Skip to main content

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

  • Protocol
  • First Online:
Nitrogen Metabolism in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2057))

  • 1206 Accesses

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Huber DM, Watson RD (1974) Nitrogen form and plant disease. Annu Rev Phytopathol 12:139–165

    Article  CAS  Google Scholar 

  2. Giles ME, Morley NJ, Baggs EM, Daniell TJ (2012) Soil nitrate reducing processes–drivers, mechanisms for spatial variation, and significance for nitrous oxide production. Front Microbiol 3:407

    Article  CAS  Google Scholar 

  3. Simek M, Cooper JE (2002) The influence of soil pH on denitrification: progress towards the understanding of this interaction over the last 50 years. Eur J Soil Sci 53:345–354

    Article  CAS  Google Scholar 

  4. Gebauer RL, Tenhunen JD, Reynolds JF (1996) Soil aeration in relation to soil physical properties, nitrogen availability, and root characteristics within an arctic watershed. Plant Soil 178:37–48

    Article  CAS  Google Scholar 

  5. Veresoglou SD, Voulgari OK, Sen R, Mamolos AP, Veresoglou DS (2011) Effects of nitrogen and phosphorus fertilization on soil pH-plant productivity relationships in upland grasslands of northern Greece. Pedosphere 21:750–752

    Article  CAS  Google Scholar 

  6. Lam HM, Coschigano KT, Oliveira IC, Melo-Oliveira R, Coruzzi GM (1996) The molecular-genetics of nitrogen assimilation into amino acids in higher plants. Annu Rev Plant Biol 47:569–593

    Article  CAS  Google Scholar 

  7. Wang P, Wang Z, Pan Q, Sun X, Chen H, Chen F, Mi G (2019) Growth of maize on mixed nitrate and ammonium promotes auxin synthesis and biomass accumulation. J Exp Bot. https://doi.org/10.1093/jxb/erz047

  8. Gupta KJ, Brotman Y, Segu S, Zeier T, Zeier J, Persijn ST, Cristescu SM, Harren FJM, Bauwe H, Fernie AR (2013) The form of nitrogen nutrition affects resistance against Pseudomonas syringae pv. phaseolicola in tobacco. J Exp Bot 64:553–568

    Article  CAS  Google Scholar 

  9. Snoeijers SS, Pérez-García A, Joosten MH, De Wit PJ (2000) The effect of nitrogen on disease development and gene expression in bacterial and fungal plant pathogens. Eur J Plant Pathol 106:493–506

    Article  CAS  Google Scholar 

  10. Wany A, Gupta KJ (2018) Reactive oxygen species, nitric oxide production and antioxidant gene expression during development of aerenchyma formation in wheat. Plant Signal Behav 13:3002–3017

    Article  Google Scholar 

  11. Lopez-Berges MS, Rispail N, Prados-Rosales RC, Di Pietro A (2010) A nitrogen response pathway regulates virulence in plant pathogenic fungi: role of TOR and the bZIP protein Mea B. Plant Signal Behav 5:1623–1625

    Article  CAS  Google Scholar 

  12. Gupta KJ, Fernie AR, Kaiser WM, van Dongen JT (2011) On the origins of nitric oxide. Trends Plant Sci 16:160–168

    Article  CAS  Google Scholar 

  13. Planchet E, Gupta KJ, Sonoda M, Kaiser WM (2005) Nitric oxide emission from tobacco leaves and cell suspensions: rate limiting factors and evidence for the involvement of mitochondrial electron transport. Plant J 41:732–743

    Article  CAS  Google Scholar 

  14. Podgorska A, Ostaszewska M, Gardeström P, Rasmusson AG, SZAL B (2015) In comparison with nitrate nutrition, ammonium nutrition increases growth of the frostbite1 Arabidopsis mutant. Plant Cell Environ 38:224–237

    Article  CAS  Google Scholar 

  15. Delledonne M, Xia Y, Dixon RA, Lamb C (1998) Nitric oxide functions as a signal in plant disease resistance. Nature 394:585

    Article  CAS  Google Scholar 

  16. Wany A, Gupta A, Kumari A, Mishra S, Singh N, Pandey S, Vanvari R, Igamberdiev A, Ferine A, Gupta KJ (2018) Nitrate nutrition influences multiple factors in order to increase energy efficiency under hypoxia in Arabidopsis. Ann Bot. https://doi.org/10.1093/aob/mcy202

Download references

Acknowledgments

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.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kapuganti Jagadis Gupta .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

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

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Singh, N., Singh, P., Pathak, P.K., Gupta, K.J. (2020). Using Different Forms of Nitrogen to Study Hypersensitive Response Elicited by Avirulent Pseudomonas syringae . In: Gupta, K. (eds) Nitrogen Metabolism in Plants. Methods in Molecular Biology, vol 2057. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9790-9_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9790-9_8

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9789-3

  • Online ISBN: 978-1-4939-9790-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics