Elevated CO2 concentration [e(CO2)] often promotes plant growth with a decrease in tissue N concentration. In this study, three experiments, two under hydroponic and one in well-watered soil, including various levels or patterns of CO2, humidity, and N supply were conducted on wheat (Triticum aestivum L.) to explore the mechanisms of e[CO2]-induced N deficiency (ECIND). Under hydroponic conditions, N uptake remained constant even as transpiration was limited 40% by raising air relative humidity and only was reduced about 20% by supplying N during nighttime rather than daytime with a reduction of 85% in transpiration. Compared to ambient CO2 concentration, whether under hydroponic or well-watered soil conditions, and whether transpiration was kept stable or decreased to 12%, e[CO2] consistently led to more N uptake and higher biomass, while lower N concentration was observed in aboveground organs, especially leaves, as long as N supply was insufficient. These results show that, due to compensation caused by active uptake, N uptake can be uncoupled from water uptake under well-watered conditions, and changes in transpiration therefore do not account for ECIND. Similar or lower tissue NO3--N concentration under e[CO2] indicated that NO3- assimilation was not limited and could therefore also be eliminated as a major cause of ECIND under our conditions. Active uptake has the potential to bridge the gap between N taken up passively and plant demand, but is limited by the energy required to drive it. Compared to ambient CO2 concentration, the increase in N uptake under e[CO2] failed to match the increase of carbohydrates, leading to N dilution in plant tissues, the apparent dominant mechanism explaining ECIND. Lower N concentration in leaves rather than roots under e[CO2] validated that ECIND was at least partially also related to changes in resource allocation, apparently to maintain root uptake activity and prevent more serious N deficiency.
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