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

Bin Li; Camilla Knudsen Baden; Natascha Kristine Krahl Hansen; Kirsten Jørgensen; Rubini Maya Kannangara; Søren Bak; Adam Matthew Takos; Frederik Rook; Steen Honoré Hansen; Birger Lindberg Møller; Christian Janfelt; Nanna Bjarnholt

doi:10.1111/tpj.12183

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

Bin Li, Camilla Knudsen Baden, Natascha Kristine Krahl Hansen, Kirsten Jørgensen, Rubini Maya Kannangara, Søren Bak, Adam Matthew Takos, Frederik Rook, Steen Honoré Hansen, Birger Lindberg Møller, Christian Janfelt, Nanna Bjarnholt

53 Citations (Scopus)

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 language	English
Article number	74
Journal	Plant Journal
Volume	74
Issue number	6
Pages (from-to)	1059-1071
ISSN	0960-7412
DOIs	https://doi.org/10.1111/tpj.12183
Publication status	Published - Jun 2013

Access to Document

10.1111/tpj.12183

tpj.12183Final published version, 1.99 MB

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/tpj.12183

Cite this

Li, B., Baden, C. K., Hansen, N. K. K., Jørgensen, K., Kannangara, R. M., Bak, S., Takos, A. M., 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. [74]. https://doi.org/10.1111/tpj.12183

Li, B, Baden, CK, Hansen, NKK, Jørgensen, K, Kannangara, RM, Bak, S, Takos, AM, Rook, F, Hansen, SH, Møller, BL , Janfelt, C & Bjarnholt, N 2013, 'Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging', Plant Journal, vol. 74, no. 6, 74, pp. 1059-1071. https://doi.org/10.1111/tpj.12183

@article{e14f80af1a034c44a03f8a0150630bf8,

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.",

author = "Bin Li and Baden, {Camilla Knudsen} and Hansen, {Natascha Kristine Krahl} and Kirsten J{\o}rgensen and Kannangara, {Rubini Maya} and S{\o}ren Bak and Takos, {Adam Matthew} and Frederik Rook and Hansen, {Steen Honor{\'e}} and M{\o}ller, {Birger Lindberg} and Christian Janfelt and Nanna Bjarnholt",

note = "{\textcopyright} 2013 The Authors. The Plant Journal {\textcopyright} 2013 Blackwell Publishing Ltd.",

year = "2013",

month = jun,

doi = "10.1111/tpj.12183",

language = "English",

volume = "74",

pages = "1059--1071",

journal = "Plant Journal",

issn = "0960-7412",

publisher = "Wiley-Blackwell",

number = "6",

}

TY - JOUR

T1 - Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging

AU - Li, Bin

AU - Baden, Camilla Knudsen

AU - Hansen, Natascha Kristine Krahl

AU - Jørgensen, Kirsten

AU - Kannangara, Rubini Maya

AU - Bak, Søren

AU - Takos, Adam Matthew

AU - Rook, Frederik

AU - Hansen, Steen Honoré

AU - Møller, Birger Lindberg

AU - Janfelt, Christian

AU - Bjarnholt, Nanna

PY - 2013/6

Y1 - 2013/6

N2 - 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.

AB - 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.

U2 - 10.1111/tpj.12183

DO - 10.1111/tpj.12183

M3 - Journal article

C2 - 23551340

SN - 0960-7412

VL - 74

SP - 1059

EP - 1071

JO - Plant Journal

JF - Plant Journal

IS - 6

M1 - 74

ER -

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

Abstract

Access to Document

Fingerprint

Cite this