Pot experiments were carried out to study the effects of urea and controlled-release nitrogen fertilizer (CRNF) application on soil fertility and nitrogen fertilizer use efficiency (NUE) and growth of rape in Southern China, and it could provide reference for the application and dissemination of CRNF in rape production. An experiment with a randomized complete block design was conducted to investigate the effects of urea and CRNF on rape biomass and yield, soil inorganic nitrogen at different growth stages, nitrogen uptake of rape, physiological characteristics of rape, soil microbial nitrogen and enzyme activity using high yielding variety "Xiangyou15". According to the standard amount of nitrogen application in rape field, four treatments were set up in this study, conventional urea (Urea), controlled-release nitrogen fertilizer 1(CRNF1), controlled-release nitrogen fertilizer 2 (CRNF2) and no nitrogen fertilizer (CK). The result showed that compared with the conventional urea treatment, CRNF significantly increased the biomass of rape by 11.2%~20.1% at flowering and harvest stage, the NO₃-N concentration of rape at flowering and harvest stage in the CRNF1 treatment was significantly increased by 43.2% and 61.8%, respectively, and the NH⁴⁺-N concentration in soil in CRNF2 treatment was obviously increased by 18.7% and 64.1%, respectively, at flowering and harvest stage, which ensured the supply of soil nitrogen at the late growth stage of rape. Compared with the Urea treatment, total nitrogen uptake in rape was significantly increased by CRNF at the bolting and flowering stage. In eventually, the nitrogen use efficiency (NUE) was increased by 23.1%~60.2% and the agronomic nitrogen use efficiency (NAE) raised by 19.1%~30.5%. CRNF1 treatment significantly increased the SPDA value and total chlorophyll content by 6.5% and 10.1%, respectively, at the late growth stage of rape. Compared with the Urea treatment, CRNF1 treatment significantly increased the soil microbial nitrogen by 142.5% at the late growth stage of rape. In addition, the activity of soil urease, FDA hydrolase in the CRNF application treatment were significantly increased by 8.4%~12.9% and 24.5%~32.4%, respectively, at the late growth stage of rape. These results demonstrate that under the condition of total nitrogen application rate unchanged, application of CRNF mainly increased soil available nitrogen content at later growth stages, improved photosynthesis, enhanced soil microbial biomass and enzyme activity, thereby increased nitrogen uptake of rape, increased NUE and dry matter accumulation of rape, and increased grain yield.