XTH acts at the microfibril-matrix interface during cell elongation.
Sulphorhodamine-labelled oligosaccharides of xyloglucan are incorporated into the cell wall of Arabidopsis and tobacco roots, and of cultured Nicotiana tabacum cells by the transglucosylase (XET) action of XTHs. In the cell wall of diffusely growing cells, the subcellular pattern of XET action revealed a 'fibrillar' pattern, different from the xyloglucan localization. The fibrillar fluorescence pattern had no net orientation in spherical cultured cells. It changed to transverse to the long axis when the cells started to elongate, a feature mirroring the rearrangements of cortical microtubules and the accompanying cellulose deposition. Interference with the polymerization of microtubules and with cellulose deposition inhibited this strong and 'fibrillar'-organized XET-action, whereas interference with actin-polymerization only decreased the intensity of enzyme action. Epidermal cells of a mutant with reduced cellulose synthesis also had low XET action. Root hairs (tip-growing cells) exhibited high XET-action over all their length, but lacked the specific parallel pattern. In both diffuse- and tip-growing cell types extraction of the incorporated fluorescent xyloglucans by a xyloglucan-specific endoglucanase reduced the fluorescence, but the 'fibrillar' appearance in diffuse growing cells was not eliminated. These results show that XTHs act on the xyloglucans attached to cellulose microfibrils. After incorporation of the fluorescent oligosaccharides, the xyloglucans decorate the cellulose microfibrils and become inaccessible to hydrolytic enzymes.
Vissenberg
Kris
K
University of Antwerp (Drie Eiken Campus), Department of Biology, Plant Physiology and Morphology, Universiteitsplein 1, 2610 Wilrijk, Belgium. kris.vissenberg@ua.ac.be
Fry
Stephen C
SC
Pauly
Markus
M
Höfte
Herman
H
Verbelen
Jean-Pierre
JP
D017360:Arabidopsis / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D048429:Cell Size; D002473:Cell Wall / Q000187:drug effects / Q000502:physiology; D004136:Dinitrobenzenes / Q000494:pharmacology; D005936:Glucans / Q000378:metabolism; D016695:Glycosyltransferases / Q000378:metabolism*; D020894:Microfibrils / Q000502:physiology*; D009570:Nitriles / Q000494:pharmacology; D018517:Plant Roots / Q000201:enzymology; D013424:Sulfanilamides / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines; D014026:Nicotiana / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure
D019086:Bridged Bicyclo Compounds, Heterocyclic; D004136:Dinitrobenzenes; D005936:Glucans; D009570:Nitriles; D013424:Sulfanilamides; D013844:Thiazoles; D053778:Thiazolidines; C012465:oryzalin; D016695:Glycosyltransferases; C473049:xyloglucan - xyloglucosyltransferase; C037068:latrunculin B; C055869:dichlobanil