Abstract
The use of plant protoplast transient expression system has facilitated the discovery and dissection of many signal transduction pathways in response to hormones, metabolites, and stresses. Recently, Arabidopsis protoplasts also have been used successfully to study plant innate immune responses triggered by pathogen-derived elicitors. Here, we describe the detailed protocols for studying innate immune responses, including cell death and early defense gene regulation activated by two types of elicitors, pathogen-associated molecular patterns and bacterial type III effectors in Arabidopsis protoplasts. This cellbased system simplifies the complex pathogen-plant interactions to pure individual signals and synchronized cell-autonomous responses. The application of this novel approach provides high temporal and spatial resolution to enhance our understanding of the distinct and overlapping signaling events in pathogen-associated molecular pattern- and bacterial type III effector-activated immune responses at the molecular and cellular level.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Gomez-Gomez, L. and Boller, T. (2000) FLS2: an LRR receptor-like kinase involved in the perception of the bacterial elicitor flagellin in Arabidopsis. Mol. Cell 5, 1003–1011.
Flor, H. H. (1971) Current status of the gene-for-gene concept. Annu. Rev. Phytopathol. 9, 275–296.
Dangl, J. L. and Jones, J. D. (2001) Plant pathogens and integrated defence responses to infection. Nature 411, 826–833.
Galan, J. E. and Collmer, A. (1999) Type III secretion machines: bacterial devices for protein delivery into host cells. Science 284, 1322–1328.
Martin, G. B., Bogdanove, A. J., and Sessa, G. (2003) Understanding the functions of plant disease resistance proteins. Annu. Rev. Plant Biol. 54, 23–61.
Belkhadir, Y., Subramaniam, R., and Dangl, J. L. (2004) Plant disease resistance protein signaling: NBS-LRR proteins and their partners. Curr. Opin. Plant Biol. 7, 391–399.
Glazebrook, J. (2001) Genes controlling expression of defense responses in Arabidopsis: 2001 status. Curr. Opin. Plant Biol. 4, 301–308.
McDowell, J. M. and Dangl, J. L. (2000) Signal transduction in the plant immune response. Trends Biochem Sci. 25, 79–82.
Espinosa, A. and Alfano, J. R. (2004) Disabling surveillance: bacterial type III secretion system effectors that suppress innate immunity. Cell. Microbiol. 6, 1027–1040.
Tao, Y., Xie, Z., Chen, W., et al. (2003) Quantitative nature of Arabidopsis responses during compatible and incompatible interactions with the bacterial pathogen Pseudomonas syringae. Plant Cell 15, 317–330.
Tena, G., Asai, T., Chiu, W.-L., and Sheen, J. (2001) Plant MAP kinase signaling cascades. Curr. Opin. Plant Biol. 4, 392–400.
Sheen, J. (2001) Signal transduction in maize and Arabidopsis mesophyll protoplasts. Plant Physiol. 127, 1466–1475.
Asai, T., Stone, J. M., Heard, J. E., et al. (2000) Fumonisin B1-induced cell death in Arabidopsis protoplasts requires jasmonate-, ethylene-, and salicylate-dependent signaling pathways. Plant Cell 12, 1823–1836.
Wu, Y., Wood, M. D., and Katagiri, F. (2003) Direct delivery of bacterial avirulence proteins into resistant Arabidopsis protoplasts leads to hypersensitive cell death. Plant J. 33, 131–137.
Asai, T., Tena, G., Plotnikova, J., et al. (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415, 977–983.
Kovtun, Y., Chiu, W.-L., Zeng, W., and Sheen, J. (1998) Suppression of auxin signal transduction by a MAPK cascade in higher plants. Nature 395, 716–720.
Yanagisawa, S., Yoo, S., and Sheen, J. (2003) Differential regulation of EIN3 stability by glucose and ethylene signalling in plants. Nature 425, 521–525.
Kovtun, Y., Chiu, W.-L., Tena, G., and Sheen, J. (2000) Functional analysis of oxidative stress-activated MAPK cascade in plants. Proc. Natl. Acad. Sci. USA 97, 2940–2945.
Felix, G., Duran, J., Volko, S., and Boller, T. (1999) Plants have a sensitive perception system for the most conserved domain of bacterial flagellin. Plant J. 18, 265–276.
Acknowledgments
This work was supported by the National Science Foundation and the National Institute of Health.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc.
About this protocol
Cite this protocol
He, P., Shan, L., Sheen, J. (2007). The Use of Protoplasts to Study Innate Immune Responses. In: Ronald, P.C. (eds) Plant-Pathogen Interactions. Methods in Molecular Biology, vol 354. Humana Press. https://doi.org/10.1385/1-59259-966-4:1
Download citation
DOI: https://doi.org/10.1385/1-59259-966-4:1
Publisher Name: Humana Press
Print ISBN: 978-1-58829-448-7
Online ISBN: 978-1-59259-966-0
eBook Packages: Springer Protocols