TRANSGENIC PLANTS OF AGROSTIS-ALBA OBTAINED BY ELECTROPORATION-MEDIATED DIRECT GENE-TRANSFER INTO PROTOPLASTS

A system for genetic transformation of Agrostis alba plants by electro poration-mediated DNA transfer to protoplasts is described. The npt II gene was used as a selectable marker. Selection with 20 mg/l G418 (ge neticin) yielded a total of over 50 resistant cell colonies from three independent experiments. Overall frequency of resistant colony format ion was 1-3 x 10(-6) based on the number of protoplasts plated and 1-2 x 10(-5) based on the number of cell colonies recovered. Subsequent s ubcultures led to the development of plants with an apparently normal morphology. DNA analysis (PCR and Southern hybridization) and enzymati c analysis showed that the G418 resistant plants carried the transgene and expressed it. This is the first successful genetic transformation of an economically important temperate grass, Agrostis.