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JXB Advance Access published online on July 12, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri225
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved.
Received January 18, 2005
Accepted May 13, 2005

RESEARCH PAPER

Molecular cloning and characterization of an almond 9-hydroperoxide lyase, a new CYP74 targeted to lipid bodies*

Giovanni Mita 1, Angela Quarta 1, Pasqua Fasano 1, Angelo De Paolis 1, Gian Pietro Di Sansebastiano 2, Carla Perrotta 2, Rina Iannacone 3, Eric Belfield 4, Richard Hughes 4, Nicolas Tsesmetzis 4, Rod Casey 4, and Angelo Santino 1*

1 Institute of Sciences of Food Production CNR Section of Lecce, via Monteroni, I-73100 Italy
2 Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università di Lecce, via Monteroni, I-73100, Lecce, Italy
3 Metapontum Agrobios, S.S. Jonica 106, Km 448.2, I-75010, Metaponto (MT), Italy
4 John Innes Centre, Norwich Research Park, Colney, Norwich NR3 7UH, UK

* To whom correspondence should be addressed.
Angelo Santino, E-mail: angelo.santino{at}ispa.cnr.it


  Abstract

Oxylipin metabolism represents one of many defence mechanisms employed by plants. It begins with the oxygenation of polyunsaturated fatty acids by lipoxygenases to form fatty acid hydroperoxides that are substrates for several enzymes, including specialized cytochrome P450s known as CYP74s. The targeting of a new CYP74, a 9-hydroperoxide lyase (HPL) from almonds, to the endomembrane system and lipid bodies, both as enzyme activity in almond seeds and as GFP fusions transiently expressed in tobacco protoplasts, is described. Such association of a CYP74 with lipid bodies has not been reported previously. Also described are the properties of a 9-HPL gene, the developmental regulation of its expression, the production and characterization of recombinant 9-HPL in Escherichia coli, and the developmental correlation between gene expression, enzyme activity, and the appearance of volatile C9 aldehydes from HPL action.

Keywords: Almond; hydroperoxide lyase; lipid body; oxylipins; seed development.
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