烟粉虱—寄主植物—双生病毒互作及芥子油苷在三者互作中的作用

发布时间：2018-03-07 18:47

  本文选题：烟粉虱　切入点：中国番茄黄化曲叶病毒　出处：《浙江大学》2015年博士论文　论文类型：学位论文

【摘要】：烟粉虱(Bemesia tabaci)是由至少36个隐种组成的隐种复合体,其中的一些隐种如MEAM1和MED由于其繁殖能力强、寄主适应范围广、抗药性强等特点近年来随着贸易活动的往来快速地入侵到世界各地。它们以持久性可循环的方式传播双生病毒,并与其进化出独特的关系。烟粉虱的暴发通常伴随着双生病毒病的流行。双生病毒对寄主植物的基因表达和生理功能的调控可能影响烟粉虱的种群动态,进而又影响病毒本身的传播。茉莉酸信号通路(JA signaling)及其调节的次生代谢物质(如芥子油苷和萜类化合物等)在植物应对病原物及昆虫取食过程中起着重要作用。本文先从烟粉虱的角度出发探讨了植物JA防御途径对烟粉虱适合度的影响及其基因表达的调控；然后探讨了重要农业经济作物番茄在应对烟粉虱取食、病毒感染以及两者共同侵染时的基因表达变化；最后以模式植物拟南芥为材料,探讨烟粉虱及病毒致病因子pC1在拟南芥芥子油苷生成中的影响以及这种影响对烟粉虱适合度的作用。研究结果如下：1) JA防御途径对烟粉虱基因表达及适合度的影响通过第二代测序技术,从转录水平上揭示了MEMA1烟粉虱应对植物JA防御反应的分子机制。烟粉虱在JA激活的烟草上取食3天后,共有242条基因上调,450条基因下调。对这些基因的分析结果表明,烟粉虱的初级代谢过程如糖代谢、氨基酸代谢、脂代谢等等都被显著地下调；与能量代谢有关的基因都被显著下调,说明能量代谢很可能受到抑制。然而与烟粉虱解毒相关的基因都显著地上调,说明烟粉虱解毒活动增强。另外,结果显示,与烟粉虱细胞周期、神经性疾病等有关的基因都受到显著地调节。这些结果表明烟粉虱主动调节其基因表达来降低植物防御对其的伤害,却也可能仍然受到植物抗性物质的毒害作用,导致烟粉虱在JA途径激活的烟草上死亡率升高。2)番茄应对烟粉虱取食、病毒感染以及烟粉虱-病毒共同侵染的分子机制通过第二代测序技术,分析了番茄在受到烟粉虱取食、病毒感染以及烟粉虱-病毒共同侵染条件下的基因表达。三种处理得到的差异表达基因分别为324条、686条和996条,共同感染对番茄基因表达的影响最为明显,不同处理间调节的基因有部分重叠也有各处理特异差异表达的基因,并且重叠的基因大多都有着相同的调节模式。病毒感染及共同感染显著地抑制了番茄的光合作用及苯丙素和类黄酮代谢过程,可能与病毒感染引起番茄植物黄化有关。同时,许多与细胞壁及碳代谢有关的基因都受到显著地调节。番茄免疫反应相关的许多抗性基因以及病程相关基因被显著上调,说明番茄可能通过基础免疫以及基因对基因抗性来识别和抵御烟粉虱取食和病毒感染。在对植物激素的分析中,发现JA途径相关基因受到病毒感染和共同侵染时的抑制,而与生长有关的激素如生长素、细胞分裂素、脱落酸、赤霉素等受到更为显著地调节,这些可能与病毒感染及昆虫取食引起植物生理异常如矮化、叶片卷曲等有关。另外有许多差异表达基因没有功能注释或者其在植物-昆虫-病毒互作中的研究很少,如组蛋白相关基因。3)次生代谢物芥子油苷在烟粉虱-植物-病毒三者互作中的作用通过对烟粉虱处理以及转pC1基因植株芥子油苷含量的测定,发现烟粉虱的取食能显著诱导拟南芥芥子油苷的生成,而pC1基因能显著地抑制拟南芥芥子油苷的生成。莲座叶中脂肪族芥子油苷含量在生长初期占主要部分,随着拟南芥的生长而逐渐降低,而吲哚族芥子油苷含量保持稳定并在后期成为叶片中主要的芥子油苷。生物测定结果显示,芥子油甘含量降低没有显著提高烟粉虱产卵量,但吲哚族芥子油苷的过量表达能显著影响烟粉虱的产卵量及发育速率；βC1基因通过影响JA途径而提高烟粉虱的适合度,次生代谢物芥子油苷及萜类等物质都可能参与到该过程。综上所述,本研究分别探索了番茄应对烟粉虱取食、病毒感染以及烟粉虱-病毒共同侵染时的分子机制以及植物JA防御途径对烟粉虱基因表达的影响;并且探讨了次生代谢物芥子油苷在烟粉虱-植物-病毒三者互作中的作用。这些研究结果有助于我们更好地理解这三者的关系,并为进一步研究抗性基因和研发防治二者共同为害的新方法提供线索。
[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.

【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：S433;S432.41
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本文编号：1580499