Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Identification of the lipoxygenase gene family from Vitis vinifera and biochemical characterisation of two 13-lipoxygenases expressed in grape berries of Sauvignon Blanc

Andriy Podolyan A , Jackie White A , Brian Jordan A and Chris Winefield A B

A Department of Wine, Food and Molecular Biosciences, Faculty of Agriculture and Life Sciences, Lincoln University, PO Box 84, Lincoln 7647, Canterbury, New Zealand.

B Corresponding author. Email: chris.winefield@lincoln.ac.nz

Functional Plant Biology 37(8) 767-784 https://doi.org/10.1071/FP09271
Submitted: 9 November 2009  Accepted: 12 April 2010   Published: 26 July 2010

Abstract

Lipoxygenases (LOXs) are a group of non-haem iron-containing dioxygenases that catalyse oxygenation of polyunsaturated fatty acids (PUFAs) and lipids, and initiate the formation of biologically active compounds known as oxylipins. Several plant oxylipins comprise important flavours and aromas in food and beverages. Analysis of the grape (Vitis vinifera L.) genome revealed that the grape LOX family consists of 18 individual members. Phylogenetic analysis places all except one of the identified grape LOXs into either a type II 13-LOX cluster or the type I 9-LOX cluster. Four LOX genes (VvLOXA, VvLOXO, VvLOXC, VvLOXD), representative of the major LOX groupings observed in the phylogenetic analyses, were selected for analysis of patterns of transcript abundance in berry tissues. VvLOXA and VvLOXO represent putative 13-LOXs, while VvLOXC represents a putative 9-LOX. VvLOXD represents a unique LOX that differs significantly from other characterised plant LOXs. All four LOXs exhibited a complex pattern of gene expression. Across all developmental stages, VvLOXA was the most abundant LOX and was expressed predominantly in berry skins. The expression pattern of VvLOXC and -D are more evenly distributed between seeds, pulp and skin, while VvLOXO is mostly expressed in the seed. Mechanical wounding and infection of berries with Botrytis cinerea Pers.: Fr resulted in rapid accumulation of VvLOXC and -O transcripts. VvLOXA expression decreased in diseased berries. Biochemical analysis of VvLOXA and -O recombinant proteins confirmed that these LOX genes encode functional 13-LOXs that exhibit different pH and temperature optima. Both enzymes showed activity with linoleic, linolenic and arachidonic acids.

Additional keywords: berry development, grapevine, green leaf volatiles, LOX, oxylipins, plant–pathogen interaction.


Acknowledgements

The authors thank Dr Mike Trought for his invaluable intellectual input into the viticultural aspects of this study. We also thank Pernod Ricard, New Zealand, for their generous use of commercial vines within the Booker vineyard on the Brancott estate for our field materials, and Marlborough Wine Research staff for help in sample collecting. This work was funded by the New Zealand Foundation for Research, Science and Technology, contract number UOAX0404.


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