The extraction of hydrogen peroxide (H2O2), superoxide anion (O2−) and malondialdehyde (MDA) were carried out as described by Wang et al. [20 (link)]. Weigh 0.5 g leaf or 1.0 g root fresh sample, grind the fresh sample into homogenate on ice with extraction solution, the supernatant was extracted by centrifugation for the determination of above parameters and antioxidant enzyme activity.
The H2O2 content was measured by hydrogen peroxide detection kit (Beyotime Institute of Biotechnology, Shanghai, China).
The O2− release rate was assayed by hydroxylamine method [45 (link)]. After mixed reaction system, the mixture was kept at 25 °C for 20 min and the OD value was measured under OD530 nm of spectrophotometer.
The MDA content was determined according to Tan et al. [49 (link)]. In total, 4 mL TCA-TBA mixture solution was added with 2 mL extraction, boiling water bath for 20 min, centrifugation at 4000× g for 10 min, supernatant was taken and read at OD450 nm, OD532 nm and OD600 nm under a visible light spectrophotometer.
The H2O2 content was measured by hydrogen peroxide detection kit (Beyotime Institute of Biotechnology, Shanghai, China).
The O2− release rate was assayed by hydroxylamine method [45 (link)]. After mixed reaction system, the mixture was kept at 25 °C for 20 min and the OD value was measured under OD530 nm of spectrophotometer.
The MDA content was determined according to Tan et al. [49 (link)]. In total, 4 mL TCA-TBA mixture solution was added with 2 mL extraction, boiling water bath for 20 min, centrifugation at 4000× g for 10 min, supernatant was taken and read at OD450 nm, OD532 nm and OD600 nm under a visible light spectrophotometer.
Free full text: Click here
