Genetic Transformation of Wheat (triticum Aestivum L.) Anther Culture- Derived Embryos by Electroporation

This report describes the delivery of plasmid DNA into wheat anther culture derived embryos. Initial electroporation experiments were conducted using enzyme (NT-1) pretreatment, 500 F capacitance, and plasmid pAM2100, which carries the herbicide resistance gene, bar, and the screenable marker gene, Gus. Electroporated embryos were allowed to recover 10 days on regeneration medium and then transferred to regeneration medium containing 0, 10, 50, and 100 ppm phoshinotricin (PPT). Twenty eight albino and 13 green plants were regenerated on control (0 ppm PPT) selection medium from 640 embryos treated with pAM2100. The regeneration frequency was similar to embryos electroporated without pAM2100 demonstrating that electroporation was not deleterious for regeneration. Four albino plants were regenerated on selection medium (10, 50, and 100 ppm PPT). One plant was screened and shown to be transgenic by Southern blot analysis of a bar gene. plant, or microbial systems. It has the advantages of being simple, effective, and applicable to a wide range of plant species, higher DNA delivery rate and can be used to deliver DNA into large numbers of cells within a very short time. The greatest electroporation success among cereal crops has been in transformation of maize (Zea mays L.; 5) and rice protoplasts (Oriza sativa L.; 6, 7, 8). The most severe drawback of electroporation-mediated gene transfer in plant research is only useful using protoplasts, which is strongly genotype dependent, and regeneration from protoplast has been difficult for most cereal genotypes. Thus, plant regeneration from wheat (Triticum aestivum L.) protoplasts has had very limited success (9, 10, 11). Transformation methods which do not use protoplasts are being investigated to avoid the drawbacks