The effects of short-term boron deficiency on several aspects (growth, biomass allocation, metabolite concentrations, gene expression, enzyme activities) related with nitrate assimilation were studied in tobacco (Nicotiana tabacum L.) plants in order to know the early changes caused by this mineral deficiency. For this purpose, plants were grown hydroponically in a nutrient solution supplemented with 10 microM boron and then transferred to a boron-free medium for 1-5 days. Nitrate concentration decreased in both leaves and roots under boron deficiency, which was not observed in control plants. This correlated with the lower net nitrate uptake rate found in boron-deficient plants when compared to boron-sufficient ones. Results suggest that boron deficiency decreases net nitrate uptake by declining the activity of nitrate transporters rather than affecting their transcript levels. This is supported by a drop in the levels of root PMA2 transcript during the boron deficient treatment, which could lead to a decrease in the plasma membrane H+-ATPase activity necessary to get protons out of cell for the cotransport with nitrate inwards. In addition, boron deficiency led to an increase in root Asn content and a decline in glutamine synthetase activity when compared to control plants, which suggest that this mineral deficiency may promote ammonium assimilation via asparagine synthetase in tobacco roots.