Symbiotic Bacteria and the Fitness of Oriental Fruit Fly,Bac

发布时间：2021-03-13 02:21

　　橘小实蝇Bactrocera dorsalis（双翅目,实蝇科）是一种世界范围内分布的重大果蔬害虫,目前化学防治仍是控制橘小实蝇暴发危害的主要手段。随着害虫抗药性的发展,以及人们对生态环境及健康问题的持续关注,迫切需要发展可持续的橘小实蝇绿色防控策略,其中包括昆虫不育技术（SIT）及其相关生物防治技术。然而,经过辐射后产生的不育雄虫与野外雄虫相比,面临着交配竞争力低下的缺陷,并且大规模饲养的不育雄虫与野外雄虫相比,活力更弱。因此,本论文以橘小实蝇为研究对象,采用生化手段结合16S r RNA基因分析,分离鉴定其肠道及生殖器官的共生菌菌群种类及多样性,通过在饲料中添加共生菌研究细菌对橘小实蝇适合度的影响;同时,通过向辐射不育的橘小实蝇雄虫饲料中添加益生菌,探究益生菌对不育雄虫交配竞争能力及雌虫再交配行为的影响。此外,我们收集了中国和泰国的橘小实蝇（Bactrocera dorsalis）、南亚实蝇（B.tau）、木瓜实蝇（B.papayae）、杨桃实蝇（B.carambolae）四种实蝇害虫的样品,利用PCR技术检测内共生菌Wolbachia,Cardinium和Spiroplasma的... 

【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校

【文章页数】：115 页

【学位级别】：博士

【文章目录】：
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


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