Genomic Approaches to Unravel Virulence and Resistance Determinants of Vector-transmitted Viruses in Tomato

揭示番茄媒介传播病毒毒力和抗性决定因素的基因组方法

• 批准号: 1339185
• 负责人: Savithramma Dinesh-Kumar
• 金额: $ 164.02万
• 依托单位: University of California-Davis
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1339185&HistoricalAwards=false
• 关键词: Genomic; Approaches; Unravel; Virulence; Resistance

PI: S.P. Dinesh-Kumar (University of California-Davis)CoPIs: Robert Gilbertson, Diane Ullman, and Ilias Tagkopoulos (University of California-Davis)Tomato is the second most important vegetable crop in the world. Tomato production in the US and worldwide is threatened by two devastating insect-transmitted viruses - tomato spotted wilt virus (TSWV) and tomato yellow leaf curl virus (TYLCV). Management of these viruses is difficult and often relies heavily on the use of insecticides to control the vectors. Insecticidal control is often ineffective, has negative human health and environmental effects, and can lead to selection of insecticide-resistant forms of the vectors. In tomato, the Ty1 locus for TYLCV and the Sw5 locus for TSWV are major genetic resistance sources, and both have been introgressed into commercial tomato varieties. However, very little is known about the mechanisms involved in Ty1- and Sw5-mediated resistance. This project will provide new insights into the nature of and interplay between mechanisms of plant resistance and susceptibility to infection by insect-transmitted viruses. These studies will provide insights into whether similar or different pathways are targeted by RNA (TSWV) versus DNA (TYLCV) viruses during infection and/or host defense. The project will then use the results from these studies to optimize and validate a new approach called miRNA-induced gene silencing (MIGS) that can be used to assess gene function in tomato. If successful, the outcomes of this project will likely lead to a new generation of antiviral strategies against these viruses. The project will also provide a 10-week training program every year for undergraduate students from Sacramento City College and high school students from Davis Senior High School.The genetic resistance mediated by Sw5 and Ty1 loci introgressed from wild tomato relatives into cultivated tomato provides the best source of protection against TSWV and TYLCV, respectively. The investigators of this project hypothesize that Sw5- and Ty1-mediated recognition of TSWV and TYLCV trigger early and discernable transcriptional changes that involve common and specific immune signaling networks. In addition, infection of these viruses on susceptible tomato genotypes induces transcriptional changes that promote virus virulence. To gain insights into the transcriptional changes, the investigators will use RNAseq approach to identify differentially expressed transcripts during TSWV and TYLCV resistance and susceptible responses in tomato. Computational methods will be used to analyze RNASeq data to identify key common and specific genes that are differentially regulated during TSWV and TYLCV resistance and susceptible interaction. The project will optimize and validate the MIGS approach for gene function studies in transgenic tomato by targeting known candidate genes. The optimized MIGS approach will be used to evaluate function of selected differentially expressed genes during TYLCV and TSWV resistance and susceptible interactions in tomato. Results from the project will likely provide insights into designing new generation of strategies to control not only TYLCV and TSWV but also other important plant pathogens. All data and biological resources will be accessible to the public. All RNAseq data will be deposited and available through GEO. All MIGS vector and candidate target gene constructs will be available upon request and through Addgene (https://www.addgene.org/) and ABRC (http://www.biosci.ohio-state.edu/~plantbio/Facilities/abrc/abrchome.htm). Seeds from transgenic tomato lines will be available through the Tomato Genetics Resource Center at UC Davis (http://tgrc.ucdavis.edu/).

PI：S.P. Dinesh-Kumar（加州大学戴维斯分校）CoPI：Robert Gilbertson、Diane Ullman 和 Ilias Tagkopoulos（加州大学戴维斯分校）番茄是世界上第二重要的蔬菜作物。美国和世界范围内的番茄生产受到两种毁灭性昆虫传播病毒的威胁：番茄斑萎病毒（TSWV）和番茄黄曲叶病毒（TYLCV）。这些病毒的管理很困难，并且通常严重依赖使用杀虫剂来控制媒介。杀虫剂控制通常是无效的，对人类健康和环境产生负面影响，并可能导致媒介产生抗杀虫剂形式。在番茄中，TYLCV 的 Ty1 基因座和 TSWV 的 Sw5 基因座是主要的遗传抗性来源，两者均已渗入到商业番茄品种中。然而，人们对 Ty1 和 Sw5 介导的耐药性的机制知之甚少。该项目将为植物抗性和昆虫传播病毒感染易感性机制的性质和相互作用提供新的见解。这些研究将深入了解 RNA (TSWV) 与 DNA (TYLCV) 病毒在感染和/或宿主防御过程中是否靶向相似或不同的途径。然后，该项目将利用这些研究的结果来优化和验证一种称为 miRNA 诱导基因沉默 (MIGS) 的新方法，该方法可用于评估番茄中的基因功能。 如果成功，该项目的成果可能会催生针对这些病毒的新一代抗病毒策略。该项目还将每年为萨克拉门托城市学院的本科生和戴维斯高中的高中生提供为期10周的培训计划。由野生番茄近缘种渗入到栽培番茄中的Sw5和Ty1位点介导的遗传抗性提供了最好的培训分别针对 TSWV 和 TYLCV 的保护源。该项目的研究人员假设 Sw5 和 Ty1 介导的 TSWV 和 TYLCV 识别触发了涉及常见和特异性免疫信号网络的早期可辨别的转录变化。此外，这些病毒对易感番茄基因型的感染会诱导转录变化，从而增强病毒毒力。为了深入了解转录变化，研究人员将使用 RNAseq 方法来识别番茄中 TSWV 和 TYLCV 抗性和易感反应期间差异表达的转录本。计算方法将用于分析 RNASeq 数据，以确定在 TSWV 和 TYLCV 耐药性和易感相互作用过程中受到差异调节的关键常见和特定基因。该项目将通过针对已知候选基因来优化和验证用于转基因番茄基因功能研究的 MIGS 方法。优化的 MIGS 方法将用于评估番茄中 TYLCV 和 TSWV 抗性和易感相互作用过程中选定差异表达基因的功能。该项目的结果可能会为设计新一代策略提供见解，不仅可以控制 TYLCV 和 TSWV，还可以控制其他重要的植物病原体。所有数据和生物资源都将向公众开放。所有 RNAseq 数据都将通过 GEO 存储和获取。所有 MIGS 载体和候选目标基因构建体将根据要求并通过 Addgene (https://www.addgene.org/) 和 ABRC (http://www.biosci.ohio-state.edu/~plantbio/Facilities/) 提供abrc/abrchome.htm）。 转基因番茄品系的种子可通过加州大学戴维斯分校番茄遗传学资源中心 (http://tgrc.ucdavis.edu/) 获得。