Methods for Measuring Nitrate Reductase, Nitrite Levels, and Nitric Oxide from Plant Tissues

  • Aakanksha WanyEmail author
  • Pradeep Kumar Pathak
  • Kapuganti Jagadis Gupta
Part of the Methods in Molecular Biology book series (MIMB, volume 2057)


Nitrogen (N) is one of the most important nutrients which exist in both inorganic and organic forms. Plants assimilate inorganic form of N [nitrate (NO3), nitrite (NO2) or ammonium (NH4+)] and incorporate into amino acids. The metabolism of N involves a series of events such as sensing, uptake, and assimilation. The initial stage is sensing, triggered by nitrate or ammonium signals initiating signal transduction processes in N metabolism. The assimilation pathway initiates with NO3/NH4+ transport to roots via specific high and low affinity (HATs and LATs) nitrate transporters or directly via ammonium transporters (AMTs). In cytosol the NO3 is reduced to NO2 by cytosolic nitrate reductase (NR) and the produced NO2 is further reduced to NH4+ by nitrite reductase (NiR) in plastids. NR has capability to reduce NO2 to nitric oxide (NO) under specific conditions such as hypoxia, low pH, and pathogen infection. The produced NO acts as a signal for wide range of processes such as plant growth development and stress. Here, we provide methods to measure NR activity, NO2 levels, and NO production in plant tissues.

Key words

DAF-FM Nitrate Nitrate reductase Nitrite Nitric oxide 


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Copyright information

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

Authors and Affiliations

  • Aakanksha Wany
    • 1
    Email author
  • Pradeep Kumar Pathak
    • 1
  • Kapuganti Jagadis Gupta
    • 1
  1. 1.National Institute of Plant Genome ResearchNew DelhiIndia

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