MRI: Acquisition of a 600 MHz NMR with cryoprobe for Integration with Existing UPLC-MS-SPE to Enable High Throughput Chemical Annotation in Plant Metabolomics

MRI：使用冷冻探针采集 600 MHz NMR，与现有 UPLC-MS-SPE 集成，以实现植物代谢组学中的高通量化学注释

• 批准号: 1126719
• 负责人: Lloyd Sumner
• 金额: $ 105.76万
• 依托单位: Noble Research Institute, LLC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1126719&HistoricalAwards=false
• 关键词: MRI; Acquisition; 600; MHz; NMR

A multi-institutional team of interdisciplinary researchers with a mutual focus upon the large-scale study of plant metabolism, biochemistry, and molecular biology (i.e. metabolomics) have joined together to acquire a 600 MHz nuclear magnetic resonance (NMR) spectrometer equipped with a cryoflowprobe. NMR is a powerful tool for the chemical characterization of the thousands of diverse metabolites found in all living organisms. Identifying and chemically characterizing metabolites is a fundamental step in understanding their biological, physiological, and ecological roles. The NMR instrument will be combined with existing ultra high-performance liquid chromatography coupled to parallel mass spectrometry detection and solid phase extraction (UPLC-MS-SPE) which will enable semi-automated metabolite purification and concentration prior to NMR analysis. Access to this instrument will enable and greatly enhance productive, cutting-edge, and emerging plant programs centralized in southeastern Oklahoma. The requested instrumentation will be located and supported by the Samuel Roberts Noble Foundation, a nonprofit research institute whose mission focuses upon the enhancement of humanity through advancements in agriculture. A significant proportion of the group?s metabolomics prior efforts focused on mass spectrometry based approaches and integrated functional genomics of the model legume Medicago truncatula. However, the scope of current and future studies have substantially expanded to include other plant species such as Arabidopsis thaliana (MCB 0520140 Arabidopsis 2010: MCB 0520283 Arabidopsis 2010), alfalfa, tomato, tobacco, hops, cotton, horse/hedge apple, switchgrass biofuels, fescue and related alkaloid producing plant fungal endophytes, and devastating fungal plant pathogens such as cotton root rot. As the diversity of the group?s programs and staff expands, so does the number of unique metabolites. Currently, it is estimated that there are over 200,000 metabolites throughout the plant kingdom, and over half of these still remain unidentified. Accordingly, the participating groups have identified a critical and urgent need for the acquisition and incorporation of NMR technology into their metabolomics and plant biochemistry technology base to advance their ability to systematically identify, structurally characterize, and comparatively profile plant metabolites. Many of the currently funded studies are focused on high anatomically resolved experiments and limited quantities of spatially distinct plant organs and cell types (i.e. pollen, trichomes, border cells). As a result, biological materials and corresponding metabolites are of very low abundance. Thus, a NMR equipped with a cryoflowprobe and coupled to on-line mass directed solid-phase extraction provides critical sensitivity enhancements that are absolutely necessary for obtaining structural data for minute quantities of metabolites in a semi-automated mode to enhance throughput. The NMR instrumentation will advance the understanding of fundamental plant metabolism and biochemistry through the identification of important novel metabolites in key model and crop species. It will also increase the biological context of our metabolomics programs through increased annotation and depth-of-coverage which will proportionately enhance gene discoveries, functional annotation of genes, and more efficient metabolic engineering of beneficial plant traits. The enabling instrumental resource will be used to recruit one new permanent staff member. Long term support for this instrument and staff will be provided by the Noble Foundation. Funding will also enhance the recruitment of future faculty, retention of current faculty, the international competitiveness and leadership of the participants in plant biochemistry and metabolomics, and used to leverage future state and federal research funding for plant biochemistry and agriculture. The proposed instrumentation will be eagerly incorporated into the existing and productive educational and training programs at all the participating institutes which serve a large number of diverse postdoctoral, graduate, undergraduate, and high school students. These specifically include: multiple hands-on, in-depth, week-long instrumental training courses of national recognition such as our metabolomics and proteomics user training programs; undergraduate instrumental analyses courses; high school Science Carnivals; an instrumental "Nut & Volt" interest group; the prestigious Noble Scholars Intern Program; and high school cooperative programs with the Southern Oklahoma Technology Center, Ardmore High School AP Chemistry, Ardmore High School Science Club, and Oklahoma Upward Bound.

一个由跨学科研究人员组成的多机构团队，共同关注植物代谢，生物化学和分子生物学（即代谢组学）的大规模研究，共同获得了配备cryoflowprobe的600 MHz核磁共振（NMR）光谱仪。 核磁共振是一种强大的工具，用于对所有生物体中发现的数千种不同代谢物进行化学表征。 识别和化学表征代谢物是了解其生物，生理和生态作用的基本步骤。 NMR仪器将与现有的超高效液相色谱结合，并与平行质谱检测和固相萃取（UPLC-MS-SPE）相结合，这将使NMR分析之前的半自动代谢物纯化和浓缩成为可能。 使用该工具将使集中在俄克拉荷马州东南部的生产性、尖端和新兴工厂项目成为可能，并大大增强这些项目。 所要求的仪器将由塞缪尔·罗伯茨诺贝尔基金会（Samuel Roberts Noble Foundation）提供定位和支持，该基金会是一家非营利性研究机构，其使命重点是通过农业进步提高人类。一个很大比例的群体？他的代谢组学先前的工作集中在基于质谱的方法和模式豆科植物蒺藜苜蓿的整合功能基因组学。然而，目前和未来的研究范围已大大扩大，包括其他植物物种，如拟南芥（MCB 0520140 Arabidopsis 2010：MCB 0520283 Arabidopsis 2010），苜蓿，番茄，烟草，啤酒花，棉花，马/绿篱苹果，柳枝稷生物燃料，牛毛草和相关的产生生物碱的植物真菌内生菌，和毁灭性的真菌植物病原体如棉花根腐病。 因为群体的多样性？的项目和人员的扩大，独特的代谢物的数量也在增加。 目前，据估计，整个植物界有超过20万种代谢物，其中一半以上仍然没有被识别。 因此，参与小组已经确定了一个关键和迫切需要的收购和纳入NMR技术到他们的代谢组学和植物生物化学技术基础，以提高他们的能力，系统地识别，结构表征，并比较剖析植物代谢物。 目前资助的许多研究都集中在高度解剖解析的实验和有限数量的空间不同的植物器官和细胞类型（即花粉，毛状体，边缘细胞）。 因此，生物材料和相应代谢物的丰度非常低。 因此，配备了cryoflowprobe和耦合到在线质量定向固相萃取的NMR提供了关键的灵敏度增强，这是绝对必要的获得微量代谢物的结构数据，在半自动化模式，以提高吞吐量。 NMR仪器将通过鉴定关键模型和作物物种中重要的新型代谢物来促进对基本植物代谢和生物化学的理解。 它还将通过增加注释和覆盖深度来增加我们的代谢组学计划的生物背景，这将成比例地增强基因发现，基因的功能注释以及有益植物性状的更有效的代谢工程。这一有利的工具性资源将用于征聘一名新的长期工作人员。 诺贝尔基金会将为该仪器和工作人员提供长期支持。 资金还将加强未来教师的招聘，现有教师的保留，植物生物化学和代谢组学参与者的国际竞争力和领导力，并用于利用未来的州和联邦植物生物化学和农业研究资金。 拟议的仪器将急切地纳入现有的和生产性的教育和培训计划，在所有参与机构，服务于大量不同的博士后，研究生，本科生和高中学生。 具体包括：国家认可的多个动手，深入，为期一周的仪器培训课程，如我们的代谢组学和蛋白质组学用户培训计划;本科仪器分析课程;高中科学嘉年华;仪器“NutVolt”兴趣小组;着名的诺贝尔学者实习计划&;与南俄克拉荷马州技术中心、阿德莫尔高中AP化学、阿德莫尔高中科学俱乐部的高中合作项目，还有俄克拉荷马州的“向上跳跃”