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Published on July 9, 2004; 10.1104/pp.104.044354


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Received April 9, 2004
Returned for revision May 21, 2004
Accepted May 22, 2004

Cloning and Functional Characterization of a Phospholipid:Diacylglycerol Acyltransferase from Arabidopsis

Ulf Ståhl , Anders S. Carlsson *, Marit Lenman , Anders Dahlqvist , Bangquan Huang , Walentyna Banas , Antoni Banas , and Sten Stymne

Department of Plant Biology and Forest Genetics, Uppsala Genetic Centre, Swedish University of Agricultural Sciences, S-750 07 Uppsala, Sweden
Department of Crop Science, Swedish University of Agricultural Sciences, S-230 53 Alnarp, Sweden
Scandinavian Biotechnology Research (ScanBi) AB, S-230 53 Alnarp, Sweden
College of Life Science, Hubei University, Wuhan, 430062, P.R. China
Institute of Biology, University of Podlasie, 08-110, Siedlce, Poland

* Corresponding author; email: anders.carlsson{at}vv.slu.se.

A new pathway for triacylglycerol biosynthesis involving a phospholipid:diacylglycerol acyltransferase (PDAT) was recently described (Dahlqvist et al., 2000). The LRO1 gene that encodes the PDAT was identified in yeast (Saccharomyces cerevisiae) and shown to have homology with animal lecithin:cholesterol acyltransferase. A search of the Arabidopsis genome database identified the protein encoded by the At5g13640 gene as the closest homolog to the yeast PDAT (28% amino acid identity). The cDNA of At5g13640 (AtPDAT gene) was overexpressed in Arabidopsis behind the cauliflower mosaic virus promoter. Microsomal preparations of roots and leaves from overexpressers had PDAT activities that correlated with expression levels of the gene, thus demonstrating that this gene encoded PDAT (AtPDAT). The AtPDAT utilized different phospholipids as acyl donor and accepted acyl groups ranging from C10 to C22. The rate of activity was highly dependent on acyl composition with highest activities for acyl groups containing several double bonds, epoxy, or hydroxy groups. The enzyme utilized both sn-positions of phosphatidylcholine but had a 3-fold preference for the sn-2 position. The fatty acid and lipid composition as well as the amounts of lipids per fresh weight in Arabidopsis plants overexpressing AtPDAT were not significantly different from the wild type. Microsomal preparations of roots from a T-DNA insertion mutant in the AtPDAT gene had barely detectable capacity to transfer acyl groups from phospholipids to added diacylglycerols. However, these microsomes were still able to carry out triacylglycerol synthesis by a diacylglycerol:diacylglycerol acyltransferase reaction at the same rate as microsomal preparations from wild type.




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