Mechanism and optimized conditions for PEG mediated DNA transfection into plant protoplasts

Experimental conditions influencing DNA uptake efficiency by maize protoplasts in polyethyleneglycol (PEG) mediated transfection experiments have been studied systematically. The data provide evidence that the extracellular DNA is precipitated efficiently by combined action of PEG together with divalent cations and DNA is taken up by the plant protoplasts in the precipitated form. The particle size is strongly effected by the pH of the PEG solution. At optimal pH 6– 6.5 a very fine and homogenous precipitate forms in presence of Ca²⁺ and Mg²⁺ ions and is efficiently incorporated by maize and rice protoplasts.Abbreviations CAT Chloramphenicolacetyl transferase - DAPI 4,6-Diamidino-2-Phenylindole - HEPES N-2-Hydroxyethylpiperazine-N-2- ethanesulfonic acid - MES 2(N-Morpholino)ethanesul fonic acid - PEG Polyethyleneglycol - SW seawater