Effects of nitrogen application on source-sink relationship, dry matter accumulation and yield of Yangliangyou-6 and Xinliangyou-6 were studied. The total nitrogen fertilizer was 600 kg/hm² urea and applied with several modes. The results indicated that the postponing N application reduced the tiller number and increased panicle bearing tiller rate compared with total nitrogen applied as basal fertilizer. This type of nitrogen application also increased the effective leaf area of full heading date, coordinated the source-sink relationship, and increased the dry matter accumulation of middle period and final stage and the yield of rice significantly. Among the tested nitrogen application modes, the treatment with a ratio of basal fertilizer: tillering fertilizer: spikelet-promoting fertilizer: spikelet-protecting fertilizer: grain fertilizer at 3︰1︰1︰1︰0 obtained the highest grain yield, and the yields of Yangliangyou-6 and Xinliangyou-6 were increased by 22.86% and 27.15%. While that with the ratio at 1︰1︰0︰0︰0 had the lowest grain yield, and the yields of the two varieties of rice were decreased by 7.69% and 6.32%.