Plant gene delivery is challenging due to the presence of plant cell walls. Conventional means such as Agrobacterium infection, biolistic particle bombardment, electroporation, or polyethylene glycol attachment are often characterized by high cost, labor extensiveness, and a significant perturbation to the growth of cells. We have succeeded in delivering GFP-encoding plasmid DNA to turfgrass cells using poly(amidoamine) dendrimers. Our new scheme utilizes the physiochemical properties as well as the nanosize of the poly(amidoamine) dendrimer for direct and noninvasive gene delivery. The GFP gene was expressed in the plant cells as observed by confocal fluorescence microscopy. The transfection efficiency may be further improved by optimizing the pH of the cell culture medium and the molar ratio of the dendrimer to DNA. The use of the current delivery system can be extended to virtually all plant species having successful regeneration systems in place.

Abstract:We report our single-molecule fluorescence microscopy and molecular dynamics simulation studies on the interaction of poly(amidoamine) dendrimer and squalane hydrocarbon in aqueous solution. Our spectrophotometry measurements indicate that this interaction increases with the pH of the solvent. Our simulations show that squalane resides primarily on the perimeter of the dendrimer at low to neutral pH, but becomes encapsulated by the dendrimer at high pH. Using single-molecule fluorescence microscopy, we have identified that the binding between PAMAM and squalane is reversible. At a pH value of 8, the approaching, binding, and characteristic times of a single fluorescently-labeled dendrimer to squalane are 0.5 s, 7.5 s, and 0.5 s, respectively. Both our spectrophotometry measurements and simulations show that the interaction between PAMAM and squalane is stronger for lower generation dendrimers. This study facilitates our understanding of using dendritic and hyperbranched polymers for gas hydrate prevention in the petroleum industry.