Agrobacterium-Mediated Transient Expression in Nicotiana benthamiana

For agroinfiltration in Nicotiana benthamiana, Agrobacterium tumefaciens strain AGL1 was transformed with the binary constructs by electroporation. Agrobacterium strains carrying the construct were grown in 5 ml liquid LB-medium supplemented with adequate antibiotic for 24 h. Cells were harvested by centrifugation, washed twice in 10 ml of 10 mM MgCl₂ and re-suspended at OD₆₀₀ 0.5 in infiltration medium (10 mM MgCl₂, 10 mM MES pH 5.6). Agroinfiltration was performed with 1 ml needleless syringe in 4–5 week-old N. benthamiana leaves.