Abstract: The temporal and spatial distribution of N accumulation in flue-cured tobacco was investigated and the relationship of N accumulation in stalk and leaves with N nutrition status of tobacco plant was studied in field at different N fertilizer levels by using cv. K326 as experimental material. The results showed that the N accumulation amount in stalk and leaves was upper layer> middle layer> lower layer. The N accumulation amount in all organs and layers increased first and then decreased with the proceeding of growth and development, and the maximum N accumulation appeared obviously earlier in leaves than in stalk at each layer. The N accumulation amount in plant extremely significantly positively correlated to the N accumulation amount in leaves and significantly positively correlated to the N accumulation amount in stalk, which extremely significantly positively correlated to the N accumulation amount in middle layer leaves and significantly positively correlated to the N accumulation amount in upper layer leaves, while its relationship with that in lower layer leaves was not significant. It indicated that the N accumulation amount in middle layer leaves reflected the N accumulation amount in whole plant in general.