Involvement of lipoxygenase-dependent production of fatty acid hydroperoxides in the development of the hypersensitive cell death induced by cryptogein on tobacco leaves.

Lipid peroxidation was investigated in relation with the hypersensitive reaction in cryptogein-elicited tobacco leaves. A massive production of free polyunsaturated fatty acid (PUFA) hydroperoxides dependent on a 9-lipoxygenase (LOX) activity was characterized during the development of leaf necrosis. The process occurred after a lag phase of 12 h, was accompanied by the concomitant increase of 9-LOX activity, and preceded by a transient accumulation of LOX transcripts. Free radical-mediated lipid peroxidation represented 10% of the process. Inhibition and activation of the LOX pathway was shown to inhibit or to activate cell death, and evidence was provided that fatty acid hydroperoxides are able to mimic leaf necrotic symptoms. Within 24 h, about 50% of leaf PUFAs were consumed, chloroplast lipids being the major source of PUFAs. The results minimize the direct participation of active oxygen species from the oxidative burst in membrane lipid peroxidation. They suggest, furthermore, the involvement of lipase activity to provide the free PUFA substrates for LOX. The LOX-dependent peroxidative pathway, responsible for tissue necrosis, appears as being one of the features of hypersensitive programmed cell death.

ESR spin trapping analysis of gamma induced radicals in sucrose: II.

Radicals induced by gamma-irradiation of sucrose, in the solid state at different temperatures and in aqueous solution, have been investigated by the spin trapping method. Electron spin resonance (ESR) combined with high performance liquid chromatography (HPLC), followed by spectral analysis with a simulation program (Voyons) revealed seven main radical species. A comparative study of the ESR signals from spin trapped gamma-induced radicals in some glycosides, disaccharides, 13C specifically labelled carbohydrates, as well as in several deoxysucroses and fructans, led to the assignment of a chemical structure to five out of the seven sucrose-nitroxide adducts previously evidenced. Sucrose is shown to be a conceivable model for the study of fructans gamma-radiolysis mechanism in aqueous solution.

E.S.R. spin trapping analysis of gamma induced radicals in sucrose.

Radicals induced by gamma-irradiation in sucrose, in the solid state at different temperatures and in aqueous solution, have been studied by the spin-trapping method. Electron spin resonance (ESR) combined with high performance liquid chromatography (HPLC) then spectra analysis with a simulation program (VOYONS), revealed seven main radical species. Their nitrogen and hydrogen splitting constants were compared with those obtained from fructose and glucose units. Assignments of chemical structures are discussed for three radical species.