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Published on May 5, 2006; 10.1104/pp.106.081679

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Received April 7, 2006
Returned for revision April 26, 2006
Accepted April 26, 2006

A NOVEL LIPOXYGENASE IN PEA ROOTS: ITS FUNCTION IN WOUNDING AND BIOTIC STRESS

Pasqua Veronico *, Donato Giannino , M. Teresa Melillo , Antonella Leone , Aurelio Reyes , Malcolm W. Kennedy , and Teresa Bleve-Zacheo

Institute of Plant Protection-Bari Section, CNR, Via Amendola 165/A, 70126 Bari, Italy
Institute of Biology and Agricultural Biotechnology-Rome Section, CNR, Via Salaria km 29,300. Monterotondo Scalo, Rome, Italy
Institute of Science of Food Production-Lecce Section, CNR, Via Monteroni, 73100 Lecce, Italy
Dunn Human Nutrition Unit, Medical Research Council (MRC), Wellcome Trust/MRC Building, Hills Road, CB2 2XY Cambridge, UK
Division of Environmental and Evolutionary Biology, Institute of Biomedical and Life Sciences, Graham Kerr Building, University of Glasgow, Glasgow G12 8QQ, UK

The genome of the pea (Pisum sativum L.) contains genes encoding a family of distinct lipoxygenases (LOX). Among these, LOXN2 showed eight exons encoding a 93.7 kDa enzyme, harboring two C-terminal deletions and an unusual Arg/Thr-Tyr motif in the domain considered to control the substrate specificity. LOXN2, when over-expressed in yeast, exhibited normal enzyme activity with an optimum at pH 4.5, and a dual positional specificity by releasing a 3:1 ratio of C-9 and C-13 oxidized products. The predicted LOXN2 structure lacked a loop present in soybean LOX-1, in a position consistent with control of both the degree of substrate access to the catalytic site, and for LOXN2's dual positional specificity. The LOXN2 gene resulted to be tightly conserved in the Progress 9 and MG103738 genotypes, respectively susceptible and resistant to the root cyst nematode Heterodera goettingiana. LOXN2 transcription was monitored in roots after mechanical injury and during nematode infection. The message peaked at 3 and 24 hr after wounding in both genotypes and was more abundant in the resistant than in the susceptible pea. In nematode-infected roots, transcription of several LOX genes was triggered, but LOXN2 which was repressed in both genotypes. In situ hybridization revealed that LOXN2 message was widespread in the cortex and endodermis of healthy roots, but specifically localized at high level in the cells bordering the nematode-induced syncytia of infected roots. However, LOXN2 transcript signal was particularly intense in collapsing syncytia of MG103738 roots, suggesting LOXN2 involvement in late mechanisms of host resistance.




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