Abstract: 【Objective】This study investigated the effects of different cultivation conditions on the physiological characteristics of Pleurotus citrinopileatus and polysaccharide content in fruiting bodies,which provided reference for polysaccharide content from P. citrinopileatus. 【Method】Taking P. citrinopileatus flat-top 8-31 as material, it was treated with a single component of cultivation material（corn cob, wood chip, straw） and treated with different illumination（white light, purple light, dark treatment, red light, blue light）, with mixed formulation and natural light as control（CK1 and CK2） and the polysaccharide was extracted by hot water extraction method, and its polysaccharide content was determined by phenol-sulfuric acid method. The effects of different cultivation material treatments and light treatments on physiological characteristics and polysaccharide content of P. citrinopileatus were analyzed. 【Result】The order of total yield and biological efficiency of different cultivation materials was CK1 ＞ wood chip ＞ corn cob ＞ straw. The extraction rate of crude polysaccharide was corn cob（8.64%） = wood chip（8.64%） ＞ straw（8.52%） ＞ CK1（8.00%）, and the polysaccharide content was as follows: corn cob（0.216 mg/mL） = wood chip（0.216 mg/mL） ＞ straw（0.213 mg/mL） ＞ CK1（0.200 mg/mL）. The fruit bodies of P. citrinopileatus were light yellow under different cultivation materials. The order of total output and biological efficiency of P. citrinopileatus under different light treatments was CK2 ＞ blue light treatment group ＞ purple light treatment group ＞ white light treatment group ＞ dark treatment group＞ red light treatment group. The order of crude polysaccharide extraction rate was red light treatment group（10.80%） ＞ white light treatment group（8.24%） ＞ CK2（8.20%） ＞ blue light treatment group（8.16%） ＞ dark treatment group（7.48%） ＞ purple light treatment group（6.72%）. The order of polysaccharide content was red light treatment group（0.270 mg/mL） ＞ white light treatment group（0.206 mg/mL） ＞ CK2（0.205 mg/mL） ＞ blue light treatment group（0.204 mg/mL） ＞ dark treatment group（0.187 mg/mL） ＞ purple light treatment group（0.168 mg/mL）. The fruit body of blue light treatment was yellow, the fruiting bodies treated with white light and purple light were pale yellow, and those treated with red light and dark treatment were white. The extraction rates of crude polysaccharides from different fruiting body parts were as follows: stipe ＞ cap ＞ pili; the order of polysaccharide content was as follows: stipe（0.362 mg/mL） ＞ cap（0.235 mg/mL） ＞ pili（0.200 mg/mL）. 【Conclusion】Different cultivation materials have no obvious effects on the color of P. citrinopileatus fruiting body, while different light treatments affect the color of P. citrinopileatus fruiting body. Different cultivation materials and light treatments all affect the physiological characteristics and polysaccharide content of P. citrinopileatus. Formulating comprehensive nutrient-rich cultivation materials can increase the output of P. citrinopileatus. Single component（corn cob, wood chip） cultivation materials or red light irradiation have positive promoting effects on the polysaccharides synthesis and accumulation of P. citrinopileatus.