本文選題：煙粉虱　切入點：中國番茄黃化曲葉病毒　出處：《浙江大學》2015年博士論文　論文類型：學位論文

【摘要】：煙粉虱(Bemesia tabaci)是由至少36個隱種組成的隱種復合體,其中的一些隱種如MEAM1和MED由于其繁殖能力強、寄主適應范圍廣、抗藥性強等特點近年來隨著貿易活動的往來快速地入侵到世界各地。它們以持久性可循環(huán)的方式傳播雙生病毒,并與其進化出獨特的關系。煙粉虱的暴發(fā)通常伴隨著雙生病毒病的流行。雙生病毒對寄主植物的基因表達和生理功能的調控可能影響煙粉虱的種群動態(tài),進而又影響病毒本身的傳播。茉莉酸信號通路(JA signaling)及其調節(jié)的次生代謝物質(如芥子油苷和萜類化合物等)在植物應對病原物及昆蟲取食過程中起著重要作用。本文先從煙粉虱的角度出發(fā)探討了植物JA防御途徑對煙粉虱適合度的影響及其基因表達的調控；然后探討了重要農業(yè)經濟作物番茄在應對煙粉虱取食、病毒感染以及兩者共同侵染時的基因表達變化；最后以模式植物擬南芥為材料,探討煙粉虱及病毒致病因子pC1在擬南芥芥子油苷生成中的影響以及這種影響對煙粉虱適合度的作用。研究結果如下：1) JA防御途徑對煙粉虱基因表達及適合度的影響通過第二代測序技術,從轉錄水平上揭示了MEMA1煙粉虱應對植物JA防御反應的分子機制。煙粉虱在JA激活的煙草上取食3天后,共有242條基因上調,450條基因下調。對這些基因的分析結果表明,煙粉虱的初級代謝過程如糖代謝、氨基酸代謝、脂代謝等等都被顯著地下調；與能量代謝有關的基因都被顯著下調,說明能量代謝很可能受到抑制。然而與煙粉虱解毒相關的基因都顯著地上調,說明煙粉虱解毒活動增強。另外,結果顯示,與煙粉虱細胞周期、神經性疾病等有關的基因都受到顯著地調節(jié)。這些結果表明煙粉虱主動調節(jié)其基因表達來降低植物防御對其的傷害,卻也可能仍然受到植物抗性物質的毒害作用,導致煙粉虱在JA途徑激活的煙草上死亡率升高。2)番茄應對煙粉虱取食、病毒感染以及煙粉虱-病毒共同侵染的分子機制通過第二代測序技術,分析了番茄在受到煙粉虱取食、病毒感染以及煙粉虱-病毒共同侵染條件下的基因表達。三種處理得到的差異表達基因分別為324條、686條和996條,共同感染對番茄基因表達的影響最為明顯,不同處理間調節(jié)的基因有部分重疊也有各處理特異差異表達的基因,并且重疊的基因大多都有著相同的調節(jié)模式。病毒感染及共同感染顯著地抑制了番茄的光合作用及苯丙素和類黃酮代謝過程,可能與病毒感染引起番茄植物黃化有關。同時,許多與細胞壁及碳代謝有關的基因都受到顯著地調節(jié)。番茄免疫反應相關的許多抗性基因以及病程相關基因被顯著上調,說明番茄可能通過基礎免疫以及基因對基因抗性來識別和抵御煙粉虱取食和病毒感染。在對植物激素的分析中,發(fā)現(xiàn)JA途徑相關基因受到病毒感染和共同侵染時的抑制,而與生長有關的激素如生長素、細胞分裂素、脫落酸、赤霉素等受到更為顯著地調節(jié),這些可能與病毒感染及昆蟲取食引起植物生理異常如矮化、葉片卷曲等有關。另外有許多差異表達基因沒有功能注釋或者其在植物-昆蟲-病毒互作中的研究很少,如組蛋白相關基因。3)次生代謝物芥子油苷在煙粉虱-植物-病毒三者互作中的作用通過對煙粉虱處理以及轉pC1基因植株芥子油苷含量的測定,發(fā)現(xiàn)煙粉虱的取食能顯著誘導擬南芥芥子油苷的生成,而pC1基因能顯著地抑制擬南芥芥子油苷的生成。蓮座葉中脂肪族芥子油苷含量在生長初期占主要部分,隨著擬南芥的生長而逐漸降低,而吲哚族芥子油苷含量保持穩(wěn)定并在后期成為葉片中主要的芥子油苷。生物測定結果顯示,芥子油甘含量降低沒有顯著提高煙粉虱產卵量,但吲哚族芥子油苷的過量表達能顯著影響煙粉虱的產卵量及發(fā)育速率；βC1基因通過影響JA途徑而提高煙粉虱的適合度,次生代謝物芥子油苷及萜類等物質都可能參與到該過程。綜上所述,本研究分別探索了番茄應對煙粉虱取食、病毒感染以及煙粉虱-病毒共同侵染時的分子機制以及植物JA防御途徑對煙粉虱基因表達的影響;并且探討了次生代謝物芥子油苷在煙粉虱-植物-病毒三者互作中的作用。這些研究結果有助于我們更好地理解這三者的關系,并為進一步研究抗性基因和研發(fā)防治二者共同為害的新方法提供線索。
[Abstract]:Whitefly (Bemesia tabaci) is composed of at least 36 cryptic species composed of cryptic species complex, some cryptic species such as MEAM1 and MED because of its strong breeding ability, adapt to a wide range of host resistance, strong in recent years with trade exchanges rapidly all over the world. They invaded in persistence cycle the transmission of geminiviruses, and developed a unique relationship. The outbreak of Bemisia tabaci is usually accompanied by a geminivirus disease epidemic. The regulation of gene expression on host plants and the physiological function of geminiviruses may affect the population dynamics of Bemisia tabaci, then influence the spread of the virus itself. The jasmonic acid signaling pathway (JA signaling) and the regulation of secondary metabolites (such as mustard oil glycosides and terpenoids etc.) plays an important role in plant response to pathogen and insect feeding process. This paper first from the point of view of Bemisia tabaci Regulation of plant defense pathways JA effects on the fitness of Bemisia tabaci and its gene expression; and then discusses the important agricultural crops of Tomato in response to whitefly infestation, virus infection and expression of both infected genes; finally in Arabidopsis thaliana, to investigate the effects of whitefly and virus virulence factor pC1 in Arabidopsis mustard oil from in the generation of fitness function and the influence of the whitefly. The results are as follows: 1) JA pathway on the expression of defense genes and whitefly fitness effects through the second generation sequencing technology, to reveal the molecular mechanism of MEMA1 JA to the plant defense response of Bemisia tabaci from the transcription level. The whitefly in JA activation feeding on tobacco for 3 days, a total of 242 genes were up-regulated and 450 genes were down regulated. The analysis results of these genes showed that primary metabolic processes such as glucose metabolism of Bemisia tabaci, Amino acid metabolism, lipid metabolism and so on are significantly reduced; associated with energy metabolism genes were significantly down regulated, energy metabolism is likely to be inhibited. However, associated with the whitefly detoxification genes are significantly up-regulated, indicating the whitefly detoxification activity increased. In addition, the results showed that B.tabaci neural cell cycle. Disease related genes were significantly regulated. These results suggest that the active regulation of the gene expression of Bemisia tabaci to reduce the damage to the plant defense, but also may still be toxic substances of plant resistance, leading to activation of the JA pathway in B.tabaci tobacco increased mortality.2) with tomato whitefly infestation. Virus infection and virus infection of whitefly common molecular mechanism through the second generation sequencing technology, analyzed by tomato whitefly infestation, virus infection and whitefly virus together The expression of the gene expression under the condition of infection. The differences between the three genes respectively 324, 686 and 996, a common infection effect on the expression of gene of tomato is the most obvious, among different treatments regulated genes overlap also expressed the specific genes, and most of them have overlapping genes adjust the same pattern. Virus infection and co infection was significantly inhibited by photosynthesis and tomato and benzene flavonoids metabolism process, may cause the tomato plant chlorosis related to viral infection. At the same time, many related to the cell wall and carbon metabolism genes are subject to significant regulation. Many resistance genes related to the immune response of tomato and pathogenesis related genes were significantly up-regulated, indicating the tomato may through basic immunization and gene for gene resistance to identify and resist feeding and virus infection. In plant tobacco whitefly induced In the analysis, found that the inhibition of JA gene related pathway by viral infections and common infection, and growth related hormones such as auxin, cytokinins, abscisic acid, gibberellic acid has more significant adjustment, these may be associated with viral infection and insect herbivory caused by plant physiological abnormalities such as dwarfing, leaf curl and so on also. There are many differences between the expression of no functional annotation or rarely in plant insect - virus interaction of genes, such as histone.3 gene) effects of secondary metabolites of glucosinolates in whitefly - plant - three virus interactions in the determination of whitefly and pC1 transgenic plants glucosinolate content the produce of B.tabaci can induce the glucosinolates in Arabidopsis thaliana, while pC1 gene can significantly inhibit the formation of glucosinolates in Arabidopsis thaliana. Containing aliphatic glucosinolates in rosette leaves Accounts for the main part in the early stage of growth, with the growth of Arabidopsis thaliana was decreased, while the glycoside content of indole glucosinolate oil remain stable and become the main glucosinolates in leaves in the later stage. The bioassay results showed that mustard oil content decreased not significantly increased whitefly oviposition, but indole glucosinolate overexpression can and the growth rate significantly affected the fecundity of B.tabaci; beta C1 gene through JA pathway and improve the effect of fitness of Bemisia tabaci, secondary metabolites of glucosinolates and terpenoid substances may be involved in the process. In summary, this study explored separately tomato whitefly feeding response, the molecular mechanism of virus infection and whitefly when the virus infection and the common pathway of plant defense on the expression of JA gene in Bemisia tabaci; and discusses the secondary metabolites in mustard oil glycosides of whitefly virus Plant interaction three These findings help us to better understand the relationship between the three, and provide clues for further research on the resistance genes and the new ways to develop and control two common hazards.

【學位授予單位】：浙江大學
【學位級別】：博士
【學位授予年份】：2015
【分類號】：S433;S432.41
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本文編號：1580499