Abstract: 【OBJECTIVE】:the paper was studied the callus induction, the foundation and culture of the suspension, and isolatation of the protoplast of anther of dongzao. 【METHODS】 the anther of dongzao was used as material to find the best culture medium of foundation and cultivation of the suspension and the protoplast isolation conditions. 【RESULTS】it is good to indicate callus when the culture medium which was added TDZ0.2mg/L、NAA0.5 mg/L and PVP2.0g/L was used. The better culture medium of callus multiplication was 1/2MS+TDZ0.4 mg/L +NAA0.2 mg/L. The liquid culture medium that was 1/2MS+TDZ0.4 mg/L +NAA0.2 mg/L was used to the suspension. When the the suspension was used to isolation, the optimal condition was 0.6M Mannital +0.1%MES+20~25 g/L Cellulase “onozuka” R-10，the isolation time was 16h.