Inactive fluorescently labeled xylanase as a novel probe for microscopic analysis of arabinoxylan containing cereal cell walls

Emmie Dornez, Sven Cuyvers, Ulla Holopainen, Emilia Nordlund, Kaisa Poutanen, Jan A Delcour, Christophe M Courtin (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

28 Citations (Scopus)

Abstract

A new technique to visualize cereal cell walls by fluorescence microscopy was developed. The novel staining technique is based on an inactive fluorescently labeled xylanase binding to arabinoxylan (AX), an important polysaccharide in grain cell walls in terms of the technological and physiological functionalities of grain. The xylanase probe could stain AX in the seed coat, nucellar epidermis, aleurone layer, and starchy endosperm, but not the highly substituted AX of the pericarp layer. The advantage of this new staining technique over the existing immunolabeling techniques is that the staining procedure is clearly faster and less laborious, and uses a smaller probe that can easily be produced by marking a well characterized enzyme with a fluorescent label. In the future, the here proposed technology can be used to develop probes having specificity also for cell wall components other than AX and thus to study plant cell walls further through fluorescence microscopy.
Original languageEnglish
Pages (from-to)6369-6375
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Volume59
Issue number12
DOIs
Publication statusPublished - Jun 2011
MoE publication typeA1 Journal article-refereed

Fingerprint

arabinoxylan
xylanases
Cell Wall
Cells
cell walls
Fluorescence microscopy
Staining and Labeling
fluorescence microscopy
Fluorescence Microscopy
aleurone layer
Endosperm
Plant Cells
cell wall components
Cellular Structures
epidermis (plant)
pericarp
Epidermis
endosperm
Polysaccharides
dyes

Keywords

  • 3-beta-Xylosidase
  • 3-beta-Xylosidase: chemistry
  • Cell Wall
  • Cell Wall: chemistry
  • Cell Wall: metabolism
  • Cereals
  • Cereals: chemistry
  • Cereals: metabolism
  • Fluorescence
  • Fluorescence: instrumentation
  • Fluorescence: methods
  • Microscopy
  • Protein Binding
  • Xylan Endo-1
  • Xylans
  • Xylans: chemistry
  • Xylans: metabolism

Cite this

@article{8ad3a61a43eb4feaaea5ee0253f0de0a,
title = "Inactive fluorescently labeled xylanase as a novel probe for microscopic analysis of arabinoxylan containing cereal cell walls",
abstract = "A new technique to visualize cereal cell walls by fluorescence microscopy was developed. The novel staining technique is based on an inactive fluorescently labeled xylanase binding to arabinoxylan (AX), an important polysaccharide in grain cell walls in terms of the technological and physiological functionalities of grain. The xylanase probe could stain AX in the seed coat, nucellar epidermis, aleurone layer, and starchy endosperm, but not the highly substituted AX of the pericarp layer. The advantage of this new staining technique over the existing immunolabeling techniques is that the staining procedure is clearly faster and less laborious, and uses a smaller probe that can easily be produced by marking a well characterized enzyme with a fluorescent label. In the future, the here proposed technology can be used to develop probes having specificity also for cell wall components other than AX and thus to study plant cell walls further through fluorescence microscopy.",
keywords = "3-beta-Xylosidase, 3-beta-Xylosidase: chemistry, Cell Wall, Cell Wall: chemistry, Cell Wall: metabolism, Cereals, Cereals: chemistry, Cereals: metabolism, Fluorescence, Fluorescence: instrumentation, Fluorescence: methods, Microscopy, Protein Binding, Xylan Endo-1, Xylans, Xylans: chemistry, Xylans: metabolism",
author = "Emmie Dornez and Sven Cuyvers and Ulla Holopainen and Emilia Nordlund and Kaisa Poutanen and Delcour, {Jan A} and Courtin, {Christophe M}",
year = "2011",
month = "6",
doi = "10.1021/jf200746g",
language = "English",
volume = "59",
pages = "6369--6375",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
publisher = "American Chemical Society ACS",
number = "12",

}

Inactive fluorescently labeled xylanase as a novel probe for microscopic analysis of arabinoxylan containing cereal cell walls. / Dornez, Emmie; Cuyvers, Sven; Holopainen, Ulla; Nordlund, Emilia; Poutanen, Kaisa; Delcour, Jan A; Courtin, Christophe M (Corresponding Author).

In: Journal of Agricultural and Food Chemistry, Vol. 59, No. 12, 06.2011, p. 6369-6375.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Inactive fluorescently labeled xylanase as a novel probe for microscopic analysis of arabinoxylan containing cereal cell walls

AU - Dornez, Emmie

AU - Cuyvers, Sven

AU - Holopainen, Ulla

AU - Nordlund, Emilia

AU - Poutanen, Kaisa

AU - Delcour, Jan A

AU - Courtin, Christophe M

PY - 2011/6

Y1 - 2011/6

N2 - A new technique to visualize cereal cell walls by fluorescence microscopy was developed. The novel staining technique is based on an inactive fluorescently labeled xylanase binding to arabinoxylan (AX), an important polysaccharide in grain cell walls in terms of the technological and physiological functionalities of grain. The xylanase probe could stain AX in the seed coat, nucellar epidermis, aleurone layer, and starchy endosperm, but not the highly substituted AX of the pericarp layer. The advantage of this new staining technique over the existing immunolabeling techniques is that the staining procedure is clearly faster and less laborious, and uses a smaller probe that can easily be produced by marking a well characterized enzyme with a fluorescent label. In the future, the here proposed technology can be used to develop probes having specificity also for cell wall components other than AX and thus to study plant cell walls further through fluorescence microscopy.

AB - A new technique to visualize cereal cell walls by fluorescence microscopy was developed. The novel staining technique is based on an inactive fluorescently labeled xylanase binding to arabinoxylan (AX), an important polysaccharide in grain cell walls in terms of the technological and physiological functionalities of grain. The xylanase probe could stain AX in the seed coat, nucellar epidermis, aleurone layer, and starchy endosperm, but not the highly substituted AX of the pericarp layer. The advantage of this new staining technique over the existing immunolabeling techniques is that the staining procedure is clearly faster and less laborious, and uses a smaller probe that can easily be produced by marking a well characterized enzyme with a fluorescent label. In the future, the here proposed technology can be used to develop probes having specificity also for cell wall components other than AX and thus to study plant cell walls further through fluorescence microscopy.

KW - 3-beta-Xylosidase

KW - 3-beta-Xylosidase: chemistry

KW - Cell Wall

KW - Cell Wall: chemistry

KW - Cell Wall: metabolism

KW - Cereals

KW - Cereals: chemistry

KW - Cereals: metabolism

KW - Fluorescence

KW - Fluorescence: instrumentation

KW - Fluorescence: methods

KW - Microscopy

KW - Protein Binding

KW - Xylan Endo-1

KW - Xylans

KW - Xylans: chemistry

KW - Xylans: metabolism

U2 - 10.1021/jf200746g

DO - 10.1021/jf200746g

M3 - Article

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SN - 0021-8561

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