Aquatic macrophytes grow in an environment where nitrogen can be available at quantitatively significant concentrations around both roots and shoots. Because of the generally higher concentration of NO 3 − in the bulk water compared to the sediment, and because of the energetic advantage of reducing NO 3 − in shoots compared to roots, aquatic plants were expected primarily to reduce NO 3 − in shoots. To test this hypothesis, nitrate reductase activity (NRA) of roots and shoots was measured in vitro on 18 aquatic macrophytes, representing 12 species. The plants were collected at seven locations with bulk water [NO 3 −] ranging from 0 to 550 μM and interstitial water [NO 3 −] ranging from 2 to 400 μM. All plants exhibited low NRA (<2 μmol NO 2 − g −1 DW h −1) in both roots and shoots, except for the amphibious species, Cardamine amare, where shoot NRA reached 24 μmol NO 2 − g −1 DW h −1. Higher NRA was however inducible, increasing 2–17 times in five selected species when induced with 500 μM NO 3 −. In 11 out of 17 plants with measurable NRA, the shoot:root NRA ratio was <1, showing that despite the proposed advantages of shoot NO 3 − reduction, root reduction is apparently still of importance in aquatic plants.