Mixed cultivation with legumes may alleviate the nitrogen (N) limitation of monoculture Eucalyptus. However, how leaf functional traits respond to N in mixed cultivation with legumes and how they affect tree growth are unclear. Thus, this study investigated the response of leaf functional traits of Eucalyptus urophylla × Eucalyptus grandis (E. urophylla × E. grandis) and Dalbergia odorifera (D. odorifera) to mixed culture and N application, as well as the regulatory pathways of key traits on seedling growth. In this study, a pot-controlled experiment was set up, and seedling growth indicators, leaf physiology, morphological parameters, and N content were collected and analyzed after 180 days of N application treatment. The results indicated that mixed culture improved the N absorption and photosynthetic rate of E. urophylla × E. grandis, further promoting seedling growth but inhibiting the photosynthetic process of D. odorifera, reducing its growth and biomass. Redundancy analysis and path analysis revealed that leaf nitrogen content, pigment content, and photosynthesis-related physiological indicators were the traits most directly related to seedling growth and biomass accumulation, with the net photosynthetic rate explaining 50.9% and 55.8% of the variation in growth indicators for E. urophylla × E. grandis and D. odorifera, respectively. Additionally, leaf morphological traits are related to the trade-off strategy exhibited by E. urophylla × E. grandis and D. odorifera based on N competition. This study demonstrated that physiological traits related to photosynthesis are reliable predictors of N nutrition and tree growth in mixed stands, while leaf morphological traits reflect the resource trade-off strategies of different tree species.