Determining the ROS and the Antioxidant Status of Leaves During Cold Acclimation

  • Andras Bittner
  • Thomas Griebel
  • Jörn van Buer
  • Ilona Juszczak-Debosz
  • Margarete BaierEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2156)


Cold slows down Calvin cycle activity stronger than photosynthetic electron transport, which supports production of reactive oxygen species (ROS). Even under extreme temperature conditions, most ROS are detoxified by the combined action of low-molecular weight antioxidants and antioxidant enzymes. Subsequent regeneration of the low-molecular weight antioxidants by NAD(P)H and thioredoxin/thiol–dependent pathways relaxes the electron pressure in the photosynthetic electron transport chain. In general, the chloroplast antioxidant system protects plants from severe damage of enzymes, metabolites, and cellular structures by both ROS detoxification and antioxidant recycling. Various methods have been developed to quantify ROS and antioxidant levels in photosynthetic tissues. Here, we summarize a series of exceptionally fast and easily applicable methods that show local ROS accumulation and provide information on the overall availability of reducing sugars, mainly ascorbate, and of thiols.

Key words

ROS H2O2 Superoxide Thiols Reducing sugars Ascorbate Digital imaging Quantification 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Andras Bittner
    • 1
  • Thomas Griebel
    • 1
  • Jörn van Buer
    • 1
  • Ilona Juszczak-Debosz
    • 1
    • 2
  • Margarete Baier
    • 1
    Email author
  1. 1.Plant Physiology, Dahlem Center of Plant SciencesFreie Universität BerlinBerlinGermany
  2. 2.Molecular PhysiologyRheinische Friedrich-Wilhelms-Universität BonnBonnGermany

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