Abstract
Sequestration of a transcription factor in a cellular membrane and releasing it on demand is an additional layer of gene regulation that is considered a rapid mode to reprogram a gene expression cascade when a plasma membrane stress signal is perceived. Better understanding of the dynamic exchange of membrane-bound transcription factors (MTFs) during biotic stress requires the development of a simple, efficient, and quick assay system. Here we report an Agrobacterium-based transient transformation method to assay the localization of fluorescent protein-tagged MTFs in tomato leaf epidermal peels that are subsequently infected with a pathogenic fungus. Essentially, our method mimics natural infection and facilitates the realistic monitoring of MTF movement during activation of a signaling event.
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References
Seo PJ (2014) Recent advances in plant membrane-bound transcription factor research: emphasis on intracellular movement. J Int Plant Biol 56(4):334–342
Kim SY, Kim SG, Kim YS et al (2007) Exploring membrane-associated NAC transcription factors in Arabidopsis: implications for membrane biology in genome regulation. Nucleic Acids Res 35(1):203–213
Heazlewood JL, Tonti-Filippini J, Verboom RE et al (2005) Combining experimental and predicted datasets for determination of the subcellular location of proteins in Arabidopsis. Plant Physiol 139(2):598–609
Chou KC, Shen HB (2010) Plant-mPLoc: a top-down strategy to augment the power for predicting plant protein subcellular localization. PLoS One 5(6):e11335. https://doi.org/10.1371/journal.pone.0011335
Lee MW, Yang Y (2006) Transient expression assay by agroinfiltration of leaves. In: Julio Salinas J, Sanchez-Serrano JJ (eds) Arabidopsis protocols. Methods in molecular biology, vol 323. Humana Press, Totowa, New Jersey, pp 225–229
Trick HN, Finer JJ (1997) SAAT: Sonication-assisted Agrobacterium-mediated transformation. Transgenic Res 6(5):329–336
Liu Y, Yang H, Sakanishi A (2006) Ultrasound: mechanical gene transfer into plant cells by sonoporation. Biotechnol Adv 24(1):1–16
Bhattacharjee P, Das R, Mandal A et al (2017) Functional characterization of tomato membrane-bound NAC transcription factors. Plant Mol Biol 93:511–532
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Chowdhury, S., Bhattacharjee, P., Basak, S., Chowdhury, S., Kundu, P. (2019). Method to Study Dynamics of Membrane-Bound Plant Transcription Factors During Biotic Interactions in Tomato. In: Gassmann, W. (eds) Plant Innate Immunity. Methods in Molecular Biology, vol 1991. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9458-8_7
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DOI: https://doi.org/10.1007/978-1-4939-9458-8_7
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-4939-9458-8
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