Abstract: Through field experiment conducted in Hexi corridor of Gansu Province, this paper discussed the effects of root separation on plant nitrogen uptakes and soil nitrate nitrogen accumulation in wheat/maize intercropping. The results showed that the nitrogen uptakes of wheat and maize with root partition were lower than those of without root partition. After wheat harvest, soil nitrate nitrogen concentration in wheat strip with root separation was the same as without root separation; however, soil nitrogen accumulation in maize strip without root separation was much higher than that of with root partition in wheat/maize intercropping. After maize harvest, soil nitrogen accumulation in wheat strip with root separation was higher than that of without root separation; yet, the soil nitrate nitrogen accumulation in maize strip with root separation was lower than that of without root separation. Crop nitrogen uptake and soil nitrate nitrogen concentration existed no significant difference between root partition and no root in wheat/no maize, no wheat /maize intercropping.