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Introduction of Foreign DNA into Yellow-Poplar Protoplasts by Electroporation

Plantlets of yellow-poplar can be regenerated from protoplasts by somatic embryogenesis. This culture system could allow the production of genetically altered trees following DNA uptake into protoplasts. Two bacterial marker genes, ß-glucuronidase (GUS) and neomycin phosphotransferase (NPT II), are being used to monitor yellow-poplar transformation. Plasmids bearing one or both of these genes were introduced into yellow-poplar protoplasts by electroporation or polyethylene glycol-mediated uptake. The protoplasts had been isolated from embryogenic suspension cultures at different time-points after transfer to fresh medium. In addition to the physical parameters of electroporation, such as voltage, capacitance, and pulse length, an important factor in delivery of DNA into cell nuclei was the interval between subculture and exposure to DNA. Assays for GUS activity indicate that the highest level of transient expression was in protoplasts isolated from 20- day old cells.


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Author(s): H. Dayton Wilde, Richard B. Meagher, Scott A. Merkle

Publication: Tree Improvement and Genetics - Southern Forest Tree Improvement Conference - 1989

Section: General Session: Biotechnology