H2O2 amounts were deciphered as previously explained [147 (link)]. Freeze dried samples were crushed and, then, the powdered sample (0.3 g) was homogenized in ice bath with 5 mL 0.1% trichloroacetic acid (TCA). The homogenate was centrifuged at 12,000 rpm for 20 min. The supernatant (0.5 mL) was combined with 10 mM potassium phosphate buffer (0.5 mL, pH 7.0) and 1 M potassium iodide (1 mL). The mixture was stored at 25 °C for 20 min and its intensity was recorded at 390 nm using a spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA).
The thiobarbituric acid (TBA) test was used to estimate lipid peroxidation in leaves [148 (link)]. The reaction mixture comprised 0.5 mL of 0.1% TCA extract that was added to 1 mL of 0.5% TBA (prepared in 20% TCA). The mixture was incubated in boiling water (95 °C, 30 min), followed by cooling in ice bath (10 min). Afterwards, the mixture was centrifuged (12,000 rpm, 5 min) and the supernatant absorbance was read at 532 and 600 nm (T60 UV-Vis, PG Instruments Ltd., Wibtoft, UK).
Plant materials were analyzed for their fatty acids content with reference to a formerly published method [149 (link)]. Gas chromatography–mass spectrometry analysis was performed on an Agilent Model 7890A series (Agilent, Dover, DE, USA).
The thiobarbituric acid (TBA) test was used to estimate lipid peroxidation in leaves [148 (link)]. The reaction mixture comprised 0.5 mL of 0.1% TCA extract that was added to 1 mL of 0.5% TBA (prepared in 20% TCA). The mixture was incubated in boiling water (95 °C, 30 min), followed by cooling in ice bath (10 min). Afterwards, the mixture was centrifuged (12,000 rpm, 5 min) and the supernatant absorbance was read at 532 and 600 nm (T60 UV-Vis, PG Instruments Ltd., Wibtoft, UK).
Plant materials were analyzed for their fatty acids content with reference to a formerly published method [149 (link)]. Gas chromatography–mass spectrometry analysis was performed on an Agilent Model 7890A series (Agilent, Dover, DE, USA).
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