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.