The study tested the regeneration of various chrysanthemum cultivars by establishing its growth cultured in vitro, then use in vitro cultured petiole or leaf as regeneration explants. The different mix of plant growth regulator treatments were added at medium, and investigated for shoot regeneration. The results showed that regeneration ability varied among tested cultivars, however no difference shown on regeneration rate between leaf and petiole types of explants in the same cultivar. Later apply three antibiotics carbenicillin, cefotaxime and timentin with various concentrations to precede antibiotic test with the ”Huang-Xiu-Fang” petiole explants of better regeneration. All three antibiotics had no obvious suppressing influence on shoot regeneration, and among them, the treatment of 200 mg/L timentin had highest regeneration rate.