Fig. 3
Optimisation of culture medium for N. benthamiana HR. a Growth curves of N. benthamiana (clone 7) and S. lycopersicum (clone 11) HR liquid cultures expressing scFvFc 2G8. Growth was followed for 28 days after HR inoculum (200 mg). b Quantification by ELISA of scFvFc 2G8 secreted by N. benthamiana #7 in MS culture medium supplemented with KNO3 10 g/L, and various concentrations of NAA. Different time intervals (T: 4, 7 and 9 days) from NAA administration have been considered. Values represent average ± SE (n = 3). Statistically significant difference (P < 0.05, Student’s t test) at 3 mg/L of NAA at T9 with respect to the other concentrations is marked by an asterisk. c Comparison by silver stained SDS-PAGE of proteins rhizosecreted by N. benthamiana HR C- at different time intervals from growth regulators addition. For each sample, a volume of culture medium normalized with root biomass (25 mg) was loaded. Lane 1: MS without growth regulators; lane 2: optimized MS supplemented with KNO3 10 g/L, NAA 3 mg/L; lane 3: MS supplemented with KNO3 12 g/L, NAA 19 mg/L, and PVP 1.5%. M: Prestained Protein Marker Sharpmass VI (EuroClone)