烟粉虱隐种内共生菌种类多样性分析及其优势共生菌对烟粉虱生长发育的影响
发布时间：2019-01-12 09:42
【摘要】：昆虫的内共生菌具有调节宿主生殖能力、协助宿主营养代谢、提高其温度适应性以及增强宿主的抗药性等功能,对昆虫种群的动态平衡起着关键性的作用。本论文利用宏基因组学的手段测定并分析了不同烟粉虱隐种内共生菌的种类和数量的差异,从基因水平分析预测了烟粉虱体内优势内共生菌的功能,并通过种群筛选进一步研究了其体内的优势菌对烟粉虱生长发育和个体发育的影响。主要研究结果如下：(1)不同隐种所含内共生菌种类和丰富度存在显著差异。烟粉虱不同隐种的宏基因组数据分析结果表明,只存在烟粉虱MEAM1隐种的共生菌鉴定到种级水平15个,属级水平26个以及科级水平19个；只存在烟粉虱MED隐种的共生菌鉴定到种级水平52个,属级水平60个以及科级水平37个；烟粉虱MEAM1隐种和MED隐种共有的共生菌分类单位鉴定到种水平42个,属水平77个以及科水平60个；这两个隐种内共生菌鉴定到总计包括(不含有重叠部分)种级水平109个属级水平163个以及科级水平116个。相对于MEA M1隐种,烟粉虱MED隐种中Pseudomonas属的种类如Pseudomonas mendocina 以及 Rickettisa 属的种类如 Rickettisn felis, Rickettisa bellii 为优势种。(2)成功筛选/分离出含R.bellii和不含R.bellii菌的烟粉虱种群。通过对全国大部分地区的烟粉虱种群内共生菌情况进行检测,并进行单独一雌一雄纯化培养五代后,获得了含有Ricketttsa bellii菌和不含有Rickettisa bellii菌的烟粉虱种群,经分子鉴定为MED隐种。(3)明确了R.bellii菌对烟粉虱个体发育和生长发育的影响。通过对实验种群生物学研究发现,含有R.bellii菌的烟粉虱从卵到初羽化阶段具有发育历期短、产卵量多、存活时间长、后代雌性个体多的生殖优势。对不同发育时期的烟粉虱雌虫解剖发现,在烟粉虱卵子发育过程中,初羽化阶段,不含有R.bellii菌的烟粉虱中未成熟卵子数量高于含有R.bellii菌的烟粉虱,但羽化第二天后,含有R.bellii菌的烟粉虱在卵子发育上更具有优势。(4)不同发育历期的烟粉虱体内的R.bellii菌拷贝数动态变化。以不同发育历期的烟粉虱MED隐种的DNA为模板,利用绝对定量PCR技术检测了供试烟粉虱中R.bellii菌的拷贝数。结果表明R.bellii菌的拷贝数在伪蛹期前没有太大变化,在伪蛹期到初羽化发生显著性增长,随后下降,但第六天又开始发生显著上升趋势。本论文通过宏基因组数据的分析,明确了不同隐种间的内共生菌种类差异及优势内共生菌,建立了特异性侵染烟粉虱的优势菌的烟粉虱种群,明确了优势种菌对烟粉虱发育历期、产卵量、存活率、抱卵量和后代性比等影响。为明确内共生菌如何影响烟粉虱生殖发育和扩散提供了理论依据,也为揭示内共生菌调控宿主适应环境能力奠定了基础。
[Abstract]:Insect endosymbionts play a key role in the homeostasis of insect populations by regulating their reproductive capacity, assisting their nutritional metabolism, improving their temperature adaptability and enhancing their host's resistance to insecticides. In this paper, we determined and analyzed the difference of species and quantity of coexisting bacteria in different Bemisia Tabaci by means of macrogenomics, and predicted the function of dominant symbiotic bacteria in Bemisia Tabaci from gene level. The effects of dominant bacteria on the growth and ontogeny of whitefly were further studied by population screening. The main results were as follows: (1) there were significant differences in the species and richness of endosymbiosis among different cryptic species. The results of macro genome data analysis of Bemisia Tabaci showed that only the symbiotic bacteria with MEAM1 cryptic species were identified to 15 species, 26 genera and 19 families. The symbiotic bacteria containing only MED cryptic species of Bemisia Tabaci were identified to have 52 species level, 60 genus level and 37 family level. The symbiotic bacteria taxonomic units shared by MEAM1 and MED species of whitefly identified 42 species with 77 genera and 60 families. A total of 163 species (109 genera) and 116 families (including 109 genera and 116 families) were identified by the two endosymbionts. Compared with MEA M1 cryptic species, the species of Pseudomonas genus in MED species of whitefly, such as Pseudomonas mendocina, and Rickettisa genus, such as Rickettisn felis, Rickettisa bellii, were the dominant species. (2) Bemisia Tabaci populations containing R.bellii and R.bellii bacteria were successfully screened and isolated. The symbiotic bacteria of Bemisia Tabaci population in most parts of the country were detected and purified for five generations with a single female and male. The Bemisia Tabaci populations containing and without Rickettisa bellii bacteria were obtained. (3) the effect of R.bellii on the individual development and growth of Bemisia Tabaci was confirmed. By studying the biology of the experimental population, it was found that the Bemisia Tabaci containing R.bellii had the advantages of short developmental period, more spawning, longer survival time and more female offspring from egg to emergence stage. Dissection of female Bemisia Tabaci at different developmental stages showed that during egg development, the number of immature eggs of Bemisia Tabaci without R.bellii was higher than that of Bemisia Tabaci with R.bellii, but after the second day of emergence, the number of immature eggs was higher than that of Bemisia Tabaci with R.bellii. Bemisia Tabaci containing R.bellii bacteria had more advantages in egg development. (4) dynamic changes of R.bellii copy number in different developmental stages of Bemisia Tabaci. The copy number of R.bellii bacteria in the tested whitefly was determined by absolute quantitative PCR using DNA of the covert species of whitefly MED in different developmental stages as a template. The results showed that the copy number of R.bellii did not change significantly before the pupa stage, but increased significantly from the pupa stage to the first emergence stage, then decreased, but began to increase significantly on the sixth day. Based on the analysis of macro genome data, the species differences and dominant endosymbionts among different cryptophore species were determined, and the population of Bemisia Tabaci, which specifically infected the dominant bacillus of Bemisia Tabaci, was established. The effects of dominant species on the developmental period, egg production, survival rate, egg holding and sex ratio of Bemisia Tabaci were determined. It provides a theoretical basis for understanding how endosymbiosis bacteria affect the reproductive development and diffusion of Bemisia Tabaci, and lays a foundation for revealing the ability of endosymbionts to regulate host adaptation to the environment.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433;Q93
本文编号：2407630
[Abstract]:Insect endosymbionts play a key role in the homeostasis of insect populations by regulating their reproductive capacity, assisting their nutritional metabolism, improving their temperature adaptability and enhancing their host's resistance to insecticides. In this paper, we determined and analyzed the difference of species and quantity of coexisting bacteria in different Bemisia Tabaci by means of macrogenomics, and predicted the function of dominant symbiotic bacteria in Bemisia Tabaci from gene level. The effects of dominant bacteria on the growth and ontogeny of whitefly were further studied by population screening. The main results were as follows: (1) there were significant differences in the species and richness of endosymbiosis among different cryptic species. The results of macro genome data analysis of Bemisia Tabaci showed that only the symbiotic bacteria with MEAM1 cryptic species were identified to 15 species, 26 genera and 19 families. The symbiotic bacteria containing only MED cryptic species of Bemisia Tabaci were identified to have 52 species level, 60 genus level and 37 family level. The symbiotic bacteria taxonomic units shared by MEAM1 and MED species of whitefly identified 42 species with 77 genera and 60 families. A total of 163 species (109 genera) and 116 families (including 109 genera and 116 families) were identified by the two endosymbionts. Compared with MEA M1 cryptic species, the species of Pseudomonas genus in MED species of whitefly, such as Pseudomonas mendocina, and Rickettisa genus, such as Rickettisn felis, Rickettisa bellii, were the dominant species. (2) Bemisia Tabaci populations containing R.bellii and R.bellii bacteria were successfully screened and isolated. The symbiotic bacteria of Bemisia Tabaci population in most parts of the country were detected and purified for five generations with a single female and male. The Bemisia Tabaci populations containing and without Rickettisa bellii bacteria were obtained. (3) the effect of R.bellii on the individual development and growth of Bemisia Tabaci was confirmed. By studying the biology of the experimental population, it was found that the Bemisia Tabaci containing R.bellii had the advantages of short developmental period, more spawning, longer survival time and more female offspring from egg to emergence stage. Dissection of female Bemisia Tabaci at different developmental stages showed that during egg development, the number of immature eggs of Bemisia Tabaci without R.bellii was higher than that of Bemisia Tabaci with R.bellii, but after the second day of emergence, the number of immature eggs was higher than that of Bemisia Tabaci with R.bellii. Bemisia Tabaci containing R.bellii bacteria had more advantages in egg development. (4) dynamic changes of R.bellii copy number in different developmental stages of Bemisia Tabaci. The copy number of R.bellii bacteria in the tested whitefly was determined by absolute quantitative PCR using DNA of the covert species of whitefly MED in different developmental stages as a template. The results showed that the copy number of R.bellii did not change significantly before the pupa stage, but increased significantly from the pupa stage to the first emergence stage, then decreased, but began to increase significantly on the sixth day. Based on the analysis of macro genome data, the species differences and dominant endosymbionts among different cryptophore species were determined, and the population of Bemisia Tabaci, which specifically infected the dominant bacillus of Bemisia Tabaci, was established. The effects of dominant species on the developmental period, egg production, survival rate, egg holding and sex ratio of Bemisia Tabaci were determined. It provides a theoretical basis for understanding how endosymbiosis bacteria affect the reproductive development and diffusion of Bemisia Tabaci, and lays a foundation for revealing the ability of endosymbionts to regulate host adaptation to the environment.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433;Q93
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