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. 2020 Oct 1;10:16231. doi: 10.1038/s41598-020-72464-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Biochemical characterization of AREB1-OX plants under drought. (A) Chlorophyll levels. The levels of chlorophyll A, B, and total chlorophyll were determined in ethanolic extracts during water deprivation by spectrophotometry at 645 nm and 663 nm. Total chlorophyll was expressed as µg/mg of tissue. (B) TBA-reactive compounds. In several stresses, plant cells produce ROS. During drought, the collapse of the photosynthetic apparatus enhances the effects of tissue oxidation, culminating in higher levels of malondialdehyde, a TBA-reactive compound produced during lipid peroxidation. (C) DAB-mediated hydrogen peroxide detection in leaves. Stressed leaves were stained with DAB and distained in an ethanol bath for H₂O₂ detection. (D) Soluble sugar quantification. The soluble sugars were spectrophotometrically quantified in ethanolic extracts by the DNS method using a glucose-based standard curve. Biochemical analyses were conducted with 3 biological and 2 technical replicates. The bars indicate 5% standard error, and asterisks indicate t-test statistical significance under 95% confidence.