title: Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging.
abstract: 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.
authors.affiliation: Department of Pharmacy, University of Copenhagen, Universitetsparken 2, DK-2100, Copenhagen, Denmark.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.forename: Camilla
authors.initials: C
authors.lastname: Knudsen
authors.forename: Natascha Krahl
authors.initials: NK
authors.lastname: Hansen
authors.forename: Kirsten
authors.initials: K
authors.lastname: Jørgensen
authors.forename: Rubini
authors.initials: R
authors.lastname: Kannangara
authors.forename: Søren
authors.initials: S
authors.lastname: Bak
authors.forename: Adam
authors.initials: A
authors.lastname: Takos
authors.forename: Fred
authors.initials: F
authors.lastname: Rook
authors.forename: Steen H
authors.initials: SH
authors.lastname: Hansen
authors.forename: Birger Lindberg
authors.initials: BL
authors.lastname: Møller
authors.forename: Christian
authors.initials: C
authors.lastname: Janfelt
authors.forename: Nanna
authors.initials: N
authors.lastname: Bjarnholt
mesh_terms: D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D005960:Glucosides / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D000070116:Lotus / Q000737:chemistry / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D002365:Manihot / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D013058:Mass Spectrometry; D009154:Mutation; D009570:Nitriles / Q000737:chemistry / Q000378:metabolism; D018515:Plant Leaves / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D035281:Plant Tubers / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D036226:Seedlings / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D045868:Sorghum / Q000737:chemistry / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D001617:beta-Glucosidase / Q000378:metabolism
chemical_list: D005960:Glucosides; D009570:Nitriles; D003577:Cytochrome P-450 Enzyme System; C492216:cytochrome P-450 79D3, Lotus japonicus; D001617:beta-Glucosidase; C011220:dhurrin