Hydroponic Cultivation of Nicotiana benthamiana: A Stepwise Protocol for Biomass Production in Molecular Farming

This protocol outlines an efficient hydroponic system for cultivating Nicotiana benthamiana plants as a platform for recombinant protein production. Two hydroponic systems, Deep Water Culture (DWC) and Ebb and Flow (E & F), were compared with conventional tissue culture methods, focusing on plant growth and leaf biomass accumulation. The finding show that hydroponic cultivation yields a ~20-fold increase in leaf biomass per plant compared to tissue culture, with no significant difference in yield between DWC and E&F systems. However, the E&F system offers improved aeration and reduced risk of root diseases, making it more suitable for long-term cultivation. This scalable cultivation pipeline, when combined with stable genetic transformation and clonal propagation via tissue culture, provides a robust and sustainable strategy for high-yield molecular farming using N. benthamiana .