Employment of Cytokinin Vectors for Marker-Free and Backbone-Free Transformation

Marker-free methods of plant transformation sacrifice the advantages of a selectable marker during regeneration or add work after regeneration to remove the marker. On the positive side, there is no stably integrated marker gene in the plant genome to present regulatory hurdles or potential biosafety hazards once the plant is released to the environment. A marker-free method that is simple and adaptable to multiple crop species—even asexually propagated species—is presented herein. This method employs an engineered vector that utilizes the isopentenyltransferase (ipt ) to drive the regeneration of intragenic cells containing the gene(s) of interest. The ipt gene also acts as a marker to screen against events where the vector backbone is stably integrated.