N2 Fixation Rates Research Articles

Unusual marine cyanobacteria/haptophyte symbiosis relies on N2 fixation even in N-rich environments

The microbial fixation of N2 is the largest source of biologically available nitrogen (N) to the oceans. However, it is the most energetically expensive N-acquisition process and is believed inhibited when less energetically expensive forms, like dissolved inorganic N (DIN), are available. Curiously, the cosmopolitan N2-fixing UCYN-A/haptophyte symbiosis grows in DIN-replete waters, but the sensitivity of their N2 fixation to DIN is unknown. We used stable isotope incubations, catalyzed reporter deposition fluorescence in-situ hybridization (CARD-FISH), and nanoscale secondary ion mass spectrometry (nanoSIMS), to investigate the N source used by the haptophyte host and sensitivity of UCYN-A N2 fixation in DIN-replete waters. We demonstrate that under our experimental conditions, the haptophyte hosts of two UCYN-A sublineages do not assimilate nitrate (NO3−) and meet little of their N demands via ammonium (NH4+) uptake. Instead the UCYN-A/haptophyte symbiosis relies on UCYN-A N2 fixation to supply large portions of the haptophyte’s N requirements, even under DIN-replete conditions. Furthermore, UCYN-A N2 fixation rates, and haptophyte host carbon fixation rates, were at times stimulated by NO3− additions in N-limited waters suggesting a link between the activities of the bulk phytoplankton assemblage and the UCYN-A/haptophyte symbiosis. The results suggest N2 fixation may be an evolutionarily viable strategy for diazotroph–eukaryote symbioses, even in N-rich coastal or high latitude waters.

N2 Fixation and New Insights Into Nitrification From the Ice-Edge to the Equator in the South Pacific Ocean

Nitrogen (N) is an essential element for life and controls the magnitude of primary productivity in the ocean. In order to describe the microorganisms that catalyze N transformations in surface waters in the South Pacific Ocean, we collected high-resolution biotic and abiotic data along a 7000 km transect, from the Antarctic ice edge to the equator. The transect, conducted between late Austral autumn and early winter 2016, covered major oceanographic features such as the polar front (PF), the subtropical front (STF) and the Pacific equatorial divergence (PED). We measured N2 fixation and nitrification rates and quantified the relative abundances of diazotrophs and nitrifiers in a region where few to no rate measurements are available. Even though N2 fixation rates are usually below detection limits in cold environments, we were able to measure this N pathway at 7/10 stations in the cold and nutrient rich waters near the PF. This result highlights that N2 fixation rates continue to be measured outside the well-known subtropical regions. The majority of the mid to high N2 fixation rates (>~20nmol L-1 d-1), however, still occurred in the expected tropical and subtropical regions. High throughput sequence analyses of the dinitrogenase reductase gene (nifH) revealed that the nifH Cluster I dominated the diazotroph diversity throughout the transect. nifH gene richness did not show a latitudinal trend, nor was it significantly correlated with N2 fixation rates. Nitrification rates above the mixed layer in the Southern Ocean ranged between 56 and 1440 nmol L-1 d-1. Our data showed a decoupling between carbon and N assimilation (NO3- and NH4+ assimilation rates) in winter in the South Pacific Ocean. Phytoplankton community structure showed clear changes across the PF, the STF and the PED, defining clear biomes. Overall, these findings provide a better understanding of the ecosystem functionality in the South Pacific Ocean across key oceanographic biomes.

The responses of moss-associated nitrogen fixation and belowground microbial community to chronic Mo and P supplements in subarctic dry heaths

Although nitrogen (N) fixation by moss-associated bacteria is the main source of new N in N-limited ecosystems like arctic tundra, we do not know which nutrient, molybdenum (Mo) or phosphorus (P), is rate-limiting for sustaining this process in the long-term. Further, how moss-associated N2 fixation impacts the belowground microbial regulation of decomposition remains unresolved. Moss-associated N2 fixation and soil microbial process rates, abundance and community structure were assessed in long-term P and Mo field additions in the Subarctic during three years. We found tendencies for stimulation of moss-associated N2 fixation by Mo in the short term, by P in the long-term, and tendencies for a stimulation of soil microbial activity by P. However, large variation in microbial activity within and below the moss exceeded any systematic variation induced by the field treatments. Our findings suggest that soil microbial activity is not limited by N at our site, and that Mo and P only occasionally limit N2 fixation during a growing season. Since increasing CO2 concentrations can induce nutrient limitation, the here reported transient limitation can easily shift into a chronic one with significant implications for ecosystem productivity and biogeochemistry.

Heterotrophic diazotrophs in a eutrophic temperate bay (Jiaozhou Bay) broadens the domain of N2 fixation in China's coastal waters

N2 fixation in coastal waters represents an overlooked process in the global marine nitrogen cycle, yet the mechanisms and controls are still poorly understood. This study first conducted simultaneous measurements of N2 fixation rates, primary production, chlorophyll a (Chl a) concentration, organic carbon concentration, and the composition of nitrogen-fixing genes (nifH) in a eutrophic temperate bay (Jiaozhou Bay, the western Yellow Sea) from 2015 to 2017. The activity of N2 fixation (not detected–1.53 nmol N L −1 h −1) were observed at most sampling stations, suggesting that the N-replete conditions in Jiaozhou Bay did not prevent N2 fixation. The major N2 fixers in JB were heterotrophic, while N2 fixation were mainly contributed by > 10 μm fraction, indicating the possible active N2 fixation of heterotrophic diazotrophs on large particles or planktonic aggregates. Positive correlations between N2 fixation and organic carbon concentration were observed in spring. Further estimates of the level of phytoplankton-derived organic carbon and its potential relationship with N2 fixation suggest that phytoplankton processes may stimulate N2 fixation in Jiaozhou Bay by increasing the bioavailability of organic carbon or/and providing suitable environmental conditions for N2 fixation. Our results first infer the co-regulation of physical factors and biological processes on N2 fixation. As Jiaozhou Bay has been subject to anthropogenic pollution of inorganic nutrients, our results provide insights into the dominant controls on N2 fixation and relevant biogeochemical processes in coastal waters under anthropogenic disturbance.

Dinitrogen fixation rates in the Bay of Bengal during summer monsoon

Biological dinitrogen (N2) fixation exerts an important control on oceanic primary production by providing bioavailable form of nitrogen (such as ammonium) to photosynthetic microorganisms. N2 fixation is dominant in nutrient poor and warm surface waters. The Bay of Bengal is one such region where no measurements of phototrophic N2 fixation rates exist. The surface water of the Bay of Bengal is generally nitrate-poor and warm due to prevailing stratification and thus, could favour N2 fixation. We commenced the first N2 fixation study in the photic zone of the Bay of Bengal using 15N2 gas tracer incubation experiment during summer monsoon 2018. We collected seawater samples from four depths (covering the mixed layer depth of up to 75 m) at eight stations. N2 fixation rates varied from 4 to 75 μmol N m−2 d−1. The contribution of N2 fixation to primary production was negligible (<1%). However, the upper bound of observed N2 fixation rates is higher than the rates measured in other oceanic regimes, such as the Eastern Tropical South Pacific, the Tropical Northwest Atlantic, and the Equatorial and Southern Indian Ocean.

Microbial nitrogen fixation and methane oxidation are strongly enhanced by light in Sphagnum mosses

Peatlands have acted as C-sinks for millennia, storing large amounts of carbon, of which a significant amount is yearly released as methane (CH4). Sphagnum mosses are a key genus in many peat ecosystems and these mosses live in close association with methane-oxidizing and nitrogen-fixing microorganisms. To disentangle mechanisms which may control Sphagnum-associated methane-oxidation and nitrogen-fixation, we applied four treatments to Sphagnum mosses from a pristine peatland in Finland: nitrogen fertilization, phosphorus fertilization, CH4 addition and light. N and P fertilization resulted in nutrient accumulation in the moss tissue, but did not increase Sphagnum growth. While net CO2 fixation rates remained unaffected in the N and P treatment, net CH4 emissions decreased because of enhanced CH4 oxidation. CH4 addition did not affect Sphagnum performance in the present set-up. Light, however, clearly stimulated the activity of associated nitrogen-fixing and methane-oxidizing microorganisms, increasing N2 fixation rates threefold and CH4 oxidation rates fivefold. This underlines the strong connection between Sphagnum and associated N2 fixation and CH4 oxidation. It furthermore indicates that phototrophy is a strong control of microbial activity, which can be directly or indirectly.

A critical review of the 15N2 tracer method to measure diazotrophic production in pelagic ecosystems

AbstractDinitrogen (N2) fixation is an important source of biologically reactive nitrogen (N) to the global ocean. The magnitude of this flux, however, remains uncertain, in part because N2 fixation rates have been estimated following divergent protocols and because associated levels of uncertainty are seldom reported—confounding comparison and extrapolation of rate measurements. A growing number of reports of relatively low but potentially significant rates of N2 fixation in regions such as oxygen minimum zones, the mesopelagic water column of the tropical and subtropical oceans, and polar waters further highlights the need for standardized methodological protocols for measurements of N2 fixation rates and for calculations of detection limits and propagated error terms. To this end, we examine current protocols of the 15N2 tracer method used for estimating diazotrophic rates, present results of experiments testing the validity of specific practices, and describe established metrics for reporting detection limits. We put forth a set of recommendations for best practices to estimate N2 fixation rates using 15N2 tracer, with the goal of fostering transparency in reporting sources of uncertainty in estimates, and to render N2 fixation rate estimates intercomparable among studies.

Latitudinal constraints on the abundance and activity of the cyanobacterium UCYN‐A and other marine diazotrophs in the North Pacific

AbstractThe number of marine environments known to harbor dinitrogen (N2)‐fixing (diazotrophic) microorganisms is increasing, prompting a reassessment of the biogeography of marine diazotrophs and N2fixation rates (NFRs). Here, we investigate the diversity, abundance, and activity of diazotrophic microorganisms in the North Pacific Subtropical Gyre (NPSG), a diazotrophic habitat, and the North Pacific Transition Zone (NPTZ), a region characterized by strong physical, chemical, and biological gradients. Samples were collected on two springtime meridional cruises during 2016 and 2017, spanning from 23.5°N to 41.4°N along 158°W. We observed an abrupt decrease in diazotrophic abundances near the southern edge of the NPTZ, which coincided with a salinity front and with a ∼10‐fold increase inSynechococcusabundance, but without a concomitant change in phosphate or nitrate concentrations. In NPSG waters south of this diazotrophic boundary,nifHgenes and NFRs were consistently detected and diazotrophic communities were dominated by UCYN‐A, an uncultivated, symbiotic cyanobacterium (2.8 × 103to 1.0 × 106nifHgene copies L−1). There was a significant positive relationship between quantitative polymerase chain reaction‐derived UCYN‐AnifHgene abundances and community NFRs in the NPSG, suggesting a large contribution of UCYN‐A to community NFRs. In the NPTZ waters to the north, NFRs were low or undetected andnifHgenes were rare, with the few detected sequences represented by UCYN‐A and noncyanobacterial diazotrophs. The patterns we observed in UCYN‐A abundance in the context of local biogeochemistry suggest that the environmental controls of this organism may differ from those of cultivated marine cyanobacterial diazotrophs.

Nitrogen and phosphorus control carbon sequestration in agricultural ecosystems: modelling carbon, nitrogen, and phosphorus balances at the Breton Plots with ecosys under historical and future climates

Perennial legumes in crop rotations increase soil C sequestration from greater productivity with N2 fixation. Here, we corroborated increases in soil organic carbon (SOC) and harvests modelled in 5 yr wheat–oats–barley–alfalfa/brome–alfalfa/brome (5Y) vs. 2 yr wheat–fallow (WF) rotations with those measured from 1929 to 2018. Harvest and SOC gains of 100–150 g C m−2 yr−1 and 15–25 g C m−2 yr−1 were modelled and measured in 5Y vs. WF rotations with different fertilizer and manure amendments. Modelled gains were closely related to annualized rates of N2 fixation by alfalfa of 8–10 g N m−2 yr−1. However, N2 fixation also drove increases in modelled N2O emissions of ca. 0.06 g N m−2 yr−1, which partially offset gains in SOC. Gains in harvest, SOC, and N2O emissions of 60–90 g C m−2 yr−1, 15 g C m−2 yr−1, and 0.05 g N m−2 yr−1 were modelled and measured in both rotations with amendments of N + P relative to unamended treatments. Harvest and SOC gains were smaller, and leaching and N2O losses larger, with amendments of N without P. After 100 yr of RCP 8.5 climate change, harvests in WF changed little from those in baseline runs, whereas those in 5Y rose with N + P because of increased N2 fixation. SOC declined in WF with all amendments and could only be raised in 5Y with N + P amendments. These model findings indicated the importance of N2 fixation and P amendments in determining responses of agroecosystem productivity and C sequestration to climate change.

Light availability modulates the effects of warming in a marine N&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; fixer

Abstract. Trichodesmium species, as a group of photosynthetic N2 fixers (diazotrophs), play an important role in the marine biogeochemical cycles of nitrogen and carbon, especially in oligotrophic waters. How ongoing ocean warming may interact with light availability to affect Trichodesmium is not yet clear. We grew Trichodesmium erythraeum IMS 101 at three temperature levels of 23, 27, and 31∘C under growth-limiting and growth-saturating light levels of 50 and 160 µmol quanta m−2 s−1, respectively, for at least 10 generations and then measured physiological performance, including the specific growth rate, N2 fixation rate, and photosynthesis. Light availability significantly modulated the growth response of Trichodesmium to temperature, with the specific growth rate peaking at ∼27∘C under the light-saturating conditions, while growth of light-limited cultures was non-responsive across the tested temperatures (23, 27, and 31∘C). Short-term thermal responses for N2 fixation indicated that both high growth temperature and light intensity increased the optimum temperature (Topt) for N2 fixation and decreased its susceptibility to supra-optimal temperatures (deactivation energy – Eh). Simultaneously, all light-limited cultures with low Topt and high Eh were unable to sustain N2 fixation during short-term exposure to high temperatures (33–34∘C) that are not lethal for the cells grown under light-saturating conditions. Our results imply that Trichodesmium spp. growing under low light levels while distributed deep in the euphotic zone or under cloudy weather conditions might be less sensitive to long-term temperature changes that occur on the timescale of multiple generations but are more susceptible to abrupt (less than one generation time span) temperature changes, such as those induced by cyclones and heat waves.

No nitrogen fixation in the Bay of Bengal?

Abstract. The Bay of Bengal (BoB) has long stood as a biogeochemical enigma, with subsurface waters containing extremely low, but persistent, concentrations of oxygen in the nanomolar range which – for some, yet unconstrained, reason – are prevented from becoming anoxic. One reason for this may be the low productivity of the BoB waters due to nutrient limitation and the resulting lack of respiration of organic material at intermediate waters. Thus, the parameters determining primary production are key in understanding what prevents the BoB from developing anoxia. Primary productivity in the sunlit surface layers of tropical oceans is mostly limited by the supply of reactive nitrogen through upwelling, riverine flux, atmospheric deposition, and biological dinitrogen (N2) fixation. In the BoB, a stable stratification limits nutrient supply via upwelling in the open waters, and riverine or atmospheric fluxes have been shown to support only less than one-quarter of the nitrogen for primary production. This leaves a large uncertainty for most of the BoB's nitrogen input, suggesting a potential role of N2 fixation in those waters. Here, we present a survey of N2 fixation and carbon fixation in the BoB during the winter monsoon season. We detected a community of N2 fixers comparable to other oxygen minimum zone (OMZ) regions, with only a few cyanobacterial clades and a broad diversity of non-phototrophic N2 fixers present throughout the water column (samples collected between 10 and 560 m water depth). While similar communities of N2 fixers were shown to actively fix N2 in other OMZs, N2 fixation rates were below the detection limit in our samples covering the water column between the deep chlorophyll maximum and the OMZ. Consistent with this, no N2 fixation signal was visible in δ15N signatures. We suggest that the absence of N2 fixation may be a consequence of a micronutrient limitation or of an O2 sensitivity of the OMZ diazotrophs in the BoB. Exploring how the onset of N2 fixation by cyanobacteria compared to non-phototrophic N2 fixers would impact on OMZ O2 concentrations, a simple model exercise was carried out. We observed that both photic-zone-based and OMZ-based N2 fixation are very sensitive to even minimal changes in water column stratification, with stronger mixing increasing organic matter production and export, which can exhaust remaining O2 traces in the BoB.

Nitrogen pools in soil covered by biological soil crusts of different successional stages in a temperate desert in Central Asia

Biological soil crusts (BSCs) are a primary source of nitrogen (N) in deserts through N-fixation. N-fixation rates may be determined by the species present within BSCs, and environmental factors like temperature and moisture, which vary seasonally. These same factors may also govern dynamics of within-soil N transformations. Few studies have explored the dynamics of different N forms in response to seasonal microclimate variations in BSCs, especially across both the growing season and the snow-covered season (winter). In this study, monthly changes in multiple soil N pools across a full year and beneath three soil cover types – bare soil, cyanobacteria BSC, and lichen BSC – were assessed across a full year in the Gurbantunggut Desert, Central Asia. We focused on multiple organic and inorganic N pools including: total N, alkali-hydrolyzable N, DON, inorganic N (NH4+-N, NO3−-N), free amino acids, and microbial biomass N. We found that different N forms had divergent trends during the year, peaked in different months, and showed non-synchronous responses related to seasonal temperature and moisture patterns. Most N forms were most abundant in lichen BSC > cyanobacteria BSC > bare soil, although significant differences were only observed in a few months. In the growing season, available N forms such as soil DON and inorganic N were related to both water and temperature. In winter, available N was strongly related to temperature variability prior to the sampling dates. The high NH4+-N/NO3−-N in the early growing season suggest that N fixation or N mineralization were highest at this time. Our results indicate non-synchronous availability of different N forms, and elevated N availability beneath lichen BSCs, which may maintain the diverse N uptake requirements for different plant species.

Carbon Transfer from the Host Diatom Enables Fast Growth and High Rate of N2 Fixation by Symbiotic Heterocystous Cyanobacteria.

Diatom–diazotroph associations (DDAs) are symbioses where trichome-forming cyanobacteria support the host diatom with fixed nitrogen through dinitrogen (N2) fixation. It is inferred that the growth of the trichomes is also supported by the host, but the support mechanism has not been fully quantified. Here, we develop a coarse-grained, cellular model of the symbiosis between Hemiaulus and Richelia (one of the major DDAs), which shows that carbon (C) transfer from the diatom enables a faster growth and N2 fixation rate by the trichomes. The model predicts that the rate of N2 fixation is 5.5 times that of the hypothetical case without nitrogen (N) transfer to the host diatom. The model estimates that 25% of fixed C from the host diatom is transferred to the symbiotic trichomes to support the high rate of N2 fixation. In turn, 82% of N fixed by the trichomes ends up in the host. Modeled C fixation from the vegetative cells in the trichomes supports only one-third of their total C needs. Even if we ignore the C cost for N2 fixation and for N transfer to the host, the total C cost of the trichomes is higher than the C supply by their own photosynthesis. Having more trichomes in a single host diatom decreases the demand for N2 fixation per trichome and thus decreases their cost of C. However, even with five trichomes, which is about the highest observed for Hemiaulus and Richelia symbiosis, the model still predicts a significant C transfer from the diatom host. These results help quantitatively explain the observed high rates of growth and N2 fixation in symbiotic trichomes relative to other aquatic diazotrophs.

Dissolved organic matter stimulates N2 fixation and nifH gene expression in Trichodesmium.

Mixotrophy, the combination of heterotrophic and autotrophic nutrition modes, is emerging as the rule rather than the exception in marine photosynthetic plankton. Trichodesmium, a prominent diazotroph ubiquitous in the (sub)tropical oceans, is generally considered to obtain energy via autotrophy. While the ability of Trichodesmium to use dissolved organic phosphorus when deprived of inorganic phosphorus sources is well known, the extent to which this important cyanobacterium may benefit from other dissolved organic matter (DOM) resources is unknown. Here we provide evidence of carbon-, nitrogen- and phosphorus-rich DOM molecules enhancing N2 fixation rates and nifH gene expression in natural Trichodesmium colonies collected at two stations in the western tropical South Pacific. Sampling at a third station located in the oligotrophic South Pacific Gyre revealed no Trichodesmium but showed presence of UCYN-B, although no nifH expression was detected. Our results suggest that Trichodesmium behaves mixotrophically in response to certain environmental conditions, providing them with metabolic plasticity and adding up to the view that mixotrophy is widespread among marine microbes.

Effect of mesoscale eddies on diazotroph community structure and nitrogen fixation rates in the South China Sea

Mesoscale eddies play a key role in ocean biogeochemistry, yet their effects on dinitrogen (N2) fixation are not well understood. Using quantitative polymerase chain reaction (qPCR) and dinitrogen-15 (15N2) tracer methods, we examined the abundance of nifH genes in N2-fixing microorganisms and N2 fixation rates in the upper water column (0–100 m) of mesoscale eddies in the northeastern South China Sea (SCS). High N2 fixation rates affected by an anticyclonic eddy were associated with high abundances of unicellular nitrogen-fixing cyanobacteria in groups A and B (UCYN-A, UCYN-B), while low nitrogen fixation was found in both cyclonic eddy and reference stations. The estimated N2 fixation rates met 9% ± 5% of the nitrogen demand for primary production affected by the anticyclonic eddy. Our results indicate that mesoscale eddies can affect nitrogen fixation through affecting nutrient availability and eddy transport, and play a crucial role in regulating N2 fixation in the oligotrophic SCS.

Nutrient stoichiometry (N:P) controls nitrogen fixation and distribution of diazotrophs in a tropical eutrophic estuary

Nitrogen fixation and its ecological regulation are poorly understood in the tropical estuaries, which are highly influenced by anthropogenic disturbances. In this study, we investigated the role of nutrient stoichiometry in the diversity, abundance and activity of N2-fixing bacterial community and their seasonal variations in the water column of a tropical eutrophic estuary (Cochin estuary). The N2 fixation rates in the estuary ranged from 0.1 to 2.0 nmol N2 l−1 h−1, with higher activity during post-monsoon and lower during monsoon. The rates are appeared to be primarily controlled by dissolved inorganic nitrogen and phosphorous (N:P) ratio. Clone library analysis of nitrogenase (nifH) gene revealed that the major N2 fixing phylotypes belong to Cluster I and Cluster III diazotrophs. The overall findings of this study suggest that monsoon induced seasonal changes in nutrient stoichiometry control the distribution and activity of diazotrophs in a tropical estuary.

In situ synthesis of Bi2MoO6@C@attapulgite photocatalyst for enhanced photocatalytic nitrogen fixation ability under simulated solar irradiation

In this article, a ternary carbon modified Bi2MoO6 @ attapulgite composite (BMO@C@ATP) was fabricated by in situ self-assembly methodology using glucose as the carbon source. The photocatalytic activity of BMO@C@ATP in nitrogen fixation reaction was then evaluated. The studies showed that BMO@C@ATP composite exhibited higher catalytic performance after carbon-doping, comparing with the BMO and BMO@ATP. The N2 fixation rate can reach up to 83.09 μmol L−1 h−1 in the presence of BMO@C@ATP, which is 2.67 times higher than that of Bi2MoO6 photocatalyst (31.11 μmol L−1 h−1). The enhanced catalytic rate of the ternary composite may be attributed to its excellent light trapping performance, large surface area, and efficient separation of carriers. Moreover, the lattice substitution of C atoms serves as the conductive electron bridge, which can accumulate electrons and promote the NH3 generation over Bi2MoO6 photocatalyst. In general, the present BMO@C@ATP photocatalytic system provides a simple and economical method for nitrogen fixation process, which shows great application prospects in the future.

N2 fixation in urbanization area rivers: spatial-temporal variations and influencing factors.

While nitrogen (N2) fixation is an important process in nitrogen (N) biogeochemical cycling, supplying a significant portion of the N in natural ecosystems, few quantitative constraints exist concerning its contribution to the N enrichment and export from river ecosystems. This study estimates the N2 fixation rates of urban rivers in the Yangtze Estuary area using acetylene reduction. The results demonstrate that the prominent spatiotemporal variability of river N2 fixation rates is driven by various environmental factors. River N2 fixation rates are significantly higher in the summer (90.57 ± 14.60 ngN·L-1·h-1) than in the winter (57.98 ± 15.73 ngN·L-1·h-1). Spatially, rivers draining urban and suburban areas have higher N2 fixation rates than those draining rural areas. The N2 fixation rates are positively correlated with the N2 fixing cyanobacteria density, water temperature, light, and the water phosphorus (P) concentration, but they are negatively correlated with the dissolved N concentration (NH4+-N and NO3--N). The N2 fixation rates annually range from 53.20 to 89.24 ngN·L-1·h-1 for all of the sampling rivers, which is equivalent to a depth integrated (0-0.6m) N input of 0.163-0.274 gN·m-2·a-1. The determined annual N input via N2 fixation is generally higher than that of marine systems, but it is lower than that of eutrophic lakes. This study provides robust evidence that N2 fixation can supply a substantial portion of the N input to human-impacted river ecosystems, which has not been sufficiently accounted for when determining the N mass balance of riverine ecosystems. A high N2 fixation rate may increase the ratio of N to P input to river systems, and therefore render P the limiting factor in aquatic eutrophication.

Quantifying Oxygen Management and Temperature and Light Dependencies of Nitrogen Fixation by Crocosphaera watsonii.

Crocosphaera is a major dinitrogen (N2)-fixing microorganism, providing bioavailable nitrogen (N) to marine ecosystems. The N2-fixing enzyme nitrogenase is deactivated by oxygen (O2), which is abundant in marine environments. Using a cellular scale model of Crocosphaera sp. and laboratory data, we quantify the role of three O2 management strategies by Crocosphaera sp.: size adjustment, reduced O2 diffusivity, and respiratory protection. Our model predicts that Crocosphaera cells increase their size under high O2 Using transmission electron microscopy, we show that starch granules and thylakoid membranes are located near the cytoplasmic membranes, forming a barrier for O2 The model indicates a critical role for respiration in protecting the rate of N2 fixation. Moreover, the rise in respiration rates and the decline in ambient O2 with temperature strengthen this mechanism in warmer water, providing a physiological rationale for the observed niche of Crocosphaera at temperatures exceeding 20°C. Our new measurements of the sensitivity to light intensity show that the rate of N2 fixation reaches saturation at a lower light intensity (∼100 μmol m-2 s-1) than photosynthesis and that both are similarly inhibited by light intensities of >500 μmol m-2 s-1 This suggests an explanation for the maximum population of Crocosphaera occurring slightly below the ocean surface.IMPORTANCECrocosphaera is one of the major N2-fixing microorganisms in the open ocean. On a global scale, the process of N2 fixation is important in balancing the N budget, but the factors governing the rate of N2 fixation remain poorly resolved. Here, we combine a mechanistic model and both previous and present laboratory studies of Crocosphaera to quantify how chemical factors such as C, N, Fe, and O2 and physical factors such as temperature and light affect N2 fixation. Our study shows that Crocosphaera combines multiple mechanisms to reduce intracellular O2 to protect the O2-sensitive N2-fixing enzyme. Our model, however, indicates that these protections are insufficient at low temperature due to reduced respiration and the rate of N2 fixation becomes severely limited. This provides a physiological explanation for why the geographic distribution of Crocosphaera is confined to the warm low-latitude ocean.

Denitrification Aligns with N2 Fixation in Red Sea Corals

Denitrification may potentially alleviate excess nitrogen (N) availability in coral holobionts to maintain a favourable N to phosphorous ratio in the coral tissue. However, little is known about the abundance and activity of denitrifiers in the coral holobiont. The present study used the nirS marker gene as a proxy for denitrification potential along with measurements of denitrification rates in a comparative coral taxonomic framework from the Red Sea: Acropora hemprichii, Millepora dichotoma, and Pleuractis granulosa. Relative nirS gene copy numbers associated with the tissues of these common corals were assessed and compared with denitrification rates on the holobiont level. In addition, dinitrogen (N2) fixation rates, Symbiodiniaceae cell density, and oxygen evolution were assessed to provide an environmental context for denitrification. We found that relative abundances of the nirS gene were 16- and 17-fold higher in A. hemprichii compared to M. dichotoma and P. granulosa, respectively. In concordance, highest denitrification rates were measured in A. hemprichii, followed by M. dichotoma and P. granulosa. Denitrification rates were positively correlated with N2 fixation rates and Symbiodiniaceae cell densities. Our results suggest that denitrification may counterbalance the N input from N2 fixation in the coral holobiont, and we hypothesize that these processes may be limited by photosynthates released by the Symbiodiniaceae.