Abstract
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.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Pieterse CM, Leon-Reyes A, Van der Ent S, Van Wees SC (2009) Networking by small-molecule hormones in plant immunity. Nat Chem Biol 5:308–316
Robert-Seilaniantz A, Grant M, Jones JDG (2011) Hormone crosstalk in plant disease and defense: more than just salisylate-jasmonate antagonism. Annu Rev Phytopathol 49:317–343
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 5:1793–1814
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 66:4885–4896
Kazan K, Lyons R (2014) Intervention of phytohormone pathways by pathogen effectors. Plant Cell 26:1–26
Hwang I, Sheen J, Müller B (2012) Cytokinin signaling networks. Annu Rev Plant Biol 63:353–380
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 19:284–295
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 157:815–830
Naseem M, Wölfling M, Dandekar T (2014) Cytokinin for immunity beyond growth, galls and green-islands. Trends Plant Sci 19:481–484
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 8:2335–2351
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
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 302:1–12
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 112:41–55
Bonfig KB, Berger S, Fatima T, González MC, Roitsch T (2006) Metabolic control of seedling development by invertases. Funct Plant Biol 34:508–516
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 4:941–944
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 14:749–762
Acknowledgments
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Naseem, M., Shams, S., Roitsch, T. (2017). Modulating the Levels of Plant Hormone Cytokinins at the Host-Pathogen Interface. In: Dandekar, T., Naseem, M. (eds) Auxins and Cytokinins in Plant Biology. Methods in Molecular Biology, vol 1569. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6831-2_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6831-2_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6829-9
Online ISBN: 978-1-4939-6831-2
eBook Packages: Springer Protocols