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（加利福尼亚大学戴维大学）COPIS：Robert Gilbertson，Diane Ullman和Ilias Tagkopoulos（加利福尼亚戴维斯大学）番茄是世界上第二重要的植物作物。美国和全球的番茄生产受到两种毁灭性昆虫传播病毒的威胁 - 番茄斑点病毒（TSWV）和番茄黄叶卷发病毒（Tylcv）。这些病毒的管理很困难，并且通常严重依赖于使用杀虫剂来控制载体。杀虫剂的控制通常无效，人类健康和环境影响负面影响，并且可能导致对媒介的抗杀虫剂形式的选择。在番茄中，用于TylCV的TY1基因座和TSWV的SW5基因座是主要的遗传抗性来源，两者都被渗入了商业番茄品种。但是，关于TY1和SW5介导的抗性所涉及的机制知之甚少。该项目将提供有关植物耐药性机制与昆虫传播病毒感染的敏感性之间的性质和相互作用的新见解。这些研究将提供有关感染和/或宿主防御期间RNA（TSWV）与DNA（TYLCV）病毒的相似或不同途径的见解。然后，该项目将使用这些研究的结果来优化和验证一种称为miRNA诱导的基因沉默（MIG）的新方法，该方法可用于评估番茄中的基因功能。 如果成功的话，该项目的结果可能会导致针对这些病毒的新一代抗病毒策略。该项目还将为萨克拉曼多市学院的本科生和戴维斯高中的高中学生提供一项为期10周的培训计划。由SW5和Ty1 Loci介导的遗传抗药性分别为养殖番茄介导的遗传抗性，分别提供了针对TSWV和TYLCV的最佳保护源。该项目的研究人员假设SW5和TY1介导的TSWV和TYLCV的识别触发了涉及常见和特定的免疫信号网络的早期和可见的转录变化。此外，这些病毒在易感番茄基因型上的感染会引起转录变化，从而促进病毒毒力。为了洞悉转录变化，研究人员将使用RNASEQ方法来识别TSWV和TYLCV抗性和番茄中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，以识别在TSWV和TYLCV抗性和易感相互作用期间受差异调节的关键常见和特定基因。该项目将通过靶向已知候选基因来优化和验证转基因番茄基因功能研究的MIG方法。优化的MIG方法将用于评估TYLCV和TSWV抗性期间选定差异表达的基因的功能以及番茄中易感性的相互作用。该项目的结果可能会提供有关设计新一代策略的见解，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被公众访问。所有RNASEQ数据将被存入并通过GEO获得。所有MIGS矢量和候选靶基因构建体将根据要求以及Addgene（https://www.addgene.org/）和ABRC（http://www.biosci.ohio.ohio-state.edu/~plantbio/~plantbio/tplantbio/Faciolitions/faciolitions/abrc/abrc/abrc/abrcrchome.htm）。 转基因番茄系的种子将通过加州大学戴维斯分校（http://tgrc.ucdavis.edu/）的番茄遗传资源中心获得。