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Molecular Plant Advance Access originally published online on August 24, 2009
Molecular Plant 2009 2(5):1025-1039; doi:10.1093/mp/ssp064
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© The Author 2009. Published by the Molecular Plant Shanghai Editorial Office in association with Oxford University Press on behalf of CSPP and IPPE, SIBS, CAS.

The CELLULOSE-SYNTHASE LIKE C (CSLC) Family of Barley Includes Members that Are Integral Membrane Proteins Targeted to the Plasma Membrane

Fenny M. Dwivanya,d, Dina Yuliaa,e, Rachel A. Burtonb, Neil J. Shirleyb, Sarah M. Wilsona, Geoffrey B. Fincherb, Antony Bacica,c, Ed Newbigina,1 and Monika S. Doblina

a Plant Cell Biology Research Centre, School of Botany, University of Melbourne Victoria 3010, Australia
b Australian Centre for Plant Functional Genomics, School of Agriculture and Wine, University of Adelaide, South Australia 5064, Australia
c Australian Centre for Plant Functional Genomics, School of Botany, University of Melbourne Victoria 3010 Australia
d Present address: Department of Biology, Institut Teknologi Bandung, Bandung, Indonesia
e Present address: Stem Cell and Cancer Institute, Jl. Jend. Ahmad Yani No.2, Pulo Mas, Jakarta 13210, Indonesia

1 To whom correspondence should be addressed. E-mail edwardjn{at}unimelb.edu.au, fax 61-3-9347-1071, tel. 61-3-8344-4871.

The CELLULOSE SYNTHASE-LIKE C (CSLC) family is an ancient lineage within the CELLULOSE SYNTHASE/CELLULOSE SYNTHASE-LIKE (CESA/CSL) polysaccharide synthase superfamily that is thought to have arisen before the divergence of mosses and vascular plants. As studies in the flowering plant Arabidopsis have suggested synthesis of the (1,4)-β-glucan backbone of xyloglucan (XyG), a wall polysaccharide that tethers adjacent cellulose microfibrils to each other, as a probable function for the CSLCs, CSLC function was investigated in barley (Hordeum vulgare L.), a species with low amounts of XyG in its walls. Four barley CSLC genes were identified (designated HvCSLC1–4). Phylogenetic analysis reveals three well supported clades of CSLCs in flowering plants, with barley having representatives in two of these clades. The four barley CSLCs were expressed in various tissues, with in situ PCR detecting transcripts in all cell types of the coleoptile and root, including cells with primary and secondary cell walls. Co-expression analysis showed that HvCSLC3 was coordinately expressed with putative XyG xylosyltransferase genes. Both immuno-EM and membrane fractionation showed that HvCSLC2 was located in the plasma membrane of barley suspension-cultured cells and was not in internal membranes such as endoplasmic reticulum or Golgi apparatus. Based on our current knowledge of the sub-cellular locations of polysaccharide synthesis, we conclude that the CSLC family probably contains more than one type of polysaccharide synthase.

Key Words: Cellulose synthase-like family C • plant cell wall biosynthesis • xyloglucan • cellulose • glycosyltransferase


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