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Localization of Nitric Oxide in Wheat Roots by DAF Fluorescence

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Plant Nitric Oxide

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1424))

Abstract

Nitric oxide is a free radical signal molecule. Various methods are available for measurement of NO. Out of all methods, fluorescent probes to localize NO is very widely used method. Diaminofluorescein in diacetate form (DAF-2DA) is most widely probe for NO measurement. This method is based on application of 4,5-diaminofluorescein diacetate (DAF-2DA) which is actively diffused into cells, once taken up by cells cytoplasmic esterases cleave the acetate groups to generate 4,5-diaminofluorescein; DAF-2. The generated DAF-2 can readily react with N2O3, which is an oxidation product of NO to generate the highly fluorescent DAF-2T (triazolofluorescein). There are various advantages and disadvantages associated with this method, but to its advantage in diffusion closely to NO producing sites, it is widely used for localization studies. Here, we describe method to make sections of the roots and localization of NO in roots subjected to hypoxic stress.

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Authors and Affiliations

  1. National Institute of Plant Genome Research, Aruna Asaf Ali Marg, 10531, New Delhi, 110067, India

    Aakanksha Wany & Kapuganti Jagadis Gupta

Authors
  1. Aakanksha Wany
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  2. Kapuganti Jagadis Gupta
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Correspondence to Kapuganti Jagadis Gupta .

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Editors and Affiliations

  1. National Institute of Plant Genome Resea, New Delhi, India

    Kapuganti Jagadis Gupta

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Wany, A., Gupta, K.J. (2016). Localization of Nitric Oxide in Wheat Roots by DAF Fluorescence. In: Gupta, K. (eds) Plant Nitric Oxide. Methods in Molecular Biology, vol 1424. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3600-7_4

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  • DOI: https://doi.org/10.1007/978-1-4939-3600-7_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3598-7

  • Online ISBN: 978-1-4939-3600-7

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