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A simple colorimetric method for determination of hydrogen peroxide in plant tissues

Plant Growth Regulation volume 49, pages 113–118 (2006)Cite this article

Abstract

A simple colorimetric method for determination of hydrogen peroxide in plant materials is described. The method is based on hydrogen peroxide producing a stable red product in reaction with 4-aminoantipyrine and phenol in the presence of peroxidase. Plant tissues was ground with trichloroacetic acid (5% w/v) and extracts were adjusted to pH 8.4 with ammonia solution. Activated charcoal was added to the homogenate to remove pigments, antioxidants and other interfering substances. The colorimetric reagent (pH 5.6) consisted of 4-aminoantipyrine, phenol, and peroxidase. With this method, we have determined the hydrogen peroxide concentration in leaves of eight species which ranged from 0.2 to 0.8 µmol g−1 FW. Changes in hydrogen peroxide concentration of Stylosanthes guianensis in response to heat stress are also analyzed using this method.

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Acknowledgements

The project was funded by grants from Guangdong Science and Technology Projects (2003C201018), Guangdong Provincial Natural Science Foundation (04105978, 5300846), and National Natural Science Foundation (30571283).

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Authors and Affiliations

  1. College of Life Science, South China Agricultural University, Guangzhou, 510642, China

    Biyan Zhou, Jihua Wang, Zhenfei Guo & Huoquan Tan

  2. Physiological Laboratory for South China Fruits, College of Horticulture, South China Agricultural University, Guangzhou, 510642, China

    Biyan Zhou & Xiaochuan Zhu

Authors
  1. Biyan Zhou
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  2. Jihua Wang
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  3. Zhenfei Guo
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  4. Huoquan Tan
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  5. Xiaochuan Zhu
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Correspondence to Zhenfei Guo.

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Zhou, B., Wang, J., Guo, Z. et al. A simple colorimetric method for determination of hydrogen peroxide in plant tissues. Plant Growth Regul 49, 113–118 (2006). https://doi.org/10.1007/s10725-006-9000-2

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  • DOI: https://doi.org/10.1007/s10725-006-9000-2

Keywords

  • 4-aminoantipyrine
  • Hydrogen peroxide
  • Peroxidase
  • Plant tissue