Stably transformed callus lines from microprojectile bombardment of cell suspension cultures of wheat

We have obtained stably transformed callus lines by direct delivery of DNA into plated suspension culture cells of wheat (Triticum aestivum L.) using high velocity microprojectile bombardment. Three different reporter or selectable marker genes, jointly present or on separate plasmids, were introduced: neomycin phospho-transferase (NPTII), beta-glucuronidase (GUS) and 5-enolpyruvylshikimate phosphate (EPSP) synthase. Kanamycin was used for the selection of resistant calli, which were screened for GUS expression by a histochemical stain. Southern analysis confirmed that the NPTII, GUS and EPSP synthase genes had been stably integrated in all of the kanamycin resistant and GUS positive lines, and NPTII and EPSP synthase activities were demonstrated in the transformed calli.