In order to clarify the regulatory effects of different N fertilizer applications on root morphology, physiological characteristics and assimilate accumulation of drip irrigated spring wheat under the climatic conditions of northern Xinjiang, strong wheat Xinchun 38 (XC 38) and medium wheat Xinchun 49 (XC 49) were used as test materials and cultivated in soil columns at Nck (300 kg·hm-2), N10 (270 kg·hm-2), N20 ( 240 kg·hm-2), N30 (210 kg·hm-2) and N0 (0 kg·hm-2), and the effects of nitrogen application on root morphological characteristics, key enzymes of nitrogen metabolism, antioxidant enzymes, dry matter accumulation distribution and yield of wheat were investigated. The results showed that the root morphological characteristics, key enzymes of nitrogen metabolism, antioxidant enzymes, shoot dry matter (SDM), reproductive organs dry matter (RODM) and yield all showed an increasing and then decreasing trend with increasing N application. Among them, N20 treatment showed the best root length density (RLD), root volume density (RVD), root mass density (RMD), nitrate reductase (NR), glutamine synthetase (GS), glutamate synthetase (GOGAT), alanine aminotransferase (GPT), superoxide dismutase (SOD), peroxidase (POD), root activity (RA), SDM, RODM and yield, which were higher than the remaining treatments by 2.08%-31.06%, 6.67%-132.33%, 12.14%-104.63%, 18.73%-125.12%, 17.21%-94.82%, 12.74%-101.30%, 3.57%-21.94%, 2.72%-28.20%, 4.84%-79.06%, 15.29%-156.55%, and 2.00%-42.61%, and malondialdehyde (MDA) content was reduced by 3.36%-15.70% compared to the rest of the treatments. Correlation analysis showed that yield was significantly positively correlated with RLD, RVD, RMD, GS and GPT and highly significantly negatively correlated with MDA. Nitrogen application and variety intercropping had significant effects on RLD, GS, POD, NR, GPT. Therefore, moderate N reduction (240 kg·hm-2) under the drip irrigation pattern in Xinjiang can improve the morphological and physiological function of wheat roots, promote the distribution and transport of dry matter to reproductive organs, and facilitate yield formation.