Plant Physiology Preview
Published on September 15, 2006; 10.1104/pp.106.086785
Received July 15, 2006
Accepted September 1, 2006
CER4 Encodes an Alcohol-Forming Fatty Acyl-CoA Reductase Involved in Cuticular Wax Production in Arabidopsis
Owen Rowland , Huanquan Zheng , Shelley R. Hepworth , Patricia Lam , Reinhard Jetter , and Ljerka Kunst *
Department of Botany, University of British Columbia, 6270 University Boulevard, Vancouver, BC, V6T 1Z4, Canada
Department of Botany, University of British Columbia, 6270 University Boulevard, Vancouver, BC, V6T 1Z4, Canada; Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada, V6T 1Z1, Canada.
* Corresponding author; email: kunst{at}interchange.ubc.ca.
A waxy cuticle that serves as a protective barrier against uncontrolled water loss and environmental damage coats the aerial surfaces of land plants. It is composed of a cutin polymer matrix and waxes. Cuticular waxes are complex mixtures of very-long-chain fatty acids (VLCFAs) and their derivatives. We report here the molecular cloning and characterization of CER4, a wax biosynthetic gene from Arabidopsis thaliana. Arabidopsis cer4 mutants exhibit major decreases in stem primary alcohols and wax esters, and slightly elevated levels of aldehydes, alkanes, secondary alcohols and ketones. This phenotype suggested that CER4 encoded an alcohol-forming fatty acyl-CoA reductase (FAR). We identified eight FAR-like genes in Arabidopsis that are highly related to an alcohol-forming FAR expressed in seeds of Simmondsia chinesis (jojoba). Molecular characterization of CER4 alleles and genomic complementation revealed that one of these eight genes, At4g33790, encoded the FAR required for cuticular wax production. Expression of CER4 cDNA in yeast resulted in the accumulation of C24:0 and C26:0 primary alcohols. Fully functional GFP-tagged CER4 protein was localized to the ER in yeast cells by confocal microscopy. Analysis of gene expression by RT-PCR indicated that CER4 was expressed in leaves, stems, flowers, siliques and roots. Expression of a GUS reporter gene driven by the CER4 promoter in transgenic plants was detected in epidermal cells of leaves and stems, consistent with a dedicated role for CER4 in cuticular wax biosynthesis. CER4 was expressed in all cell types in the elongation zone of young roots. These data indicate that CER4 is an alcohol-forming FAR that has specificity for VLCFAs and is responsible for the synthesis of primary alcohols in the epidermal cells of aerial tissues and in roots.
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