A cDNA clone of wheat oxalate oxidase (OxO) under the control of the constitutive CAMV 35S promotor was expressed in tomato plants by Agrobacterium -mediated transformation. Twenty-six transgenic tomato lines were obtained and analysed. PCR experiments confirmed the incorporation of the OxO gene in all tested tomato lines. The transgenic tomato plants expressed a 124-kDa protein showing OxO activity, and were able to convert different oxalic acid (OA) concentrations in vitro . In a detached leaf assay, most of the transgenic lines showed reduced disease symptoms compared with controls, following inoculation with Botrytis cinerea . In addition, leaves of the line T15 showed a marked reduction in symptoms compared with the control following inoculation with Sclerotinia sclerotiorum .

An effective colonization of the host plant tissue by the necrotrophic fungus Sclerotinia sclerotiorum requires the secretion of the non-host specific toxin oxalic acid (OA), which is known to suppress the generation of reactive oxygen intermediates (ROI). A full-length cDNA coding for an oxalate decarboxylase (TOXDC), which converts OA into CO 2 and formate, was isolated from the basidiomycete Trametes versicolor. It was overexpressed in tobacco plants to study the role of ROI and OA in the interaction between tobacco and S. sclerotiorum. The transgenic plants contained less OA and showed a delayed colonization of S. sclerotiorum; furthermore a strong ROI accumulation and nearly no catalase activity compared to the wild type (WT) plants could be detected. In addition, inoculation experiments with transgenic catalase-deficient plants (CAT1AS) and in vitro studies showed that S. sclerotiorum copes with strong ROI stress. Our results indicate that OA supports the infection process caused by S. sclerotiorum and the fungus itself is able to tolerate high ROI concentrations.

The non-protein amino acid b-aminobutyric acid (BABA) protects plants against a wide range of pathogens. Protection of cucumber plants by BABA depends on the potentiation of pathogen specific defence responses. To contribute to the analyses of the mode of action of BABA, we established a protocol for a fast and reproducible leaf disc assay to evaluate the effect of this chemical compound on cucumbers infected with either the biotrophic downy mildew fungus Pseudoperonospora cubensis, or the necrotrophic microbial pathogen Colletotrichum lagenarium. Accumulation of callose could be found in interactions with both pathogens after BABA-treatment. Furthermore, a localized rapid cell death and the production of reactive oxygen intermediates were detected after downy mildew attack. In contrast to this, degenerated primary hyphae were found in BABA induced tissue after inoculation with C. lagenarium.

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