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.

尽管根部分泌显然代表了植物的巨大碳成本，但调节渗出的机制知之甚少。迄今为止，大多数已知分析根分泌的技术取决于样品的破坏性有机分析。过去，这些包括比色和色谱法在内的古典技术帮助科学家鉴定了各种渗出的代谢物。但是，这些技术都不可用于实时监视分泌物。开发一项技术来检查基于原位的非侵入性方法来分析实时根源分泌是科学界的当前目标。一种非侵入性的实时方法来监测根部分泌物，将有助于理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型化学相互作用。这项研究的目的是使用一种称为拉曼化学成像显微镜（RCIM）的新技术探索拟南芥根分泌。 RCIM允许基于它们的极化能力对几乎所有次级代谢产物（无质量切割）进行快速和无损的实时分析，而无需有机提取和色谱分离。该项目将研究以下特定目标：（1）对塔利亚纳曲霉根的实时分泌总体分泌，以响应非生物和生物胁迫，（2）利用不同的分泌泵抑制剂来验证AIM 1的结果，并将各种分泌物的成果和各自的分泌泵验证。从这个探索项目中获得的结果将突出显示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在僵持模式下的观察和分析，这将促进分泌的代谢产物的鉴定，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A. Thaliana产生最广泛的分泌物分析。通过该项目将开发的方法将是广泛适用的，并将为已知的二级代谢产物识别未知功能的能力，这些功能是对生物和非生物提示响应的分泌和调节的。该项目将有助于将研究和教育联系起来的PI持续活动。在实验设计和技术专业知识领域，将增强博士后研究员和一个参与该项目的本科暑期实习生的职业机会。