Chinese Bulletin of Botany >

2022 , Vol. 57 >Issue 3: 320 - 326

DOI: https://doi.org/10.11983/CBB22043

TECHNIQUES AND METHODS

Detection of Reactive Oxygen Species Using H2DCFDA Probe in Plant

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  • College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China

Received date: 2022-03-10

  Accepted date: 2022-05-11

  Online published: 2022-05-11

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Abstract

Reactive oxygen species (ROS) are a ‘double-edged sword’ in plants. On the one hand, ROS, as a signal molecule, plays pivotal roles in many aspects of life activities; on the other hand, excessive accumulation of ROS can cause oxidative damage to biological macromolecules. Accurate detection of ROS is essential to assess its intracellular redox status. Due to the characteristics of short half-life and strong reactivity of ROS components, their qualitative and quantitative analysis are difficult. It is critical to select the appropriate detection method and improve the spatiotemporal accuracy of detection for research in plant sciences and in other fields. At present, fluorescent probe analysis has attracted the attention of researchers because of its advantages of high sensitivity, good selectivity, low detection limit and strong intuition. This article introduces the detailed operation protocol and attentions for ROS detection using 2′,7′-dichlorodi-hydrofluorescein diacetate (H2DCFDA) fluorescent probe based on flow cytometry and confocal microscope. These methods can be used to detect ROS levels and distribution in model plant tissues, including Oryza sativa, Arabidopsis thaliana, Zea mays and Glycine max.

Key words： reactive oxygen species; fluorescent probe; 2',7'-dichlorodi-hydrofluorescein diacetate; plant

Cite this article

Haitao Hu, Tingting Qian, Ling Yang . Detection of Reactive Oxygen Species Using H2DCFDA Probe in Plant[J]. Chinese Bulletin of Botany, 2022 , 57(3) : 320 -326 . DOI: 10.11983/CBB22043

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