AGROBEST: A Highly Efficient Agrobacterium-Mediated Transient Expression System in Arabidopsis Seedlings

Agrobacterium-mediated transient transformation for gene expression is a simple and fast method to analyze transgene functions in plants. Agroinfiltration in leaves of Nicotiana benthamiana is a common method for transient expression. However, agroinfiltration in leaves of Arabidopsis thaliana is challenging due to the low and variable efficiency. Here, we describe procedures of a highly efficient and robust Agrobacterium-mediated transient expression system, named AGROBEST (Agrobacterium-mediated enhanced seedling transformation) for gene expression in A. thaliana seedlings. High efficiency of AGROBEST has been achieved by virulence (vir) gene pre-induction of a specific disarmed Agrobacterium tumefaciens strain C58C1(pTiB6S3ΔT)H followed by co-cultivation with Arabidopsis seedlings in an optimized medium with AB salts and buffered acidic plant culture medium. The stable acidic medium largely increases Agrobacterium-mediated transient expression levels and reduces plant defense responses, suggesting that AGROBEST enables high transient expression efficiency by compromising plant immunity. In summary, AGROBEST is a simple, fast, reliable, and robust transient expression system offering a quick and convenient method to observe protein localization, protein-protein interactions, promoter activities, and gene functional studies in Arabidopsis seedlings.