SGER: Development of a Non-invasive Method for Analysis, Mapping and Studying the Biological Interactions of the Root Secretone Using Raman Chemical Imaging Microscopy (RCIM)

虽然根分泌物清楚地代表了一个显着的碳成本的植物，调节渗出的机制知之甚少。迄今为止，大多数已知的分析根分泌物的技术都依赖于对样品的破坏性有机分析。在过去，这些经典的技术，包括比色法和色谱法，帮助科学家确定各种渗出的代谢物。然而，这些技术中没有一种可用于实时监测分泌物。开发一种技术来检查基于原位的非侵入性方法，用于分析实时根分泌物是科学界当前的目标。一种非侵入性的真实的时间监测根系分泌物的方法将有助于理解和阐明植物根系与其他根系、微生物和线虫之间的各种未知的新型化学相互作用。本研究的目的是探索拟南芥根分泌物原位使用一种新的技术称为拉曼化学成像显微镜（RCIM）。RCIM允许基于其极化率对几乎所有次级代谢物进行快速和非破坏性的实时分析（无质量截止），而不涉及有机萃取和色谱分离。本研究的主要目的是：（1）实时测定A.（2）利用不同的分泌泵抑制剂验证目标1的结果，并将代谢产物与各自的分泌泵进行分类。从这个探索性项目获得的结果将突出RCIM作为一个强大的工具，回答根际生物学的基本问题。更广泛的影响。RCIM在植物生物学研究中的独特应用是在对峙模式下进行观察和分析，这将有助于鉴定分泌的代谢物，并提供有关这些代谢物在根际相互作用中的生物学意义的数据。该项目的结果将产生最广泛的收集分泌概况A。thaliana.将通过该项目开发的方法将广泛适用，并将提供识别已知次级代谢产物的未知功能的能力，这些次级代谢产物在生物和非生物线索的作用下分泌和调节。该项目将有助于PI正在开展的将研究与教育联系起来的活动。参与该项目的一名博士后研究员和一名本科生暑期实习生的职业机会将在实验设计和技术专长领域得到加强。

Although root secretions clearly represent a significant carbon cost to the plant, the mechanisms that regulate exudation are poorly understood. Most of the techniques known to date that analyze root secretions depend upon the destructive organic analysis of samples. In the past, these classical techniques that include colorimetric and chromatography have helped scientists to identify various exuded metabolites. However, none of these techniques are useful for monitoring secretions in real-time. Developing a technology to examine in situ-based non-invasive methods for analyzing real-time root secretions is a current goal of the scientific community. A non-invasive real time method to monitor root secretions will help to understand and elucidate various unknown novel chemical interactions between plant roots with other roots, microbes and nematodes. The purpose of this research is to explore Arabidopsis thaliana root secretions in situ using a novel technique known as Raman Chemical Imaging Microscopy (RCIM). RCIM allows rapid and non-destructive real-time analysis of nearly all secondary metabolites (no mass cut-off) based on their polarizability, without involving organic extraction and chromatographic separations. This project will investigate the following specific objectives: (1) to conduct a study of total root secretions in real-time from A. thaliana roots in response to abiotic and biotic stresses, (2) to utilize different secretion pump inhibitors to validate the results of Aim 1, and assort metabolites to the respective secretion pumps. The results obtained from this exploratory project will highlight RCIM as a powerful tool for answering basic questions in rhizosphere biology. Broader impacts. The unique application of RCIM in plant biology research is the observation and analysis in a standoff mode, which will facilitate the identification of secreted metabolites and provide data pertaining to the biological significance of these metabolites in the rhizospheric interactions. The outcome of this project will generate the most extensive collection of secretion profiling in A. thaliana. The methodology that will be developed through this project will be broadly applicable and will offer the ability to identify an unknown function for known secondary metabolites that are secreted and regulated in response to biotic and abiotic cues. The project will contribute to ongoing activities of the PI linking research and education. The career opportunities for a postdoctoral fellow and one undergraduate summer intern involved in the project will be enhanced in the areas of experimental design and technical expertise.