Abstract
Mitogen-Activated Protein Kinase (MAPK) cascade is one of the main signaling components mediating abiotic and biotic stress and hormone information in plants. Plant MAPK study has been impeded with a genetic approach using a long-term phenotypic analysis in spite of the transient nature of the protein kinase signaling. Arabidopsis leaf mesophyll protoplasts provide a versatile resource for diverse cell-based assays to acquire immediate molecular and biochemical responses with transient expression of MAPK cascade components of interests. Thus, it is an attractive tool for a high-throughput functional analysis of Arabidopsis MAPK cascade signaling. However, transient expression in Arabidopsis mesophyll protoplast (TEAMP) system requires mastered skills for protoplast preparation and handling to achieve steady and stable data. Here, we have described two analytical methods for MAPK cascade signaling using TEAMP system.
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References
Doczi R, Okresz L, Romero AE, Paccanaro A, Bogre L (2012) Exploring the evolutionary path of plant MAPK networks. Trends Plant Sci 17:518–525
Smékalová V, Doskočilová A, Komis G, Šamaj J (2013) Crosstalk between secondary messengers, hormones and MAPK modules during abiotic stress signalling in plants. Biotechnol Adv 32(1):2–11
Andreasson E, Ellis B (2010) Convergence and specificity in the Arabidopsis MAPK nexus. Trends Plant Sci 15:106–113
Kiegerl S et al (2000) SIMKK, a mitogen-activated protein kinase (MAPK) kinase, is a specific activator of the salt stress-induced MAPK, SIMK. Plant Cell 12:2247–2258
Junttila MR, Li SP, Westermarck J (2008) Phosphatase-mediated crosstalk between MAPK signalling pathways in the regulation of cell survival. FASEB J 22:954–965
Qi MS, Elion EA (2005) MAP kinase pathways. J Cell Sci 118:3569–3572
MAPK Group (2002) Mitogen-activated protein kinase cascades in plants: a new nomenclature. Trends Plant Sci 7:301–308
Tena G, Asai T, Chiu WL, Sheen J (2001) Plant mitogen-activated protein kinase signaling cascades. Curr Opin Plant Biol 4:392–400
Pitzschke A, Schikora A, Hirt H (2009) MAPK cascade signalling networks in plant defence. Curr Opin Plant Biol 12:421–426
Jonak C, Okresz L, Bogre L, Hirt H (2002) Complexity, cross talk and integration of plant MAP kinase signalling. Curr Opin Plant Biol 5:415–424
Mazarei M, Al-Ahmad H, Rudis MR, Stewart CN Jr (2008) Protoplast isolation and transient gene expression in switchgrass, Panicum virgatum L. Biotechnol J 3:354–359
Yoo SD, Cho YH, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2:1565–1572
Cho YH, Yoo SD (2010) Expression of epitope-tagged proteins in Arabidopsis leaf mesophyll protoplasts. Methods Mol Biol 657:33–42
Acknowledgments
We thank Prof Jen Sheen for sharing information and encouragement on this manuscript preparation. Most of all, we deeply appreciate tremendous efforts to establish and improve the TEAMP system by many former and current members of Sheen lab at HMS/MGH and Yoo lab at Korea University. This work has been supported by grants from the National Research Foundation (NRF-2012R1A2A2A01005933), Korean RDA (PJ009106052013) and KOREA University to Sang-Dong Yoo.
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Im, J.H., Yoo, SD. (2014). Transient Expression in Arabidopsis Leaf Mesophyll Protoplast System for Cell-Based Functional Analysis of MAPK Cascades Signaling. In: Komis, G., Šamaj, J. (eds) Plant MAP Kinases. Methods in Molecular Biology, vol 1171. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0922-3_1
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DOI: https://doi.org/10.1007/978-1-4939-0922-3_1
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-0922-3
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