Abstract: A field experiment was conducted using the winter wheat (Triticum aestivum) variety Shimai 15. The source of organic nitrogen was cow manure, and four fertilization treatments were included, i.e., no N fertilizer application, single application of urea, single application of cow manure, and mixed application of urea and cow manure. The effects of different applications of inorganic and organic nitrogen on canopy apparent photosynthesis (CAP), photosynthetic rate of flag leaves (P_n), leaf area index (LAI), florescence parameters and grain yield of winter wheat were determined. The results showed that urea had the largest effect on the early growth period, as at this stage the CAP, P_n and LAI of the single application of urea were the highest, which was followed by the mixed application and the single application of cow manure. However, 10 days after anthesis, the single application of cow manure and the mixed application delayed the leaf senescence process when compared with the single application of urea. This could be due to the two treatments having higher antioxidant enzyme activity and promoting a longer green leaf duration, which could maintain a higher photosynthetic capability. What’s more, the mixed application had a better performance and got the highest grain yield. Consequently, the mixed application of organic and inorganic fertilizers could delay leaf senescence and maintain a better canopy structure and higher photosynthesis capability at the late grain filling stage, which resulted in a higher grain yield.