Quantitative Measurements of Biochemical and Molecular Markers of Oxidative Stress Signaling and Responses

Abstract

Increases in cellular oxidation are a part of most plant responses to challenging conditions and are commonly described as oxidative stress. While this phenomenon is closely related to the accumulation of reactive oxygen species, these latter compounds can be difficult to measure. Complementary measurements to assess cellular redox state are, therefore, very useful in studies of plant responses to stress. Here, we detail protocols for three complementary approaches that can be used to assess the intensity of oxidative stress. These involve quantification of marker transcripts, assays of the extractable activities of major antioxidative enzymes, and measurement of antioxidant buffers. We confirm experimentally that the data obtained by such approaches can provide reliable information on the intensity of oxidative stress.

Keywords: Antioxidants; Ascorbate; Ascorbate peroxidase; Catalase; Dehydroascorbate reductase; Glutathione; Glutathione reductase; Marker transcript; Redox state; UDP glycosyl transferase.