal weight
3.3.6 Association of weight, development time and survival
3.4 Discussion
Chapter 4: Effect of bacteria on mating competitiveness of the sterile male, Bactrocera dorsalis
4.1 Introduction
4.2 Materials and methods
4.2.1 Fly stocks and experiment design
4.2.2 Bacteria strains and the cultures
4.2.3 Effect of bacteria enriched diet on mating competitiveness of sterile males
4.2.4 Inhibition of female remating
4.2.5 Assessing probiotic effect on adult weight
4.2.6 Nutritional analyses
4.2.7 Statistical analysis
4.3 Results and analyses
4.3.1 Effects of Bacterial Inoculation on mating competitiveness of sterile males
4.3.2 Inhibition of female remating
4.3.3 Weight of adult flies
4.3.4 Biochemical analysis
4.3.4.1 Protein content
4.3.4.2 Triglyceride content
4.3.4.3 Glucose content
4.4 Discussion
Chapter 5: High-throughput next-generation of 16S r RNA gene reveals the core gut microbiome of the sterile oriental fruit fly, Bactrocera dorsalis
5.1 Introduction
5.2 Materials and methods
5.2.1 Flies
5.2.2 Gut dissection and DNA extraction
5.2.3 PCR amplification and illumina Mi Seq sequencing
5.2.4 Statistical and bioinformatics analysis
5.3 Results and analyses
5.3.1 The bacterial community
5.3.2 The bacterial composition at the phyla level
5.3.3 The bacterial composition at the family level
5.3.4 The bacterial composition at the genus level
5.4 Discussion
Chapter 6: Occurrence of the endosymbiotic bacteria across populations of Bactreocra fruit flies: Frequency and prevalence of Wolbachia, Cardinium, and Spiroplasma
6.1 Introduction
6.2 Material and methods
6.2.1 Insect sampling
6.2.2 DNA template preparation
6.2.3 PCR amplification and sequencing for reproductive symbionts
6.3 Results and analyses
6.3.1 Presence of Wolbachia
6.3.2 Presence of Cardiniun
6.3.3 Presence of Spiroplasma
6.4 Discussion
Chapter 7: General discussion and conclusion
References
Appendices
Curriculum Vitae
Acknowledgement
【參考文獻】：
期刊論文
[1]北京地區(qū)桔小實蠅的發(fā)生規(guī)律及防治措施研究[J]. 王曉梅,孫靜雙,李志強,段彥麗,張庭凱,高占月. 中國南方果樹. 2016(03)
[2]Bt殺蟲蛋白處理后二化螟幼蟲中腸細菌群落的變化[J]. 李振,韓蘭芝,劉玉娣,侯茂林. 昆蟲學報. 2016(03)
[3]不同抗藥性水平二化螟幼蟲中腸細菌群落多樣性分析[J]. 張玨鋒,何月平,陳建明. 昆蟲學報. 2013(09)
[4]Wolbachia與昆蟲精卵細胞質(zhì)不親和[J]. 袁林玲,劉晨,鄭雅,王玉鳳. 應用昆蟲學報. 2012(01)
[5]利用GARP生態(tài)位模型預測桔小實蠅（Bactrocera dorsalis）在中國的適生區(qū)域[J]. 周國梁,陳晨,葉軍,胡白石,劉鳳權(quán). 生態(tài)學報. 2007(08)
[6]桔小實蠅不育處理試驗研究初報[J]. 梁廣勤,梁帆,吳佳教,李惠河. 江西農(nóng)業(yè)大學學報. 2003(06)
本文編號：3079404
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