FIGURE 2.
Optimization of transient expression in rice. (A) Effect of co-cultivation medium and seedling growth temperature on transient expression. (B) Time-course for transient expression. Transient expression efficiency was quantified by the measuring 4-MU fluorescence of total protein extract isolated from the first leaf of rice seedlings at different time points during the co-cultivation when 0.02% Silwet® L-77 and bacteria of A600nm = 0.6 were used in the co-cultivation medium. (C) Effect of Agrobacterium growth temperature on transient expression. Agrobacterium strain transformed by pCAMBIA1304 was grown in LB medium with antibiotics at temperatures mentioned in the graph and 1 h co-cultivation in the medium with shaking was also carried out at the same temperature. (D) Effect of bacterial density on transient expression efficiency. (E) Effect of vacuum pressure on transient expression efficiency. (F) Effect of acetosyringone concentration on transient expression efficiency. (G) Effect of glucose concentration on transient expression efficiency. (H) Effect of Silwet® L-77 concentration on transient expression. n = 3 biological replicates (mean ± standard error for A–H). ***p-value ≤ 0.001, **p-value ≤ 0.01, *p-value ≤ 0.05.