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.
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Acknowledgments
Mathias Cohen thanks IDEX Paris-Saclay, France, and Ghent University, Belgium, for PhD funding. Lug Trémulot is supported by a PhD grant from the Université Paris Saclay and MESRI (Ministère de l’Enseignement Supérieur, de la Recherche, et de l’Innovation), France. Work in Graham Noctor’s laboratory is supported by the French Agence Nationale de la Recherche HIPATH project (ANR-17-CE20-0025) and by the Institut Universitaire de France (IUF). This work was supported by the FWO/FNRS Excellence of Science Research Project 30829584 award to Frank Van Breusegem.
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1 Electronic Supplementary Material
Electronic Supplemental Table 1
Model worksheet for converting absorbance changes to ascorbate and glutathione contents and redox states. As an example, actual data obtained for leaf extracts of two Arabidopsis genotypes are shown. These are the wild-type, Col-0, and the catalase-deficient cat2 mutant, which accumulates glutathione disulfide (GSSG) (XLS 55 kb)
Electronic Supplemental Table 2
Model worksheet for converting absorbance changes to activities for four antioxidative enzymes. As an example, actual data obtained for leaf extracts of two Arabidopsis genotypes are shown. These are the wild-type, Col-0, and the catalase-deficient cat2 mutant. Both were grown for 3 weeks in long days at 200 μmol/m2/s (XLS 51 kb)
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Noctor, G., Cohen, M., Trémulot, L., Van Breusegem, F., Mhamdi, A. (2023). Quantitative Measurements of Biochemical and Molecular Markers of Oxidative Stress Signaling and Responses. In: Couée, I. (eds) Plant Abiotic Stress Signaling. Methods in Molecular Biology, vol 2642. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3044-0_11
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DOI: https://doi.org/10.1007/978-1-0716-3044-0_11
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