The aim of this study was to optimize the in vitro chromosome-doubling procedure in wheat anther culture. Colchicine, at concentrations of 100–5000 mg/l, was added to the induction medium for 1–5 days. Beneficial effects were obtained with concentrations of 100 and 1000 mg/l colchicine. With time, significant reductions in embryo–like structures as well as higher doubling indices were found. Similar results were obtained with the high- and low-responding genotypes. Colchicine (100 mg/l), added 5 and 20 days after inoculation for 1 and 3 days increased the induction response, but this value was reduced when colchicine was added 10 or 15 days after inoculation. The doubling effect was similar to the control, except for a significant increase with the 3-day application 20 days after inoculation. The highest success index was reached when colchicine was added to the culture medium after 20 days.