Plant Cell Journal of Pharmacology and Experimental Therapeutics
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Published on May 11, 2007; 10.1105/tpc.106.045948

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Received July 15, 2006
Returned for revision April 1, 2007
Accepted April 18, 2007

CYP703 Is an Ancient Cytochrome P450 in Land Plants Catalyzing in-Chain Hydroxylation of Lauric Acid to Provide Building Blocks for Sporopollenin Synthesis in Pollen

Marc Morant 1, Kirsten Jørgensen 1, Hubert Schaller 2, Franck Pinot 2, Birger Lindberg Møller 1, Danièle Werck-Reichhart 2, and Søren Bak 1*

1 Plant Biochemistry Laboratory, Department of Plant Biology and Center for Molecular Plant Physiology, Faculty of Life Sciences, University of Copenhagen, DK-1871 Frederiksberg C, Copenhagen, Denmark
2 Departments of Plant Metabolic Responses and Plant Isoprenoids, Botanic Institute, Louis Pasteur University, 67083 Strasbourg Cedex, France

* To whom correspondence should be addressed. E-mail: bak{at}life.ku.dk.

CYP703 is a cytochrome P450 family specific to land plants. Typically, each plant species contains a single CYP703. Arabidopsis thaliana CYP703A2 is expressed in the anthers of developing flowers. Expression is initiated at the tetrad stage and restricted to microspores and to the tapetum cell layer. Arabidopsis CYP703A2 knockout lines showed impaired pollen development and a partial male-sterile phenotype. Scanning electron and transmission electron microscopy of pollen from the knockout plants showed impaired pollen wall development with absence of exine. The fluorescent layer around the pollen grains ascribed to the presence of phenylpropanoid units in sporopollenin was absent in the CYP703A2 knockout lines. Heterologous expression of CYP703A2 in yeast cells demonstrated that CYP703 catalyzes the conversion of medium-chain saturated fatty acids to the corresponding monohydroxylated fatty acids, with a preferential hydroxylation of lauric acid at the C-7 position. Incubation of recombinant CYP703 with methanol extracts from developing flowers confirmed that lauric acid and in-chain hydroxy lauric acids are the in planta substrate and product, respectively. These data demonstrate that in-chain hydroxy lauric acids are essential building blocks in sporopollenin synthesis and enable the formation of ester and ether linkages with phenylpropanoid units. This study identifies CYP703 as a P450 family specifically involved in pollen development.




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