Figure 1

Optimization of transient expression in the FAST assay. A, Effect of Silwet L-77 concentration on transient expression. Transient expression efficiency was quantified by the average luminescence intensity of each Arabidopsis seedling expressing a d35S::hRLuc construct when Silwet L-77 of indicated concentration and bacteria of OD600 = 0.3 were included in the cocultivation medium. The assay was carried out after 40 hr cocultivation. The negative control indicated by letter "C" was performed using Agrobacteria cells carrying no binary vector. B, Effect of bacteria density on transient expression. Transient expression efficiency was quantified by the average luminescence intensity of each Arabidopsis seedling expressing a d35S::hRLuc construct when 0.005% Silwet L-77 and bacteria of indicated density were included in the cocultivation medium. The assay was carried out after 40 hr cocultivation. C, Time-course for transient expression. Transient expression efficiency was quantified by the average luminescence intensity of each Arabidopsis seedling at different time points during the cocultivation when 0.005% Silwet L-77 and bacteria of OD600 = 0.5 were used in the cocultivation medium. D, Expression of a d35S::NLS-YFP-GUS construct in a representative Arabidopsis seedling after 36 hr cocultivation when 0.005% Silwet L-77 and bacteria of OD600 = 0.5 were used in the cocultivation medium. Scale bar = 0.3 mm. E, Western blot analysis of NLS-YFP-GUS expression in 8 transformed (NLS-YFP-GUS) or untransformed (C) Arabidopsis seedlings after 36 hr cocultivation when 0.005% Silwet L-77 and bacteria of OD600 = 0.5 were used in the cocultivation medium.