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

ABSTRACT

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 μF 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.