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

Bin Li, Camilla Knudsen, Natascha Krahl Hansen, Kirsten Jørgensen, Rubini Kannangara, Søren Bak, Adam Takos, Fred Rook, Steen H. Hansen, Birger Lindberg Møller, Christian Janfelt, Nanna Bjarnholt

Research output: Contribution to journalJournal articleResearchpeer-review

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.
Original languageEnglish
JournalPlant Journal
Volume74
Issue number6
Pages (from-to)1059-1071
ISSN0960-7412
DOIs
Publication statusPublished - 2013
Externally publishedYes

Keywords

  • Metabolite imaging
  • DESI-MS
  • Cyanogenic glucosides
  • Hydroxynitrile glucosides
  • Lotus japonicus
  • Manihot esculenta
  • Sorghum bicolor
  • Technical advance

Cite this

Li, B., Knudsen, C., Hansen, N. K., Jørgensen, K., Kannangara, R., Bak, S., Takos, A., Rook, F., Hansen, S. H., Møller, B. L., Janfelt, C., & Bjarnholt, N. (2013). Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging. Plant Journal, 74(6), 1059-1071. https://doi.org/10.1111/tpj.12183