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Plant Cell, Vol. 12, 691-706, May 2000, Copyright © 2000, American Society of Plant Physiologists

Virus-Induced Silencing of a Plant Cellulose Synthase Gene

Rachel A. Burtona, David M. Gibeauta, Antony Bacicb, Kim Findlayc, Keith Robertsc, Andrew Hamiltond, David C. Baulcombed, and Geoffrey B. Finchera
a Department of Plant Science, University of Adelaide, Waite Campus, Glen Osmond, South Australia 5064, Australia
b School of Botany, University of Melbourne, Parkville, Victoria 3052, Australia
c Department of Cell Biology, John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom
d Sainsbury Laboratory, John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom

Correspondence to: Geoffrey B. Fincher, gfincher{at}waite.adelaide.edu.au (E-mail), 61-8-8303-7109 (fax)

Specific cDNA fragments corresponding to putative cellulose synthase genes (CesA) were inserted into potato virus X vectors for functional analysis in Nicotiana benthamiana by using virus-induced gene silencing. Plants infected with one group of cDNAs had much shorter internode lengths, small leaves, and a "dwarf" phenotype. Consistent with a loss of cell wall cellulose, abnormally large and in many cases spherical cells ballooned from the undersurfaces of leaves, particularly in regions adjacent to vascular tissues. Linkage analyses of wall polysaccharides prepared from infected leaves revealed a 25% decrease in cellulose content. Transcript levels for at least one member of the CesA cellulose synthase gene family were lower in infected plants. The decrease in cellulose content in cell walls was offset by an increase in homogalacturonan, in which the degree of esterification of carboxyl groups decreased from ~50 to ~33%. The results suggest that feedback loops interconnect the cellular machinery controlling cellulose and pectin biosynthesis. On the basis of the phenotypic features of the infected plants, changes in wall composition, and the reduced abundance of CesA mRNA, we concluded that the cDNA fragments silenced one or more cellulose synthase genes.




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