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JXB Advance Access originally published online on July 12, 2005
Journal of Experimental Botany 2005 56(419):2321-2333; 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.

RESEARCH PAPER

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

Giovanni Mita1, Angela Quarta1, Pasqua Fasano1, Angelo De Paolis1, Gian Pietro Di Sansebastiano2, Carla Perrotta2, Rina Iannacone3, Eric Belfield4, Richard Hughes4, Nicolas Tsesmetzis4, Rod Casey4 and Angelo Santino1,{dagger}

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

{dagger} To whom correspondence should be addressed. Fax: +39 0 8324 22620. E-mail: angelo.santino{at}ispa.cnr.it

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.

Key words: Almond, hydroperoxide lyase, lipid body, oxylipins, seed development


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