Abstract: In an effort to optimize the tissue culture response of a tall fescue (Festuca arundinacea) regeneration system, the effects of 2,4-D on tissue culture responses were investigated using immature inflorescences, leaf tips, young stems, and young nodes as explants. Two clones (SACD and 98-19), which are self-bred dwarf clones of tall fescue were used. Immature inflorescences were the best vegetative organ for callus induction, with the highest callus induction rate of 94%. For tips of leaves, stems and nodes, the callus induction rate was 0%; the optimal concentration of 2,4-D was from 7 to 9 mg/L in MS with a callus induction rate of 83% to 94%. Significant differences were also observed between the tested clones but the interaction between 2,4-D concentrations and the clones was not significant. Subculture medium of 2,4-D 4 mg/L in MS resulted in the highest embryogenic callus induction rate (98%), highest green spot induction rate (78%) during subculture, and high green plant regeneration rate (57%) in regeneration medium of “MS+2,4-D 2 mg/L+6-BA 1 mg/L”. The rooting medium used (“1/2 MS+NAA 0.5 mg/L”), gave a rooting rate of 100%.