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.
authors.affiliation: Laboratory of Food Chemistry and Biochemistry & Leuven Food Science and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven , Kasteelpark Arenberg 20-box 2463, B-3001 Leuven, Belgium.
authors.forename: Emmie
authors.initials: E
authors.lastname: Dornez
authors.forename: Sven
authors.initials: S
authors.lastname: Cuyvers
authors.forename: Ulla
authors.initials: U
authors.lastname: Holopainen
authors.forename: Emilia
authors.initials: E
authors.lastname: Nordlund
authors.forename: Kaisa
authors.initials: K
authors.lastname: Poutanen
authors.forename: Jan A
authors.initials: JA
authors.lastname: Delcour
authors.forename: Christophe M
authors.initials: CM
authors.lastname: Courtin
mesh_terms: D002473:Cell Wall / Q000737:chemistry* / Q000378:metabolism; D002523:Edible Grain / Q000737:chemistry* / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding; D043325:Xylan Endo-1,3-beta-Xylosidase / Q000737:chemistry*; D014990:Xylans / Q000737:chemistry* / Q000378:metabolism
chemical_list: D014990:Xylans; C085118:arabinoxylan; D043325:Xylan Endo-1,3-beta-Xylosidase