Nitrous oxide (N2O) is a strong greenhouse effective gas (GHG); the primary human source of N2O is agricultural activities. Excessive nitrogen (N) fertilization of agricultural land is now widely recognized as a major contributor. In soil, the microbial processes of nitrification and denitrification are the principal sources of N2O. However, it remains poorly understood how conventional hydroponics influences GHG emission. Here, we compared GHG fluxes from soil and rockwool used for hydroponics under identical nutrient conditions. Tomato plants (Solanum lycopersicum, momotaro) were grown in soil or by hydroponics using rockwool. In situ emissions of CH4, CO2, and N2O, and the abundance of genes involved in nitrification and denitrification were measured during cultivation. Hydroponics with rockwool mitigated CO2 emission by decreasing the microbial quantity in the rhizosphere. Dilution of the nutrient solution significantly decreased N2O emission from rockwool. Although proliferation of nitrifiers or denitrifiers in the rhizosphere did not induce N2O emission, reuse or long-term use of rockwool induced a 3.8-fold increase in N2O emission. Our data suggest that hydroponics has a lower environmental impact and that adequate fertilizer application, rather than bacterial control, governs N2O fluxes in hydroponic cultivation using rockwool.