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

在现实世界条件下对植物进行表型鉴定是非常具有挑战性的。弗罗默实验室(斯坦福大学)开发了一套基因编码的生物传感器，可以报告亚细胞水平的离子或代谢物(例如离子、糖)，或者以高时间分辨率报告特定转运蛋白的活动。通常，表达这些传感器的植物使用荧光显微镜进行分析。这个项目将探索是否可以使用远程成像系统来量化植物叶片特定区域中的离子水平(这里是信号中间钙作为概念的证明)。密歇根州立大学的克莱默实验室开发了一种生长室，可以模拟和回放田间条件，并使用荧光成像系统同时显示光合作用参数。这一合作将这两个创新平台结合在一起：动态环境成像系统(DEPI)和基因编码的超灵敏荧光生物传感器技术。该项目旨在开发一种新的系统，可以以前所未有的深度监测完整植物中的遗传编码传感器，并为高通量光合作用表型鉴定提供平行选择。该项目将为建立系统和工具奠定基础，作为社区资源，有可能改变世界各地的光合作用研究计划。这一高风险项目将为拟南芥和作物的遗传变异表型鉴定奠定基础，并将遗传编码生物传感器的有用性扩大到大规模筛查。此外，该项目还将培养一名具有物理化学经验的表型鉴定博士后科学家。该项目还将培训高中生和本科生，在可能的情况下，少数族裔学生将参与这一努力。这样的成像系统将为完整植物的分子事件表型鉴定提供一个全新的工具，从而为筛选遗传变异和发现整个植物水平的新生物学提供一个新的工具。该项目将通过监测新型的超灵敏钙传感器来验证长期存在的关于钙在信号过程中的作用以及叶片在波动的环境条件和特定信号过程中的反应模式的假设，从而展示这项技术的潜力。其中一个挑战是联合植物成像系统的灵敏度。Frommer实验室开发了新型的超灵敏钙传感器，将在这里使用，这可能使我们能够同时观察数百种植物种群的整个生长周期中的钙动态。如果成功，这种方法可以扩展到其他荧光生物传感器，并应用于作物筛选。同时，这样的系统可能会在植物细胞信号和叶绿体离子动力学领域发现新的生物学。

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.