EAGER: Plant membrane on-a-chip for the genome wide studies of plant transport processes

植物细胞被质膜包围，质膜形成细胞边界，有助于维持细胞内外不同的化学环境。基于膜的分隔允许不同的过程在细胞的不同部分同时发生，并根据植物的需要进行。这些细胞过程确保植物的生长、发育和对环境胁迫的反应，并在很大程度上依赖于离子和有机分子在膜上的重新分布。大多数营养物质和代谢物不能自由地穿过细胞膜，因此特定的蛋白质，如通道、泵和共转运蛋白，被嵌入到植物的细胞膜中。随着科学家努力了解植物如何适应极端天气条件、病原体压力和污染，揭示这些运输系统是如何运行的，对于设计可持续的方法来提高作物产量和促进生态系统繁荣非常重要。在这个项目中，提出了一种装置，它可以采集带有嵌入式传输系统的植物细胞膜样本，以测试它们对各种感兴趣的条件的响应和特性。该装置由一个透明、导电的塑料表面组成，它以高度受控的方式测量穿过植物细胞膜的材料，使科学家能够探测运输系统的反应，然后将其与植物的来源和生长条件联系起来。该项目还将通过康奈尔大学的女性活动，培养高中生对农业和生命科学行业的职业选择的兴奋，这些行业涉及生物技术和在植物、食品和农业中的应用。通过嵌入其中的离子通道和转运体监测植物细胞膜上的溶质和离子的流动，这在今天是一个巨大的挑战，但对于分配未知转运体基因的功能，解决转运体底物的特异性和调控模式，将代谢途径与细胞室、植物生长和发育联系起来，以及弥合基因和表型差距，这是至关重要的。今天的技术不能满足需要的原因有很多，包括低吞吐量和缺乏敏感性，特别是对于通量比离子通道低几个数量级的转运体。在这里，提出了一种新的技术，它将平面植物膜、微流体环境控制和透明、导电的聚合物结合在一起，组成了一个“植物膜生物电子装置”。该装置能够对传送器功能进行双模(光或电)测量。这种新型的传感器设备可以高度多路传输植物运输系统上的大型数据集，这是以前不可能实现的。这样的大数据集为大数据科学方法提供了支持，使我们能够在植物如何适应遗传扰动、极端天气条件、病原体压力和其他对提高作物产量和促进生态系统繁荣至关重要的关键方面的发现和突破。该项目还将培养下一代高中生对农业和生命科学行业的职业选择感到兴奋，这些行业涉及生物技术及其在植物、食品和农业中的应用，通过康奈尔的妇女活动，让K-12女孩及其父母参与进来。该奖项由植物基因组研究计划和生理机制和生物机械计划共同资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Plant cells are surrounded by the plasma membrane that forms cellular boundaries and helps to maintain chemically distinct environments within and outside the cell. Membrane-based compartmentation allows different processes to occur simultaneously in different parts of the cell and in accordance with plant needs. These cellular processes ensure plant growth, development and response to environmental stresses and largely depend on ions and organic molecules redistribution across membranes. Most nutrients and metabolites cannot freely cross the membrane, so specific proteins, known as channels, pumps, and cotransporters, are embedded in plant membranes for this purpose. As scientists strive to understand how plants adapt to extreme weather conditions, pathogen pressures, and pollution, uncovering how these transport systems operate is important for devising sustainable approaches for improving crop yield and promoting flourishing ecosystems. In this project, a device is proposed that can take samples of plant cell membranes with embedded transport systems to test their response and properties to various conditions of interest. The device consists of a transparent, conductive plastic surface that measures the material that crosses the plant cell membrane in a highly controlled manner, allowing scientists to probe, and later connect, the responses of transport systems with the plant from which it was derived and the conditions under which that plant was grown. This project will also foster high school student excitement about career choices in agriculture and life science industries that involve biotechnology and applications in plants, food, and farming through Cornell’s WOMEN event.Monitoring the flux of solutes and ions across plant cell membranes through ion channels and transporters embedded within them, is a significant challenge today, but is fundamental for assigning function to unknown transporter genes, tackling transporter substrate specificities and mode of regulation, linking metabolic pathways to cellular compartments, plant growth and development, and bridging the genotype to phenotype gap. Today's technologies are inadequate for a number of reasons, including low throughput and lack of sensitivity, especially for transporters, which have fluxes several orders of magnitude lower than ion channels. Here, a new technology is proposed that combines planar plant membranes, microfluidic environmental control, and a transparent, electrically conducting polymer, comprising a “plant membrane bioelectronic device.” This device is capable of dual-mode (optical or electrical) measurement of transporter function. This new kind of sensor device can be highly multiplexed for collection of large data sets on plant transporter systems in a way that has not been possible before. Such large data sets feed into big data science approaches for enabling discoveries and breakthroughs in our understanding of how plants adapt to genetic perturbations, extreme weather conditions, pathogen pressures, and other critical aspects important for improving crop yield and promoting flourishing ecosystems. This project will also foster the next generation of high school students becoming excited about career choices in agriculture and life science industries that involve biotechnology and applications in plants, food, and farming through ongoing outreach activities that engage K-12 girls and their parents through Cornell’s WOMEN event.This award was co-funded by the Plant Genome Research Program and the Physiological Mechanisms and Biomechanics Program.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.