Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging

Authors

  • Bin Li,

    1. Department of Pharmacy, University of Copenhagen, Copenhagen, Denmark
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  • Camilla Knudsen,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Natascha Krahl Hansen,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Kirsten Jørgensen,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Rubini Kannangara,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Søren Bak,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Adam Takos,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Fred Rook,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Steen H. Hansen,

    1. Department of Pharmacy, University of Copenhagen, Copenhagen, Denmark
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  • Birger Lindberg Møller,

    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
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  • Christian Janfelt,

    Corresponding author
    • Department of Pharmacy, University of Copenhagen, Copenhagen, Denmark
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  • Nanna Bjarnholt

    Corresponding author
    1. Department of Plant and Environmental Sciences, University of Copenhagen, Plant Biochemistry Laboratory and Villum Research Center ‘Pro-Active Plants’, Center for Synthetic Biology, Frederiksberg C, Copenhagen, Denmark
    • Department of Pharmacy, University of Copenhagen, Copenhagen, Denmark
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For correspondence (e-mails christian.janfelt@sund.ku.dk or nnb@life.ku.dk).

Summary

In comparison with the technology platforms developed to localize transcripts and proteins, imaging tools for visualization of metabolite distributions in plant tissues are less well developed and lack versatility. This hampers our understanding of plant metabolism and dynamics. In this study, we demonstrate that desorption electrospray ionization mass spectrometry imaging (DESI-MSI) of tissue imprints on porous Teflon may be used to accurately image the distribution of even labile plant metabolites such as hydroxynitrile glucosides, which normally undergo enzymatic hydrolysis by specific β-glucosidases upon cell disruption. This fast and simple sample preparation resulted in no substantial differences in the distribution and ratios of all hydroxynitrile glucosides between leaves from wild-type Lotus japonicus and a β-glucosidase mutant plant that lacks the ability to hydrolyze certain hydroxynitrile glucosides. In wild-type, the enzymatic conversion of hydroxynitrile glucosides and the concomitant release of glucose were easily visualized when a restricted area of the leaf tissue was damaged prior to sample preparation. The gene encoding the first enzyme in hydroxynitrile glucoside biosynthesis in L. japonicus leaves, CYP79D3, was found to be highly expressed during the early stages of leaf development, and the hydroxynitrile glucoside distribution in mature leaves reflected this early expression pattern. The utility of direct DESI-MSI of plant tissue was demonstrated using cryo-sections of cassava (Manihot esculenta) tubers. The hydroxynitrile glucoside levels were highest in the outer cell layers, as verified by LC–MS analyses. The unexpected discovery of a hydroxynitrile-derived di-glycoside shows the potential of DESI-MSI to discover and guide investigations into new metabolic routes.

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