Determination of Oxidative Stress and Antioxidant Enzyme Activity for Physiological Phenotyping During Heavy Metal...

The overproduction of reactive oxygen species during abiotic stress in plants causes oxidative stress that damage the cell normal functions. For reactive oxygen species (ROS) scavenging, plants developed a defense system with antioxidant enzymes. To measure the oxidative stress and antioxidant enzymes activity spectral enzymatic analysis was used, that is material-intensive, time-consuming, and inefficient. In the present study, the four more studied and main antioxidant enzymatic assays are miniaturized in a 96-well plate system and monitored the activity of enzymes by spectrophotometry. This method has obvious advantages over the standard cuvette analysis method because the miniaturization of the 96-well microplate system decreases the amount of reaction mixture and enzymes extract, thrifts working time, and consumable costs as well.