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Plant and Cell Physiology Advance Access originally published online on February 2, 2009
Plant and Cell Physiology 2009 50(3):584-594; doi:10.1093/pcp/pcp017
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

A Cellulose Synthase-Containing Compartment Moves Rapidly Beneath Sites of Secondary Wall Synthesis

Raymond Wightman, Robin Marshall and Simon R. Turner*

Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK

*Corresponding author: E-mail, simon.turner{at}manchester.ac.uk; Fax, +44-161-275-3938.


  Abstract

The woody secondary walls of plants represent the major sites of cellulose deposition. The polymerization of cellulose occurs at the plasma membrane by the secondary wall cellulose synthase complex (CSC). In the long, cylindrical cells that make up the xylem, secondary wall deposition is confined to discrete regions of the cell, and yellow fluorescent protein (YFP)-labeled CSCs are also localized to these regions. Using fluorescence loss in photobleaching (FLIP) of complete hoops containing YFP–CSCs, we demonstrate movement of the complexes beneath the nascent secondary wall in developing xylem vessels. We have devised a method for determining particle velocities for particles moving around a cylindrical object using data from FLIP. By applying this method to the hoops of YFP–CSCs of the developing vessels, we have obtained the first estimates of speed of these complexes. These speeds are calculated to be in excess of 7 µm s–1 and are far higher than those speeds previously reported for the primary wall complex. These high speeds are unlikely to be consistent with CSC movement being attributed to cellulose synthesis alone, and suggest the existence of a highly motile compartment beneath the nascent secondary wall.

Keywords: Arabidopsis - Cellulose synthase complex - FLIP - Secondary cell wall - Xylem - YFP.

Abbreviations: CFP, cyan fluorescent protein; CSC, cellulose synthase complex; DCB, dichlorobenzonitrile; DMSO, dimethylsulfoxide; ER, endoplasmic reticulum; FLIP, fluorescence loss in photobleaching; FRAP, fluorescence recovery after photobleaching; YFP, yellow fluorescent protein.

(Received October 6, 2008; Accepted January 27, 2009)
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