Abstract: The in vitro plantlets were used as the materials,and MS+3.0 mg/L 6-BA+0.3 mg/L NAA+7 g/L agar+25 g/L glucose was used as the basal medium for studying the effects of organic additives, bottle sizes and light qualities on the accumulation of polysaccharide content during the growth of in vitro plantlets by the L₉（3⁴） orthogonal experimental design during in Anoectochilus roxburghii. The results showed that the order of polysaccharide content was light quality>additive>bottle size. The average value of banana mud in the additive was 0.291,and there was a significant difference between banana mud and apple juice and carrot juice（P<0.05）. The mean value of blue light in the light mass is 0.289, and there is a significant difference between blue light and white and red light （P<0.05）. The mean value of 6.5 cm in the bottle size was 0.279,which was significantly different from that of the 8.0 cm bottle（P<0.05）,and there was no significant difference（P>0.05）. Consolidated results showed that the best culture conditions for the accumulation of polysaccharide content was MS+6-BA 3.0 mg/L+NAA 0.3 mg/L+7 g/L agar+25 g/L glucose+50 g/L banana（pH 5.8） in the culture bottle with 6.5 cm diameter in the blue light.

Key words: Anoectochilus roxburghii, tissue culture, polysaccharide contents