To promote the commercial application dissemination of new flue-cured tobacco variety YH05, the effects of nitrogen application rates and different nitrogen form ratios (NO3--N) on the availability of nitrogen fertilizer, growth, yield and production value of YH05 were investigated by the methods of field experiment. The results showed that nitrogen application rate, different nitrogen form ratios and the interaction between the two factors not only affected nitrogen contents, but also affected nitrogen absorption and transport. Nitrogen application rate was the main influential factor on nitrogen agronomic efficiency, while apparent efficiency of nitrogen was affected by nitrogen application rate, different nitrogen forms ratio and the interaction. The three factors affected agronomic and economic characters differently. The results suggested that the optimal nitrogen application rate of YH05 was 7 g of per tobacco plant and 25% of nitrate nitrogen ratio when considering agronomic characters, economic characters and nitrogen fertilizer availability.