煙粉虱隱種內(nèi)共生菌種類多樣性分析及其優(yōu)勢共生菌對煙粉虱生長發(fā)育的影響
發(fā)布時間：2019-01-12 09:42
【摘要】：昆蟲的內(nèi)共生菌具有調(diào)節(jié)宿主生殖能力、協(xié)助宿主營養(yǎng)代謝、提高其溫度適應性以及增強宿主的抗藥性等功能,對昆蟲種群的動態(tài)平衡起著關鍵性的作用。本論文利用宏基因組學的手段測定并分析了不同煙粉虱隱種內(nèi)共生菌的種類和數(shù)量的差異,從基因水平分析預測了煙粉虱體內(nèi)優(yōu)勢內(nèi)共生菌的功能,并通過種群篩選進一步研究了其體內(nèi)的優(yōu)勢菌對煙粉虱生長發(fā)育和個體發(fā)育的影響。主要研究結(jié)果如下：(1)不同隱種所含內(nèi)共生菌種類和豐富度存在顯著差異。煙粉虱不同隱種的宏基因組數(shù)據(jù)分析結(jié)果表明,只存在煙粉虱MEAM1隱種的共生菌鑒定到種級水平15個,屬級水平26個以及科級水平19個；只存在煙粉虱MED隱種的共生菌鑒定到種級水平52個,屬級水平60個以及科級水平37個；煙粉虱MEAM1隱種和MED隱種共有的共生菌分類單位鑒定到種水平42個,屬水平77個以及科水平60個；這兩個隱種內(nèi)共生菌鑒定到總計包括(不含有重疊部分)種級水平109個屬級水平163個以及科級水平116個。相對于MEA M1隱種,煙粉虱MED隱種中Pseudomonas屬的種類如Pseudomonas mendocina 以及 Rickettisa 屬的種類如 Rickettisn felis, Rickettisa bellii 為優(yōu)勢種。(2)成功篩選/分離出含R.bellii和不含R.bellii菌的煙粉虱種群。通過對全國大部分地區(qū)的煙粉虱種群內(nèi)共生菌情況進行檢測,并進行單獨一雌一雄純化培養(yǎng)五代后,獲得了含有Ricketttsa bellii菌和不含有Rickettisa bellii菌的煙粉虱種群,經(jīng)分子鑒定為MED隱種。(3)明確了R.bellii菌對煙粉虱個體發(fā)育和生長發(fā)育的影響。通過對實驗種群生物學研究發(fā)現(xiàn),含有R.bellii菌的煙粉虱從卵到初羽化階段具有發(fā)育歷期短、產(chǎn)卵量多、存活時間長、后代雌性個體多的生殖優(yōu)勢。對不同發(fā)育時期的煙粉虱雌蟲解剖發(fā)現(xiàn),在煙粉虱卵子發(fā)育過程中,初羽化階段,不含有R.bellii菌的煙粉虱中未成熟卵子數(shù)量高于含有R.bellii菌的煙粉虱,但羽化第二天后,含有R.bellii菌的煙粉虱在卵子發(fā)育上更具有優(yōu)勢。(4)不同發(fā)育歷期的煙粉虱體內(nèi)的R.bellii菌拷貝數(shù)動態(tài)變化。以不同發(fā)育歷期的煙粉虱MED隱種的DNA為模板,利用絕對定量PCR技術檢測了供試煙粉虱中R.bellii菌的拷貝數(shù)。結(jié)果表明R.bellii菌的拷貝數(shù)在偽蛹期前沒有太大變化,在偽蛹期到初羽化發(fā)生顯著性增長,隨后下降,但第六天又開始發(fā)生顯著上升趨勢。本論文通過宏基因組數(shù)據(jù)的分析,明確了不同隱種間的內(nèi)共生菌種類差異及優(yōu)勢內(nèi)共生菌,建立了特異性侵染煙粉虱的優(yōu)勢菌的煙粉虱種群,明確了優(yōu)勢種菌對煙粉虱發(fā)育歷期、產(chǎn)卵量、存活率、抱卵量和后代性比等影響。為明確內(nèi)共生菌如何影響煙粉虱生殖發(fā)育和擴散提供了理論依據(jù),也為揭示內(nèi)共生菌調(diào)控宿主適應環(huán)境能力奠定了基礎。
[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.
【學位授予單位】：西南大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S433;Q93
本文編號：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.
【學位授予單位】：西南大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S433;Q93
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