[Objectives]The paper aimed to reveal the effects of continuous positioning of different nitrogen application levels on photosynthetic characteristics and yield of cotton. [Methods]In 2018, fixed-point field trials of 6 treatments with different nitrogen levels of 0, 90, 180, 270, 360, 450 kg·hm-2 were set up, and each treatment was represented by N0, N90, N180, N270, N360, and N450, respectively. Repeated experiments were carried out on the original position of each treatment in 2018 for two consecutive years of 2019 and 2020. The photosynthetic characteristics and yield of cotton under different nitrogen application levels were measured and their changing rules were studied. [Results]The treatment of average leaf area index(LAI)from large to small was N270, N450, N360, N180, N90, N0 during the whole growth period in 2019 and 2020. The relative chlorophyll content(SPAD), net photosynthetic rate(Pn), transpiration rate(Tr), and stomatal conductance(Gs)showed a parabolic change trend that first increased and then decreased with the increase of nitrogen application rate. All treatments with medium nitrogen(N270)were the highest, while the intercellular CO2 concentration(Ci)was the lowest. In addition, the differences of LAI、SPAD, Pn, Tr and Gs of each treatment in the positioning three years(2020)were higher than those in the positioning two years(2019). The output of each year from 2018 to 2020 showed a single peak curve of first increasing and then decreasing with the increase of nitrogen application level, and basically number of bolls per plant and the yield under the treatment of medium nitrogen N270 were the highest. The yields of N270 treatments for two and three consecutive years of nitrogen application were significantly higher than those of no fertilizer(N0), low nitrogen(N90, N180), and high nitrogen(N360, N450)treatments. The longer the positioning period, the more significant the performance would be, In 2019 the output of N270 treatment respectively increased by 17.10%, 5.11%, 3.83%, 3.83%, 5.16% compared with N0, N90, N180, N360, N450 treatments, and in 2020 it respectively increased by 41.40%, 18.30%, 8.12%, 9.08% and 8.44%. [Conclusions]The photosynthesis of cotton without continuous nitrogen application was the worst, causing serious yield reduction. Low nitrogen of 90 and 180 kg·hm-2 did not significantly increase photosynthesis, and the increase in yield was smaller. Under the conditions of high nitrogen content of 360 and 450 kg·hm-2, the continuous positioning time was the longer, the inhibition on cotton photosynthesis was the stronger, and the cotton yield was the smaller. This experiment showed that when the continuous nitrogen application rate was 270 kg·hm-2, the photosynthesis capacity of cotton was the highest, and the cotton yield could be maintained at a high level. [ABSTRACT FROM AUTHOR]