Although N, especially mixtures of NO3 and NH4, can increase the productivity of maize (Zea mays L.), it is unclear when this N needs to be available for maximum benefit. This study was conducted to determine the importance of preanthesis N form and the need for available postanthesis N on the productivity of maize. Four single-cross hybrids (B73 x LH51, LH74 x LH51, LH74 x LH82, and LHE136 x LH82) were grown in field-hydroponics and supplied N as either all NO3 or an equal mixture of NO3 and NH4 (mixed N nutrition) until anthesis, whereupon N was withdrawn from one-half of the plants. At physiological maturity, whole shoots were harvested, separated into component fractions, weighed, and analyzed for yield, yield components, and macronutrient composition. Compared with all NO3, mixed N nutrition increased grain yield of all hybrids whether the N was available full season (average of 11%) or only during vegetative growth (average of 10%). In contrast, regardless of the N form, N deprivation during grain fill decreased grain yield for three of the four hybrids (range of 6 to 19%) but increased yield (5%) for LHE136 x LH82. Changes in sink-source partitioning, kernel number, and plant N accumulation were closely related to both yield enhancement associated with mixed N nutrition, and yield reduction due to N removal. Therefore, these data show that N form, timing of N availability and genotype are all factors that interact to influence N use and productivity of maize.