• Here, the interaction of Melodoigyne incognita virulent and avirulent pathotypes with susceptible and Mi-resistant tomato (Solanum lycopersicon) has been studied. Significant differences in nematode penetration occurred 2 days postinoculation (dpi) and became stable from 3 dpi onwards. The hypersensitive cell response (HR) in resistant plants prevented the installation of the avirulent pathotype. The virulent pathotype overcame the Mi (nematode) resistance and induced feeding sites in root cells without triggering HR. • Reactive oxygen species (ROS), visualized by subcellular reduction of nitroblue tetrazolium, accumulated in nematode penetrated cells. Quantitative analyses with dichlorofluorescein indicated that the oxidative burst occurred very early with both pathotypes, with an enhanced rate in hyper-responsive cells. • Hydrogen peroxide (H₂O₂), detected by cerium chloride reaction, accumulated in the cell walls and especially in cells neighbouring HR. The apoplastic location of cerium perhydroxide indicated that either the plasma membrane or the cell wall was the primary site of the superoxide/H₂O₂ generator. • The data provide evidence, for the first time, for ROS-generated signals and their spatiotemporal expression in the host and nonhost interaction of tomato with nematodes.

The genome of pea (Pisum sativum) contains genes encoding a family of distinct lipoxygenases (LOX). Among these, LOXN2 showed eight exons encoding a 93.7-kD enzyme, harboring two C-terminal deletions and an unusual arginine/threonine-tyrosine motif in the domain considered to control the substrate specificity. LOXN2, when overexpressed 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 (Glycine max) LOX1, in a position consistent with control of the degree of substrate access to the catalytic site and for LOXN2's dual positional specificity. The LOXN2 gene was 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 h 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 except 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.

The response of resistant wheat–Aegilops ventricosa introgression line H-93-8 and its susceptible parent, Triticum aestivum H-10-15, to Ha71 Spanish population of Heterodera avenae was studied to determine the changes in peroxidase gene expression during incompatible and compatible wheat–nematode interactions. Twenty peroxidase genes were characterized from both 211 expressed sequence tags and 259 genomic DNA clones. Alignment of deduced amino acid sequences and phylogenetic clustering with peroxidases from other plant species showed that these enzymes fall into seven different groups (designated TaPrx108 to TaPrx114) which represent peroxidases secreted to the apoplast by a putative N-terminal peptide signal. TaPrx111, TaPrx112, and TaPrx113 were induced by nematode infection in both genotypes but with differing magnitude and timing. TaPrx112 and TaPrx113 groups increased more in resistant than in susceptible infected lines. In addition, in situ hybridization analyses of genes belonging to TaPrx111, TaPrx112, and TaPrx113 groups revealed a more intense signal in cells close to the vascular cylinder and parenchyma vascular cells of resistant than susceptible wheat when challenged by nematodes. These data seem to suggest that wheat apoplastic peroxidases, because of their different expression in quantity and timing, play different roles in the plant response to nematode infection.