This project uses funds to add an integrated mass spectrometry-based tissue imaging system, including a tissue sectioning station and a new generation MALDI-based quadrupole-time of flight mass spectrometry (Q-TOFMS) system that incorporates integrated ion mobility spectrometry, to the M.J. Murdock Metabolomics Laboratory (MML, www.murdockmetabolomics.wsu.edu, co-directed by three of the PIs) at Washington State University. Addition of this integrated instrumentation system to the MML enables researchers in the IBC and the WSU campus as a whole to acquire tissue imaging data that incorporates not only high mass accuracy and high resolution MS and MS/MS-based identification of proteins and metabolites, but also ion mobility spectrometric characterization of such compounds, increasing confidence in compound identification and quantification, while reducing noise in data acquisition. A critical missing link in past research has been the ability to identify, at the cellular or sub-tissue level, where biochemical processes occur or how pathways are regulated, especially in complex organs such as plant stems, roots, rhizomes and leaves. Acquisition of this integrated instrumentation enables fundamental mechanistic questions to be addressed that have remained unanswered for decades due to inadequate spatial resolution of previous analytical approaches. The integrated tissue imaging system will be housed in a newly renovated lab within the MML designed to accommodate such instrumentation, and expands the MML's current strengths in metabolomics and proteomics, furthering the goal of developing the MML as a world premier plant metabolomics facility.
Plants produce a fantastic diversity of compounds and substances that are used in pollination, plant defense, communication within individual plants and between plants and other organisms, among many other functions. Many of these highly specialized substances, such as flavors, fragrances, gums, resins, toxins, structural components, and nutritional and medicinal compounds, are produced in very few cell types or even in individual cells in plants. The ability to detect and measure various metabolites and the proteins that produce them in distinct single cell types in plant tissues has long been near impossible, even though this ability is vital in the long run to obtain a complete understanding of plant metabolism. It is generally considered that such knowledge is critical to future efforts to improve plants for human use. Regulation of these processes at the individual cellular level in plants is poorly understood, largely as a result of inability to detect and measure metabolites in specific plant cells. The new tissue imaging technology allows for detection and measurement of metabolites and proteins in specific cells in complex biological tissues such as stems, roots, rhizomes, flowers and leaves, and other specialized cell types in plants, or other tissues in other organisms (fungi, animals, insects, etc.). This new technology thus enables researchers to test hypotheses about important and fundamental biological processes related to medicinal plant metabolism, biofuel production, plant-insect interactions and basic plant biochemistry, among other areas. Data acquired with this instrument system will be part of the research projects of graduate students, undergraduate students and post-doctoral fellows in the fields of plant biochemistry, chemistry, interdisciplinary omics, and metabolic engineering. In addition, the new instrument system will be integrated into two specific efforts to attract current and future students to the fields of plant biochemistry and metabolomics particularly and to science in general and to provide educational advantages to the students at WSU. First, plant tissue imaging will be incorporated into the Advanced Plant Biochemistry course taught by CoPI Lange. Second, PI Gang will participate in CoPI Weller's "Pumping-Up the Math and Science Pipeline: Grade School to College" program that focuses on Native American and Hispanic populations on the Colville Reservation (largest in the Pacific Northwest) and in non-tribal regions surrounding Omak, WA, which hosts high school seniors for summer research internships.
