Soil nitrogen balance resulting from N fixation and rhizodeposition by the symbiotic association Anthyllis vulneraria/Mesorhizobium metallidurans grown in highly polluted Zn,Pb and Cd mine tailings

Background and aims

The association of the legume Anthyllis vulneraria and the grass Festuca arvernensis, was found to be very efficient for the phytostabilisation of highly multi-metal contaminated mine tailings. Our objective was to quantify the contribution of Anthyllis inoculated with its symbiotic bacteria Mesorhizobium metallidurans to the soil N pool and to test whether a starter nitrogen fertilization may improve symbiotic nitrogen fixation and the growth of Festuca.

Methods

Plants of Festuca and of Anthyllis inoculated with M. metallidurans were grown separately during eight months in pots filled with mine contaminated soil. Estimation of the N fluxes was realized using ¹⁵?N isotopic methods.

Results

Starter N fertilization (28 kg N ha^?1) improved symbiotic N₂ fixation and the growth of both species. Belowground N balance (N rhizodeposition – soil N uptake) of the non-fertilized Anthyllis at maturity was negative (?30.6 kg N ha^?1). However, the amount of N derived from fixation, including above- and belowground parts, was 78.6 kg N ha^?1, demonstrating the ability of this symbiotic association to improve soil N content after senescence.

Conclusions

i) soil N enrichment by the N₂-fixing symbiotic association occurs after plant senescence, when decaying leaves and shoots are incorporated into the soil; ii) application of a starter fertilization is an efficient solution to improve phytostabilisation of highly contaminated sites.     

Nitrogen contributions from faba bean (Vicia faba L.) reliant on soil rhizobia or inoculation

Background and Aims

Understanding the impact of soil rhizobial populations and inoculant rhizobia in supplying sufficient nodulation is crucial to optimising N₂ fixation by legume crops. This study explored the impact of different rates of inoculant rhizobia and contrasting soil rhizobia on nodulation and N₂ fixation in faba bean (Vicia faba L.).

Methods

Faba beans were inoculated with one of seven rates of rhizobial inoculation, from no inoculant to 100 times the normal rate of inoculation, sown at two field sites, with or without soil rhizobia present, and their nodulation and N₂ fixation assessed.

Results

At the site without soil rhizobia, inoculation increased nodule number and increased N₂ fixation from 21 to 129 kg shoot N ha^?1, while N₂ fixation increased from 132 to 218 kg shoot N ha^?1 at the site with high background soil rhizobia. At the site without soil rhizobia, inoculation increased concentrations of shoot N from 14 to 24 mg g^?1, grain N from 32 to 45 mg g^?1, and grain yields by 1.0 Mg (metric tonne) ha^?1. Differences in nodulation influenced the contributions of fixed N to the system, which varied from the net removal of 20 kg N ha^?1 from the system in the absence of rhizobia, to a net maximum input of 199 kg N ha^?1 from legume shoot and root residues, after accounting for removal of N in grain harvest.

Conclusions

The impact of inoculation and soil rhizobia strongly influenced grain yield, grain N concentration and the potential contributions of legume cropping to soil N fertility. In soil with resident rhizobia, N₂ fixation was improved only with the highest inoculation rate.     

Nitrous oxide emissions and nitrate leaching from a rain-fed wheat-maize rotation in the Sichuan Basin, China

Aims

A 3-year field experiment (October 2004–October 2007) was conducted to quantify N₂O fluxes and determine the regulating factors from rain-fed, N fertilized wheat-maize rotation in the Sichuan Basin, China.

Methods

Static chamber-GC techniques were used to measure soil N₂O fluxes in three treatments (three replicates per treatment): CK (no fertilizer); N150 (300 kg N fertilizer ha^?1 yr^?1 or 150 kg N?ha^?1 per crop); N250 (500 kg N fertilizer ha^?1 yr^?1 kg or 250 kg N?ha^?1 per crop). Nitrate (NO ₃ ^? ) leaching losses were measured at nearby sites using free-drained lysimeters.

Results

The annual N₂O fluxes from the N fertilized treatments were in the range of 1.9 to 6.7 kg N?ha^?1 yr^?1 corresponding to an N₂O emission factor ranging from 0.12 % to 1.06 % (mean value: 0.61 %). The relationship between monthly soil N₂O fluxes and NO ₃ ^- leaching losses can be described by a significant exponential decaying function.

Conclusions

The N₂O emission factor obtained in our study was somewhat lower than the current IPCC default emission factor (1 %). Nitrate leaching, through removal of topsoil NO ₃ ^? , is an underrated regulating factor of soil N₂O fluxes from cropland, especially in the regions where high NO ₃ ^- leaching losses occur.     

Nitrogen fixation and transfer in grass-clover leys under organic and conventional cropping systems

Background and aim

Symbiotic dinitrogen (N₂) fixation is the most important external N source in organic systems. Our objective was to compare symbiotic N₂ fixation of clover grown in organically and conventionally cropped grass-clover leys, while taking into account nutrient supply gradients.

Methods

We studied leys of a 30-year-old field experiment over 2 years in order to compare organic and conventional systems at two fertilization levels. Using ¹⁵N natural abundance methods, we determined the proportion of N derived from the atmosphere (PNdfa), the amount of Ndfa (ANdfa), and the transfer of clover N to grasses for both red clover (Trifolium pratense L.) and white clover (Trifolium repens L.).

Results

In all treatments and both years, PNdfa was high (83 to 91 %), indicating that the N₂ fixation process is not constrained, even not in the strongly nutrient deficient non-fertilized control treatment. Annual ANdfa in harvested clover biomass ranged from 6 to 16 g?N m^?2. At typical fertilizer input levels, lower sward yield in organic than those in conventional treatments had no effect on ANdfa because of organic treatments had greater clover proportions. In two-year-old leys, on average, 51 % of N taken up by grasses was transferred from clover.

Conclusion

Both, organically and conventionally cropped grass-clover leys profited from symbiotic N₂ fixation, with high PNdfa, and important transfer of clover N to grasses, provided sufficient potassium- and phosphorus-availability to sustain clover biomass production.     

Nitrous oxide emissions during the non-rice growing seasons of two subtropical rice-based rotation systems in southwest China

Background and aims

High nitrous oxide (N₂O) emissions may occur during the non-rice growing season of Chinese rice-upland crop rotation systems. However, our understanding of N₂O emission during this season is poor due to a scarcity of available field N₂O measurements.

Methods

Using the static manual chamber-GC technique, seasonal N₂O emissions during the non-rice growing season were simultaneously measured at two adjacent rice-wheat and rice-rapeseed fields in southwest China for three consecutive annual rotation cycles (May 2005 to May 2008).

Results

Compared to the control, N fertilizer applications significantly enhanced soil N₂O emissions from both wheat and rapeseed systems. Seasonal cumulative N₂O fluxes from wheat systems were on average 2.6 kg N ha^?1 for the recommended practice (RP [150 kg N ha^?1]) and 5.0 kg N ha^?1 for the conventional practice (CP [250 kg N ha^?1]). Lower N₂O emissions were observed from the adjacent rapeseed systems. Average cumulative seasonal N₂O fluxes from rapeseed were 1.5 and 2.2 kg N ha^?1 for the RP and CP treatments, respectively. The first 3 weeks after N fertilization were the “hot moment” of N₂O emissions for both the wheat and rapeseed systems. The lowest yield-scaled N₂O fluxes for wheat were obtained at the RP treatment (mean: 0.81 kg N Mg^?1) while for rapeseed the CP treatment produced the lowest yield-scaled fluxes (mean: 0.79 kg N Mg^?1). On average, the direct N₂O emission factors (EF_d) for the wheat system (1.76 %) were over two times higher than for the rapeseed system (0.73 %).

Conclusions

Intercropping of rapeseed tends to result in lower N₂O emissions than wheat for rice-upland crop rotation systems of southwest China, indicating that either the N fertilization or the cropping system need to be considered not only for improving the estimate of regional and/or national N₂O fluxes but also for proposing the climate-smart agricultural management practice to reduce N₂O emissions from agricultural soils.     

Plant and soil responses of an alpine steppe on the Tibetan Plateau to multi-level nitrogen addition

Background

Although plant growth in alpine steppes on the Tibetan Plateau has been suggested to be sensitive to nitrogen (N) addition, the N limitation conditions of alpine steppes remain uncertain.

Methods

After 2 years of fertilization with NH₄NO₃ at six rates (0, 10, 20, 40, 80 and 160 kg N ha^?1 yr^?1), the responses of plant and soil parameters as well as N₂O fluxes were measured.

Results

At the vegetation level, N addition resulted in an increase in the aboveground N pool from 0.5?±?0.1 g m^?2 in the control plots to 1.9?±?0.2 g m^?2 in the plots at the highest N input rate. The aboveground C pool, biomass N concentration, foliar δ¹⁵N, soil NO₃ ^?-N and N₂O flux were also increased by N addition. However, as the N fertilization rate increased from 10 kg N ha^?1 yr^?1 to 160 kg N ha^?1 yr^?1, the N-use efficiency decreased from 12.3?±?4.6 kg C kg N^?1 to 1.6?±?0.2 kg C kg N^?1, and the N-uptake efficiency decreased from 43.2?±?9.7 % to 9.1?±?1.1 %. Biomass N:P ratios increased from 14.4?±?2.6 in the control plots to 20.5?±?0.8 in the plots with the highest N input rate. Biomass N:P ratios, N-uptake efficiency and N-use efficiency flattened out at 40 kg N ha^?1 yr^?1. Above this level, soil NO₃ ^?-N began to accumulate. The seasonal average N₂O flux of growing season nonlinearly increased with increased N fertilization rate and linearly increased with the weighted average foliar δ¹⁵N. At the species level, N uptake responses to relative N availability were species-specific. Biomass N concentration of seven out of the eight non-legume species increased significantly with N fertilization rates, while Kobresia macrantha and the one legume species (Oxytropics glacialis) remained stable. Both the non-legume and the legume species showed significant ¹⁵N enrichment with increasing N fertilization rate. All non-legume species showed significant increased N:P ratios with increased N fertilization rate, but not the legume species.

Conclusions

Our findings suggest that the Tibetan alpine steppes might be N-saturated above a critical N load of 40 kg N ha^?1 yr^?1. For the entire Tibetan Plateau (ca. 2.57 million km²), a low N deposition rate (10 kg N ha^?1 yr^?1) could enhance plant growth, and stimulate aboveground N and C storage by at least 1.1?±?0.3 Tg N yr^?1 and 31.5?±?11.8 Tg C yr^?1, respectively. The non-legume species was N-limited, but the legume species was not limited by N.     

Contribution of 15N-labelled leaf litter to N turnover, nitrous oxide emissions and N sequestration in a beech forest during eleven years

Aims

Decomposition of leaf litterfall plays a major role for nitrogen (N) dynamics in soils. However, little is known as to which extent beech leaf litter contributes to N turnover and nitrous oxide (N₂O) emissions within one decade after litterfall.

Methods

In 1997, we exchanged recently fallen leaf litter by ¹⁵N-labelled litter in a beech stand (Fagus sylvatica) at the Solling, Germany. Measurements were conducted 2–3 and 10–11 years after litter exchange.

Results

Two years after litter exchange, 92 % of added ¹⁵N was recovered in the surface 10 cm of the soil. The labelled N was primarily found in the upper part of the F layer of the moder type humus. Eleven years after litter exchange, 73 % of the added ¹⁵N was lost and the remaining 27 % was mainly recovered in the lower part of the F layer indicating N sequestration. The remaining leaf litter N was subject to measurable N mineralisation (2–3 % of litter N) and N₂O production (0.02 %). Between 0.3 % (eleventh year) and 0.6 % (second year) of total annual N₂O emissions were attributed to beech leaf litter of a single year.

Conclusions

Most of the annual N₂O emissions (1.33–1.54 kg N ha^?1 yr^?1) were probably derived from older soil N pools.     

Combination of nitrogen deposition and ultraviolet-B radiation decreased litter decomposition in subtropical China

Aims

The interactive effects of enhanced nitrogen (N) deposition and ultraviolet-B (UV-B) radiation on litter decomposition are still unknown. The aims are to test whether the interactive effects of the two environmental factors on litter decomposition and nutrient loss are stronger than that of each factor alone.

Methods

Experiment included five treatments: elevated UV-B radiation (UV-B, 10 % enhancement), low N addition (N1, 30 kg N ha^?1 year^?1), high N addition (N2, 60 kg N ha^?1 year^?1), the two combined treatments of the two factors (UV-B+N1 and UV-B+N2), and an unmanipulated control.

Results

The annual decomposition rates under combination of UV-B and N addition significantly decreased compared with that under UV-B and N additions for Pinus massoniana, and did also compared with that under UV-B but did not significantly differ with N additions for Cyclobalanopsis glauca. Negative effects of N additions alone on lignin degradation and P loss were partly offset but negative effect on N loss was further amplified when was combined with UV-B.

Conclusions

The combination of N deposition and UV-B radiation on litter decomposition and nutrient loss was significantly different from that of each factor alone without a general response pattern of decomposition, and was regulated by litter chemistry.     

Changes in stable nitrogen and carbon isotope ratios of plants and soil across a boreal forest fire chronosequence

Background and Aim

Nitrogen (N) and carbon (C) isotopic signatures (δ¹⁵N and δ¹³C) serve as powerful tools for understanding temporal changes in ecosystem processes, but how these signatures change across boreal forest chronosequences is poorly understood.

Methods

The δ¹⁵N, δ¹³C, and C/N ratio of foliage of eight dominant plant species, including trees, understory shrubs, and a moss, as well as humus, were examined across a 361 years fire-driven chronosequence in boreal forest in northern Sweden.

Results

The δ¹³C and C/N ratio of plants and humus increased along the chronosequence, suggesting increasing plant stress through N limitation. Despite increasing biological N fixation by cyanobacteria associated with feather mosses, δ¹⁵N showed an overall decline, and δ¹⁵N of the feather moss and associated vascular plants diverged over time from that of atmospheric N₂.

Conclusions

Across this chronosequence the N fixed by cyanobacteria is unlikely to be used by mosses and vascular plants without first undergoing mineralization and mycorrhizal transport, which would cause a change in δ¹⁵N signature due to isotopic fractionation. The decreasing trend of δ¹⁵N suggests that as the chronosequence proceeds, the plants may become more dependent on N transferred from mycorrhizal fungi or from N deposition.     

Nitrogen deposition promotes ecosystem carbon accumulation by reducing soil carbon emission in a subtropical forest

Background and aims

Tropical and subtropical forests are experiencing high levels of atmospheric nitrogen (N) deposition, but the responses of such forests ecosystems to N deposition remain poorly understood.

Methods

We conducted an 8-year field experiment examining the effect of experimental N deposition on plant growth, soil carbon dioxide efflux, and net ecosystem production (NEP) in a subtropical Chinese fir forest. The quantities of N added were 0 (control), 60, 120, and 240 kg ha^?1 year^?1.

Results

NEP was lowest under ambient conditions and highest with 240 kg of N ha^?1 year^?1 treatment. The net increase in ecosystem carbon (C) storage ranged from 9.2 to 16.4 kg C per kg N added in comparison with control. In addition, N deposition treatments significantly decreased heterotrophic respiration (by 0.69–1.85 t C ha^?1 year^?1) and did not affect plant biomass. The nitrogen concentrations were higher in needles than that in fine roots.

Conclusions

Our findings suggest that the young Chinese fir forest is carbon source and N deposition would sequester additional atmospheric CO₂ at high levels N input, mainly due to reduced soil CO₂ emission rather than increased plant growth, and the amount of sequestered C depended on the rate of N deposition.     

Assessment of nitrate leaching loss on a yield-scaled basis from maize and wheat cropping systems

Background and aims

It is so far a gap in knowledge to assess nitrate (NO₃ ^?) leaching loss linking with crop yield for a given cereal cropping system.

Methods

We conducted a meta-analysis on 32 published studies reporting both NO₃ ^? leaching losses and crop yields in the maize (N?=?20) and wheat (N?=?12) systems.

Results

On average, 22 % and 15 % of applied fertilizer N to wheat and maize systems worldwide are leached in the form of NO₃ ^?, respectively. The average area-scaled NO₃ ^- leaching loss for maize (57.4 kg N ha^?1) was approx. two times higher than for wheat (29.0 kg N ha^?1). While, if scaled to crop yields, the average yield-scaled NO₃ ^? losses were comparable between maize (5.40 kg N Mg^?1) and wheat (5.41 kg N Mg^?1) systems. Across all sites, the lowest yield-scaled NO₃ ^? leaching losses were observed at slightly suboptimal fertilization rates, corresponding to 90 % and 96 % of maximum maize or wheat yields, respectively.

Conclusions

Our findings suggest that small adjustments of agricultural N management practices can effectively reduce yield-scaled NO₃ ^? leaching losses. However, further targeted field experiments are still needed to identify at regional scale best agricultural management practices for reducing yield-scaled NO₃ ^? leaching losses in maize and wheat systems.     

N2 Fixation in Feather Mosses is a Sensitive Indicator of N Deposition in Boreal Forests

Nitrogen (N) fixation in the feather moss–cyanobacteria association represents a major N source in boreal forests which experience low levels of N deposition; however, little is known about the effects of anthropogenic N inputs on the rate of fixation of atmospheric N₂ in mosses and the succeeding effects on soil nutrient concentrations and microbial community composition. We collected soil samples and moss shoots of Pleurozium schreberi at six distances along busy and remote roads in northern Sweden to assess the influence of road-derived N inputs on N₂ fixation in moss, soil nutrient concentrations and microbial communities. Soil nutrients were similar between busy and remote roads; N₂ fixation was higher in mosses along the remote roads than along the busy roads and increased with increasing distance from busy roads up to rates of N₂ fixation similar to remote roads. Throughfall N was higher in sites adjacent to the busy roads but showed no distance effect. Soil microbial phospholipid fatty acid (PLFA) composition exhibited a weak pattern regarding road type. Concentrations of bacterial and total PLFAs decreased with increasing distance from busy roads, whereas fungal PLFAs showed no distance effect. Our results show that N₂ fixation in feather mosses is highly affected by N deposition, here derived from roads in northern Sweden. Moreover, as other measured factors showed only weak differences between the road types, atmospheric N₂ fixation in feather mosses represents a highly sensitive indicator for increased N loads to natural systems.     

Nitrogen deposition and soil carbon content affect nitrogen mineralization during primary succession in acid inland drift sand vegetation

Background and aims

Two inland dunes in the Netherlands receiving low (24) and high (41 kg N ha^?1 yr^?1) nitrogen (N) deposition were compared for N dynamics and microbial activity to investigate the potential effect of N on succession rate of the vegetation and loss of pioneer habitats.

Methods

Primary succession stages were sampled, including bare sand, and vegetation dominated by Polytrichum piliferum, Campylopus introflexus, lichens and grasses respectively, representing a series of vegetation types in undisturbed drift sand sites with succession starting on bare sand containing virtually no organic matter. Microbial characteristics and potential N mineralization were analysed in a laboratory experiment.

Results

Organic matter accumulated during succession, resulting in a lower pH and in higher microbial biomass (bacteria and fungi), respiration and net N mineralization. The increase in respiration and N mineralization was largely due to the development of an ectorganic layer in the middle stages of succession. The observed effects of N deposition were (1) decrease of microbial biomass, (2) higher net N mineralization per m², (3) higher levels of free nitrogen in the soil, and (4) a higher microbial N:P ratio.

Conclusions

Elevated N deposition leads to higher N availability which may cause accelerated succession.     

N2 fixation of pea hypernodulating mutants is more tolerant to root pruning than that of wild type

Background and aims

As a legume, pea plant has the ability to symbiotically fix N₂. However, symbiotic N₂ fixation is very sensitive to environmental stresses that affect plant growth, and there is little knowledge on the impact of root pruning on N₂ fixation and plant growth.

Methods

In this study, we removed half of the nodulated roots of pea wild-type Frisson and hypernodulating mutants P64, P118, and P121. Dinitrogen fixation was measured using ¹⁵N labeling and carbon assimilation and partitioning between plant organs using ¹³C labeling.

Results

Root pruning decreased N₂ fixation by ?46 to ?79 % in wild-type and mutants. Pea mutant P118 had a lower decrease of specific activity of N₂ fixation (?17 %) than both wild-type and other mutants (?36 to ?62 %). For all genotypes, root pruning increased root and nodule sinks strengths for carbon. For P118 and for P121, this was associated to higher nodule growth than for control plants, as measured 8 days after root pruning.

Conclusion

This is the first analysis of N₂-fixing plant response to root pruning. Importantly, we showed that some hypernodulating mutant pea lines (P118 and to a lesser extent P121) withstood this stress better than wild-type did.     

Seasonal variation in nitrogen fixation and effects of climate change in a subarctic heath

Background and aims

Nitrogen fixation associated with cryptogams is potentially very important in arctic and subarctic terrestrial ecosystems, as it is a source of new nitrogen (N) into these highly N limited systems. Moss-, lichen- and legume-associated N₂ fixation was studied with high frequency (every second week) during spring, summer, autumn and early winter to uncover the seasonal variation in input of atmospheric N₂ to a subarctic heath with an altered climate.

Methods

We estimated N₂ fixation from ethylene production by acetylene reduction assay in situ in a field experiment with the treatments: long- vs. short-term summer warming using plastic tents and litter addition (simulating expansion of the birch forest).

Results

N₂ fixation activity was measured from late April to mid November and 33 % of all N₂ was fixed outside the vascular plant growing season (Jun–Aug). This substantial amount underlines the importance of N₂ fixation in the cold period. Warming increased N₂ fixation two- to fivefold during late spring. However, long-term summer warming tended to decrease N₂ fixation outside the treatment (tents present) period. Litter alone did not alter N₂ fixation but in combination with warming N₂ fixation increased, probably because N₂ fixation became phosphorus limited under higher temperatures, which was alleviated by the P supply from the litter.

Conclusion

In subarctic heath, the current N₂ fixation period extends far beyond the vascular plant growing season. Climate warming and indirect effects such as vegetation changes affect the process of N₂ fixation in different directions and thereby complicate predictions of future N cycling.     

Nitrogen fertilisation reduces grass-induced N2 fixation of tree seedlings from semi-arid savannas

Aims

Coexistence of trees and grasses in nutrient-poor arid savannas may result in competition for soil N. While grasses may be more effective than woody plants in acquiring N from the soil, some leguminous woody species rely on N₂ fixation. We assessed the role of N₂ fixation in the N-budget of Acacia mellifera seedlings by varying N supply and grass competition.

Methods

The contribution of N₂ fixation to the N-budget of Acacia mellifera seedlings with varying N supply and grass competition was determined by measuring growth, nutrient concentrations, and ¹⁵N values.

Results

Tree seedlings were 4-fold taller and had 20-fold more biomass in the absence of grass. Tree foliar δ¹⁵N was lower with (?0.25?±?0.2‰, n?=?9) than without grasses (5.2?±?0.1‰, n?=?64). The contribution of N₂-fixation to the N budget decreased with increasing N supply. Greater reliance on N₂-fixation by trees in the presence of grasses did not result in greater biomass accumulation or tissue [N] relative to tree seedlings grown without grass competition. Tree seedlings competing with grass had significantly more negative δ¹³C (?29.5?±?0.6‰) than seedlings without grass competition (?28.8‰?±?0.5‰).

Conclusions

Induction of N₂-fixation by grass may have resulted from competition for nutrients. N₂-fixation enables tree seedlings to compensate for limited soil N and survive grass competition at a critical and vulnerable developmental stage of germination and establishment.     

Liebig’s law of the minimum applied to a greenhouse gas: alleviation of P-limitation reduces soil N2O emission

Background and aims

Emission of the greenhouse gas (GHG) nitrous oxide (N₂O) are strongly affected by nitrogen (N) fertilizer application rates. However, the role of other nutrients through stoichiometric relations with N has hardly been studied. We tested whether phosphorus (P) availability affects N₂O emission. We hypothesized that alleviation of plant P-limitation reduces N₂O emission through lowering soil mineral N concentrations.

Methods

We tested our hypothesis in a pot experiment with maize (Zea mays L.) growing on a P-limiting soil/sand mixture. Treatment factors included P and N fertilization and inoculation with Arbuscular Mycorrhizal Fungi (AMF; which can increase P uptake).

Results

Both N and P fertilization, as well as their interaction significantly (P?<?0.01) affected N₂O emission. Highest N₂O emissions (2.38 kg N₂O-N ha^?1) were measured at highest N application rates without P fertilization or AMF. At the highest N application rate, N₂O fluxes were lowest (0.71 kg N₂O-N ha^?1) with both P fertilization and AMF. The N₂O emission factors decreased with 50 % when P fertilization was applied.

Conclusions

Our results illustrate the importance of the judicious use of all nutrients to minimize N₂O emission, and thereby further underline the intimate link between sound agronomic practice and prudent soil GHG management.     

Atmospheric nitrogen inputs and losses along an urbanization gradient from Boston to Harvard Forest,MA

Urbanization alters nitrogen (N) cycling, but the spatiotemporal distribution and impact of these alterations on ecosystems are not well-quantified. We measured atmospheric inorganic N inputs and soil leaching losses along an urbanization gradient from Boston, MA to Harvard Forest in Petersham, MA. Atmospheric N inputs at urban sites (12.3 ± 1.5 kg N ha^?1 year^?1) were significantly greater than non-urban (5.7 ± 0.5 kg N ha^?1 year^?1) sites with NH₄ ⁺ (median value of 77 ± 4 %) contributing thrice as much as NO₃ ^?. Proximity to urban core correlated positively with NH₄ ⁺ (R² = 0.57, p = 0.02) and total inorganic N inputs (R² = 0.61, p = 0.01); on-road CO₂ emissions correlated positively with NO ₃ ^? inputs (R² = 0.74, p = 0.003). Inorganic N leaching rates correlated positively with atmospheric N input rates (R² = 0.61, p = 0.01), but did not differ significantly between urban and non-urban sites (p > 0.05). Our empirical measurements of atmospheric N inputs are greater for urban areas and less for rural areas compared to modeled regional estimates of N deposition. Five of the nine sites had NO ₃ ^? leached that came almost entirely from nitrification, indicating that the NO₃ ^? in leachate came from biological processes rather than directly passing through the soil. A significant proportion (17–100 %) of NO ₃ ^? leached from the other four sites came directly from the atmosphere. Surprisingly, the four sites where atmospheric sources made up the largest proportion of leachate NO₃ ^? also had relatively low N leaching rates, suggesting that atmospheric N inputs added to terrestrial ecosystems can move to multiple sinks and losses simultaneously, rather than being lost via leaching only after abiotic and biotic sinks have become saturated. This study improves our understanding of atmospheric N deposition and leaching in urban ecosystems, and highlights the need to incorporate urbanization effects in N deposition models.     

N2-fixation by methanotrophs sustains carbon and nitrogen accumulation in pristine peatlands

Symbiotic relationships between N₂-fixing prokaryotes and their autotrophic hosts are essential in nitrogen (N)-limited ecosystems, yet the importance of this association in pristine boreal peatlands, which store 25 % of the world’s soil (C), has been overlooked. External inputs of N to bogs are predominantly atmospheric, and given that regions of boreal Canada anchor some of the lowest rates found globally (~1 kg N ha^?1 year^?1), biomass production is thought to be limited primarily by N. Despite historically low N deposition, we show that boreal bogs have accumulated approximately 12–25 times more N than can be explained by atmospheric inputs. Here we demonstrate high rates of biological N₂-fixation in prokaryotes associated with Sphagnum mosses that can fully account for the missing input of N needed to sustain high rates of C sequestration. Additionally, N amendment experiments in the field did not increase Sphagnum production, indicating that mosses are not limited by N. Lastly, by examining the composition and abundance of N₂-fixing prokaryotes by quantifying gene expression of 16S rRNA and nitrogenase-encoding nifH, we show that rates of N₂-fixation are driven by the substantial contribution from methanotrophs, and not from cyanobacteria. We conclude biological N₂-fixation drives high sequestration of C in pristine peatlands, and may play an important role in moderating fluxes of methane, one of the most important greenhouse gases produced in peatlands. Understanding the mechanistic controls on biological N₂-fixation is crucial for assessing the fate of peatland carbon stocks under scenarios of climate change and enhanced anthropogenic N deposition.     

Nitrogen fixation and methanotrophy in forest mosses along a N deposition gradient

Nitrogen deposition has decreased the plant-associated nitrogen (N₂) fixation when measured using the indirect acetylene reduction assay (ARA). However, nitrogen deposition can also lead to changes in the diversity of moss symbionts, e.g. affect methanotrophic N₂ fixation, which is not measured by ARA. To test this hypothesis we compared ARA with the direct stable isotope method (¹⁵N₂ incorporation) and studied methanotrophy in two mosses, Hylocomium splendens and Pleurozium schreberi, collected from seven forest sites along a boreal latitudinal N deposition transect. We recognized that the two independent N₂ fixation measures gave corresponding results with the conversion factor of 3.3, but the ¹⁵N₂ method was more sensitive for finding a signal of low N₂ fixation activity. Methane carbon fixation associated with mosses was under the detection limit (<2 nmol C g⁻¹ h⁻¹). N₂ fixation rates were more pronounced in the mosses with higher C/N ratio, and in the green upper parts of the shoot than in the lower brownish parts. Sequencing of nifH genes revealed that dominating diazotrophs were affiliated to cyanobacterial genera Nostoc and Nodularia, but methanotrophic diazotrophs were not found in the nifH libraries. We conclude that the suppression of N₂ fixation along the deposition gradient was consistent regardless of the measurement technique, and microbial community changes toward methanotrophic or otherwise acetylene-sensitive N₂ fixation could not explain this trend.