Agrobacterium tumefaciens-mediated transient transformation of Arabidopsis thaliana leaves
MANGANO, SILVINA; GONZALEZ, CINTIA DANIELA; PETRUCCELLI, SILVANA
METHODS IN MOLECULAR BIOLOGY (CLIFTON, N.J.)
Lugar: New York; Año: 2014 vol. 1062 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 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.