EAGER: Lipids on the move

植物对地球上的所有生命都是必不可少的：它们提供我们呼吸的氧气、药物、工业和建筑的原材料、燃料、营养和食物。在全球范围内，62%的收获作物用于人类食品，3%用于生产用于再植的种子、生物能源和工业产品，35%用于动物饲料。联合国的优先目标包括“消除饥饿，实现粮食安全，改善营养，促进可持续农业”。这不是一个微不足道的目标，因为植物实际上是“扎根于现场”，无法逃脱不利的条件。了解植物如何感知环境变化，将这些变化信号传递给植物的其他部分并相应地调整发育是至关重要的。脂质是一种油和脂溶性分子，在植物和动物的发育中起着重要作用，并作为环境条件变化的信号。因此，了解植物脂质的功能及其运输机制不仅有助于对植物发育过程的科学理解，还将转化为切实改善食物的可持续性和质量：了解脂质如何影响植物生长、种子和果实的发育，将带来更高的产量和更多的食物。-了解脂质如何影响植物在不断变化的环境（寒冷、冰冻、干旱、营养不足的土壤、昆虫取食等）中的生存，将减少作物损失。脂质是植物的基本成分。它们为代谢过程提供能量，是细胞膜成分，并作为细胞内信号。简单的脂质，如氧化脂质是生物应激的远距离信号，表明其他脂质也起着系统信号的作用。鉴于移动信号对非生物胁迫和发育的协调以及对植物生存和粮食安全的根本重要性，有必要明确证明远距离磷脂信号及其组成部分的存在。PI的实验室已经在韧皮部分泌物中发现了脂质磷脂酸（PA）和PA结合蛋白（PLAFP），并提出韧皮部脂质结合蛋白，特别是PLAFP和PA，在环境胁迫下的发育和长距离信号传导中起作用。为了验证这一假设，实验室将使用转基因方法、嫁接、western blot、共聚焦显微镜、免疫金标记和NightOWL成像相结合的方法来研究PLAFP蛋白的定位和运动。-监测野生型、PLAFP过表达和PLAFP敲除植物的脂质运动。质谱将确认放射性标记的可移动化合物的身份。-确定PLAFP-PA复合物的结合机制和特异性及其运动，使用“红灯亮”光遗传开关和一种新颖的FRET方法相结合，在体外和植物内可视化蛋白质-脂质相互作用。一旦建立，该系统将为研究任何系统中的蛋白质-配体相互作用提供工具。这项工作将由一名博士后研究员、一名研究生和一名本科生协助。他们将能够参加与德国杜塞尔多夫海因里希-海涅大学的国际合作/交流。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Plants are essential for all life on earth: they provide the oxygen we breathe, medicines, raw materials for industry and construction, fuel, nutrition, and food. Globally, 62% of harvested crop is used for human food, 3% to produce seeds for replanting, bioenergy, and industrial products, and 35% for animal feed. The priority goals of the United Nations include "End hunger, achieve food security, improve nutrition, and promote sustainable agriculture". This is not a trivial goal as plants are quite literally "rooted to the spot" and cannot escape adverse conditions. It is essential to understand how plants sense environmental changes, signal these changes to other parts of the plant and adjust development accordingly. Lipids are oil and fat-soluble molecules that play an important role in plant and animal development and serve as signals for changing environmental conditions. As a result, understanding the function of plant lipids and the mechanism of their transport will serve not only the scientific understanding of plant developmental processes but will also translate into tangible improvements of food sustainability and quality: - Understanding how lipids affect plant growth and seed and fruit development will lead to higher yields and more food. - Understanding how lipids affect plant survival in changing environments (cold, freezing, drought, nutrient-deficient soil, insect feeding etc.) will reduce crop losses. Lipids are essential components of plants. They provide energy for metabolic processes, are membrane components, and serve as intracellular signals. Simple lipids such as oxylipins are long-distance signals of biotic stress suggesting other lipids function as systemic signals as well. Given the fundamental importance of mobile signals for the coordination of abiotic stress and development, and thus for plant survival and food security, it is imperative to unequivocally demonstrate the existence of long-distance phospholipid-signaling and its components. The PI's lab has identified the lipid phosphatidic acid (PA) and a PA-binding protein (PLAFP) in phloem exudates and proposed that phloem lipid-binding proteins, particularly PLAFP and PA act in development and in long-distance signaling in response to environmental stress. To test this hypothesis, the lab will - Investigate localization and movement of the protein PLAFP using a combination of transgenic approaches, grafting, western blot, confocal microscopy, immunogold labeling and NightOWL imaging. - Monitor lipid movement in wild type, PLAFP-overexpression and PLAFP knock-down plants. MS will confirm the identity of the radiolabeled mobile compound. - Determine binding mechanisms and specificity of the PLAFP-PA complex and its movement using a combination of a "Red-Light-ON" optogenetic switch with a novel FRET approach that visualizes protein-lipid interaction in vitro and in-planta. Once established this system will provide a tool to study protein-ligand interactions in any system.The work will be assisted by a postdoctoral researcher, a graduate student, and an undergraduate. They will be able to participate in an international collaboration/exchange with the Heinrich-Heine University Duesseldorf, Germany.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.