Leaf N:P ratio does not predict productivity trends across natural terrestrial ecosystems.

Nitrogen (N) and phosphorus (P) are crucial nutrients for regulating plant growth. The classic growth rate hypothesis (GRH) proposes that fast-growing organisms have lower N:P ratios, and it is promising to predict net primary productivity (NPP) using the leaf N:P ratio at the community level (N:PCom ). However, whether leaf N:P ratio can predict NPP in natural ecosystems on a large scale remains nebulous. Here, we systematically calculated leaf N:PCom (community biomass-weighted mean and species arithmetic mean) using the consistently measured data of 2192 plant species-site combinations and productivity (biomass-based aboveground NPP and flux-based NPP) in 66 natural ecosystems in China. Unexpectedly, leaf N:PCom hardly predicted productivity in natural ecosystems due to their weak correlation, although significantly negative or positive relationships across different ecosystems were observed. The ambiguous relationship between leaf N:P and species dominance reflected a luxury consumption of N and P in turnover and structure in natural communities, unlike what GRH suggests. Climate, soil, and leaf nutrients (rather than N:P) influenced productivity, which highlighted the importance of external environment and nutrient constrains. Our findings pose a major challenge for leaf N:PCom as a direct parameter in productivity models and further question the direct application of classic hypotheses in short-term experiments or model species to long-term and complex natural ecosystems.

Legume-microbiome interactions unlock mineral nutrients in regrowing tropical forests.

Legume trees form an abundant and functionally important component of tropical forests worldwide with N2-fixing symbioses linked to enhanced growth and recruitment in early secondary succession. However, it remains unclear how N2-fixers meet the high demands for inorganic nutrients imposed by rapid biomass accumulation on nutrient-poor tropical soils. Here, we show that N2-fixing trees in secondary Neotropical forests triggered twofold higher in situ weathering of fresh primary silicates compared to non-N2-fixing trees and induced locally enhanced nutrient cycling by the soil microbiome community. Shotgun metagenomic data from weathered minerals support the role of enhanced nitrogen and carbon cycling in increasing acidity and weathering. Metagenomic and marker gene analyses further revealed increased microbial potential beneath N2-fixers for anaerobic iron reduction, a process regulating the pool of phosphorus bound to iron-bearing soil minerals. We find that the Fe(III)-reducing gene pool in soil is dominated by acidophilic Acidobacteria, including a highly abundant genus of previously undescribed bacteria, Candidatus Acidoferrum, genus novus. The resulting dependence of the Fe-cycling gene pool to pH determines the high iron-reducing potential encoded in the metagenome of the more acidic soils of N2-fixers and their nonfixing neighbors. We infer that by promoting the activities of a specialized local microbiome through changes in soil pH and C:N ratios, N2-fixing trees can influence the wider biogeochemical functioning of tropical forest ecosystems in a manner that enhances their ability to assimilate and store atmospheric carbon.

Rapid nitrogen fixation by canopy microbiome in tropical forest determined by both phosphorus and molybdenum.

Biological nitrogen fixation is critical for the nitrogen cycle of tropical forests, yet we know little about the factors that control the microbial nitrogen fixers that colonize the microbiome of leaves and branches that make up a forest canopy. Forest canopies are especially prone to nutrient limitation because they are (1) disconnected from soil nutrient pools and (2) often subject to leaching. Earlier studies have suggested a role of phosphorus and molybdenum in controlling biological N-fixation rates, but experimental confirmation has hitherto been unavailable. Here we present the results of a manipulation of canopy nutrient availability. Our findings demonstrate a primary role of phosphorus in constraining overall N fixation by canopy cyanobacteria, but also a secondary role of molybdenum in determining per-cell fixation rates. A conservative evaluation suggests that canopy fixation can contribute to significant N fluxes at the ecosystem level, especially as bursts following atmospheric inputs of nutrient-rich dust.

Phosphatase activity and nitrogen fixation reflect species differences, not nutrient trading or nutrient balance, across tropical rainforest trees.

A fundamental biogeochemical paradox is that nitrogen-rich tropical forests contain abundant nitrogen-fixing trees, which support a globally significant tropical carbon sink. One explanation for this pattern holds that nitrogen-fixing trees can overcome phosphorus limitation in tropical forests by synthesizing phosphatase enzymes to acquire soil organic phosphorus, but empirical evidence remains scarce. We evaluated whether nitrogen fixation and phosphatase activity are linked across 97 trees from seven species, and tested two hypotheses for explaining investment in nutrient strategies: trading nitrogen-for-phosphorus or balancing nutrient demand. Both strategies varied across species but were not explained by nitrogen-for-phosphorus trading or nutrient balance. This indicates that (1) studies of these nutrient strategies require broad sampling within and across species, (2) factors other than nutrient trading must be invoked to resolve the paradox of tropical nitrogen fixation, and (3) nitrogen-fixing trees cannot provide a positive nitrogen-phosphorus-carbon feedback to alleviate nutrient limitation of the tropical carbon sink.

Phosphorus and species regulate N2 fixation by herbaceous legumes in longleaf pine savannas.

Longleaf pine savannas house a diverse community of herbaceous N2-fixing legume species that have the potential to replenish nitrogen (N) losses from fire. Whether legumes fill this role depends on the factors that regulate symbiotic fixation, including soil nutrients such as phosphorus (P) and molybdenum (Mo) and the growth and fixation strategies of different species. In greenhouse experiments, we determined how these factors influence fixation for seven species of legumes grown in pure field soil from two different regions of the southeastern US longleaf pine ecosystem. We first added P and Mo individually and in combination, and found that P alone constrained fixation. Phosphorus primarily influenced fixation by regulating legume growth. Second, we added N to plants and found that species either downregulated fixation (facultative strategy) or maintained fixation at a constant rate (obligate strategy). Species varied nearly fourfold in fixation rate, reflecting differences in growth rate, taxonomy and fixation strategy. However, fixation responded strongly to P addition across all species in our study, suggesting that the P cycle regulates N inputs by herbaceous legumes.

Taxonomic identity determines N2 fixation by canopy trees across lowland tropical forests.

Legumes capable of fixing atmospheric N2 are abundant and diverse in many tropical forests, but the factors determining ecological patterns in fixation are unresolved. A long-standing idea is that fixation depends on soil nutrients (N, P or Mo), but recent evidence shows that fixation may also differ among N2-fixing species. We sampled canopy-height trees across five species and one species group of N2-fixers along a landscape P gradient, and manipulated P and Mo to seedlings in a shadehouse. Our results identify taxonomy as the major determinant of fixation, with P (and possibly Mo) only influencing fixation following tree-fall disturbances. While 44% of trees did not fix N2, other trees fixed at high rates, with two species functioning as superfixers across the landscape. Our results raise the possibility that fixation is determined by biodiversity, evolutionary history and species-specific traits (tree growth rate, canopy stature and response to disturbance) in the tropical biome.

Nitrogen and phosphorus limitation over long-term ecosystem development in terrestrial ecosystems.

Nutrient limitation to net primary production (NPP) displays a diversity of patterns as ecosystems develop over a range of timescales. For example, some ecosystems transition from N limitation on young soils to P limitation on geologically old soils, whereas others appear to remain N limited. Under what conditions should N limitation and P limitation prevail? When do transitions between N and P limitation occur? We analyzed transient dynamics of multiple timescales in an ecosystem model to investigate these questions. Post-disturbance dynamics in our model are controlled by a cascade of rates, from plant uptake (very fast) to litter turnover (fast) to plant mortality (intermediate) to plant-unavailable nutrient loss (slow) to weathering (very slow). Young ecosystems are N limited when symbiotic N fixation (SNF) is constrained and P weathering inputs are high relative to atmospheric N deposition and plant N:P demand, but P limited under opposite conditions. In the absence of SNF, N limitation is likely to worsen through succession (decades to centuries) because P is mineralized faster than N. Over long timescales (centuries and longer) this preferential P mineralization increases the N:P ratio of soil organic matter, leading to greater losses of plant-unavailable N versus P relative to plant N:P demand. These loss dynamics favor N limitation on older soils despite the rising organic matter N:P ratio. However, weathering depletion favors P limitation on older soils when continual P inputs (e.g., dust deposition) are low, so nutrient limitation at the terminal equilibrium depends on the balance of these input and loss effects. If NPP switches from N to P limitation over long time periods, the transition time depends most strongly on the P weathering rate. At all timescales SNF has the capacity to overcome N limitation, so nutrient limitation depends critically on limits to SNF.

Molybdenum and phosphorus interact to constrain asymbiotic nitrogen fixation in tropical forests.

Biological di-nitrogen fixation (N(2)) is the dominant natural source of new nitrogen to land ecosystems. Phosphorus (P) is thought to limit N(2) fixation in many tropical soils, yet both molybdenum (Mo) and P are crucial for the nitrogenase reaction (which catalyzes N(2) conversion to ammonia) and cell growth. We have limited understanding of how and when fixation is constrained by these nutrients in nature. Here we show in tropical forests of lowland Panama that the limiting element on asymbiotic N(2) fixation shifts along a broad landscape gradient in soil P, where Mo limits fixation in P-rich soils while Mo and P co-limit in P-poor soils. In no circumstance did P alone limit fixation. We provide and experimentally test a mechanism that explains how Mo and P can interact to constrain asymbiotic N(2) fixation. Fixation is uniformly favored in surface organic soil horizons--a niche characterized by exceedingly low levels of available Mo relative to P. We show that soil organic matter acts to reduce molybdate over phosphate bioavailability, which, in turn, promotes Mo limitation in sites where P is sufficient. Our findings show that asymbiotic N(2) fixation is constrained by the relative availability and dynamics of Mo and P in soils. This conceptual framework can explain shifts in limitation status across broad landscape gradients in soil fertility and implies that fixation depends on Mo and P in ways that are more complex than previously thought.

Potassium, phosphorus, or nitrogen limit root allocation, tree growth, or litter production in a lowland tropical forest.

We maintained a factorial nitrogen (N), phosphorus (P), and potassium (K) addition experiment for 11 years in a humid lowland forest growing on a relatively fertile soil in Panama to evaluate potential nutrient limitation of tree growth rates, fine-litter production, and fine-root biomass. We replicated the eight factorial treatments four times using 32 plots of 40 x 40 m each. The addition of K was associated with significant decreases in stand-level fine-root biomass and, in a companion study of seedlings, decreases in allocation to roots and increases in height growth rates. The addition of K and N together was associated with significant increases in growth rates of saplings and poles (1-10 cm in diameter at breast height) and a further marginally significant decrease in stand-level fine-root biomass. The addition of P was associated with a marginally significant (P = 0.058) increase in fine-litter production that was consistent across all litter fractions. Our experiment provides evidence that N, P, and K all limit forest plants growing on a relatively fertile soil in the lowland tropics, with the strongest evidence for limitation by K among seedlings, saplings, and poles.

Increased plant growth from nitrogen addition should conserve phosphorus in terrestrial ecosystems.

Inputs of available nitrogen (N) to ecosystems have grown over the recent past. There is limited general understanding of how increased N inputs affect the cycling and retention of other potentially limiting nutrients. Using a plant-soil nutrient model, and by explicitly coupling N and phosphorus (P) in plant biomass, we examine the impact of increasing N supply on the ecosystem cycling and retention of P, assuming that the main impact of N is to increase plant growth. We find divergent responses in the P cycle depending on the specific pathway by which nutrients are lost from the ecosystem. Retention of P is promoted if the relative propensity for loss of plant available P is greater than that for the loss of less readily available organic P. This is the first theoretical demonstration that the coupled response of ecosystem-scale nutrient cycles critically depends on the form of nutrient loss. P retention might be lessened, or reversed, depending on the kinetics and size of a buffering reactive P pool. These properties determine the reactive pool's ability to supply available P. Parameterization of the model across a range of forest ecosystems spanning various environmental and climatic conditions indicates that enhanced plant growth due to increased N should trigger increased P conservation within ecosystems while leading to more dissolved organic P loss. We discuss how the magnitude and direction of the effect of N may also depend on other processes.