EAGER:Novel approaches to study green leaf volatile perception

EAGER：研究绿叶挥发性感知的新方法

• 批准号: 2051699
• 负责人: Johannes Stratmann
• 金额: $ 29.98万
• 依托单位: University of South Carolina at Columbia
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-15 至 2024-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2051699&HistoricalAwards=false
• 关键词: EAGER; Novel; approaches; study; green

Plants that are injured by herbivores emit volatile organic compounds (VOC) into the airspace which can be perceived by neighboring plants. These plants interpret the volatile compounds as alarm signals and start to upregulate defense responses against their enemies to be prepared for an impending attack. Effects of volatile compounds as inducers of defenses are well characterized, but it is unknown how plants perceive them. In general, alarm signals are perceived by dedicated receptor proteins. Most herbivore-induced volatile compounds are very small and difficult to track, which impedes the identification of their receptors. This project will develop a novel approach for labeling volatile compounds with very small tags that can be used to track the compounds and thus identify their receptors. The tagging strategy can be adapted for other small infochemicals and thus transform our knowledge of communication in plants. Identification of receptor proteins for volatile alarm signals may help to devise novel strategies for sustainable crop production by relying on innate plant defenses rather than agrochemicals. Aspects of this project will also be incorporated into an Ecology and Evolution laboratory class. In addition, undergraduates from groups underrepresented in STEM disciplines will be actively recruited to provide extra-curricular experiential learning opportunities. These students will work on VOC signaling related to this project. This will develop their practical, analytical, and communication skills, which prepares them for professional careers. In addition, two graduate students will be trained in biology and chemistry to provide a cutting-edge interdisciplinary education in chemical ecology and molecular biochemistry.Many aspects of plant communication are mediated by volatile organic compounds such as green leaf volatiles (GLVs). When plants are injured by herbivores, they emit GLVs, which can be perceived not only by distant parts of the wounded plants (within-plant communication) but also by neighboring plants (interplant communication). Perception of these GLVs is interpreted by receiver tissues as a sign of imminent danger and results in priming or upregulation of defense responses. The objective of this EAGER proposal is to advance novel approaches to identify GLV receptor proteins that initiate defense signaling. Very small non-proteinaceous ligands like GLVs (containing only six or eight carbons) are difficult to tag and to track. This proposal takes a novel approach by using ‘click chemistry’ to couple bioactive minitagged GLVs to reporter tags in order to identify receptor proteins for GLVs. Reporters that emit fluorescent light will be used to localize the receptor-GLV-reporter complex, and biotin-based reporter tags will be used to purify this complex and to isolate a GLV receptor. Additionally, GLV-induced rapid phosphorylation-based signaling events will be determined by a phosphoproteomics approach to identify early GLV-responsive signaling proteins such as GLV receptors. Although the role of GLVs in plant communication is well documented, it is completely unknown how they are perceived by plants. This is a significant knowledge gap that hampers our understanding of GLV signaling and function. Identification and characterization of GLV receptors would transform our understanding of plant communication at the molecular and ecological level.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.

被食草动物伤害的植物会释放挥发性有机化合物（VOC）到空气中，这些化合物可以被邻近的植物感知。这些植物将挥发性化合物解释为警报信号，并开始上调对敌人的防御反应，为即将到来的攻击做好准备。挥发性化合物作为防御诱导剂的作用已经得到了很好的表征，但植物如何感知它们还不清楚。一般来说，警报信号被专用的受体蛋白感知。大多数植食性昆虫诱导的挥发性化合物非常小，难以追踪，这阻碍了其受体的鉴定。该项目将开发一种新的方法，用非常小的标签标记挥发性化合物，可用于跟踪化合物，从而识别其受体。标记策略可以适用于其他小的信息化学物质，从而改变我们对植物通讯的认识。识别挥发性警报信号的受体蛋白可能有助于设计新的策略，通过依赖先天植物防御而不是农用化学品来实现可持续的作物生产。该项目的各个方面也将纳入生态学和进化实验室课程。此外，将积极招募STEM学科代表性不足的群体的本科生，以提供课外体验式学习机会。这些学生将从事与本项目相关的VOC信号。这将培养他们的实践，分析和沟通技能，为他们的职业生涯做好准备。此外，还将培养两名生物学和化学专业的研究生，提供化学生态学和分子生物化学的前沿交叉学科教育，植物通讯的许多方面都是由挥发性有机化合物如绿色叶挥发物（GLV）介导的。当植物被食草动物伤害时，它们会释放GLV，不仅可以被受伤植物的远处部分（植物内通信），而且可以被邻近植物（植物间通信）感知。这些GLV的感知被受体组织解释为即将发生危险的信号，并导致防御反应的启动或上调。EAGER的目标是提出新的方法来鉴定启动防御信号的GLV受体蛋白。非常小的非蛋白质配体，如GLV（仅含有6或8个碳原子），很难标记和跟踪。该提议采用了一种新的方法，通过使用“点击化学”将生物活性的微型标记的GLV偶联到报告标签，以鉴定GLV的受体蛋白。发射荧光的报告基因将用于定位受体-GLV-报告基因复合物，基于生物素的报告基因标签将用于纯化该复合物并分离GLV受体。此外，GLV诱导的基于快速磷酸化的信号传导事件将通过磷酸化蛋白质组学方法来确定，以鉴定早期GLV响应性信号传导蛋白，如GLV受体。虽然GLV在植物通讯中的作用是有据可查的，但植物如何感知它们是完全未知的。这是一个重大的知识差距，阻碍了我们对GLV信号转导和功能的理解。GLV受体的鉴定和表征将在分子和生态水平上改变我们对植物通讯的理解。该奖项反映了NSF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。