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首页> 外文学位 >Characterization of Begomoviruses from the Dominican Republic and Peru and Functional Analysis of the Genes of the First New World Monopartite Begomovirus.
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Characterization of Begomoviruses from the Dominican Republic and Peru and Functional Analysis of the Genes of the First New World Monopartite Begomovirus.

机译：来自多米尼加共和国和秘鲁的乞eg病毒的表征和第一批新世界单方乞B病毒基因的功能分析。

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摘要

Begomoviruses are an emergent and economically important group of single stranded DNA viruses in tropical and subtropical regions worldwide. In the second chapter the biological and molecular properties of three strains of the bipartite begomovirus Jatropha mosaic virus (JMV) from the Dominican Republic (DO) were determined. Jatropha plants inoculated with the infectious clones of these JMV strains developed yellow mosaic symptoms, which fulfilled Koch's postulates. For this disease. Furthermore, these JMV strains also infected the crop plants, tobacco and common bean, revealing a possible link between weed- and crop-infecting begomoviruses. Finally, asymmetry in replication and infectivity of pseudorecombinants formed between the DNA components of these strains was consistent with a high degree of genetic diversity, and suggested a long history of local evolution of JMV in the DO. In the third chapter, the first bona fide New World monopartite begomovirus Tomato leaf deformation virus (ToLDeV) was characterized, and shown to be the causal agent of tomato leaf curl disease in Peru and Ecuador based on multimeric clones of the genomic DNA of three ToLDeV genotypes, delivered via agroinoculation, inducing leaf curl symptoms in tomato indistinguishable from those observed in the tomato fields. Biological properties of ToLDeV were similar to those of Old World monopartite tomato-infecting begomoviruses, including lack of sap-transmissibility, phloem limitation, a resistance phenotype in tomato germplasm with the Ty-1 gene and functional properties of the V1 (capsid protein) and C4 genes. Differences in symptom phenotypes induced by the ToLDeV genotypes in tomato (e.g., whether a recovery phenotype was observed or not) and N. benthamiana plants were associated with a highly divergent left intergenic region and C4 gene. Finally, evidence is presented that the ToLDeV genotypes emerged from the DNA-A component of a New World bipartite progenitor via parallel evolution, mutation and recombination. In the fourth chapter, the functions of the ToLDeV C2 (TrAP) and C4 genes were investigated by mutational analysis and ectopic expression via two viral vectors with distinct tissue tropism (phloem-limited and non-phloem limited). ToLDeV C2 mutants replicated in N. benthamiana leaf discs, but the infectivity was host-specific. The capacity of the TrAP and C4 viral proteins to suppress gene silencing was evaluated. Neither TrAP nor C4 protein suppressed local silencing; however, C4 acted as a strong suppressor of long-distance silencing. Both C2 and C4 showed some capacity to suppress transcriptional gene silencing based on restoring GFP expression in transcriptionally silenced 16c-TGS plants. Moreover, a high level of cytosine methylation in the left intergenic region of ToLDeV was associated with tomato plants showing the recovery phenotype following infection with ToLDeV. Finally, subcellular localization studies revealed that the TrAP protein localized to the nucleus, whereas C4 was localized to the plasma membrane.
机译：在世界范围内的热带和亚热带地区，Begomovirus病毒是一种新兴且具有经济意义的单链DNA病毒。在第二章中，确定了来自多米尼加共和国（DO）的三联双歧Begomovirus麻风树花叶病毒（JMV）的生物学和分子特性。接种了这些JMV菌株感染性克隆的麻风树属植物出现了黄色花叶病症状，符合科赫的假设。对于这种疾病。此外，这些JMV菌株还感染了农作物，烟草和普通豆，揭示了杂草感染和农作物感染的bemomovirus可能存在联系。最后，在这些菌株的DNA组分之间形成的假重组体的复制和感染性的不对称性与高度遗传多样性相一致，并表明DO中JMV的局部进化历史悠久。在第三章中，描述了第一个真正的新世界单方begomovirus番茄叶片变形病毒（ToLDeV），并基于三个ToLDeV基因组DNA的多聚体克隆显示出秘鲁和厄瓜多尔番茄叶片卷曲病的病原体。通过农杆菌接种传递的基因型在番茄中诱导的叶片卷曲症状与在番茄田中观察到的症状没有区别。 ToLDeV的生物学特性与旧世界感染番茄的单倍体病毒相似，包括缺乏汁液的传递性，韧皮部限制，番茄种质中具有Ty-1基因和V1（衣壳蛋白）功能特性的抗性表型和C4基因。番茄中ToLDeV基因型引起的症状表型差异（例如是否观察到恢复表型）与本氏烟草植物的差异与左发基因间区域和C4基因高度相关。最后，有证据表明，ToLDeV基因型是通过平行进化，突变和重组从新大陆二分祖细胞的DNA-A组分中产生的。在第四章中，通过突变分析和异位表达，通过两种具有不同组织嗜性的病毒载体（韧皮部限制和非韧皮部限制）研究了ToLDeV C2（TrAP）和C4基因的功能。 ToLDeV C2突变体在本氏烟草叶盘中复制，但感染性是宿主特异性的。评估了TrAP和C4病毒蛋白抑制基因沉默的能力。 TrAP和C4蛋白均不能抑制局部沉默。但是，C4是长距离沉默的强大抑制器。 C2和C4均显示出一些功能，这些功能可通过恢复沉默的16c-TGS植物中的GFP表达来抑制转录基因沉默。此外，ToLDeV左基因间区域的高水平胞嘧啶甲基化与番茄植株有关，番茄植株在被ToLDeV感染后表现出恢复表型。最后，亚细胞定位研究表明TrAP蛋白定位于细胞核，而C4定位于质膜。

著录项

  • 作者

    Melgarejo Gutierrez, Tomas Adan.;

  • 作者单位

    University of California, Davis.;

  • 授予单位 University of California, Davis.;
  • 学科 Agriculture Plant Pathology.;Biology Virology.
  • 学位 Ph.D.
  • 年度 2014
  • 页码 174 p.
  • 总页数 174
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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