A correct determination of nitrogen (N) fertilization thresholds in wheat that is based on objective yield produces efficient use of this nutrient. Nitrogen fertilization recommendations for traditional wheat require determination of nitrate (NO 3 − )-N availability at 60 cm deep at planting time. However, this methodology is complicated, expensive, and time-consuming; thus, the determination of NO 3 − -N level at a lesser depth and at a different time would be desirable. The goals of this work were to determine available N in soil thresholds for traditional and French germplasm wheats and the feasibility of diagnosing N requirements by measuring NO 3 − -N at 40 cm deep, at planting or tillering times, in the southeastern Pampas. The experiments were factorial combinations of N rates and fertilization times (planting and tillering) at different sites and years during 2002-2006. Nitrogen fertilization significantly increased grain yield and protein content. French varieties presented greater grain yield (23%), lower protein content (11%), and greater yield per N unit, indicating greater N-use efficiency (NUE) than traditional varieties. A similar relationship was determined between grain yield and available N at both sampling depths. This might be explained by the strong association between NO 3 − -N content at 60 and 40 cm deep at both sampling dates. Maximum yield and available N determined at 60 or 40 cm soil deep showed that thresholds were lower for tillering than for planting, regardless of the genotype (152 and 174 kg of available N, respectively). Available N thresholds for 95% of maximum yield were less at 0-40 cm deep than at 0-60 cm deep (10 and 14 kg N ha −1 for traditional and French genotypes, respectively). The results of this experiment suggest the possibility of diagnosing N requirements for wheat by measuring NO 3 − -N content at 40 cm deep, instead of the usual 60 cm, for both traditional and French genotypes.