A strategy that optimization of medium compositions for maximum biomass followed by feeding of sucrose for maximum polysaccharide synthesis was developed for enhancing polysaccharide production in suspension culture of protocorm-like bodies (PLBs) of Dendrobium huoshanense C.Z. Tang et S.J. Cheng. In growth stage, the original half-strength MS medium was optimized with carbon sources, nitrogen sources and metal ion combinations. The effects of different carbon sources on PLBs growth were remarkable and sucrose at 35gl −1 was the most suitable. Sole nitrate nitrogen of 30mmoll −1 was the best for PLBs growth. Metal ions (Ca 2+ , Fe 2+ , Mn 2+ and Zn 2+ ) showed different influences on PLBs growth. The optimal concentration of Ca 2+ , Fe 2+ , Mn 2+ and Zn 2+ was 4.5mmoll −1 , 0.1mmoll −1 , 0.5mmoll −1 and 0.06mmoll −1 , respectively. In the optimized medium (sucrose, nitrate, Ca 2+ , Fe 2+ , Mn 2+ and Zn 2+ concentration as described above, the other component concentration seen in half-strength MS), 33.9gDWl −1 PLBs were harvested after 30 days of culture and biomass increase was improved 245% as compared with that in the original medium. In production stage, polysaccharide synthesis was significantly improved by the feeding sucrose. The maximum polysaccharide production (22gl −1 ) was obtained in the case of 50gl −1 sucrose feeding at day 30 of culture, which was about 109-fold higher than that in the original medium without feeding of sucrose.