Evidence shows that many tree species can take up organic nitrogen (N) in the form of free amino acids from soils, but few studies have been conducted to compare organic and inorganic N uptake patterns in temperate and tropical tree species in relation to mycorrhizal status and successional state. We labeled intact tree roots by brief 15N exposures using field hydroponic experiments in a temperate forest and a tropical forest in China. A total of 21 dominant tree species were investigated, 8 in the temperate forest and 13 in the tropical forest. All investigated tree species showed highest uptake rates for NH4+ (ammonium), followed by glycine and NO3- (nitrate). Uptake of NH4+ by temperate trees averaged 12.8 μg N g-1 dry weight (d.w.) root h-1, while those by tropical trees averaged 6.8 μg N g-1 d.w. root h-1. Glycine uptake rates averaged 3.1 μg N g-1 d.w. root h-1 for temperate trees and 2.4 μg N g-1 d.w. root h-1 for tropical trees. NO3- uptake was the lowest (averaging 0.8 μg N g-1 d.w. root h-1 for temperate trees and 1.2 μg N g-1 d.w. root h-1 for tropical trees). Uptake of NH4+ accounted for 76% of the total uptake of all three N forms in the temperate forest and 64% in the tropical forest. Temperate tree species had similar glycine uptake rates as tropical trees, with the contribution being slightly lower (20% in the temperate forest and 23% in the tropical forest). All tree species investigated in the temperate forest were ectomycorrhizal and all species but one in the tropical forest were arbuscular mycorrhizal (AM). Ectomycorrhizal trees showed significantly higher NH4+ and lower NO3- uptake rates than AM trees. Mycorrhizal colonization rates significantly affected uptake rates and contributions of NO3- or NH4+, but depended on forest types. We conclude that tree species in both temperate and tropical forests preferred to take up NH4+, with organic N as the second most important N source. These findings suggest that temperate and tropical forests demonstrate similar N uptake patterns although they differ in physiology of trees and soil biogeochemical processes., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)