One-time fertilization with controlled-release urea (CRU) is a research hotspot for its lower labor cost and stability of nitrogen (N) supply for rice growth. Yet the fertilizer formulation needs to be further improved to better adjust the N supplement to meet the demand of rice plants and obtain a higher grain yield. Therefore, the effects of novel fertilizer formulations composed of CRU, urease inhibitor (UI) and nitrification inhibitor (NI) on the rice growth and photosynthetic characteristics as well as high-yield formation were tested through a two-year field experiment. The result indicated that the combined use of CRU and UI treatment can achieve higher yields than with CRU at the same N application level. Meanwhile, with a 20% reduction of N use, one-time application of CRU + UI can obtain the same high yield as the conventional split application of urea. Compared with conventional fertilization and CRU treatment, the CRU + UI treatment had suitable leaf area and biomass accumulation at the vegetative growth stage and high effective stem tiller rate. More post-anthesis dry matter accumulation, higher net photosynthesis rate and low senescence rate were guaranteed for its high yield and nitrogen agronomic efficiency.