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Molecular Characterization of an Arabidopsis Gene Encoding Hydroperoxide Lyase, a Cytochrome P-450 That Is Wound Inducible1

Nicholas J. Bate2, 3, *, Sobhana Sivasankar2, Claire Moxon, John M.C. Riley, John E. Thompson, and Steven J. Rothstein

Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario, Canada N1G 2W1 (N.J.B., S.S., C.M., S.J.R.); and Department of Biology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1 (J.M.C.R., J.E.T.)

Hydroperoxide lyase (HPL) cleaves lipid hydroperoxides to produce volatile flavor molecules and also potential signal molecules. We have characterized a gene from Arabidopsis that is homologous to a recently cloned HPL from green pepper (Capsicum annuum). The deduced protein sequence indicates that this gene encodes a cytochrome P-450 with a structure similar to that of allene oxide synthase. The gene was cloned into an expression vector and expressed in Escherichia coli to demonstrate HPL activity. Significant HPL activity was evident when 13S-hydroperoxy-9(Z),11(E),15(Z)-octadecatrienoic acid was used as the substrate, whereas activity with 13S-hydroperoxy-9(Z),11(E)-octadecadienoic acid was approximately 10-fold lower. Analysis of headspace volatiles by gas chromatography-mass spectrometry, after addition of the substrate to E. coli extracts expressing the protein, confirmed enzyme-activity data, since cis-3-hexenal was produced by the enzymatic activity of the encoded protein, whereas hexanal production was limited. Molecular characterization of this gene indicates that it is expressed at high levels in floral tissue and is wound inducible but, unlike allene oxide synthase, it is not induced by treatment with methyl jasmonate.

1   This work was supported by the Natural Sciences and Engineering Research Council of Canada.
2   These authors contributed equally to this publication.
3   Present address: Molecular Genetics Section, Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, Saskatchewan, Canada S7N 0X2.
*   Corresponding author; e-mail baten{at}em.agr.ca; fax 1-306-956-7247.

Plant Physiol. (1998) 117: 1393-1400
Copyright Clearance Center:   0032-0889/98/117//08
© 1998 American Society of Plant Physiologists




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