Object To establish an effective plantlet regeneration system of Aconitum carmichaeli Debx. for the purpose to obtain a large number of high quality seedling in a short time. Methods Leaves in vitro were tried as the explants and cultivated in different media with the various portion of hormones. Results The medium of MS+BA 1.0 mg/L+KT 0.5 mg/L+NAA 0.2 mg/L was the most suitable one for the induction of shoots; the medium of MS+BA 1.5 mg/L+NAA 0.1 mg/L was beneficial to the propagation of shoots; the medium of MS+BA 0.5 mg/L+NAA 0.1 mg/L was good for the elongation of shoots; and the medium of 1/2 MS without any hormone was suitable for the induction of roots. Conclusion Rapid propagation of A. carmichaeli could be achieved by tissue culture and this will lead to the possibility for its seedling in the industrial production.