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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

CYP86B1 Is Required for Very Long Chain -Hydroxyacid and ,-Dicarboxylic Acid Synthesis in Root and Seed Suberin Polyester^1,[W],[OA]

Institut de Biologie Moléculaire des Plantes, CNRS-Université Louis Pasteur Unité Propre de Recherche 2357, Département Réseaux Métaboliques Végétaux, F–67083 Strasbourg cedex, France (V.C., I.B., D.M., H.S., F.P.); and Institut für Zelluläre und Molekulare Botanik, Abteilung Ökophysiologie, Universität Bonn, 53115 Bonn, Germany (P.D., L.S., R.F.)

Suberin composition of various plants including Arabidopsis (Arabidopsis thaliana) has shown the presence of very long chain fatty acid derivatives C20 in addition to the C16 and C18 series. Phylogenetic studies and plant genome mining have led to the identification of putative aliphatic hydroxylases belonging to the CYP86B subfamily of cytochrome P450 monooxygenases. In Arabidopsis, this subfamily is represented by CYP86B1 and CYP86B2, which share about 45% identity with CYP86A1, a fatty acid -hydroxylase implicated in root suberin monomer synthesis. Here, we show that CYP86B1 is located to the endoplasmic reticulum and is highly expressed in roots. Indeed, CYP86B1 promoter-driven β-glucuronidase expression indicated strong reporter activities at known sites of suberin production such as the endodermis. These observations, together with the fact that proteins of the CYP86B type are widespread among plant species, suggested a role of CYP86B1 in suberin biogenesis. To investigate the involvement of CYP86B1 in suberin biogenesis, we characterized an allelic series of cyp86B1 mutants of which two strong alleles were knockouts and two weak ones were RNA interference-silenced lines. These root aliphatic plant hydroxylase lines had a root and a seed coat aliphatic polyester composition in which C22- and C24-hydroxyacids and ,-dicarboxylic acids were strongly reduced. However, these changes did not affect seed coat permeability and ion content in leaves. The presumed precursors, C22 and C24 fatty acids, accumulated in the suberin polyester. These results demonstrate that CYP86B1 is a very long chain fatty acid hydroxylase specifically involved in polyester monomer biosynthesis during the course of plant development.

² These authors contributed equally to the article.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Vincent Compagnon (vincent.compagnon{at}ibmp-ulp.u-strasbg.fr).

^[W] The online version of this article contains Web-only data.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.109.141408

^* Corresponding author; e-mail franck.pinot{at}ibmp-ulp.u-strasbg.fr.

Received May 14, 2009; accepted May 27, 2009; published June 12, 2009.

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