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

34 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

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

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