A Highly-Efficient Agrobacterium-Mediated Method for Transient Gene Expression and Functional Studies in the Leaves of Arabidopsis and Various Other Plant Species

Abstract While the use of stable transformation technology has led to great insight into gene function, its application in high-throughput studies remains arduous. Although techniques such as agro-infiltration have been widely used in species such as Nicotiana benthamiana for the rapid detection of gene expression and protein interaction analysis, this technique does not work efficiently in the other plant species, including Arabidopsis thaliana. As an efficient high-throughput transient expression system is currently lacking in the model plant species Arabidopsis thaliana, we describe a method that is characterized by high efficiency, reproducibility and suitability for a variety of functional proteins in a number of plant species, including Arabidopsis thaliana, Brassica oleracea, Capsella rubella, Thellungiella salsuginea, Thellungiella halophila, Solanum tuberosum, Capsicum annuum and Nicotiana benthamiana. Efficiency of this method was independently verified in three independent research facilities pointing to the robustness of this technique. Furthermore, in addition to demonstrating the utility of this technique in a range of species, we also present a case study wherein we assess protein-protein interactions in the sucrose biosynthesis pathway in Arabidopsis.