TRTech-PGR: Mass Spectrometry Imaging of in vivo Isotope labeling

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Understanding plant metabolism at the molecular level in response to pathogens or hostile growth conditions is crucial for the bioengineering of crop plants with the growing challenges imposed by globalization and climate change. The new biotechnological tools to be developed in this project will allow plant biologists to visualize cellular level metabolic changes in both temporal and spatial dimensions. This project is focused on developing tools to understand dynamic plant metabolism at single-cell resolution; however, the potential impact extends beyond plant biology across many scientific disciplines, including microbial biotechnology, precision medicine, and environmental applications. In addition to training graduate and undergraduate students in this interdisciplinary field between biological and physical sciences, a middle or high school teacher will be recruited to this project each year through the Research Experiences for Teachers program at Iowa State University. Teachers will work on an environment-related duckweed project, which they can continue after going back to the classroom and develop their own projects.The technology which will be developed is mass spectrometry imaging (MSI) of in vivo isotope labeling, referred to as MSIi. In vivo isotope labeling can trace metabolic activities and provide temporal information of metabolic changes. Isotope labeled metabolite distributions can then be visualized by MSI directly on tissues at cellular resolution. Hydroponic cultures will be optimized for in vivo isotope labeling of three model plants, duckweed, Arabidopsis, and maize. MSI data of plant tissues labeled with 2H, 15N, or 13C will be used to identify unknown metabolites directly on tissues and visualize metabolite flux in selected biological systems. To expedite the data analysis in an automatic and robust fashion, software tools will be developed by adding multiple functionalities to the existing open-source software, MSiReader, such as correcting isotope measurement errors, generating isotopologue images and abundances, visualizing labeling efficiency, and enabling untargeted labeling search. Once successful, the new technology can be expanded to many new directions, including fluxomics at single-cell resolution, high-precision metabolic bioengineering, and cell- or tissue-specific dynamic stress responses.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.