Agrobacterium tumefaciens-Mediated Transient Transformation of Arabidopsis thaliana Leaves

Abstract

Transient assays provide a convenient alternative to stable transformation. Compared to the generation of stably transformed plants, agroinfi ltration is more rapid, and samples can be analyzed a few days after inocu-lation. Nevertheless, at difference of tobacco and other plant species, Arabidopsis thaliana remains recalci-trant to routine transient assays. In this chapter, we describe a transient expression assay using simple infi ltration of intact Arabidopsis leaves with Agrobacterium tumefaciens carrying a plasmid expressing a reporter fl uorescent protein. In this protocol, Agrobacterium aggressiveness was increased by a prolonged treatment in an induction medium defi cient in nutrients and containing acetosyringone. Besides, Arabidopsisplants were cultivated in intermediate photoperiod (12 h light–12 h dark) to promote leaf growth.