Nitrogen retention across a gradient of ¹⁵N additions to an unpolluted temperate forest soil in Chile

Accelerated nitrogen (N) inputs can drive nonlinear changes in N cycling, retention, and loss in forest ecosystems. Nitrogen processing in soils is critical to understanding these changes, since soils typically are the largest N sink in forests. To elucidate soil mechanisms that underlie shifts in N cycling across a wide gradient of N supply, we added ¹⁵NH₄ ¹⁵NO₃ at nine treatment levels ranging in geometric sequence from 0.2 kg to 640 kg N·ha^-1·yr^-1 to an unpolluted old-growth temperate forest in southern Chile. We recovered roughly half of ¹⁵N tracers in 0-25 cm of soil, primarily in the surface 10 cm. Low to moderate rates of N supply failed to stimulate N leaching, which suggests that most unrecovered ¹⁵N was transferred from soils to unmeasured sinks above ground. However, soil solution losses of nitrate increased sharply at inputs >160 kg N·ha^-1·yr ^-1, corresponding to a threshold of elevated soil N availability and declining ¹⁵N retention in soil. Soil organic matter (<5.6 mm) dominated tracer retention at low rates of N input, but coarse roots and particulate organic matter became increasingly important at higher N supply. Coarse roots and particulate organic matter together accounted for 38% of recovered ¹⁵N in soils at the highest N inputs and may explain a substantial fraction of the "missing N" often reported in studies of fates of N inputs to forests. Contrary to expectations, N additions did not stimulate gross N cycling, potential nitrification, or ammonium oxidizer populations. Our results indicate that the nonlinearity in N retention and loss resulted directly from excessive N supply relative to sinks, independent of plant-soil-microbial feedbacks. However, N additions did induce a sharp decrease in microbial biomass C:N that is predicted by N saturation theory, and which could increase long-term N storage in soil organic matter by lowering the critical C:N ratio for net N mineralization. All measured sinks accumulated ¹⁵N tracers across the full gradient of N supply, suggesting that short-term nonlinearity in N retention resulted from saturation of uptake kinetics, not uptake capacity, in plant, soil, and microbial pools.