Abstract
Transient assays provide a convenient alternative to stable transformation. Compared to the generation of stably transformed plants, agroinfiltration is more rapid, and samples can be analyzed a few days after inoculation. Nevertheless, at difference of tobacco and other plant species, Arabidopsis thaliana remains recalcitrant to routine transient assays. In this chapter, we describe a transient expression assay using simple infiltration of intact Arabidopsis leaves with Agrobacterium tumefaciens carrying a plasmid expressing a reporter fluorescent protein. In this protocol, Agrobacterium aggressiveness was increased by a prolonged treatment in an induction medium deficient in nutrients and containing acetosyringone. Besides, Arabidopsis plants were cultivated in intermediate photoperiod (12 h light–12 h dark) to promote leaf growth.
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Acknowledgements
This research was supported by the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) through the grants PICT2007-0479 and PICT2010-2366 to Petruccelli Silvana and by Universidad Nacional de La Plata (project 11X/498). Petruccelli Silvana is a member of the Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina (CONICET). Silvina Mangano is a researcher of Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas. Universidad Nacional de la Plata.
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Mangano, S., Gonzalez, C.D., Petruccelli, S. (2014). Agrobacterium tumefaciens-Mediated Transient Transformation of Arabidopsis thaliana Leaves. In: Sanchez-Serrano, J., Salinas, J. (eds) Arabidopsis Protocols. Methods in Molecular Biology, vol 1062. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-580-4_8
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DOI: https://doi.org/10.1007/978-1-62703-580-4_8
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