Fatty Acid Amide Hydrolases and Chemical Communication in Plants

植物利用小分子作为信号来同步它们的生长、发育和对环境的反应。该项目旨在了解一组进化保守的植物酶如何利用化学信号来调节生长并影响其微生物环境。 具体的研究目标包括发现新的酶及其作为通信信号的底物分子。这项研究的广泛应用可能会提供新的策略，通过操纵作物和土壤系统中的植物-微生物相互作用来提高农业产量。此外，拟议的研究活动通过北得克萨斯大学的研究课程与本科生培训机会相结合，该大学是一所为少数群体服务的大型公立大学。拟南芥脂肪酸酰胺水解酶（AtFAAH）的三维结构与各种各样的酰胺底物复合，有助于确定参与底物相互作用的关键残基。通过比较来自87种植物的FAAH序列，结合口袋中这些关键残基的保守差异揭示了两组不同的FAAH，进一步扩展了这些酶可能识别的小分子底物的类型。基于这些新的，结构驱动的推论，和表现出的滥交AtFAAH（组I），该提案将调查FAAH蛋白质的作用更广泛的相互作用的植物与他们的微生物环境和植物生长的内部调制。总体假设是FAAH通过小分子酰胺的水解作用于通信轴，以监视生物环境并调节植物生长和发育。该假设将在两种植物物种与多个FAAH基因和已知的微生物相互作用，涉及FAAH基板通过三个目标进行测试。1)表征FAAH抑制对两种G.植物生长和微生物组的影响。hirsutum和M.蒺藜2)通过代谢组学平台评估FAAH改变植物的代谢物变化。3)通过离体脂质组学分析和体外酶测定检查I组和II组FAAH的底物范围。这项研究提案的目标将更广泛地与32名本科生的培训机会相结合，通过在北德克萨斯大学提供两次课堂本科生研究体验（CURE）。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。

Plants use small molecules as signals to synchronize their growth, development and responses to their environment. This project seeks to understand how an evolutionarily-conserved group of plant enzymes utilizes chemical signals to regulate growth and to influence their microbial environment. Specific research aims include the discovery of new enzymes and their substrate molecules that act as communication signals. Broader applications of this research may offer new strategies to enhance agricultural outputs by manipulating plant-microbe interactions in crop and soil systems. In addition, the proposed research activities are integrated with undergraduate training opportunities through a research course at the University of North Texas, a large minority-serving, public university. The three-dimensional structure of the Arabidopsis fatty acid amide hydrolase (AtFAAH) complexed with a diverse range of acylamide substrates helped to identify key residues involved in substrate interactions. By comparing FAAH sequences from 87 plant species, conserved differences in these key residues in the binding pocket revealed two distinct groups of FAAHs, expanding further the types of small molecule substrates that might be recognized by these enzymes. Based on these new, structure-driven inferences, and the demonstrated promiscuity of AtFAAH (group I), this proposal will investigate the role of FAAH proteins more broadly in the interaction of plants with their microbial environment and in the internal modulation of plant growth. The overarching hypothesis is that FAAH acts in an axis of communication through hydrolysis of small molecule acylamides to both surveil the biotic environment and adjust plant growth and development. The hypothesis will be tested in two plant species with multiple FAAH genes and with known microbial interactions that involve FAAH substrates through three aims. 1) Characterize the effects of FAAH suppression on plant growth and microbiomes of both G. hirsutum and M. truncatula. 2) Evaluate the metabolite changes in FAAH-altered plants through metabolomics platforms. 3) Examine substrate ranges for group I and group II FAAH through ex vivo lipidomics analysis and in vitro enzyme assays. The objectives of this research proposal will be integrated more broadly with training opportunities for 32 undergraduate students through two offerings of a classroom undergraduate research experience (CURE) at the University of North Texas.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.