Abstract: To establish the cell suspension culture of soybean,LongYe 01-491 was used as materials,and its hypocotyl was induced to the callus. Different concentrations of the precursors (sodium acetate and L-phenylalanine),and fungi elicitors (Penicillium and Aspergillus niger) were added into the cell suspension culture to investigate their effects on the accumulation of isoflavones in cell suspension culture. The results showed that when 100 μmol/L L-phe was added to the suspended cells on the 9th day,the yield of soybean isoflavones was up to (12.52±0.51) mg/flask which was 3.23 times of the control group;When 80 μmol/L NaAc was added to the suspended cells on the 0th day,the yield of soybean isoflavones was up to (5.55±0.36) mg/flask which was 1.43 times of the control group;When 0.5 mL elicitor of Penicillium was added to the suspended cells on the 9th day,the yield of soybean isoflavones was up to (10.20±0.17) mg/flask,which was 2.56 times of the control group;When 0.5 mL elicitor of Aspergillus niger was added to the suspended cells at the 9th day,the yield of soybean isoflavones was up to (7.25±0.30) mg/flask,which was 1.82 times of the control group. In conclusion,the addition of precursors (sodium acetate and L-phenylalanine) and the fungal elicitors (Penicillium and Aspergillus niger) can increase the accumulation of soybean isoflavones in suspended culture of soybean.