Impact of seasonal change on dissimilatory nitrate reduction to ammonium (DNRA) triggering the retention of nitrogen in lake.

Nitrogen (N) reduction processes including denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are critical for the eutrophication in the lake water. However, the understanding about the dominant pathways of N cycling keep limited due to the high complexity of N cycle processes in lacustrine environment. The N fractions in sediments collected from Shijiuhu Lake were measured using high-resolution (HR)-Peeper technique and chemical extraction method in varied seasons. The abundance and microbial community compositions of functional genes involved in various N-cycling processes were also obtained using high-throughput sequencing. The results showed that NH4+ concentrations in the pore water remarkably increased from the upper layer toward the deeper layer and from winter to spring. This trend suggested that higher temperature facilitated the accumulation of NH4+ in the water. Decreased NO3- concentrations were also detected at deeper sediment layers and higher temperature, indicating the intensification of N reduction on anaerobic conditions. The NH4+-N concentrations reduced in spring along with the slight change of NO3--N in solid sediment, indicating the desorption and release of mobile NH4+ from solid phase to the solution. Remarkably decreased absolute abundances of functional genes were found in spring with DNRA bacteria nrfA gene as dominant genus and Anaeromyxobacter as the most dominant bacterium (21.67 ± 1.03%). Higher absolute abundance (146.2-788.1 × 105 Copies/g) of nrfA gene relative to other genes was mainly responsible for the increase of bio-available NH4+ in the sediments. Generally, microbial DNRA pathway predominated the N reduction and retention processes in the lake sediment at higher temperature and water depth even experiencing the suppression of DNRA bacteria abundance. These results suggested the existence of ecological risk via N retention by the action of the DNRA bacteria in the sediment on the condition of higher temperature, further provided valuable information for N management of eutrophic lakes.

Synergistic Impacts of Arsenic and Antimony Co-contamination on Diazotrophic Communities.

Nitrogen (N) shortage poses a great challenge to the implementation of in situ bioremediation practices in mining-contaminated sites. Diazotrophs can fix atmospheric N2 into a bioavailable form to plants and microorganisms inhabiting adverse habitats. Increasing numbers of studies mainly focused on the diazotrophic communities in the agroecosystems, while those communities in mining areas are still not well understood. This study compared the variations of diazotrophic communities in composition and interactions in the mining areas with different extents of arsenic (As) and antimony (Sb) contamination. As and Sb co-contamination increased alpha diversities and the abundance of nifH encoding the dinitrogenase reductase, while inhibited the diazotrophic interactions and substantially changed the composition of communities. Based on the multiple lines of evidence (e.g., the enrichment analysis of diazotrophs, microbe-microbe network, and random forest regression), six diazotrophs (e.g., Sinorhizobium, Dechloromonas, Trichormus, Herbaspirillum, Desmonostoc, and Klebsiella) were identified as keystone taxa. Environment-microbe network and random forest prediction demonstrated that these keystone taxa were highly correlated with the As and Sb contamination fractions. All these results imply that the above-mentioned diazotrophs may be resistant to metal(loid)s.

Everything Is Everywhere: Physiological Responses of the Mediterranean Sea and Eastern Pacific Ocean Epiphyte Cobetia Sp. to Varying Nutrient Concentration.

Bacteria are essential in the maintenance and sustainment of marine environments (e.g., benthic systems), playing a key role in marine food webs and nutrient cycling. These microorganisms can live associated as epiphytic or endophytic populations with superior organisms with valuable ecological functions, e.g., seagrasses. Here, we isolated, identified, sequenced, and exposed two strains of the same species (i.e., identified as Cobetia sp.) from two different marine environments to different nutrient regimes using batch cultures: (1) Cobetia sp. UIB 001 from the endemic Mediterranean seagrass Posidonia oceanica and (2) Cobetia sp. 4B UA from the endemic Humboldt Current System (HCS) seagrass Heterozostera chilensis. From our physiological studies, both strains behaved as bacteria capable to cope with different nutrient and pH regimes, i.e., N, P, and Fe combined with different pH levels, both in long-term (12 days (d)) and short-term studies (4 d/96 h (h)). We showed that the isolated strains were sensitive to the N source (inorganic and organic) at low and high concentrations and low pH levels. Low availability of phosphorus (P) and Fe had a negative independent effect on growth, especially in the long-term studies. The strain UIB 001 showed a better adaptation to low nutrient concentrations, being a potential N2-fixer, reaching higher growth rates (µ) than the HCS strain. P-acquisition mechanisms were deeply investigated at the enzymatic (i.e., alkaline phosphatase activity, APA) and structural level (e.g., alkaline phosphatase D, PhoD). Finally, these results were complemented with the study of biochemical markers, i.e., reactive oxygen species (ROS). In short, we present how ecological niches (i.e., MS and HCS) might determine, select, and modify the genomic and phenotypic features of the same bacterial species (i.e., Cobetia spp.) found in different marine environments, pointing to a direct correlation between adaptability and oligotrophy of seawater.

Functional traits of poplar leaves and fine roots responses to ozone pollution under soil nitrogen addition.

Concurrent ground-level ozone (O3) pollution and anthropogenic nitrogen (N) deposition can markedly influence dynamics and productivity in forests. Most studies evaluating the functional traits responses of rapid-turnover organs to O3 have specifically examined leaves, despite fine roots are another major source of soil carbon and nutrient input in forest ecosystems. How elevated O3 levels impact fine root biomass and biochemistry remains to be resolved. This study was to assess poplar leaf and fine root biomass and biochemistry responses to five different levels of O3 pollution, while additionally examining whether four levels of soil N supplementation were sufficient to alter the impact of O3 on these two organs. Elevated O3 resulted in a more substantial reduction in fine root biomass than leaf biomass; relative to leaves, more biochemically-resistant components were present within fine root litter, which contained high concentrations of lignin, condensed tannins, and elevated C:N and lignin: N ratios that were associated with slower rates of litter decomposition. In contrast, leaves contained more labile components, including nonstructural carbohydrates and N, as well as a higher N:P ratio. Elevated O3 significantly reduced labile components and increased biochemically-resistant components in leaves, whereas they had minimal impact on fine root biochemistry. This suggests that O3 pollution has the potential to delay leaf litter decomposition and associated nutrient cycling. N addition largely failed to affect the impact of elevated O3 levels on leaves or fine root chemistry, suggesting that soil N supplementation is not a suitable approach to combating the impact of O3 pollution on key functional traits of poplars. These results indicate that the significant differences in the responses of leaves and fine roots to O3 pollution will result in marked changes in the relative belowground roles of these two litter sources within forest ecosystems, and such changes will independently of nitrogen load.

Plants and mycorrhizal symbionts acquire substantial soil nitrogen from gaseous ammonia transport.

Nitrogen (N) is an essential nutrient that limits plant growth in many ecosystems. Here we investigate an overlooked component of the terrestrial N cycle - subsurface ammonia (NH3 ) gas transport and its contribution to plant and mycorrhizal N acquisition. We used controlled mesocosms, soil incubations, stable isotopes, and imaging to investigate edaphic drivers of NH3 gas efflux, track lateral subsurface N transport originating from 15 NH3 gas or 15 N-enriched organic matter, and assess plant and mycorrhizal N assimilation from this gaseous transport pathway. NH3 is released from soil organic matter, travels belowground, and contributes to root and fungal N content. Abiotic soil properties (pH and texture) influence the quantity of NH3 available for subsurface transport. Mutualisms with arbuscular mycorrhizal (AM) fungi can substantially increase plant NH3 -N uptake. The grass Brachypodium distachyon acquired 6-9% of total plant N from organic matter-N that traveled as a gas belowground. Colonization by the AM fungus Rhizophagus irregularis was associated with a two-fold increase in total plant N acquisition from subsurface NH3 gas. NH3 gas transport and uptake pathways may be fundamentally different from those of more commonly studied soil N species and warrant further research.

Amino acids dominate diffusive nitrogen fluxes across soil depths in acidic tussock tundra.

Organic nitrogen (N) is abundant in soils, but early conceptual frameworks considered it nonessential for plant growth. It is now well recognised that plants have the potential to take up organic N. However, it is still unclear whether plants supplement their N requirements by taking up organic N in situ: at what rate is organic N diffusing towards roots and are plants taking it up? We combined microdialysis with live-root uptake experiments to measure amino acid speciation and diffusion rates towards roots of Eriophorum vaginatum. Amino acid diffusion rates (321 ng N cm-2  h-1 ) were c. 3× higher than those for inorganic N. Positively charged amino acids made up 68% of the N diffusing through soils compared with neutral and negatively charged amino acids. Live-root uptake experiments confirmed that amino acids are taken up by plants (up to 1 µg N g-1  min-1 potential net uptake). Amino acids must be considered when forecasting plant-available N, especially when they dominate the N supply, and when acidity favours proteolysis over net N mineralisation. Determining amino acid production pathways and supply rates will become increasingly important in projecting the extent and consequences of shrub expansion, especially considering the higher C : N ratio of plants relative to soil.

Recently photoassimilated carbon and fungus-delivered nitrogen are spatially correlated in the ectomycorrhizal tissue of Fagus sylvatica.

Ectomycorrhizal plants trade plant-assimilated carbon for soil nutrients with their fungal partners. The underlying mechanisms, however, are not fully understood. Here we investigate the exchange of carbon for nitrogen in the ectomycorrhizal symbiosis of Fagus sylvatica across different spatial scales from the root system to the cellular level. We provided 15 N-labelled nitrogen to mycorrhizal hyphae associated with one half of the root system of young beech trees, while exposing plants to a 13 CO2 atmosphere. We analysed the short-term distribution of 13 C and 15 N in the root system with isotope-ratio mass spectrometry, and at the cellular scale within a mycorrhizal root tip with nanoscale secondary ion mass spectrometry (NanoSIMS). At the root system scale, plants did not allocate more 13 C to root parts that received more 15 N. Nanoscale secondary ion mass spectrometry imaging, however, revealed a highly heterogenous, and spatially significantly correlated distribution of 13 C and 15 N at the cellular scale. Our results indicate that, on a coarse scale, plants do not allocate a larger proportion of photoassimilated C to root parts associated with N-delivering ectomycorrhizal fungi. Within the ectomycorrhizal tissue, however, recently plant-assimilated C and fungus-delivered N were spatially strongly coupled. Here, NanoSIMS visualisation provides an initial insight into the regulation of ectomycorrhizal C and N exchange at the microscale.

Canopy and understory nitrogen addition have different effects on fine root dynamics in a temperate forest: implications for soil carbon storage.

Elucidating the effects of atmospheric nitrogen (N) deposition on fine root dynamics and the potential underlying mechanisms is required to understand the changes in belowground and aboveground carbon storage. However, research on these effects in forests has mostly involved direct understory addition of N and has ignored canopy interception and processing of N. Here, we conducted a field experiment comparing the effects of canopy addition of N (CAN) with those of understory addition of N (UAN) at three N-addition rates (0, 25 and 50 kg N ha-1 yr-1 ) on fine root dynamics in a temperate deciduous forest. Fine root production and biomass were significantly higher with CAN than with UAN. At the same N-addition rate, increases in fine root production with CAN were at least two-fold greater than with UAN. At the high N-addition rate and relative to the control, fine root biomass was significantly increased by CAN (by 23.5%) but was significantly decreased by UAN (by 12.2%). Our results indicate that traditional UAN may underestimate the responses of fine root dynamics to atmospheric N deposition in forest ecosystems. Canopy N processes should be considered for more realistic assessments of the effects of atmospheric N deposition in forests.

Homoploid F1 hybrids and segmental allotetraploids of japonica and indica rice subspecies show similar and enhanced tolerance to nitrogen deficiency than parental lines.

It remains unclear whether the merger of two divergent genomes by hybridization at the homoploid level or coupled with whole-genome duplication (WGD; allopolyploidy) can result in plants having better tolerance to stress conditions. In this study, we compared phenotypic performance and gene expression in the two diploid subspecies of rice (Oryza sativa subsp. japonica and indica), their reciprocal F1 hybrids, and in segmental allotetraploids under normal and nitrogen (N)-deficient conditions. We found that F1 hybrids and tetraploids showed higher and similar levels of tolerance to N deficiency than either parent. In parallel, total expression levels of 18 relevant functional genes were less perturbed by N deficiency in the F1 hybrids and tetraploids than in the parents. This was consistent with stable intrinsic partitioning of allelic/homoeologous expression defined by parental legacy in the homoploid F1 hybrids/tetraploids between the two conditions. The results suggest that genetic additivity at both the homoploid and allopolyploidy level might lead to similar beneficial phenotypic responses to nitrogen stress compared with the parents. The lack of synergistic responses to N limitation concomitant with WGD, relative to that exhibited by F1 hybrids, adds new empirical evidence in support of the emerging hypothesis that hybridization by itself can play a significant role in plant adaptive evolution in times of stress.

Effect of two types of adsorption sites of activated carbon fibers on nitrate ion adsorption.

In this study, the activation with ZnCl2 and heat treatment at 950 °C were performed to polyacrylonitrile-based carbon fiber and phenol resin to prepare two types of activated carbon fibers (ACFs) with different amounts of quaternary nitrogen (N-Q). In order to investigate the effect of chemical properties of ACFs on nitrate adsorption, various experiments and assess were conducted on these samples. The pHpzc of Py-7.0Z4-9.5HT10 and PhR-6.0Z4-9.5HT30 was about 7.1 and 7.0, respectively, and was no significant difference in the results. The C-π sites had a large effect on the amount of nitrate adsorption, and its adsorption capacity was greatest when the solution pH was about 3. However, the adsorption affinity (Ke) of C-π sites was weaker than that of N-Q, and the contribution for adsorption was small when the solution was low concentration or neutral, while the N-Q was the dominant adsorption site of ACFs. It is expected that the N-Q affected the adsorption kinetics, and the higher the amount of N-Q, the faster the adsorption rate. Py-7.0Z4-9.5HT10 had a large amount of C-π sites and N-Q. Therefore, the equilibrium adsorption amount (Qe) of Py-7.0Z4-9.5HT10 was 1.02 mmol/g at solution pH 3, and the adsorption kinetics were also fast.

Synergistic effects of nitrogen and CO2 enrichment on alpine grassland biomass and community structure.

Global environmental change is altering the Earth's ecosystems. However, much research has focused on ecosystem-level responses, and we know substantially less about community-level responses to global change stressors. Here we conducted a 6-yr field experiment in a high-altitude (4600 m asl) alpine grassland on the Tibetan Plateau to explore the effects of nitrogen (N) addition and rising atmospheric CO2 concentration on plant communities. Our results showed that N and CO2 enrichment had synergistic effects on alpine grassland communities. Adding nitrogen or CO2 alone did not alter total community biomass, species diversity or community composition, whereas adding both resources together increased community biomass, reduced species diversity and altered community composition. The observed decline in species diversity under simultaneous N and CO2 enrichment was associated with greater community biomass and lower soil water content, and driven by the loss of species characterised simultaneously by tall stature and small specific leaf area. Our findings point to the co-limitation of alpine plant community biomass and structure by nitrogen and CO2 , emphasising the need for future studies to consider multiple aspects of global environmental change together to gain a more complete understanding of their ecological consequences.

Whole-plant optimality predicts changes in leaf nitrogen under variable CO2 and nutrient availability.

Vegetation nutrient limitation is essential for understanding ecosystem responses to global change. In particular, leaf nitrogen (N) is known to be plastic under changed nutrient limitation. However, models can often not capture these observed changes, leading to erroneous predictions of whole-ecosystem stocks and fluxes. We hypothesise that an optimality approach can improve representation of leaf N content compared to existing empirical approaches. Unlike previous optimality-based approaches, which adjust foliar N concentrations based on canopy carbon export, we use a maximisation criterion based on whole-plant growth, and allow for a lagged response of foliar N to this maximisation criterion to account for the limited plasticity of this plant trait. We test these model variants at a range of Free-Air CO2 Enrichment and N fertilisation experimental sites. We show that a model based solely on canopy carbon export fails to reproduce observed patterns and predicts decreasing leaf N content with increased N availability. However, an optimal model which maximises total plant growth can correctly reproduce the observed patterns. The optimality model we present here is a whole-plant approach which reproduces biologically realistic changes in leaf N and can thereby improve ecosystem-level predictions under transient conditions.

Nitrate Ion Adsorption Characteristics of Activated Carbon Fibers with and without Quaternary Nitrogen Effective for Anion Adsorption.

By comparing the two types of activated carbon fibers (ACFs), characteristics of adsorption sites for nitrate ion other than quaternary nitrogen (N-Q) were investigated. Using phenol resin as precursors, activation with ZnCl2 was performed, and then heat treatment at 950 °C was carried out to prepare ACFs without N-Q, while ACFs with N-Q was prepared in the same method using polyacrylonitrile-based carbon fiber as precursors. We assessed the amount of functional groups, elemental composition, porous properties, and model of unit crystal size of graphene. For both ACFs with N-Q and without N-Q, equilibrium adsorption amount was not always simply proportional to surface area, but to the average number of benzene rings (Bz-rings) of graphene universally. PhR-5.0Z4 had only 20 benzene rings per graphene unit, but after heat treatment at 950 °C, the number drastically increased to 1088 (PhR-5.0Z4-9.5HT30). However, when the ACFs contained a large amount of oxygen, the number of Bz-rings was limited to 792 (PhR-5.0Z4-Ox-9.5HT30) even after heat treatment at 950 °C, and did not increase sufficiently. Cπ sites are more susceptible to oxygen inhibition than N-Q in adsorbing nitrate ions. For ACFs having Cπ sites as main adsorption sites, the heat treatment at 950 °C without oxidation can enhance the nitrate ion adsorption capacity.

How much nitrogen does Africa need to feed itself by 2050?

Nitrogen (N) fertilizers are very important for global food self-sufficiency (FSS), particularly for Africa, where the N input in agriculture is very low. This is the first work which studies and calculates the amount of N fertilizer that each country in Africa needs to feed itself by 2050. In this study, we used five different scenarios of inorganic fertilizer N (IFN) use and human diets to calculate the amount of N fertilizer needed to achieve FSS in Africa by 2050 and analyze the changes in N budget; N losses and N use efficiency (NUE). These scenarios include 1) business as usual (BAU), 2) equitable diet (EqD; self-sufficiency), 3) an IFN input 20% less than the EqD (S1), 4) an IFN input 40% less than the EqD (S2), and 5) a 20% increase in IFN input relative to the EqD (S3). Under the BAU scenario, production trends continue as they have over the past five decades, including an unhealthy human diet. In the EqD scenario, the priority is to meet the local demand for both animal and plant proteins with a healthy human diet. Under the EqD scenario, increasing the total N input from 35 kg N ha-1 yr-1 to 181 kg N ha-1 yr-1 during 2016-2050 is needed to achieve FSS in Africa. This increase in N fertilizer use represents unprecedented N inputs to African terrestrial ecosystems - at least 52 Tg N yr-1 - which would lead to inevitable increases in N losses. We also found that the NUE would decrease from 63% during 2010-2016 to 50% by 2050, whereas the total N surplus would increase from 13 kg N ha-1 yr-1 to 90 kg N ha-1 yr-1 by 2050. The estimated gaseous emissions would increase from 8 kg N ha-1 yr-1 to 61 kg N ha-1 yr-1 by 2050. Our findings conclude that, it is very important to consider the high N losses in Africa if the EqD scenario is applied. The S1 and S2 scenarios result in much less environmental N loss, and better NUE compared with the EqD scenario. Therefore, based on these findings we can recommend the implementation of the S2 scenario with an IFN dose of 77 kg N ha-1 yr-1, in parallel with the use of modern agricultural techniques and the increased use of organic inputs.

Different strategies of nitrogen acquisition in two tropical seagrasses under nitrogen enrichment.

Tropical marine seagrasses live in environments with low nutrient concentrations. However, as land development intensifies along tropical coastlines, the marine environment in which these organisms grow is becoming more nutrient-rich. Nitrogen (N) uptake, assimilation, translocation and storage under a diversity of N sources in enriched conditions were investigated in two tropical seagrass species, Cymodocea serrulata and Thalassia hemprichii, from an oligotrophic marine environment. Both seagrasses were able to take up different inorganic and organic N sources through their above- and belowground tissues when enriched with high N concentrations. The uptake rates of T. hemprichii were generally higher than C. serrulata in leaves and rhizome, whereas root uptake was systematically higher in C. serrulata. Acropetal and basipetal translocation was observed in both species. Reduction and assimilation of N, measured in terms of their nitrate reductase and glutamine synthetase activity, were correlated with nitrate and ammonium uptake rates, respectively. Cymodocea serrulata showed a tendency to immediately use the available N, whereas T. hemprichii allocated more N in assimilation and storage investment. The responses of these seagrasses to N-enrichment demonstrate their ability to adapt to over-enrichment by varying N sources in the first step of the eutrophication process.

Unaltered phenology but increased production of ectomycorrhizal roots of Pinus elliottii under 4 years of nitrogen addition.

Timing (phenology) and amount (production) are two integral facets of root growth, and their shifts have profound influences on below-ground resource acquisition. However, the environmental control and the effects of nitrogen (N) deposition on the production and phenology of ectomycorrhizal (ECM) roots remain unclear. Using a 4 yr minirhizotron experiment, we explored the control of the production and three phenophases (initiation, peak, and cessation of growth) of ECM roots in two soil layers and investigated their dynamic responses to N addition in a seasonally dry subtropical Pinus elliottii forest. We found a stronger control of water availability on the production and a stronger control of temperature on the phenology of ECM roots under ambient conditions. Temperature was correlated positively with initiation and negatively with cessation, especially in the shallow layer. N addition did not affect the phenology of ECM roots but increased their production by modifying N and phosphorus (P) stoichiometry in the soil and foliage. Our findings suggest a greater sensitivity of production than phenology of ECM roots to N addition. The increased production of ECM roots under N addition could be driven by N-induced P limitation or some combination of below-ground resources (P, N, water).

Plant-microbe competition: does injection of isotopes of C and N into the rhizosphere effectively characterise plant use of soil N?

Despite considerable attention over the last 25 yr, the importance of early protein breakdown products to plant nitrogen (N) nutrition remains uncertain. We used rhizosphere injection of 15 N-, 13 C- and 14 C-labelled inorganic N and amino acid (l-alanine), with chase periods from 1 min to 24 h, to investigate the duration of competition for amino acid between roots (Triticum aestivum) and soil microorganisms. We further investigated how microbial modification of l-alanine influenced plant carbon (C) and N recovery. From recovery of C isotopes, intact alanine uptake was 0.2-1.3% of added. Soil microbes appeared to remove alanine from soil solution within 1 min and release enough NH4+ to account for all plant 15 N recovery (over 24 h) within 5 min. Microbially generated inorganic or keto acid C accounted for < 25% of the lowest estimate of intact alanine uptake. Co-location of C and N labels appears a reasonable measure of intact uptake. Potential interference from microbially modified C is probably modest, but may increase with chase period. Similarly, competition for l-alanine is complete within a few minutes in soil, whereas NO3- added at the same rate is available for > 24 h, indicating that long chase periods bias outcomes and fail to accurately simulate soil processes.

Newly depolymerized large organic N contributes directly to amino acid uptake in young maize plants.

The contribution of large molecular size organic nitrogen (N) to plant N uptake is unclear. Soils with and without maize, at three pH levels, were treated with (carbon-14 and -13 (14 C, 13 C), 15 N) triple-labelled > 100 kDa organic N. After 48 h, soil and maize were sampled for bulk and compound specific isotope analysis to study the turnover in soil and plant 13 C and 15 N uptake. Mineralization of > 100 kDa organic N increased with higher pH only in soil without maize. The > 100 kDa organic N disappeared rapidly in soils with and without maize, but surprisingly more > 100 kDa organic N derived amino acids remained in soil with than without maize - most likely in the microbial biomass. Total 15 N uptake in maize increased with higher soil pH. The organic N uptake was estimated to account for 20-30% of the total 15 N uptake. Organic N uptake was confirmed by the presence of 13 C-labelled amino acids in maize roots. The study suggests that the importance of plant organic N uptake increases when N is derived from complex molecules such as proteins compared to studies using single amino acids as N source, and that rhizosphere microorganisms increase anabolic utilization of organic N compared to microorganisms in the bulk soil.

Exploring the role of ectomycorrhizal fungi in soil carbon dynamics.

The extent to which ectomycorrhizal (ECM) fungi enable plants to access organic nitrogen (N) bound in soil organic matter (SOM) and transfer this growth-limiting nutrient to their plant host, has important implications for our understanding of plant-fungal interactions, and the cycling and storage of carbon (C) and N in terrestrial ecosystems. Empirical evidence currently supports a range of perspectives, suggesting that ECM vary in their ability to provide their host with N bound in SOM, and that this capacity can both positively and negatively influence soil C storage. To help resolve the multiplicity of observations, we gathered a group of researchers to explore the role of ECM fungi in soil C dynamics, and propose new directions that hold promise to resolve competing hypotheses and contrasting observations. In this Viewpoint, we summarize these deliberations and identify areas of inquiry that hold promise for increasing our understanding of these fundamental and widespread plant symbionts and their role in ecosystem-level biogeochemistry.

Nitrogen turnover and N2O/N2 ratio of three contrasting tropical soils amended with biochar.

Biochar has been reported to reduce emission of nitrous oxide (N2O) from soils, but the mechanisms responsible remain fragmentary. For example, it is unclear how biochar effects on N2O emissions are mediated through biochar effects on soil gross N turnover rates. Hence, we conducted an incubation study with three contrasting agricultural soils from Kenya (an Acrisol cultivated for 10-years (Acrisol10); an Acrisol cultivated for over 100-years (Acrisol100); a Ferralsol cultivated for over 100 years (Ferralsol)). The soils were amended with biochar at either 2% or 4% w/w. The 15N pool dilution technique was used to quantify gross N mineralization and nitrification and microbial consumption of extractable N over a 20-day incubation period at 25 °C and 70% water holding capacity of the soil, accompanied by N2O emissions measurements. Direct measurements of N2 emissions were conducted using the helium gas flow soil core method. N2O emissions varied across soils with higher emissions in Acrisols than in Ferralsols. Addition of 2% biochar reduced N2O emissions in all soils by 53 to 78% with no significant further reduction induced by addition at 4%. Biochar effects on soil nitrate concentrations were highly variable across soils, ranging from a reduction, no effect and an increase. Biochar addition stimulated gross N mineralization in Acrisol-10 and Acrisol-100 soils at both addition rates with no effect observed for the Ferralsol. In contrast, gross nitrification was stimulated in only one soil but only at a 4% application rate. Also, biochar effects on increased NH4 + immobilization and NO3 -consumption strongly varied across the three investigated soils. The variable and bidirectional biochar effects on gross N turnover in conjunction with the unambiguous and consistent reduction of N2O emissions suggested that the inhibiting effect of biochar on soil N2O emission seemed to be decoupled from gross microbial N turnover processes. With biochar application, N2 emissions were about an order of magnitude higher for Acrisol-10 soils compared to Acrisol-100 and Ferralsol-100 soils. Our N2O and N2 flux data thus support an explanation of direct promotion of gross N2O reduction by biochar rather than effects on soil extractable N dynamics. Effects of biochar on soil extractable N and gross N turnover, however, might be highly variable across different soils as found here for three typical agricultural soils of Kenya.