Modulating the Levels of Plant Hormone Cytokinins at the Host-Pathogen Interface

  • Muhammad NaseemEmail author
  • Shabana Shams
  • Thomas Roitsch
Part of the Methods in Molecular Biology book series (MIMB, volume 1569)


Cytokinins are adenine and non-adenine derived heterogeneous class of regulatory molecules that participate in almost every aspect of plant biology. They also affect plant defense responses as well as help microbial pathogens to establish pathogenesis. The functional approaches that ensure desired and subtle modulations in the levels of plant cytokinins are highly instrumental in assessing their functions in plant immunity. Here, we describe a detailed working protocol regarding the enhanced production of cytokinins from plants that harbor isopentenyltransferase (IPT) enzyme gene under the control of 4xJERE (jasmonic acid and elicitor-responsive element) pathogen-inducible promoter. Our devised expression system is a context-dependent solution when it comes to investigating host-pathogen interactions under the modulated conditions of plant cytokinins.

Key words

Cytokinins Pathogen-inducible promoter Host-pathogen interaction Transient plant transformation 



We are thankful to Anwar Hussain and Markus Kirschke for their help in analyzing tobacco samples for cytokinin determination. We appreciate DAAD (Deutscher Akademischer Austauschdienst) for providing doctoral fellowship to MN. This work was supported by funding from the Czech Ministry of Education, Youth and Sports of CR within the National Sustainability Program I (NPU I), grant number LO1415, to T.R.


  1. 1.
    Pieterse CM, Leon-Reyes A, Van der Ent S, Van Wees SC (2009) Networking by small-molecule hormones in plant immunity. Nat Chem Biol 5:308–316CrossRefPubMedGoogle Scholar
  2. 2.
    Robert-Seilaniantz A, Grant M, Jones JDG (2011) Hormone crosstalk in plant disease and defense: more than just salisylate-jasmonate antagonism. Annu Rev Phytopathol 49:317–343CrossRefPubMedGoogle Scholar
  3. 3.
    Naseem M, Philippi N, Hussain A, Wangorsch G, Ahmed N, Dandekar T (2012) Integrated systems view on networking by hormones in Arabidopsis immunity reveals multiple crosstalk for cytokinins. Plant Cell 5:1793–1814CrossRefGoogle Scholar
  4. 4.
    Naseem M, Kaltdorf M, Dandekar T (2015) The nexus between growth and defence signalling: auxin and cytokinin modulate plant immune response pathways. J Exp Bot 66:4885–4896CrossRefPubMedGoogle Scholar
  5. 5.
    Kazan K, Lyons R (2014) Intervention of phytohormone pathways by pathogen effectors. Plant Cell 26:1–26CrossRefGoogle Scholar
  6. 6.
    Hwang I, Sheen J, Müller B (2012) Cytokinin signaling networks. Annu Rev Plant Biol 63:353–380CrossRefPubMedGoogle Scholar
  7. 7.
    Choi J, Huh SN, Kojima M, Sakakibara H, Paek KH, Hwang I (2010) The cytokinin-activated transcription factor ARR2 promotes plant immunity via TGA3/NPR-1-dependent salicylic acid signalling in Arabidopsis. Dev Cell 19:284–295CrossRefPubMedGoogle Scholar
  8. 8.
    Grosskinsky DK, Naseem M, Abdelmohsen UR, Plickert N, Engelke T, Griebel T, Zeier J, Novák O, Strnad M, Pfeifhofer H, van der Graaff E, Simon U, Roitsch T (2011) Cytokinins mediate resistance against Pseudomonas syringae in tobacco through increased antimicrobial phytoalexin synthesis independent of salicylic acid signalling. Plant Physiol 157:815–830CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Naseem M, Wölfling M, Dandekar T (2014) Cytokinin for immunity beyond growth, galls and green-islands. Trends Plant Sci 19:481–484CrossRefPubMedGoogle Scholar
  10. 10.
    Hinsch J, Vrabka J, Oeser B, Novák O, Galuszka P, Tudzynski P (2015) De novo biosynthesis of cytokinins in the biotrophic fungus Claviceps purpurea. Environ Microbiol 8:2335–2351Google Scholar
  11. 11.
    Radhika V, Ueda N, Tsuboi Y, Kojima M, Kikuchi J, Kudo T, Sakakibara H (2015) Methylated cytokinins from the phytopathogen Rhodococcus fascians mimic plant hormone activity. Plant Physiol. doi: 10.1104/pp.15.00787 PubMedPubMedCentralGoogle Scholar
  12. 12.
    Sardesai N, Lee LY, Chen H, Yi H, Olbricht GR, Stirnberg A, Jeffries J, Xiong K, Doerge RW, Gelvin SB (2013) Cytokinins secreted by Agrobacterium promote transformation by repressing a plant myb transcription factor. Sci Signal 302:1–12Google Scholar
  13. 13.
    Novák J, Pavlů J, Novák O, Nožková-Hlaváčková V, Špundová M, Hlavinka J, Koukalová Š, Skalák J, Černý M, Brzobohatý B (2013) High cytokinin levels induce a hypersensitive-like response in tobacco. Ann Bot 112:41–55CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Bonfig KB, Berger S, Fatima T, González MC, Roitsch T (2006) Metabolic control of seedling development by invertases. Funct Plant Biol 34:508–516CrossRefGoogle Scholar
  15. 15.
    Patro S, Kumar D, Ranjan R, Maiti IB, Dey N (2012) The development of efficient plant promoters for transgene expression employing plant virus promoters. Mol Plant 4:941–944CrossRefGoogle Scholar
  16. 16.
    Rushton PJ, Reinstädler A, ReinLipka V, Lippok B, Somssich IE (2002) Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen- and wound-induced signaling. Plant Cell 14:749–762CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Muhammad Naseem
    • 1
    • 2
    Email author
  • Shabana Shams
    • 3
  • Thomas Roitsch
    • 4
    • 5
  1. 1.Deptartment of Bioinformatics, Biocenter University of Würzburg Am Hubland Würzburg GermanyJulius-Maximilians-Universität WürzburgWürzburgGermany
  2. 2.Department of Molecular Biology and GeneticsBoğaziçi UniversityBebekTurkey
  3. 3.Department of Animal SciencesQuaid-i-Azam UniversityIslamabadPakistan
  4. 4.Department of Plant and Environmental Sciences, Copenhagen Plant Science CentreUniversity of CopenhagenTaastrupDenmark
  5. 5.Global Change Research CentreCzech Globe AS CRDrásovCzech Republic

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