To investigate the effects of nitrogen application levels and intercropping with sweet potato on the growth, leaf photosynthetic characteristics, and nitrogen metabolism enzyme activities of flue-cured tobacco, a pot experiment simulating field planting was conducted using flue-cured tobacco variety LY1306 and sweet potato variety Pushu 32. The experiment included two planting patterns (A1, flue-cured tobacco monoculture; A2, flue-cured tobacco and sweet potato intercropping) and four nitrogen application levels (N0, no nitrogen; N1, 0.64 g·pot-1; N2, 0.80 g·pot-1; N3, 0.96 g·pot-1), resulting in a total of 8 treatments. The differences in agronomic traits, root morphological parameters, leaf photosynthetic characteristics, and nitrogen metabolism enzyme activities of flue-cured tobacco under different treatments were studied. The results showed that on 60 days after transplanting, under the same nitrogen lapplication evel, compared with monocultured flue-cured tobacco, intercropped flue-cured tobacco exhibited increases in plant height, maximum leaf area, aboveground dry weight, root dry weight, root tip number, root volume, SPAD value, and net photosynthetic rate (Pn), by 1.93%-10.24%, 5.01%-14.23%, 23.65%-45.83%, 32.43%-53.75%, 4.62%-16.74%, 5.92%-27.82%, 0.42%-6.28%, and 9.39%-32.55%, respectively. All these above measured parameters reached their maximum values at the N2 level. Although the values at the N1 level were lower than those at the N3 level, there were no significant differences between them. The activities of nitrate reductase (NR) and glutamine synthetase (GS) in the leaves of intercropped tobacco were 3.93%-19.13% and 3.03%-5.74% higher than those in monocultured flue cured tobacco, respectively, and both enzyme activities gradually increased with increasing nitrogen application rate, reaching their highest values at the N3 level. Under the conditions of this experiment, both nitrogen application level and planting pattern had significant or highly significant effects on the agronomic traits, photosynthetic parameters, and nitrogen metabolism enzyme activities of flue-cured tobacco. However, their interaction of two factors had a significant or highly significant effect only on root tip number, root volume, intercellular CO2 concentration (Ci), and transpiration rate (Tr). Overall, under the tobacco-sweet potato intercropping system in the rainfed areas of western Henan, a nitrogen application rate of 48-60 kg·hm-2 was sufficient to meet the normal growth and development requirements.