NSF Postdoctoral Fellowship in Biology FY 2019: The Effects of Tomato Domestication on the Circadian Clock and its Interaction with Resistance to Phytophthora infestans

2019 财年 NSF 生物学博士后奖学金：番茄驯化对昼夜节律的影响及其与致病疫霉抗性的相互作用

• 批准号: 1907077
• 负责人: Ben Mansfeld
• 金额: $ 21.6万
• 依托单位: Mansfeld, Ben Nathan
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1907077&HistoricalAwards=false
• 关键词: NSF; Postdoctoral; Fellowship; Biology; FY

This action funds an NSF National Plant Genome Initiative Postdoctoral Research Fellowship in Biology for FY 2019. The fellowship supports a research and training plan in a host laboratory for the Fellow who also presents a plan to broaden participation in biology. The title of the research and training plan for this fellowship to Ben Mansfeld is "The Effects of Tomato Domestication on the Circadian Clock and its Interaction with Resistance to Phytophthora infestans". The host institution for the fellowship is the Donald Danforth Plant Science Center and the sponsoring scientist is Dr. Rebecca Bart.Tomato is the second most important vegetable crop in the world, the fruit of which is eaten fresh or consumed in several processed products such as soups and sauces. Tomatoes are rich with important vitamins, minerals as well as antioxidants with potential anti-cancer and other healthful capabilities. The origin of the tomato plant is in equatorial countries in South America. As such, over millions of years, tomato plants have evolved a biological clock adapted to the day length around the equator. However, as humans domesticated tomatoes over the last few thousand years, they also traveled with the seeds, breeding tomatoes to be more successful crop plants in high latitude regions. Through this process, the tomato biological clock was altered to fit the new long days in these regions of the planet. The process of altering a crop's biological clock through domestication is not unique to tomato and may be quite common; however, because of its important role in regulating how the plant protects itself from disease and insects, an altered biological clock may thus increase a plant's susceptibility to disease. This research aims to use tomato as a model to better understand how crop domestication affected the biological clock and how this, in turn, may have affected the plant's innate ability to defend itself from disease. The overarching societal goal for this research is to contribute to increased crop defenses at a basic level, helping growers reduce their inputs and produce cheaper, sustainable food for a growing population. Training though this project will also further the Fellow's skills in plant genetics, genomics and plant-pathogen interactions. Broader impacts include mentoring high school and undergraduate students interested in learning more about bioinformatics, plant disease and crop improvement.This research will explore the hypothesis that alterations to the biological clock have affected cultivated tomato responses to infection, specifically by Phytophthora infestans, the pathogen that causes tomato and potato late blight. First, a collection of wild and domesticated tomato lines will be screened for their circadian rhythm traits, using an automatic camera system. A high-throughput bioassay will be then be used to screen if these lines show a different response to infection by P. infestans, at different times (a "gated" response). Transgenic tomato lines with altered clock rhythms will also be generated and screened to test the effects of mis-regulation of clock genes on the gated response to infection. The two lines showing strongest difference in gated response will be used in a 72-hour gene expression study, in which leaves will be inoculated with the pathogen at dawn and dusk and sampled every 4 hours for RNA sequencing. Weighted gene co-expression networks will then be used to compare the transcriptional responses of wild and domesticated tomato and identify hub genes at the center of network co-expression modules that differ between lines. Finally, to map loci contributing to differences in wild and domesticated tomato, a segregating population will be phenotyped and quantitative trait loci mapped using a bulk segregant approach. Candidate genes will be validated using CRISPR-based mutagenesis. All the data generated in this research will be made accessible through an online web application as well as through public repositories such as the NCBI's Short Read Archive (SRA), Dryad (https://datadryad.org), and the Sol Genomics Network (SGN; https://solgenomics.net).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这项行动为2019财年的NSF国家植物基因组计划生物学博士后研究奖学金提供资金。该研究金支持研究员在东道实验室的研究和培训计划，研究员还提出了扩大生物学参与的计划。本·曼斯菲尔德的研究和培训计划的标题是“番茄驯化对昼夜节律钟的影响及其与抗疫霉菌的相互作用”。该奖学金的主办机构是唐纳德丹佛斯植物科学中心，赞助科学家是丽贝卡巴特博士。番茄是世界上第二大重要的蔬菜作物，其果实可鲜食或在几种加工产品中食用，如汤和酱汁。西红柿富含重要的维生素，矿物质以及具有潜在抗癌和其他健康能力的抗氧化剂。番茄植物的起源是在南美洲的赤道国家。因此，数百万年来，番茄植物已经进化出一种适应赤道周围白昼长度的生物钟。然而，在过去的几千年里，随着人类驯化西红柿，他们也带着种子旅行，在高纬度地区培育西红柿成为更成功的作物。通过这个过程，番茄的生物钟被改变，以适应地球上这些地区新的长日照。通过驯化改变作物生物钟的过程并不是番茄独有的，可能相当普遍;然而，由于它在调节植物如何保护自己免受疾病和昆虫侵害方面的重要作用，改变的生物钟可能因此增加植物对疾病的易感性。这项研究旨在以番茄为模型，更好地了解作物驯化如何影响生物钟，以及这反过来又如何影响植物防御疾病的先天能力。这项研究的总体社会目标是在基本水平上增加作物防御，帮助种植者减少投入，为不断增长的人口生产更便宜，可持续的食物。本项目的培训还将进一步提高研究员在植物遗传学、基因组学和植物-病原体相互作用方面的技能。更广泛的影响包括指导有兴趣学习更多关于生物信息学、植物疾病和作物改良的高中和本科生。这项研究将探讨生物钟的改变影响了栽培番茄对感染的反应的假设，特别是引起番茄和马铃薯晚疫病的病原体致病疫霉。首先，将使用自动照相系统筛选野生和驯化番茄品系的昼夜节律特征。然后将使用高通量生物测定来筛选这些细胞系是否在不同时间对致病疫霉感染显示出不同的应答（“门控”应答）。还将产生和筛选具有改变的时钟节律的转基因番茄系，以测试时钟基因的误调节对感染的门控反应的影响。在门控响应中显示最强差异的两个品系将用于72小时基因表达研究，其中叶片将在黎明和黄昏用病原体接种，并且每4小时取样用于RNA测序。加权基因共表达网络，然后将用于比较野生和驯化番茄的转录反应，并确定枢纽基因在网络共表达模块的中心，不同的线。最后，为了定位野生和驯化番茄差异的基因座，将对分离群体进行表型分析，并使用大量分离体方法定位数量性状基因座。将使用基于CRISPR的诱变来验证候选基因。在这项研究中产生的所有数据将通过在线网络应用程序以及通过公共存储库，如NCBI的短读档案（SRA），Dryad（https：//www.example.com）和Sol基因组学网络（SGN; https://solgenomics.net）进行访问。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。datadryad.org