Using Different Forms of Nitrogen to Study Hypersensitive Response Elicited by Avirulent Pseudomonas syringae

  • Namrata Singh
  • Pooja Singh
  • Pradeep Kumar Pathak
  • Kapuganti Jagadis GuptaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2057)


Nitrate, ammonium, or a combination of both is the form of N available for nitrogen assimilation from soil by the plants. Nitrogen is an important and integral part of amino acids, nucleotides, and defense molecules. Hence it is very important to study the role of nitrate and ammonium nutrition in plant defense via hypersensitive response (HR). Shifting plants from ammonium nitrate Hoagland solution to nitrate Hoagland nutrition slightly enhances root length and leaf area. HR phenotype is different in nitrate and ammonium grown plants when challenged with avirulent Pseudomonas syringae DC3000 avrRpm1. HR is also associated with increased production of reactive oxygen species (ROS) and nitric oxide (NO). Hence to understand HR development it is essential to measure HR lesions, cell death, ROS, NO, and bacterial growth. Here we provide a stepwise protocol of various parameters to study HR in Arabidopsis in response to nitrate and ammonium nutrition.

Key words

Nitrate Ammonium Nitric oxide Reactive oxygen species Hypersensitive response Pseudomonas syringae 



NS is a recipient of JRF and SRF from CSIR, India. PS and PKP are recipients of UGC JRF and SRF. This work is supported by SERB Early Carrier Research Award to KJG by SERB, Govt. of India.


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

Authors and Affiliations

  • Namrata Singh
    • 1
  • Pooja Singh
    • 1
  • Pradeep Kumar Pathak
    • 1
  • Kapuganti Jagadis Gupta
    • 1
    Email author
  1. 1.National Institute of Plant Genome ResearchNew DelhiIndia

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