Genetically Transformed Maize Plants from Protoplasts

TLDR

Maize protoplasts from embryogenic A188 cultures were transformed with a plasmid carrying the NPT II gene under a 35S promoter by high‑voltage electroporation, selected on kanamycin, and regenerated into mature plants. Transformed cell lines exhibited NPT II activity and Southern blotting confirmed integration of the NPT II gene in callus and regenerated plants.

Abstract

Genetically transformed maize plants were obtained from protoplasts treated with recombinant DNA. Protoplasts that were digested from embryogenic cell suspension cultures of maize inbred A188 were combined with plasmid DNA containing a gene coding for neomycin phosphotransferase (NPT II) next to the 35S promoter region of cauliflower mosaic virus. A high voltage electrical pulse was applied to the protoplasts, which were then grown on filters placed over feeder layers of maize suspension cells (Black Mexican Sweet) and selected for growth in the presence of kanamycin. Selected cell lines showed NPT II activity. Plants were regenerated from transformed cell lines and grown to maturity. Southern analysis of DNA extracted from callus and plants indicated the presence of the NPT II gene.