PAPM EAGER: A Plant Observatory for remote sensing of biochemical reactions in vivo

PAPM EAGER：遥感体内生化反应的植物观测站

• 批准号: 1650196
• 负责人: David Kramer
• 金额: $ 29.98万
• 依托单位: Carnegie Institution of Washington
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1650196&HistoricalAwards=false
• 关键词: PAPM; EAGER; Plant; Observatory; remote

Phenotyping plants under real world conditions is highly challenging. The Frommer lab (Stanford) developed a suite of genetically encoded biosensors that report subcellular levels of ions or metabolites (e.g. ions, sugars) or that report the activity of particular transporters with high temporal resolution. Typically, plants expressing these sensors are analyzed using fluorescence microscopy. This project will explore whether ion levels can be quantified (here the signaling intermediate calcium as a proof of concept) in specific regions of plant leaves using a remote imaging system. The Kramer lab at MSU developed a growth chamber that can mimic and replay field conditions and simultaneously phenotype photosynthetic parameters using a fluorescence imaging system. This collaboration brings together these two innovative platforms: the dynamic environmental imaging system (DEPI), and genetically encoded ultrasensitive fluorescent biosensor technology. The project aims to develop a novel system that can monitor genetically encoded sensors in intact plants with unprecedented depth and the parallel option for high throughput phenotyping of photosynthesis. The project will lay the groundwork for establishing systems and tools as community resources with the potential to transform photosynthesis research programs around the world. This high-risk project will lay the basis for phenotyping genetic variants in Arabidopsis as well as crops and expand the usefulness of genetically encoded biosensors to large scale screening. In addition, this project will train a postdoctoral scientist with experience in physical chemistry in phenotyping. The project will also train high school students and undergraduate students, and where possible minority students will be engaged in this endeavor.Such an imaging system would present a completely novel tool for phenotyping molecular events in intact plants and thus present a new tool for screening genetic variants and to discover new biology at the whole plant level. The project will demonstrate the potential of this technology by monitoring novel ultrasensitive calcium sensors to test long-standing hypotheses regarding the role of calcium in signaling processes and the response patterns in leaves in fluctuating environmental conditions and specific signaling processes. One of the challenges is the sensitivity of the combined plant-imaging system. The Frommer lab developed novel ultrasensitive calcium sensors that will be used here and that may enable us to observe calcium dynamics over the whole growth cycle in populations of 100s of plants simultaneously. This approach, if successful, could be expanded to other fluorescent biosensors and implemented for crop plant screening. At the same time, such a system may uncover new biology in the areas of plant cell signaling and chloroplast ion dynamics.

现实世界中的表型植物是高度挑战的。 Frommer Lab（Stanford）开发了一套遗传编码的生物传感器，这些生物传感器报告了亚细胞水平的离子或代谢产物（例如离子，糖）或报告具有高时间分辨率的特定转运蛋白的活性。通常，使用荧光显微镜分析表达这些传感器的植物。该项目将使用远程成像系统探索是否可以在植物叶的特定区域中量化离子水平（此处信号中间钙作为概念证明）。 MSU的Kramer Lab开发了一个生长室，该生长室可以使用荧光成像系统模仿和重播场状况并同时表型光合参数。这种合作汇集了这两个创新的平台：动态环境成像系统（DEPI）和遗传编码的超敏感荧光生物传感器技术。该项目旨在开发一种新型系统，该系统可以监测具有前所未有的深度的完整植物中的遗传传感器，并且是光合作用的高吞吐量表型的平行选择。该项目将为建立系统和工具作为社区资源的基础奠定基础，并有可能改变世界各地的光合作用研究计划。这个高风险的项目将为拟南芥的表型遗传变异和农作物提供基础，并扩大遗传编码的生物传感器的有用性到大规模筛选。此外，该项目将培训一位具有表型物理化学经验的博士后科学家。该项目还将培训高中生和本科生，并在可能的少数群体中参与这项工作。类似的成像系统将为完整植物中的表型分子事件提供一个新颖的工具，因此为筛选遗传变异并在整个工厂层面发现新的生物学。该项目将通过监测新型的超敏感钙传感器来测试有关钙在信号过程中的作用以及在波动的环境条件和特定信号过程中叶片中的响应模式的长期假设来证明该技术的潜力。挑战之一是组合植物成像系统的敏感性。 Frommer Lab开发了新型的超敏感钙传感器，这些传感器将在这里使用，这可能使我们能够同时观察100s植物种群的整个生长周期中的钙动力学。如果成功的话，可以将这种方法扩展到其他荧光生物传感器，并实施用于作物植物筛查。同时，这种系统可能会在植物细胞信号传导和叶绿体离子动力学领域中发现新的生物学。