Abstract: Juglans regia'Xincuifeng' seedlings were used as research materials to compare the effects of biomass, physiological characteristics and enzyme activities under different phosphorus gradients. The results showed that phosphate fertilizer has a certain regulation effect on the growth of J. regia seedlings. Appropriate phosphorus application was beneficial to the growth and development of J. regia seedlings. Among them, the treatment of J. regia seedling leaves at 30 g/plant phosphorus concentration was the best, and the total biomass of the plant was the largest. Above or below 30 g/plant phosphorus concentration, the aboveground and underground biomass of the plant decreased. The soluble sugar and soluble protein in the leaves of J. regia seedlings reached the peak at 30 g/plant phosphorus treatment, which was 70.89 mg/g and 40.95 mg/g higher than CK. Proline, malondialdehyde and root activity all showed a trend of decreasing first and then increasing with the increase of phosphorus supply, and CK reached the maximum value. In general, the soil phosphorus supply level of 30 g/plant can meet the good growth and physiological and biochemical reactions of J. regia seedlings. Excessive or low phosphorus levels are not conducive to the growth and development of J. regia seedlings.