Agrobacterium tumefaciens mediated transient transformation of Arabidopsis thaliana leaves

MANGANO, SILVINA; GONZALEZ, CINTIA DANIELA; PETRUCCELLI, SILVANA*,

Arabidopsis Protocols, Third Edition

Humana Press

Lugar: New York; Año: 2014; p. 165 - 173

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 thaliana leaves with Agrobacterium tumefaciens carrying a plasmid expressing a reporter fluorescence protein. In this protocol Agrobacterium aggressiveness was increased by a prolonged treatment in the induction medium in presence of acetosyringone and deficient in nutrients. Besides, the Arabidopsis plants were cultivated in intermediate period light cycle (12 h light-12 h dark) to promote leaf growth.