Study on gene transfer into plant cells by electroinjection

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We have found that by use of electric field pulses we can transform intact yeast cells with plasmid DNA and transfect free mesophyll cells of tobacco with virus RNA (ELECTROINJECTION). In this project we studied the feasibility of the electro-injection to transfer foreign genes into intact cultured cells and tissues. We also studied the effects of cell walls on the electric field-mediated entry of foreign genes into and expression of the genes in the protoplast-derived cells (the cells that were obtained by the culture of protoplasts).Based on transient expression assay of plasmid DNA and virus RNA and particles, scanning electron microscopy and Southern hybridizaiton analysis, we have revealed that the regenerated cell walls at the initial stages of protoplast culture did not inhibit uptake and expression of plasmid DNA and virus RNA but that cell walls of intact cells inhibit uptake of these DNA and RNA. Direct transfer of these genetic materials into intact cultured cells and tissues by use of electric pulses was not successful under the conditions studied. These results indicate that increase of the porosity of intact cell walls (by enzymatic or mechanical treatment) is needed to introduce genetic materials into the cells by electric pulses.