Response of grain yield and milled-rice protein content to nitrogen topdress (N) timing at panicle initiation stage (PIS) is critical for quantifying real-time N requirement for target grain yield and milled-rice protein content. Two split-split-plot experiments with three replications, one in 2004 and the other in 2005, were conducted in Experimental Farm, Seoul National University, Suwon, Korea. The experiments included three N rates at tillering stage (TS), three N timing treatments at panicle initiation stage (PIS) and two rice cultivars. The N rates at TS, N timing at PIS, and rice cultivars were randomly assigned to main plot, sub plot, and sub-sub plot, respectively. Results showed that the delayed N application at PIS reduced grain yield in 2004 and increased milled-rice protein content in both years significantly at 0.05 probability level. The calculated optimum N timing at PIS from pooled data by N rates and rice cultivars in two years was at 28 days before heading (DBH). However, real-time of N timing at PIS was dependent on plant growth and N status around PIS that in turn was dependent on applied N rates at TS. The optimum N timing at PIS was at 30 DBH for no N treatments at TS while at 27 DBH for 3.6 and 7.2 kg N/10a treatments and at 27 and 29 DBH for Hwaseongbyeo and Daeanbyeo, respectively. In general, earlier applied N at PIS resulted in lower milled-rice protein content but the highest grain yield was expected to be obtained when N topdress at PIS was applied at the time when shoot N concentration started to drop below about 23 mg/g due to dilution effect after transplanting. In conclusion, the results of our experiments imply that the currently recommended N topdress time (24DBH) at PIS in Korea should be reconsidered for the higher grain yield and the better quality of rice.