Plant Physiol, January 2002, Vol. 128, pp. 150-159

Elemental Sulfur and Thiol Accumulation in Tomato and Defense against a Fungal Vascular Pathogen¹

Department of Biology and Biochemistry, University of Bath, Bath, BA2 7AY United Kingdom (J.S.W., S.A.H., R.M.C.); Agriculture and Environment Division, IACR-Rothamsted, Hertfordshire, AL5 2JQ United Kingdom (M.J.H.); and Department of Agricultural Sciences, IACR, Long Ashton Research Station, University of Bristol, Long Ashton, Bristol, BS41 9AF United Kingdom (M.H.B.)

The occurrence of fungicidal, elemental S is well documented in certain specialized prokaryotes, but has rarely been detected in eukaryotes. Elemental S was first identified in this laboratory as a novel phytoalexin in the xylem of resistant genotypes of Theobroma cacao, after infection by the vascular, fungal pathogen Verticillium dahliae. In the current work, this phenomenon is demonstrated in a resistant line of tomato, Lycopersicon esculentum, in response to V. dahliae. A novel gas chromatography-mass spectroscopy method using isotope dilution analysis with ³⁴S internal standard was developed to identify unambiguously and quantify ³²S in samples of excised xylem. Accumulation of S in vascular tissue was more rapid and much greater in the disease-resistant than in the disease-susceptible line. Levels of S detected in the resistant variety (approximately 10 µg g¹ fresh weight excised xylem) were fungitoxic to V. dahliae (spore germination was inhibited >90% at approximately 3 µg mL¹). Scanning electron microscopy-energy dispersive x-ray microanalysis confirmed accumulation of S in vascular but not in pith cells and in greater amounts and frequency in the Verticillium spp.-resistant genotype. More intensive localizations of S were occasionally detected in xylem parenchyma cells, vessel walls, vascular gels, and tyloses, structures in potential contact with and linked with defense to V. dahliae. Transient increases in concentrations of sulfate, glutathione, and Cys of vascular tissues from resistant but not susceptible lines after infection may indicate a perturbation of S metabolism induced by elemental S formation; this is discussed in terms of possible S biogenesis.