Impact of brassinosteroids and ethylene on ascorbic acid accumulation in tomato leaves.

Plant steroid hormones brassinosteroids (BRs) and the gaseous hormone ethylene (ET) alter the ascorbic acid-glutathione (AA-GSH) levels in tomato (Solanum lycopersicum L.) plants. The interaction of these hormones in regulating antioxidant metabolism is however unknown. The combined use of genetics (BR-mutants) and chemical application (BR/ET-related chemicals) shows that BRs and ET signalling pathways interact, to regulate leaf AA content and synthesis. BR-deficient (d(x)) leaves display low total AA but BR-accumulating (35S:D) leaves show normal total AA content. Leaves with either BR levels lower or higher than wild type plants showed a higher oxidised AA redox state. The activity of L-galactono-1,4-lactone dehydrogenase (L-GalLDH), the mitochondrial enzyme that catalyses the last step in AA synthesis is lower in d(x) and higher in 35S:D plants. BR-deficient mutants show higher ET production but it is restored to normal levels when BR content is increased in 35S:D plants. Suppression of ET signalling using 1-methylcyclopropene in d(x) and 35S:D plants restored leaf AA content and L-GalLDH activity, to the values observed in wild type. The suppression of ET action in d(x) and 35S:D leaves leads to the respective decreasing and increasing respiration, indicating an opposite response compared to AA synthesis. This inverse relationship is lacking in ET suppressed d(x) plants in response to external BRs. The modifications in the in vivo activity of L-GalLDH activity do not correlate with changes in the level of the enzyme. Taken together, these data suggest that ET suppresses and BRs promote AA synthesis and accumulation.

Heat shock response in tomato brassinosteroid mutants indicates that thermotolerance is independent of brassinosteroid homeostasis.

Brassinosteroids (BRs) are plant steroid hormones and, when applied exogenously, they induce physiological responses, including tolerance to heat shock (HS). How endogenous BR content and altered perception of BRs influence thermal tolerance is poorly understood. BR-induced thermotolerance in tomato seedlings with altered BR homeostasis was examined by assessing the survival, ion leakage and lipid peroxidation of seedlings from a BR-deficient mutant (extreme dwarf d(x)), a partially BR-insensitive mutant curl3(-abs) allele (curl3 altered brassinolide sensitivity) and a line overexpressing the Dwarf, BR-biosynthesis gene (35SD). We confirmed that treatment with 1 µM of epi-brassinolide (EBL) induces thermotolerance of wild type seedlings following a HS regime at 45 °C. The curl3(-abs) seedlings had the highest basal tolerance to heat, whereas the EBL-induced thermal tolerance of d(x) seedlings was greatest and responded to lower EBL concentrations. The d(x) and 35SD seedlings had similar thermal tolerance; however, they showed increased signs of oxidative stress. EBL reduced the induction of lipid peroxidation of seedlings after recovery from heat. Highest oxidative stress and peroxidase (POX) activity (EC 1.11.1.7) was in BR-deficient d(x) mutant seedlings. EBL was able of inducing POX activity but not other antioxidant enzymes; however, effects of HS on POX activity of seedlings were absent or less marked. Taking together, results indicate that thermal tolerance is independent of endogenous BR content, but HS-mediated oxidative stress depends on BR levels.