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物，3%用于生产补种种子、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“结束饥饿、实现粮食安全、改善营养和促进可持续农业”。这不是一个微不足道的目标，因为植物从字面上讲是“扎根于现场”的，无法逃脱不利条件。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，这是至关重要的。脂类是油和脂溶分子，在植物和动物的发育中发挥重要作用，并作为环境条件变化的信号。因此，了解植物脂肪的功能及其运输机制不仅有助于科学地了解植物的发育过程，而且还将转化为对食物可持续性和质量的切实改善：-了解脂肪如何影响植物生长和种子和果实发育，将导致更高的产量和更多的食物。-了解脂类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、昆虫觅食等)中的生存。将减少农作物的损失。脂类是植物的基本成分。它们为新陈代谢过程提供能量，是膜成分，并充当细胞内信号。简单的脂类，如氧磷脂，是生物压力的长距离信号，这表明其他脂类也起到了系统信号的作用。鉴于移动信号对于协调非生物胁迫和发育，从而对于植物生存和食物安全的根本重要性，有必要明确地证明长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渗出物中鉴定了脂磷脂酸(PA)和PA结合蛋白(PLAFP)，并提出韧皮部脂类结合蛋白，特别是PLAFP和PA在发育和响应环境胁迫的远程信号中发挥作用。为了验证这一假设，该实验室将使用转基因方法、嫁接、蛋白质印迹、共聚焦显微镜、免疫金标记和睡巾成像的组合来研究PLAFP蛋白的定位和运动。-监测野生型、PLAFP过表达和PLAFP基因敲除植物中的脂质运动。MS将确认标记的移动化合物的身份。-确定PLAFP-PA复合体及其运动的结合机制和特异性，使用“Red-Light-On”光遗传开关与一种新的FRET方法相结合，该方法可在体外和平面内可视化蛋白质-脂质相互作用。一旦建立，这个系统将提供一个研究任何系统中蛋白质-配体相互作用的工具。这项工作将得到一名博士后研究员、一名研究生和一名本科生的协助。他们将能够参与与德国杜塞尔多夫海因里希-海因大学的国际合作/交流。这一奖项反映了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。

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.