A gas analysis system was built to study the relationship between the reductant cost of NO(3)(-) assimilation and the measured rate of CO(2) and O(2) exchange in roots, leaves, and stems+ petioles of soybean (Glycine max L. Merr. cv Maple glen) plants. The measurements were used to calculate the diverted reductant utilization rate (DRUR = 4*[measured rate of CO(2) + measured rate of O(2)], in moles of high-energy electron [e(-)] per gram per hour) in plants in the presence (N+) and absence (N-) of NO(3)(-). The differences in DRUR between the N+ and N- treatments provided a measure of the NO(3)(-)-coupled DRUR of 25-d-old plants, whereas a (15)NO(3)(-)-enriched nutrient solution was used to obtain an independent measure of the rate of NO(3)(-) assimilation. The measured reductant cost for the whole plant was 9.6 e(-) per N assimilated, a value within the theoretical range of four to 10 e(-) per N assimilated. The results predicted that shoots accounted for about 55% of the whole-plant NO(3)(-) assimilation over the entire day, with shoots dominating in the light, and roots in the dark. The gas analysis approach described here holds promise as a powerful, noninvasive tool to study the regulation of NO(3)(-) assimilation in plant tissue.