Production of transgenic tall fescue and red fescue plants by particle bombardment of mature seed-derived highly regenerative tissues

Highly regenerative tissues of tall fescue and red fescue produced from mature seed-derived embryogenic callus were induced and proliferated on medium containing 2,4-dichlorophenoxyacetic acid (4.5 or 9.0 muM), 6-benzylaminopurine (0, 0.044, 0.44 or 2.2 muM) and cupric sulfate (0.1 or 5.0 muM) under dim-light conditions (10 to 30 muE m(-2) s(-1) 16 h light). Tall fescue tissues were transformed with three plasmids containing the genes for hygromycin phosphotransferase (hpr), phosphinothricin acetyltransferase (bar) and beta -glucuronidase (uidA; gus), and red fescue with three plasmids containing hpt, uidA and a synthetic green fluorescent protein gene [sgfp (S65T)]. DNA from To plants of eight independently transformed lines from tall fescue and 11 from red fescue were analyzed by PCR and DNA blot hybridization. The co-expression frequency of all three transgenes [hpt/bar/uidA or hpt/uidA/sgfp (S65T)] in transgenic tall fescue and red fescue plants was 25-27%; for two transgenes [hpt/bar or hpt/uidA for tall fescue and hpt/uidA or hpt/sgfp(S65T) for red fescue], the co-expression frequency was 50-75%.