The procedure of Velikova, et al.30 (link) was used to estimate H2O2 content using a standard curve of known concentrations and expressed as µg g−1 FW.
MDA content was determined according to the methods described by Heath and Packer31 (link). One milliliter of extract was added to 2 mL of a reaction solution containing 20% (v/v) trichloroacetic acid and 0.5% (v/v) thiobarbituric acid. The solution was placed in a water bath at 95 °C for 30 minutes before transferring to an ice water bath. The solution was centrifuged at 10,000 rpm for 10 minutes and the absorbance of the supernatant recorded at 532 and 600 nm.
The method of Lutts32 (link) was used to estimate electrolyte leakage (EL). Electrical conductivity (L1) was recorded with a conductivity meter (PCS Testr35). The samples were then autoclaved at 120 °C for 20 minutes and electrical conductivity recorded (L2) after equilibration at 25 °C.
The method of Yang33 (link) was used to determine cell viability in plant roots. Cell viability assays can be performed with fluorescein diacetate (FDA). Fluorescence was measured at 488–494 nm using a microscope (Nikon A1R Confocal Laser Scanning Microscope, Japan).
For the cell non-viability assay, the method of Truernit and Haseloff34 (link) was used. Fluorescence was recorded at 535–617 nm using a confocal microscope (Nikon A1R Confocal Laser Scanning Microscope, Japan).
MDA content was determined according to the methods described by Heath and Packer31 (link). One milliliter of extract was added to 2 mL of a reaction solution containing 20% (v/v) trichloroacetic acid and 0.5% (v/v) thiobarbituric acid. The solution was placed in a water bath at 95 °C for 30 minutes before transferring to an ice water bath. The solution was centrifuged at 10,000 rpm for 10 minutes and the absorbance of the supernatant recorded at 532 and 600 nm.
The method of Lutts32 (link) was used to estimate electrolyte leakage (EL). Electrical conductivity (L1) was recorded with a conductivity meter (PCS Testr35). The samples were then autoclaved at 120 °C for 20 minutes and electrical conductivity recorded (L2) after equilibration at 25 °C.
The method of Yang33 (link) was used to determine cell viability in plant roots. Cell viability assays can be performed with fluorescein diacetate (FDA). Fluorescence was measured at 488–494 nm using a microscope (Nikon A1R Confocal Laser Scanning Microscope, Japan).
For the cell non-viability assay, the method of Truernit and Haseloff34 (link) was used. Fluorescence was recorded at 535–617 nm using a confocal microscope (Nikon A1R Confocal Laser Scanning Microscope, Japan).
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