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)

SGER：开发一种使用拉曼化学成像显微镜 (RCIM) 分析、绘图和研究根分泌物生物相互作用的非侵入性方法

• 批准号: 0713774
• 负责人: Harsh Bais
• 金额: --
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2009-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0713774&HistoricalAwards=false
• 关键词: SGER; Development; Non; invasive; Method

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.

尽管根分泌物显然代表着植物的一大碳成本，但调节分泌物的机制却鲜为人知。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。过去，包括比色法和层析法在内的这些经典技术曾帮助科学家识别各种渗出的代谢物。然而，这些技术都不能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实时分析根分泌物的方法是科学界目前的目标。一种非侵入性的实时监测根分泌物的方法将有助于了解和阐明植物根与其他根、微生物和线虫之间的各种未知的新的化学相互作用。本研究的目的是利用一种被称为拉曼化学成像显微镜(RCIM)的新技术来原位探索拟南芥的根分泌物。RCIM允许基于其极化性对几乎所有次级代谢物进行快速且非破坏性的实时分析(无质量截断)，而不涉及有机提取和层析分离。本项目将研究以下具体目标：(1)实时研究刺参根系总分泌物对非生物和生物胁迫的响应；(2)利用不同的分泌泵抑制剂来验证AIM 1的结果，并将代谢物分类到各自的分泌泵。从这一探索性项目中获得的结果将突出表明，RCIM是回答根际生物学基本问题的有力工具。更广泛的影响。RCIM在植物生物学研究中的独特应用是以对峙模式进行观察和分析，这将有助于鉴定分泌的代谢物，并提供与这些代谢物在根际相互作用中的生物学意义有关的数据。该项目的结果将在拟南芥中产生最广泛的分泌物图谱收集。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生代谢物的未知功能的能力，这些次生代谢物是响应生物和非生物信号而分泌和调节的。该项目将有助于国际和平研究所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。