Integration of nitrogen dynamics into the Noah-MP land surface model v1.1 for climate and environmental predictions

NASA Astrophysics Data System (ADS)

Cai, X.; Yang, Z.-L.; Fisher, J. B.; Zhang, X.; Barlage, M.; Chen, F.

2016-01-01

Climate and terrestrial biosphere models consider nitrogen an important factor in limiting plant carbon uptake, while operational environmental models view nitrogen as the leading pollutant causing eutrophication in water bodies. The community Noah land surface model with multi-parameterization options (Noah-MP) is unique in that it is the next-generation land surface model for the Weather Research and Forecasting meteorological model and for the operational weather/climate models in the National Centers for Environmental Prediction. In this study, we add a capability to Noah-MP to simulate nitrogen dynamics by coupling the Fixation and Uptake of Nitrogen (FUN) plant model and the Soil and Water Assessment Tool (SWAT) soil nitrogen dynamics. This model development incorporates FUN's state-of-the-art concept of carbon cost theory and SWAT's strength in representing the impacts of agricultural management on the nitrogen cycle. Parameterizations for direct root and mycorrhizal-associated nitrogen uptake, leaf retranslocation, and symbiotic biological nitrogen fixation are employed from FUN, while parameterizations for nitrogen mineralization, nitrification, immobilization, volatilization, atmospheric deposition, and leaching are based on SWAT. The coupled model is then evaluated at the Kellogg Biological Station - a Long Term Ecological Research site within the US Corn Belt. Results show that the model performs well in capturing the major nitrogen state/flux variables (e.g., soil nitrate and nitrate leaching). Furthermore, the addition of nitrogen dynamics improves the modeling of net primary productivity and evapotranspiration. The model improvement is expected to advance the capability of Noah-MP to simultaneously predict weather and water quality in fully coupled Earth system models.

Nitrate and Ammonium Induced Photosynthetic Suppression in N-Limited Selenastrum minutum.

PubMed

Elrifi, I R; Turpin, D H

1986-05-01

Nitrate-limited chemostat cultures of Selenastrum minutum Naeg. Collins (Chlorophyta) were used to determine the effects of nitrogen addition on photosynthesis, dark respiration, and dark carbon fixation. Addition of NO(3) (-) or NH(4) (+) induced a transient suppression of photosynthetic carbon fixation (70 and 40% respectively). Intracellular ribulose bisphosphate levels decreased during suppression and recovered in parallel with photosynthesis. Photosynthetic oxygen evolution was decreased by N-pulsing under saturating light (650 microeinsteins per square meter per second). Under subsaturating light intensities (<165 microeinsteins per square meter per second) NH(4) (+) addition resulted in O(2) consumption in the light which was alleviated by the presence of the tricarboxylic acid cycle inhibitor fluoroacetate. Addition of NO(3) (-) or NH(4) (+) resulted in a large stimulation of dark respiration (67 and 129%, respectively) and dark carbon fixation (360 and 2080%, respectively). The duration of N-induced perturbations was dependent on the concentration of added N. Inhibition of glutamine 2-oxoglutarate aminotransferase by azaserine alleviated all these effects. It is proposed that suppression of photosynthetic carbon fixation in response to N pulsing was the result of a competition for metabolites between the Calvin cycle and nitrogen assimilation. Carbon skeletons required for nitrogen assimilation would be derived from tricarboxylic acid cycle intermediates. To maintain tricarboxylic acid cycle activity triose phosphates would be exported from the chloroplast. This would decrease the rate of ribulose bisphosphate regeneration and consequently decrease net photosynthetic carbon accumulation. Stoichiometric calculations indicate that the Calvin cycle is one source of triose phosphates for N assimilation; however, during transient N resupply the major demand for triose phosphates must be met by starch or sucrose breakdown. The effects of N-pulsing on O(2) evolution, dark respiration, and dark C-fixation are shown to be consistent with this model.

Dependence of wheat and rice respiration on tissue nitrogen and the corresponding net carbon fixation efficiency under different rates of nitrogen application

NASA Astrophysics Data System (ADS)

Sun, Wenjuan; Huang, Yao; Chen, Shutao; Zou, Jianwen; Zheng, Xunhua

2007-02-01

To quantitatively address the role of tissue N in crop respiration under various agricultural practices, and to consequently evaluate the impact of synthetic fertilizer N application on biomass production and respiration, and hence net carbon fixation efficiency ( E ncf), pot and field experiments were carried out for an annual rotation of a rice-wheat cropping system from 2001 to 2003. The treatments of the pot experiments included fertilizer N application, sowing date and planting density. Different rates of N application were tested in the field experiments. Static opaque chambers were used for sampling the gas. The respiration as CO2 emission was detected by a gas chromatograph. A successive biomass clipping method was employed to determine the crop autotrophic respiration coefficient ( R a). Results from the pot experiments revealed a linear relationship between R a and tissue N content as R a = 4.74N-1.45 ( R 2 = 0.85, P < 0.001). Measurements and calculations from the field experiments indicated that fertilizer N application promoted not only biomass production but also increased the respiration of crops. A further investigation showed that the increase of carbon loss in terms of respiration owing to fertilizer N application exceeded that of net carbon gain in terms of aboveground biomass when fertilizer N was applied over a certain rate. Consequently, the E ncf declined as the N application rate increased.

Estimated anthropogenic nitrogen and phosphorus inputs to the land surface of the conterminous United States--1992, 1997, and 2002

USGS Publications Warehouse

Sprague, Lori A.; Gronberg, Jo Ann M.

2013-01-01

Anthropogenic inputs of nitrogen and phosphorus to each county in the conterminous United States and to the watersheds of 495 surface-water sites studied as part of the U.S. Geological Survey National Water-Quality Assessment Program were quantified for the years 1992, 1997, and 2002. Estimates of inputs of nitrogen and phosphorus from biological fixation by crops (for nitrogen only), human consumption, crop production for human consumption, animal production for human consumption, animal consumption, and crop production for animal consumption for each county are provided in a tabular dataset. These county-level estimates were allocated to the watersheds of the surface-water sites to estimate watershed-level inputs from the same sources; these estimates also are provided in a tabular dataset, together with calculated estimates of net import of food and net import of feed and previously published estimates of inputs from atmospheric deposition, fertilizer, and recoverable manure. The previously published inputs are provided for each watershed so that final estimates of total anthropogenic nutrient inputs could be calculated. Estimates of total anthropogenic inputs are presented together with previously published estimates of riverine loads of total nitrogen and total phosphorus for reference.

Nitrogen Fixation Inputs in Pasture and Early Successional Forest in the Brazilian Amazon Region: Evidence From a Claybox Mesocosm Study

NASA Astrophysics Data System (ADS)

Davidson, Eric A.; Markewitz, Daniel; de O. Figueiredo, Ricardo; de Camargo, Plínio B.

2018-02-01

The role of biological nitrogen fixation (BNF) during secondary forest succession and in tropical pastures has been investigated and debated for several decades. Here we present results of a replicated experimental study in a degraded cattle pasture of eastern Amazonia using mass balance and a 15N tracer in lined soil pit mesocosms with three treatments: (1) plant-free control plots, (2) pasture grass Brachiaria brizantha, and (3) regrowth of early successional secondary forest species. Accumulation of N in grass biomass slightly exceeded estimates of net N mineralization from the plant-free control plots but was within the margin of error, so inputs of BNF may not have been needed. In contrast, the secondary forest vegetation accumulated about 3 times as much biomass N annually as the net N mineralization estimate, suggesting at least some role for BNF. Based on isotopic and mass measurements of N-fixing species, BNF was estimated to contribute at least 27 ± 3% of mean annual plant uptake in the secondary forest regrowth vegetation plots. Although BNF is probably important for recuperation of tropical secondary forests following land use change, the majority of the N taken up by both grasses and secondary forest regrowth arose from mineralization of the stocks of soil N.

Biome-scale nitrogen fixation strategies selected by climatic constraints on nitrogen cycle.

PubMed

Sheffer, Efrat; Batterman, Sarah A; Levin, Simon A; Hedin, Lars O

2015-11-23

Dinitrogen fixation by plants (in symbiosis with root bacteria) is a major source of new nitrogen for land ecosystems(1). A long-standing puzzle(2) is that trees capable of nitrogen fixation are abundant in nitrogen-rich tropical forests, but absent or restricted to early successional stages in nitrogen-poor extra-tropical forests. This biome-scale pattern presents an evolutionary paradox(3), given that the physiological cost(4) of nitrogen fixation predicts the opposite pattern: fixers should be out-competed by non-fixers in nitrogen-rich conditions, but competitively superior in nitrogen-poor soils. Here we evaluate whether this paradox can be explained by the existence of different fixation strategies in tropical versus extra-tropical trees: facultative fixers (capable of downregulating fixation(5,6) by sanctioning mutualistic bacteria(7)) are common in the tropics, whereas obligate fixers (less able to downregulate fixation) dominate at higher latitudes. Using a game-theoretic approach, we assess the ecological and evolutionary conditions under which these fixation strategies emerge, and examine their dependence on climate-driven differences in the nitrogen cycle. We show that in the tropics, transient soil nitrogen deficits following disturbance and rapid tree growth favour a facultative strategy and the coexistence of fixers and non-fixers. In contrast, sustained nitrogen deficits following disturbance in extra-tropical forests favour an obligate fixation strategy, and cause fixers to be excluded in late successional stages. We conclude that biome-scale differences in the abundance of nitrogen fixers can be explained by the interaction between individual plant strategies and climatic constraints on the nitrogen cycle over evolutionary time.

The fate of nitrogen fixed by diazotrophs in the ocean

NASA Astrophysics Data System (ADS)

Mulholland, M. R.

2007-01-01

While we now know that N2 fixation is a significant source of new nitrogen (N) in the marine environment, little is known about the fate of this N (and associated C), despite the importance of diazotrophs to global carbon and nutrient cycles. Specifically, does N fixed during N2 fixation fuel autotrophic or heterotrophic growth and thus facilitate carbon (C) export from the euphotic zone, or does it contribute primarily to bacterial productivity and respiration in the euphotic zone? For Trichodesmium, the diazotroph we know the most about, the transfer of recently fixed N2 (and C) appears to be primarily through dissolved pools. The release of N varies among and within populations and as a result of the changing physiological state of cells and populations. The net result of trophic transfers appears to depend on the co-occurring organisms and the complexity of the colonizing community. In order to understand the impact of diazotrophy on carbon flow and export in marine systems, we need a better understanding of the trophic flow of elements in Trichodesmium-dominated communities and other diazotrophic communities under various defined physiological states. Nitrogen and carbon fixation rates themselves vary by orders of magnitude within and among studies of Trichodesmium, highlighting the difficulty in extrapolating global rates of N2 fixation from direct measurements. Because the stoichiometry of N2 and C fixation does not appear to be in balance with that of particles, and the relationship between C and N2 fixation rates is also variable, it is equally difficult to derive global rates of one from the other. This paper seeks to synthesize what is known about the fate of diazotrophic production in the environment. A better understanding of the physiology and physiological ecology of Trichodesmium and other marine diazotrophs is necessary to quantify and predict the effects of increased or decreased diazotrophy in the context of the carbon cycle and global change.

Role of boron nutrient in nodules growth and nitrogen fixation rates in soybean genotypes under water stress conditions

USDA-ARS?s Scientific Manuscript database

Although boron has a stimulatory effect on nodule growth and nitrogen fixation, mechanisms of how boron affects nodules growth and nitrogen fixation, especially under water stress, are still unknown. The stimulatory effect of boron (B) on nodules and nitrogen fixation (NF) is influenced by biotic (s...

Integration of nitrogen dynamics into the Noah-MP land surface model v1.1 for climate and environmental predictions

DOE PAGES

Cai, X.; Yang, Z. -L.; Fisher, J. B.; ...

2016-01-15

Climate and terrestrial biosphere models consider nitrogen an important factor in limiting plant carbon uptake, while operational environmental models view nitrogen as the leading pollutant causing eutrophication in water bodies. The community Noah land surface model with multi-parameterization options (Noah-MP) is unique in that it is the next-generation land surface model for the Weather Research and Forecasting meteorological model and for the operational weather/climate models in the National Centers for Environmental Prediction. Here in this study, we add a capability to Noah-MP to simulate nitrogen dynamics by coupling the Fixation and Uptake of Nitrogen (FUN) plant model and the Soilmore » and Water Assessment Tool (SWAT) soil nitrogen dynamics. This model development incorporates FUN's state-of-the-art concept of carbon cost theory and SWAT's strength in representing the impacts of agricultural management on the nitrogen cycle. Parameterizations for direct root and mycorrhizal-associated nitrogen uptake, leaf retranslocation, and symbiotic biological nitrogen fixation are employed from FUN, while parameterizations for nitrogen mineralization, nitrification, immobilization, volatilization, atmospheric deposition, and leaching are based on SWAT. The coupled model is then evaluated at the Kellogg Biological Station – a Long Term Ecological Research site within the US Corn Belt. Results show that the model performs well in capturing the major nitrogen state/flux variables (e.g., soil nitrate and nitrate leaching). Furthermore, the addition of nitrogen dynamics improves the modeling of net primary productivity and evapotranspiration. The model improvement is expected to advance the capability of Noah-MP to simultaneously predict weather and water quality in fully coupled Earth system models.« less

Integration of nitrogen dynamics into the Noah-MP land surface model v1.1 for climate and environmental predictions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cai, X.; Yang, Z. -L.; Fisher, J. B.

Climate and terrestrial biosphere models consider nitrogen an important factor in limiting plant carbon uptake, while operational environmental models view nitrogen as the leading pollutant causing eutrophication in water bodies. The community Noah land surface model with multi-parameterization options (Noah-MP) is unique in that it is the next-generation land surface model for the Weather Research and Forecasting meteorological model and for the operational weather/climate models in the National Centers for Environmental Prediction. Here in this study, we add a capability to Noah-MP to simulate nitrogen dynamics by coupling the Fixation and Uptake of Nitrogen (FUN) plant model and the Soilmore » and Water Assessment Tool (SWAT) soil nitrogen dynamics. This model development incorporates FUN's state-of-the-art concept of carbon cost theory and SWAT's strength in representing the impacts of agricultural management on the nitrogen cycle. Parameterizations for direct root and mycorrhizal-associated nitrogen uptake, leaf retranslocation, and symbiotic biological nitrogen fixation are employed from FUN, while parameterizations for nitrogen mineralization, nitrification, immobilization, volatilization, atmospheric deposition, and leaching are based on SWAT. The coupled model is then evaluated at the Kellogg Biological Station – a Long Term Ecological Research site within the US Corn Belt. Results show that the model performs well in capturing the major nitrogen state/flux variables (e.g., soil nitrate and nitrate leaching). Furthermore, the addition of nitrogen dynamics improves the modeling of net primary productivity and evapotranspiration. The model improvement is expected to advance the capability of Noah-MP to simultaneously predict weather and water quality in fully coupled Earth system models.« less

Distribution of nitrogen fixation and nitrogenase-like sequences amongst microbial genomes

PubMed Central

2012-01-01

Background The metabolic capacity for nitrogen fixation is known to be present in several prokaryotic species scattered across taxonomic groups. Experimental detection of nitrogen fixation in microbes requires species-specific conditions, making it difficult to obtain a comprehensive census of this trait. The recent and rapid increase in the availability of microbial genome sequences affords novel opportunities to re-examine the occurrence and distribution of nitrogen fixation genes. The current practice for computational prediction of nitrogen fixation is to use the presence of the nifH and/or nifD genes. Results Based on a careful comparison of the repertoire of nitrogen fixation genes in known diazotroph species we propose a new criterion for computational prediction of nitrogen fixation: the presence of a minimum set of six genes coding for structural and biosynthetic components, namely NifHDK and NifENB. Using this criterion, we conducted a comprehensive search in fully sequenced genomes and identified 149 diazotrophic species, including 82 known diazotrophs and 67 species not known to fix nitrogen. The taxonomic distribution of nitrogen fixation in Archaea was limited to the Euryarchaeota phylum; within the Bacteria domain we predict that nitrogen fixation occurs in 13 different phyla. Of these, seven phyla had not hitherto been known to contain species capable of nitrogen fixation. Our analyses also identified protein sequences that are similar to nitrogenase in organisms that do not meet the minimum-gene-set criteria. The existence of nitrogenase-like proteins lacking conserved co-factor ligands in both diazotrophs and non-diazotrophs suggests their potential for performing other, as yet unidentified, metabolic functions. Conclusions Our predictions expand the known phylogenetic diversity of nitrogen fixation, and suggest that this trait may be much more common in nature than it is currently thought. The diverse phylogenetic distribution of nitrogenase-like proteins indicates potential new roles for anciently duplicated and divergent members of this group of enzymes. PMID:22554235

High potential of nitrogen fixation in pristine, ombrotrophic bogs in Southern Patagonia

NASA Astrophysics Data System (ADS)

Knorr, Klaus-Holger; Horn, Marcus A.; Bahamonde Aguilar, Nelson A.; Borken, Werner

2015-04-01

Nitrogen (N) input in pristine peatlands occurs via natural input of inorganic N through atmospheric deposition or biological dinitrogen (N2) fixation. However, N2 fixation is to date mostly attributed to bacteria and algae associated to Sphagnum and its contribution to plant productivity and peat buildup has been often underestimated in previous studies. Based on net N storage, exceptionally low N deposition, and high abundance of vascular plants at pristine peatlands in Southern Patagonia, we hypothesized that there must be a high potential of non-symbiotic N2 fixation not limited to the occurrence of Sphagnum. To this end, we chose two ombrotrophic bogs with spots that are dominated either by Sphagnum or by vascular, cushion-forming plants and sampled peat from different depths for incubation with 15N2 to determine N2 fixation potentials. Moreover, we analyzed 15N2 fixation by a nodule-forming, endemic conifer inhabiting the peatlands. Results from 15N2 uptake were compared to the conventional approach to study N2 fixation by the acetylene reduction assay (ARA). Using 15N2 as a tracer, high non-symbiotic N2 fixation rates of 0.3-1.4 μmol N g-1 d-1 were found down to 50 cm under micro-oxic conditions (2 vol.%) in samples from both plots either covered by Sphagnum magellanicum or by vascular cushion plants. Peat N concentrations suggested a higher potential of non-symbiotic N2 fixation under cushion plants, likely because of the availability of easily decomposable organic compounds as substrates and oxic conditions in the rhizosphere. In the Sphagnum plots, high N2 fixation below 10 cm depth would rather reflect a potential fixation that may switch on during periods of low water levels when oxygen penetrates deeper into the peat. 15N natural abundance of live Sphagnum from 0-10 cm pointed to N uptake solely from atmospheric deposition and non-symbiotic N2 fixation. 15N signatures of peat from the cushion plant plots indicated additional N supply from N mineralization. Nitrogen fixation by the conifer Lepidothamnus fonkii was exceptionally high, reaching 3.1 μmol N g-1 d.w. d-1 detected in roots, stems, and green biomass. For L. fonkii, we could identify a specific association with Beijerinckiaceae as N2 fixing bacteria in the root nodules, whereas the rhizosphere peat was dominated by other diazotrophs. The ARA considerably underestimated N2 fixation and can thus not be recommended for peatland studies. Our findings suggest that non-symbiotic or associative N2 fixation overcomes N deficiency in different vegetation communities and has great significance for N cycling and peat accumulation in pristine peatlands.

QTL analysis of symbiotic nitrogen fixation in a black bean RIL population

USDA-ARS?s Scientific Manuscript database

Dry bean (Phaseolus vulgaris L) acquires nitrogen (N) from the atmosphere through symbiotic nitrogen fixation (SNF) but it has a low efficiency to fix nitrogen. The objective of this study is to map the genes controlling nitrogen fixation in common bean. A mapping population consisting of 122 recomb...

Nitrogen Fixation by Anaerobes is Stimulated by Low Oxygen and Insensitive to Combined Nitrogen in Coastal Sediments

NASA Astrophysics Data System (ADS)

Jenkins, B. D.; Spinette, R.; Jones, A.; Puggioni, G.; Ehrlich, A.; Brown, S. M.

2016-02-01

Coastal sediments are typically zones of nitrogen removal via coupled nitrification-denitrification pathways. Increasingly, there are reports of nitrogen fixation in anthropogenically impacted sediments containing ample combined nitrogen. In previous work in the estuarine sediments of Narragansett Bay, we found that anaerobes related to Desulfovibrio spp. and in the Desulfuromonadales express genes for nitrogen fixation (nifH). We also determined that nitrogen fixation rates and gene expression are elevated during periods of seasonal hypoxia. Statistical modeling shows that a combination of elevated phytoplankton biomass as with a duration of hypoxia for a week or longer lead to conditions that promote nitrogen fixation as measured by acetylene reduction. Interestingly, diazotrophs closely related to those identified in Narragansett Bay are present and active in other low oxygen systems, suggesting that expansion of hypoxic events may lead to unanticipated consequences for the benthic nitrogen cycle in many ecosystems. To determine controls on diazotrophy on the organismal level, we isolated and sequenced the genomes of two Narragansett Bay members of the Desulfovibrio. We found that these organisms are insensitive to nitrate and urea, as they are missing the genes to assimilate these nitrogen sources. However, their nitrogen fixation is suppressed by increasing concentrations of ammonium, indicating that they may be sensitive to this nitrogen source in the environment. The paradox of detectable nitrogen fixation in the background of measurable ammonium in estuarine systems is a newly emergent theme and suggests that there are complex microbial interactions and/or structure to the nutrient regimes allowing for fixation.

Biochemical Approaches to Improved Nitrogen Fixation

USDA-ARS?s Scientific Manuscript database

Improving symbiotic nitrogen fixation by legumes has emerged again as an important topic on the world scene due to the energy crisis and lack of access to nitrogen fertilizer in developing countries. We have taken a biochemical genomics approach to improving symbiotic nitrogen fixation in legumes. L...

Nitrate and Ammonium Induced Photosynthetic Suppression in N-Limited Selenastrum minutum1

PubMed Central

Elrifi, Ivor R.; Turpin, David H.

1986-01-01

Nitrate-limited chemostat cultures of Selenastrum minutum Naeg. Collins (Chlorophyta) were used to determine the effects of nitrogen addition on photosynthesis, dark respiration, and dark carbon fixation. Addition of NO3− or NH4+ induced a transient suppression of photosynthetic carbon fixation (70 and 40% respectively). Intracellular ribulose bisphosphate levels decreased during suppression and recovered in parallel with photosynthesis. Photosynthetic oxygen evolution was decreased by N-pulsing under saturating light (650 microeinsteins per square meter per second). Under subsaturating light intensities (<165 microeinsteins per square meter per second) NH4+ addition resulted in O2 consumption in the light which was alleviated by the presence of the tricarboxylic acid cycle inhibitor fluoroacetate. Addition of NO3− or NH4+ resulted in a large stimulation of dark respiration (67 and 129%, respectively) and dark carbon fixation (360 and 2080%, respectively). The duration of N-induced perturbations was dependent on the concentration of added N. Inhibition of glutamine 2-oxoglutarate aminotransferase by azaserine alleviated all these effects. It is proposed that suppression of photosynthetic carbon fixation in response to N pulsing was the result of a competition for metabolites between the Calvin cycle and nitrogen assimilation. Carbon skeletons required for nitrogen assimilation would be derived from tricarboxylic acid cycle intermediates. To maintain tricarboxylic acid cycle activity triose phosphates would be exported from the chloroplast. This would decrease the rate of ribulose bisphosphate regeneration and consequently decrease net photosynthetic carbon accumulation. Stoichiometric calculations indicate that the Calvin cycle is one source of triose phosphates for N assimilation; however, during transient N resupply the major demand for triose phosphates must be met by starch or sucrose breakdown. The effects of N-pulsing on O2 evolution, dark respiration, and dark C-fixation are shown to be consistent with this model. PMID:16664788

Nitrogen and Phosphorus Addition Affects Biological N2 Fixation and Sphagnum Moss in an Ombrotrophic Bog

NASA Astrophysics Data System (ADS)

Zivkovic, T.; Ardichvili, A.; Moore, T. R.

2016-12-01

Most of the 18 Pg nitrogen (N) accumulated in northern nutrient-poor and Sphagnum-dominated peatlands (bogs and fens) can be attributed to N2 fixation by diazotrophs either associated with the live Sphagnum or non-symbiotically in the deeper peat. Where atmospheric N deposition is low (< 0.2 g m-2 y-1), ombrotrophic bogs rely on N2 fixation as the primary source of N that sustains primary production. The industrial revolution and anthropogenic impacts in the last 200 years have resulted in larger atmospheric N deposition as ammonium (NH4) and nitrate (NO3). One effect of increased N deposition in Sphagnum is a switch from N to phosphorus (P) limitation suggested by the increase in tissue N:P>16. It is unclear how Sphagnum hosted diazotrophic activity may be affected by N deposition and thus changes in N:P ratio. We investigated the effects of long-term addition of different sources of nitrogen (0, 1.6, 3.2 and 6.4 g N m-2 y-1 as NH4Cl and NaNO3), and phosphorus (5 g P m-2 y-1 as KH2PO4) on Sphagnum nutrient status (N, P and N:P), net primary productivity (NPP) and Sphagnum-associated N2 fixation at Mer Bleue, a temperate ombrotrophic bog. Our study shows that N concentration in Sphagnum tissue increased with larger rates of N addition, with a stronger effect on Sphagnum from NH4 than NO3. The addition of P created a 3.5 fold increase in Sphagnum P content compared to controls. Sphagnum NPP decreased linearly with the rise in N:P ratio, while linear growth declined exponentially with increase in Sphagnum N content. N2 fixation significantly decreased in response to even the smallest addition of both N species. In contrast, the addition of P increased N2 fixation by up to 100 times compared to N treatments and up to 5-30 times compared to controls. The change in N2 fixation was best modeled by the N:P ratio, across all experimental treatments. Although elevated N deposition substantially decreases N2 fixation, the N:P ratio in Sphagnum may be a good predictor, likely owing to a strong P-limitation.

Nitrogen fixation in denitrified marine waters.

PubMed

Fernandez, Camila; Farías, Laura; Ulloa, Osvaldo

2011-01-01

Nitrogen fixation is an essential process that biologically transforms atmospheric dinitrogen gas to ammonia, therefore compensating for nitrogen losses occurring via denitrification and anammox. Currently, inputs and losses of nitrogen to the ocean resulting from these processes are thought to be spatially separated: nitrogen fixation takes place primarily in open ocean environments (mainly through diazotrophic cyanobacteria), whereas nitrogen losses occur in oxygen-depleted intermediate waters and sediments (mostly via denitrifying and anammox bacteria). Here we report on rates of nitrogen fixation obtained during two oceanographic cruises in 2005 and 2007 in the eastern tropical South Pacific (ETSP), a region characterized by the presence of coastal upwelling and a major permanent oxygen minimum zone (OMZ). Our results show significant rates of nitrogen fixation in the water column; however, integrated rates from the surface down to 120 m varied by ∼30 fold between cruises (7.5±4.6 versus 190±82.3 µmol m(-2) d(-1)). Moreover, rates were measured down to 400 m depth in 2007, indicating that the contribution to the integrated rates of the subsurface oxygen-deficient layer was ∼5 times higher (574±294 µmol m(-2) d(-1)) than the oxic euphotic layer (48±68 µmol m(-2) d(-1)). Concurrent molecular measurements detected the dinitrogenase reductase gene nifH in surface and subsurface waters. Phylogenetic analysis of the nifH sequences showed the presence of a diverse diazotrophic community at the time of the highest measured nitrogen fixation rates. Our results thus demonstrate the occurrence of nitrogen fixation in nutrient-rich coastal upwelling systems and, importantly, within the underlying OMZ. They also suggest that nitrogen fixation is a widespread process that can sporadically provide a supplementary source of fixed nitrogen in these regions.

Stimulation of nitrogen fixation in soddy-podzolic soils with fungi

NASA Astrophysics Data System (ADS)

Kurakov, A. V.; Prokhorov, I. S.; Kostina, N. V.; Makhova, E. G.; Sadykova, V. S.

2006-09-01

Stimulation of nitrogen fixation in soddy-podzolic soils is related to the hydrolytic activity of fungi decomposing plant polymers. It was found that the rate of nitrogen fixation upon the simultaneous inoculation of the strains of nitrogen-fixing bacteria Bacillus cereus var. mycoides and the cellulolytic fungus Trichoderma asperellum into a sterile soil enriched with cellulose or Jerusalem artichoke residues is two to four times higher than upon the inoculation of the strains of Bacillus cereus var. mycoides L1 only. The increase in the nitrogen fixation depended on the resistance of the substrates added into the soil to fungal hydrolysis. The biomass of the fungi decomposing plant polymers increased by two-four times. The nitrogen-fixing activity of the soil decreased when the growth of the fungi was inhibited with cycloheximide, which attested to a close correlation between the intensity of the nitrogen fixation and the decomposition of the plant polymers by fungi. The introduction of an antifungal antibiotic, together with starch or with plant residues, significantly (by 60-90%) decreased the rate of nitrogen fixation in the soll.

Nitrogen fixation on early Mars and other terrestrial planets: experimental demonstration of abiotic fixation reactions to nitrite and nitrate.

PubMed

Summers, David P; Khare, Bishun

2007-04-01

Understanding the abiotic fixation of nitrogen is critical to understanding planetary evolution and the potential origin of life on terrestrial planets. Nitrogen, an essential biochemical element, is certainly necessary for life as we know it to arise. The loss of atmospheric nitrogen can result in an incapacity to sustain liquid water and impact planetary habitability and hydrological processes that shape the surface. However, our current understanding of how such fixation may occur is almost entirely theoretical. This work experimentally examines the chemistry, in both gas and aqueous phases, that would occur from the formation of NO and CO by the shock heating of a model carbon dioxide/nitrogen atmosphere such as is currently thought to exist on early terrestrial planets. The results show that two pathways exist for the abiotic fixation of nitrogen from the atmosphere into the crust: one via HNO and another via NO(2). Fixation via HNO, which requires liquid water, could represent fixation on a planet with liquid water (and hence would also be a source of nitrogen for the origin of life). The pathway via NO(2) does not require liquid water and shows that fixation could occur even when liquid water has been lost from a planet's surface (for example, continuing to remove nitrogen through NO(2) reaction with ice, adsorbed water, etc.).

Symbiotic Nitrogen Fixation and the Challenges to Its Extension to Nonlegumes

PubMed Central

Mus, Florence; Crook, Matthew B.; Garcia, Kevin; Garcia Costas, Amaya; Geddes, Barney A.; Kouri, Evangelia D.; Paramasivan, Ponraj; Ryu, Min-Hyung; Oldroyd, Giles E. D.; Poole, Philip S.; Udvardi, Michael K.; Voigt, Christopher A.

2016-01-01

Access to fixed or available forms of nitrogen limits the productivity of crop plants and thus food production. Nitrogenous fertilizer production currently represents a significant expense for the efficient growth of various crops in the developed world. There are significant potential gains to be had from reducing dependence on nitrogenous fertilizers in agriculture in the developed world and in developing countries, and there is significant interest in research on biological nitrogen fixation and prospects for increasing its importance in an agricultural setting. Biological nitrogen fixation is the conversion of atmospheric N2 to NH3, a form that can be used by plants. However, the process is restricted to bacteria and archaea and does not occur in eukaryotes. Symbiotic nitrogen fixation is part of a mutualistic relationship in which plants provide a niche and fixed carbon to bacteria in exchange for fixed nitrogen. This process is restricted mainly to legumes in agricultural systems, and there is considerable interest in exploring whether similar symbioses can be developed in nonlegumes, which produce the bulk of human food. We are at a juncture at which the fundamental understanding of biological nitrogen fixation has matured to a level that we can think about engineering symbiotic relationships using synthetic biology approaches. This minireview highlights the fundamental advances in our understanding of biological nitrogen fixation in the context of a blueprint for expanding symbiotic nitrogen fixation to a greater diversity of crop plants through synthetic biology. PMID:27084023

Symbiosis revisited: phosphorus and acid buffering stimulate N2 fixation but not Sphagnum growth

NASA Astrophysics Data System (ADS)

van den Elzen, Eva; Kox, Martine A. R.; Harpenslager, Sarah F.; Hensgens, Geert; Fritz, Christian; Jetten, Mike S. M.; Ettwig, Katharina F.; Lamers, Leon P. M.

2017-03-01

In pristine Sphagnum-dominated peatlands, (di)nitrogen (N2) fixing (diazotrophic) microbial communities associated with Sphagnum mosses contribute substantially to the total nitrogen input, increasing carbon sequestration. The rates of symbiotic nitrogen fixation reported for Sphagnum peatlands, are, however, highly variable, and experimental work on regulating factors that can mechanistically explain this variation is largely lacking. For two common fen species (Sphagnum palustre and S. squarrosum) from a high nitrogen deposition area (25 kg N ha-1 yr-1), we found that diazotrophic activity (as measured by 15 - 15N2 labeling) was still present at a rate of 40 nmol N gDW-1 h-1. This was surprising, given that nitrogen fixation is a costly process. We tested the effects of phosphorus availability and buffering capacity by bicarbonate-rich water, mimicking a field situation in fens with stronger groundwater or surface water influence, as potential regulators of nitrogen fixation rates and Sphagnum performance. We expected that the addition of phosphorus, being a limiting nutrient, would stimulate both diazotrophic activity and Sphagnum growth. We indeed found that nitrogen fixation rates were doubled. Plant performance, in contrast, did not increase. Raised bicarbonate levels also enhanced nitrogen fixation, but had a strong negative impact on Sphagnum performance. These results explain the higher nitrogen fixation rates reported for minerotrophic and more nutrient-rich peatlands. In addition, nitrogen fixation was found to strongly depend on light, with rates 10 times higher in light conditions suggesting high reliance on phototrophic organisms for carbon. The contrasting effects of phosphorus and bicarbonate on Sphagnum spp. and their diazotrophic communities reveal strong differences in the optimal niche for both partners with respect to conditions and resources. This suggests a trade-off for the symbiosis of nitrogen fixing microorganisms with their Sphagnum hosts, in which a sheltered environment apparently outweighs the less favorable environmental conditions. We conclude that microbial activity is still nitrogen limited under eutrophic conditions because dissolved nitrogen is being monopolized by Sphagnum. Moreover, the fact that diazotrophic activity can significantly be upregulated by increased phosphorus addition and acid buffering, while Sphagnum spp. do not benefit, reveals remarkable differences in optimal conditions for both symbiotic partners and calls into question the regulation of nitrogen fixation by Sphagnum under these eutrophic conditions. The high nitrogen fixation rates result in high additional nitrogen loading of 6 kg ha-1 yr-1 on top of the high nitrogen deposition in these ecosystems.

PHYSIOLOGY OF NITROGEN FIXATION BY BACILLUS POLYMYXA

PubMed Central

Grau, F. H.; Wilson, P. W.

1962-01-01

Grau, F. H. (University of Wisconsin, Madison) and P. W. Wilson. Physiology of nitrogen fixation by Bacillus polymyxa. J. Bacteriol. 83:490–496. 1962.—Of 17 strains of Bacillus polymyxa tested for fixation of molecular nitrogen, 15 fixed considerable quantities (30 to 150 μg N/ml). Two strains of the closely related B. macerans did not use N2, but possibly other members of this species may do so. Confirmation of fixation was obtained by showing incorporation of N15 into cell material. Both iron and molybdenum are specifically required for fixation; without the addition of these metals to the nitrogen-free medium, the growth rate and the total nitrogen fixed were reduced about 30 to 50%. No requirement for added molybdenum could be shown when ammonia was the nitrogen source, and the absence of iron caused only a slight decrease in growth. Washed-cell suspensions of B. polymyxa containing an active hydrogenase readily incorporated N15 into cell materials when provided with mannitol, glucose, or pyruvate but not when formate was the substrate. Hydrogen is a specific inhibitor of fixation, reducing both the rate and final amount of nitrogen fixed; it did not reduce growth on ammonia. Fixation was strictly anaerobic, 1% oxygen in the gas phase being sufficient to stop fixation. Arsenate is a powerful inhibitor of fixation of N2 by washed-cell suspensions of B. polymyxa, indicating that high-energy phosphate may be significant for this process. PMID:13901244

The effects of acid rain on nitrogen fixation in Western Washington coniferous forests

Treesearch

Robert Denison; Bruce Caldwell; Bernard Bormann; Lindell Eldred; Cynthia Swanberg; Steven Anderson

1976-01-01

We investigated both the current status of nitrogen fixation in Western Washington forests, and the potential effects of acid rain on this vital process. Even the low concentrations of sulfur dioxide presently found in the Northwest are thought to have an adverse effect on nitrogen fixation by limiting the distribution of the epiphytic nitrogen-fixing lichen, ...

Nitrogen Cycling in Seagrass Beds Dominated by Thalassia testudinum and Halodule wrightii: the Role of Nitrogen Fixation and Ammonium Oxidation in Regulating Ammonium Availability

NASA Astrophysics Data System (ADS)

Capps, R.; Caffrey, J. M.; Hester, C.

2016-02-01

Seagrass meadows provide key ecosystem services including nursery and foraging grounds, storm and erosion buffers, biodiversity enhancers and global carbon and nutrient cycling. Nitrogen concentrations are often very low in coastal waters and sediments, which may limit primary productivity. Biological nitrogen fixation is a microbial process that converts dinitrogen to ammonium, which is readily taken up by seagrasses. In the oxygenated rhizospheres, diazotrophs provide the plant with ammonium and use root exudates as an energy source. Nitrogen fixation rates and nutrient concentrations differ between seagrass species and substrate types. Thalassia testudinum has a higher biomass and is a climax species than Halodule wrightii, which is a pioneer species. Nitrogen fixation rates are relatively consistent in Thalassia testudinum dominated sediments. However, it is relatively variable in sediments occupied by Halodule wrightii. Nitrogen fixation rates are higher in bare substrate compared to areas with Thalassia testudinum, which may be due to T. testudinum's greater efficiency in nutrient retention because it is a climax species. We hypothesize that seasonal shifts in nitrogen fixation will coincide with seasonal shifts in seagrass biomass due to higher nutrient requirements during peak growth and lower requirements during senescence and dormancy. The ratio of porewater ammonium to phosphate suggests that seagrass growth may be nitrogen limited as does nitrogen demand, estimated from gross primary productivity. Significant rates of ammonium oxidation in both surface and rhizosphere sediments contribute to this imbalance. Thus, nitrogen fixation may be critical in supporting plant growth.

Molecular Biology of Nitrogen Fixation

ERIC Educational Resources Information Center

Shanmugam, K. T.; Valentine, Raymond C.

1975-01-01

Reports that as a result of our increasing knowledge of the molecular biology of nitrogen fixation it might eventually be possible to increase the biological production of nitrogenous fertilizer from atmospheric nitrogen. (GS)

Chemosynthetic symbionts of marine invertebrate animals are capable of nitrogen fixation.

PubMed

Petersen, Jillian M; Kemper, Anna; Gruber-Vodicka, Harald; Cardini, Ulisse; van der Geest, Matthijs; Kleiner, Manuel; Bulgheresi, Silvia; Mußmann, Marc; Herbold, Craig; Seah, Brandon K B; Antony, Chakkiath Paul; Liu, Dan; Belitz, Alexandra; Weber, Miriam

2016-10-24

Chemosynthetic symbioses are partnerships between invertebrate animals and chemosynthetic bacteria. The latter are the primary producers, providing most of the organic carbon needed for the animal host's nutrition. We sequenced genomes of the chemosynthetic symbionts from the lucinid bivalve Loripes lucinalis and the stilbonematid nematode Laxus oneistus. The symbionts of both host species encoded nitrogen fixation genes. This is remarkable as no marine chemosynthetic symbiont was previously known to be capable of nitrogen fixation. We detected nitrogenase expression by the symbionts of lucinid clams at the transcriptomic and proteomic level. Mean stable nitrogen isotope values of Loripes lucinalis were within the range expected for fixed atmospheric nitrogen, further suggesting active nitrogen fixation by the symbionts. The ability to fix nitrogen may be widespread among chemosynthetic symbioses in oligotrophic habitats, where nitrogen availability often limits primary productivity.

Recent developments in the structural organization and regulation of nitrogen fixation genes in Herbaspirillum seropedicae.

PubMed

Pedrosa, F O; Benelli, E M; Yates, M G; Wassem, R; Monteiro, R A; Klassen, G; Steffens, M B; Souza, E M; Chubatsu, L S; Rigo, L U

2001-10-04

Herbaspirillum seropedicae is a nitrogen-fixing bacterium found in association with economically important gramineae. Regulation of nitrogen fixation involves the transcriptional activator NifA protein. The regulation of NifA protein and its truncated mutant proteins is described and compared with that of other nitrogen fixation bacteria. Nitrogen fixation control in H. seropedicae, of the beta-subgroup of Proteobacteria, has regulatory features in common with Klebsiella pneumoniae, of the gamma-subgroup, at the level of nifA expression and with rhizobia and Azospirillum brasilense, of the alpha-subgroup, at the level of control of NifA by oxygen.

A representation of the phosphorus cycle for ORCHIDEE (revision 4520)

NASA Astrophysics Data System (ADS)

Goll, Daniel S.; Vuichard, Nicolas; Maignan, Fabienne; Jornet-Puig, Albert; Sardans, Jordi; Violette, Aurelie; Peng, Shushi; Sun, Yan; Kvakic, Marko; Guimberteau, Matthieu; Guenet, Bertrand; Zaehle, Soenke; Penuelas, Josep; Janssens, Ivan; Ciais, Philippe

2017-10-01

Land surface models rarely incorporate the terrestrial phosphorus cycle and its interactions with the carbon cycle, despite the extensive scientific debate about the importance of nitrogen and phosphorus supply for future land carbon uptake. We describe a representation of the terrestrial phosphorus cycle for the ORCHIDEE land surface model, and evaluate it with data from nutrient manipulation experiments along a soil formation chronosequence in Hawaii. ORCHIDEE accounts for the influence of the nutritional state of vegetation on tissue nutrient concentrations, photosynthesis, plant growth, biomass allocation, biochemical (phosphatase-mediated) mineralization, and biological nitrogen fixation. Changes in the nutrient content (quality) of litter affect the carbon use efficiency of decomposition and in return the nutrient availability to vegetation. The model explicitly accounts for root zone depletion of phosphorus as a function of root phosphorus uptake and phosphorus transport from the soil to the root surface. The model captures the observed differences in the foliage stoichiometry of vegetation between an early (300-year) and a late (4.1 Myr) stage of soil development. The contrasting sensitivities of net primary productivity to the addition of either nitrogen, phosphorus, or both among sites are in general reproduced by the model. As observed, the model simulates a preferential stimulation of leaf level productivity when nitrogen stress is alleviated, while leaf level productivity and leaf area index are stimulated equally when phosphorus stress is alleviated. The nutrient use efficiencies in the model are lower than observed primarily due to biases in the nutrient content and turnover of woody biomass. We conclude that ORCHIDEE is able to reproduce the shift from nitrogen to phosphorus limited net primary productivity along the soil development chronosequence, as well as the contrasting responses of net primary productivity to nutrient addition.

Symbiotic nitrogen fixation in an arid ecosystem measured by sup 15 N natural abundance

DOE Office of Scientific and Technical Information (OSTI.GOV)

Johnson, G.V.

1990-05-01

Plants dependent on nitrogen fixation have an {sup 15}N abundance similar to the atmosphere, while non-nitrogen fixing plants usually are enriched in {sup 15}N and are similar to soil nitrogen values. The natural abundance of {sup 15}N in leaf tissues and soils was determined to evaluate symbiotic nitrogen fixation by several legumes and actinorhizal species in the Sevilleta Long-term Ecological Research area in central New Mexico. Comparison of {delta}{sup 15}N values for the legume Prosopis glandulosa (mesquite) to adjacent Atriplex canascens (fourwing saltbush) indicated that P. glandulosa obtained 66% of its nitrogen by fixation. The legume Hoffmanseggia jamesii was foundmore » to be utilizing soil nitrogen. The {delta}{sup 15}N values for the actinorhizal plants, Elaeagnus angustifolia and Cercocarpus montanus, while below values for soil nitrogen, did not differ from associated non-fixing plants.« less

Biological Nitrogen Fixation In Tropical Dry Forests Of Costa Rica

NASA Astrophysics Data System (ADS)

Gei, M. G.; Powers, J. S.

2012-12-01

Evidence suggests that tropical dry forests (TDF) are not nitrogen (N) deficient. This evidence includes: high losses of gaseous nitrogen during the rainy season, high ecosystem soil N stocks and high N concentrations in leaves and litterfall. Its been commonly hypothesized that biological nitrogen fixation is responsible for the high availability of N in tropical soils. However, the magnitude of this flux has rarely if ever been measured in tropical dry forests. Because of the high cost of fixing N and the ubiquity of N fixing legume trees in the TDF, at the individual tree level symbiotic fixation should be a strategy down-regulated by the plant. Our main goal was to determine the rates of and controls over symbiotic N fixation. We hypothesized that legume tree species employ a facultative strategy of nitrogen fixation and that this process responds to changes in light availability, soil moisture and nutrient supply. We tested this hypothesis both on naturally established trees in a forest and under controlled conditions in a shade house by estimating the quantities of N fixed annually using the 15N natural abundance method, counting nodules, and quantifying (field) or manipulating (shade house) the variation in important environmental variables (soil nutrients, soil moisture, and light). We found that in both in our shade house experiment and in the forest, nodulation varied among different legume species. For both settings, the 15N natural abundance approach successfully detected differences in nitrogen fixation among species. The legume species that we studied were able to regulate fixation depending on the environmental conditions. They showed to have different strategies of nitrogen fixation that follow a gradient of facultative to obligate fixation. Our data suggest that there exists a continuum of nitrogen fixation strategies among species. Any efforts to define tropical legume trees as a functional group need to incorporate this variation.

Beneficial effects of aluminum enrichment on nitrogen-fixing cyanobacteria in the South China Sea.

PubMed

Liu, Jiaxing; Zhou, Linbin; Ke, Zhixin; Li, Gang; Shi, Rongjun; Tan, Yehui

2018-04-01

Few studies focus on the effects of aluminum (Al) on marine nitrogen-fixing cyanobacteria, which play important roles in the ocean nitrogen cycling. To examine the effects of Al on the nitrogen-fixing cyanobacteria, bioassay experiments in the oligotrophic South China Sea (SCS) and culture of Crocosphaera watsonii in the laboratory were conducted. Field data showed that 200 nM Al stimulated the growth and the nitrogenase gene expression of Trichodesmium and unicellular diazotrophic cyanobacterium group A, and the nitrogen fixation rates of the whole community. Laboratory experiments demonstrated that Al stimulated the growth and nitrogen fixation of C. watsonii under phosphorus limited conditions. Both field and laboratory results indicated that Al could stimulate the growth of diazotrophs and nitrogen fixation in oligotrophic oceans such as the SCS, which is likely related to the utilization of phosphorus, implying that Al plays an important role in the ocean nitrogen and carbon cycles by influencing nitrogen fixation. Copyright © 2018 Elsevier Ltd. All rights reserved.

The relationship between nitrogen fixation and the production of HD from D2 by cell-free extracts of soya-bean nodule bacteroids

PubMed Central

Turner, G. L.; Bergersen, F. J.

1969-01-01

1. Cell-free extracts prepared from soya-bean nodule bacteroids produced HD from D2 in the presence of dithionite, an ATP-generating system and nitrogen. 2. Crude extracts of bacteroids or of Azotobacter vinelandii showed some background D2 exchange when any one of these was omitted. 3. Partial purification of bacteroid extracts diminished this background activity and gave increased D2 exchange and nitrogen fixation. 4. Although increasing pN2 stimulated both reactions, the apparent Km (N2) for nitrogen fixation was much higher than the apparent Km (N2) for D2 exchange when partially purified bacteroid extracts were used. 5. Carbon monoxide was a competitive inhibitor of nitrogen fixation by partially purified bacteroid extracts, but D2 exchange was inhibited in a non-competitive fashion. 6. These results are discussed in relation to the possible existence of enzyme-bound intermediates of nitrogen fixation. PMID:5353527

Nitrogen fixation rates associated with the invasive macroalgae Sargassum horneri around Catalina Island, CA

NASA Astrophysics Data System (ADS)

DeLiberto, A.

2016-02-01

Nitrogen fixation is an important process which allows organisms access to biologically unavailable dinitrogen gas. Bacteria, known as diazotrophs use the enzyme nitrogenase to convert N2 to NH3. These bacteria, including certain species of heterotrophic bacteria and cyanobacteria, can be symbiotically associated with marine macroalgae, facilitating nutrient cycling in oligotrophic regions. As many species within the genera Sargassum are associated with nitrogen fixation, this study hypothesized that nitrogenase activity would be associated with the benthic invasive Sargassum horneri on Catalina Island. In the past decade, Sargassum horneri, an invasive from Japan, has spread throughout the waters around Catalina Island. Using the acetylene reduction procedure using flame ionization detection, initial nitrogenase activity of S. horneri sampled from Indian Rock was observed. Nitrogen fixation rates increased with decomposition, particularly in dark/anaerobic treatments, suggesting the presence of heterotrophic bacteria. In addition, acetate additions greatly increase nitrogen fixation rates, once again indicating heterotrophic nitrogen fixing bacteria.

Sulphate reduction and nitrogen fixation rates associated with roots, rhizomes and sediments from Zostera noltii and Spartina maritima meadows.

PubMed

Nielsen, L B; Finster, K; Welsh, D T; Donelly, A; Herbert, R A; de Wit, R; Lomstein, B A

2001-01-01

Sulphate reduction rates (SRR) and nitrogen fixation rates (NFR) associated with isolated roots, rhizomes and sediment from the rhizosphere of the marine macrophytes Zostera noltii and Spartina maritima, and the presence and distribution of Bacteria on the roots and rhizomes, were investigated. Between 1% and 3% of the surface area of the roots and rhizomes of both macrophytes were colonized by Bacteria. Bacteria on the surfaces of S. maritima roots and rhizomes were evenly distributed, while the distribution of Bacteria on Z. noltii roots and rhizomes was patchy. Root- and rhizome-associated SRR and NFR were always higher than rates in the bulk sediment. In particular, nitrogen fixation associated with the roots and rhizomes was 41-650-fold higher than in the bulk sediment. Despite the fact that sulphate reduction was elevated on roots and rhizomes compared with bulk sediment, the contribution of plant-associated sulphate reduction to overall sulphate reduction was small (< or =11%). In contrast, nitrogen fixation associated with the roots and rhizomes accounted for 31% and 91% of the nitrogen fixed in the rhizosphere of Z. noltii and S. maritima respectively. In addition, plant-associated nitrogen fixation could supply 37-1,613% of the nitrogen needed by the sulphate-reducing community. Sucrose stimulated nitrogen fixation and sulphate reduction significantly in the root and rhizome compartments of both macrophytes, but not in the bulk sediment.

Low rates of nitrogen fixation in eastern tropical South Pacific surface waters

PubMed Central

Knapp, Angela N.; Casciotti, Karen L.; Berelson, William M.; Prokopenko, Maria G.; Capone, Douglas G.

2016-01-01

An extensive region of the Eastern Tropical South Pacific (ETSP) Ocean has surface waters that are nitrate-poor yet phosphate-rich. It has been proposed that this distribution of surface nutrients provides a geochemical niche favorable for N2 fixation, the primary source of nitrogen to the ocean. Here, we present results from two cruises to the ETSP where rates of N2 fixation and its contribution to export production were determined with a suite of geochemical and biological measurements. N2 fixation was only detectable using nitrogen isotopic mass balances at two of six stations, and rates ranged from 0 to 23 µmol N m−2 d−1 based on sediment trap fluxes. Whereas the fractional importance of N2 fixation did not change, the N2-fixation rates at these two stations were several-fold higher when scaled to other productivity metrics. Regardless of the choice of productivity metric these N2-fixation rates are low compared with other oligotrophic locations, and the nitrogen isotope budgets indicate that N2 fixation supports no more than 20% of export production regionally. Although euphotic zone-integrated short-term N2-fixation rates were higher, up to 100 µmol N m−2 d−1, and detected N2 fixation at all six stations, studies of nitrogenase gene abundance and expression from the same cruises align with the geochemical data and together indicate that N2 fixation is a minor source of new nitrogen to surface waters of the ETSP. This finding is consistent with the hypothesis that, despite a relative abundance of phosphate, iron may limit N2 fixation in the ETSP. PMID:26976587

Nitrogen on Mars: Insights from Curiosity

NASA Technical Reports Server (NTRS)

Stern, J. C.; Sutter, B.; Jackson, W. A.; Navarro-Gonzalez, Rafael; McKay, Chrisopher P.; Ming, W.; Archer, P. Douglas; Glavin, D. P.; Fairen, A. G.; Mahaffy, Paul R.

2017-01-01

Recent detection of nitrate on Mars indicates that nitrogen fixation processes occurred in early martian history. Data collected by the Sample Analysis at Mars (SAM) instrument on the Curiosity Rover can be integrated with Mars analog work in order to better understand the fixation and mobility of nitrogen on Mars, and thus its availability to putative biology. In particular, the relationship between nitrate and other soluble salts may help reveal the timing of nitrogen fixation and post-depositional behavior of nitrate on Mars. In addition, in situ measurements of nitrogen abundance and isotopic composition may be used to model atmospheric conditions on early Mars.

The Fixation of Nitrogen.

ERIC Educational Resources Information Center

Andrew, S. P. S.

1978-01-01

Discusses the fixation of atmospheric nitrogen in the form of ammonia as one of the foundations of modern chemical industry. The article describes ammonia production and synthesis, purifying the hydrogen-nitrogen mix, nitric acid production, and its commericial plant. (HM)

"Cold" Fixation: Reconciliation of Nitrogen Fixation Rates and Diazotroph Assemblages in the Arctic Ocean

NASA Astrophysics Data System (ADS)

Fong, A. A.; Waite, A.; Rost, B.; Richter, K. U.

2016-02-01

Measurements of biological nitrogen fixation are typically conducted in oligotrophic subtropical and tropical marine environments where concentrations of fixed inorganic nitrogen are low. To date, only a handful of nitrogen fixation studies have been conducted in high latitude marine environments, but further investigation is needed to resolve the distribution of cold ocean diazotrophic assemblages. Nitrogen fixation rates and nifH gene distributions were measured at seven stations from 5°E to 20°E, north of 81°N in the Arctic Ocean at the onset of summer 2015. Discrete water samples in ice-covered regions were collected from the sea surface to 200 m for 15N2-tracer additions and targeted nifH gene and transcript analyses. Previous work suggests that heterotrophic bacteria dominate diazotrophic communities in the Arctic Ocean. Therefore, additional nifH gene surveys of sinking particles were conducted to test for enrichment on organic matter-rich microenvironments. Together, these measurements aim to reconcile diazotrophic activity with microbial community composition, further elucidating how nitrogen fixers could impact current concepts in polar carbon and nutrient cycling.

Sugar enrichment provides evidence for a role of nitrogen fixation in coral bleaching.

PubMed

Pogoreutz, Claudia; Rädecker, Nils; Cárdenas, Anny; Gärdes, Astrid; Voolstra, Christian R; Wild, Christian

2017-09-01

The disruption of the coral-algae symbiosis (coral bleaching) due to rising sea surface temperatures has become an unprecedented global threat to coral reefs. Despite decades of research, our ability to manage mass bleaching events remains hampered by an incomplete mechanistic understanding of the processes involved. In this study, we induced a coral bleaching phenotype in the absence of heat and light stress by adding sugars. The sugar addition resulted in coral symbiotic breakdown accompanied by a fourfold increase of coral-associated microbial nitrogen fixation. Concomitantly, increased N:P ratios by the coral host and algal symbionts suggest excess availability of nitrogen and a disruption of the nitrogen limitation within the coral holobiont. As nitrogen fixation is similarly stimulated in ocean warming scenarios, here we propose a refined coral bleaching model integrating the cascading effects of stimulated microbial nitrogen fixation. This model highlights the putative role of nitrogen-fixing microbes in coral holobiont functioning and breakdown. © 2017 The Authors. Global Change Biology Published by John Wiley & Sons Ltd.

Diazotrophy in the Deep: An analysis of the distribution, magnitude, geochemical controls, and biological mediators of deep-sea benthic nitrogen fixation

NASA Astrophysics Data System (ADS)

Dekas, Anne Elizabeth

Biological nitrogen fixation (the conversion of N2 to NH3) is a critical process in the oceans, counteracting the production of N2 gas by dissimilatory bacterial metabolisms and providing a source of bioavailable nitrogen to many nitrogen-limited ecosystems. One currently poorly studied and potentially underappreciated habitat for diazotrophic organisms is the sediments of the deep-sea. Although nitrogen fixation was once thought to be negligible in non-photosynthetically driven benthic ecosystems, the present study demonstrates the occurrence and expression of a diversity of nifH genes (those necessary for nitrogen fixation), as well as a widespread ability to fix nitrogen at high rates in these locations. The following research explores the distribution, magnitude, geochemical controls, and biological mediators of nitrogen fixation at several deep-sea sediment habitats, including active methane seeps (Mound 12, Costa Rica; Eel River Basin, CA, USA; Hydrate Ridge, OR, USA; and Monterey Canyon, CA, USA), whale-fall sites (Monterey Canyon, CA), and background deep-sea sediment (off-site Mound 12 Costa Rica, off-site Hydrate Ridge, OR, USA; and Monterey Canyon, CA, USA). The first of the five chapters describes the FISH-NanoSIMS method, which we optimized for the analysis of closely associated microbial symbionts in marine sediments. The second describes an investigation of methane seep sediment from the Eel River Basin, where we recovered nifH sequences from extracted DNA, and used FISH-NanoSIMS to identify methanotrophic archaea (ANME-2) as diazotrophs, when associated with functional sulfate-reducing bacterial symbionts. The third and fourth chapters focus on the distribution and diversity of active diazotrophs (respectively) in methane seep sediment from Mound 12, Costa Rica, using a combination of 15N-labeling experiments, FISH-NanoSIMS, and RNA and DNA analysis. The fifth chapter expands the scope of the investigation by targeting diverse samples from methane seep, whale-fall, and background sediment collected along the Eastern Pacific Margin, and comparing the rates of nitrogen fixation observed to geochemical measurements collected in parallel. Together, these analyses represent the most extensive investigation of deep-sea nitrogen fixation to date, and work towards understanding the contribution of benthic nitrogen fixation to global marine nitrogen cycling.

Net Anthropogenic Nitrogen Inputs in the Seattle, WA Metropolitan Area

NASA Astrophysics Data System (ADS)

Larson, E. K.; Alberti, M.

2014-12-01

Nitrogen loading has been identified as a potential stressor to marine ecosystems of the Puget Sound in the Pacific Northwest, and the Washington State Department of Ecology has estimated that anthropogenic sources of dissolved inorganic nitrogen to the Sound are 2.7 times higher than natural loads (Mohamedali et al. 2011). The Seattle urban area, situated in the southeast of the Sound, has the largest population in the northwestern US. Heavily urbanized along the coast, the 4 counties comprising the region (Snohomish, King, Pierce, and Kitsap) also include forests and agriculture. Urban and agricultural areas tend to have substantial anthropogenic N loading due to fertilizer application, presence of N-fixing vegetation, N atmospheric deposition, and human and other animal waste. To determine the relative contribution of urban vs. rural agricultural activities to N loads from the Seattle region to the Puget Sound, we used the Net Anthropogenic Nitrogen Inputs (NANI) calculator developed by Hong et al. (2011) for the watersheds of this region. The NANI calculator uses nationally available datasets to calculate NANI as the sum of oxidized N deposition, fertilizer application, agricultural N fixation, net food and feed inputs, and net animal and human N consumption. We found that NANI ranged from approximately 100 to 1500 kg m-2 y-1, with some of the highest rates in watersheds with high impervious surface or agricultural areas with N-fixing crops or large fertilizer additions. Many of the agricultural watersheds have intervening low-NANI watershed between themselves and the coast, thus it is likely that agricultural NANI is attenuated before entering the Puget Sound. The urban areas in the region do not have these attenuating watersheds, and so are likely to be the main contributor to the observed total aquatic N yield. This information is helpful for developing policies to reduce N loading to the Sound.

Nitrogen cycling during secondary succession in Atlantic Forest of Bahia, Brazil.

PubMed

Winbourne, Joy B; Feng, Aida; Reynolds, Lovinia; Piotto, Daniel; Hastings, Meredith G; Porder, Stephen

2018-01-22

Carbon accumulation in tropical secondary forests may be limited in part by nitrogen (N) availability, but changes in N during tropical forest succession have rarely been quantified. We explored N cycle dynamics across a chronosequence of secondary tropical forests in the Mata Atlântica of Bahia, Brazil in order to understand how quickly the N cycle recuperates. We hypothesized that N fixation would decline over the course of succession as N availability and N gaseous losses increased. We measured N fixation, KCl-extractable N, net mineralization and nitrification, resin-strip sorbed N, gaseous N emissions and the soil δ 15 N in stands that were 20, 35, 50, and > 50 years old. Contrary to our initial hypothesis, we found no significant differences between stand ages in any measured variable. Our findings suggest that secondary forests in this region of the Atlantic forest reached pre-disturbance N cycling dynamics after just 20 years of succession. This result contrasts with previous study in the Amazon, where the N cycle recovered slowly after abandonment from pasture reaching pre-disturbance N cycling levels after ~50 years of succession. Our results suggest the pace of the N cycle, and perhaps tropical secondary forest, recovery, may vary regionally.

Respiration , nitrogen fixation, and mineralizable nitrogen spatial and temporal patterns within two Oregon Douglas-fir stands.

Treesearch

Sharon M. Hope; Ching-Yan. Li

1997-01-01

Substrate respiration, mineralizable nitrogen, and nitrogen fixation rates, substrate moisture,content, and temperature were measured in trenched and undisturbed plots within two western Oregon Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) stands. The stands represent two different environments and ages. Woods Creek, the site of the lower...

Nitrogen Inputs via Nitrogen Fixation in Northern Plants and Soils

NASA Astrophysics Data System (ADS)

Thorp, N. R.; Wieder, R. K.; Vile, M. A.

2015-12-01

Dominated by cold and often acidic water logged environments, mineralization of organic matter is slow in the majority of northern ecosystems. Measures of extractable ammonium and nitrate are generally low and can be undetectable in peat pore waters. Despite this apparent nitrogen limitation, many of these environments produce deep deposits of soil organic matter. Biological nitrogen fixation carried out by autotrophic and heterotrophic diazotrophs associated with cryptograms provides the majority of known nitrogen inputs in these northern ecosystems. Nitrogen fixation was assessed in a variety of northern soils within rhizospheres of dominant plant communities. We investigated the availability of this newly fixed nitrogen to the vascular plant community in nitrogen limited northern plant communities. We tracked nitrogen flow from 15N2 gas fixed in Sphagnum mosses into tissues of two native vascular plant species, boreal cranberry (Vaccinium oxycoccus) and black spruce (Picea mariana). 15N-labeled Sphagnum microcosms were grown within variable mesh size exclusion/inclusion fabrics in a nitrogen addition experiment in situ in order to investigate the role of mycorrhizal fungi in the uptake of newly fixed nitrogen. Up to 24% of daily fixed 15N label was transferred to vascular plant tissues during 2 months. Nitrogen addition resulted in decreased N2 fixation rates; however, with higher nitrogen availability there was a higher rate of 15N label uptake into the vascular plants, likely the result of increased production of dissolved organic nitrogen. Reliance on mycorrhizal networks for nitrogen acquisition was indicated by nitrogen isotope fractionation patterns. Moreover, N2 fixation activities in mosses were stimulated when vascular plants were grown in moss microcosms versus "moss only" treatments. Results indicate that bog vascular plants may derive considerable nitrogen from atmospheric N2 biologically fixed within Sphagnum mosses. This work demonstrates that diazotroph-mediated 15N labeling is a viable technique for tracking nitrogen flow without altering form and concentration of native nitrogen pools in a nitrogen limited ecosystem.

Methane-Stimulated Benthic Marine Nitrogen Fixation at Deep-Sea Methane Seeps

NASA Astrophysics Data System (ADS)

Dekas, A. E.; Orphan, V.

2011-12-01

Biological nitrogen fixation (the conversion of N2 to NH3) is a critical process in the oceans, counteracting the production of N2 gas by dissimilatory bacterial metabolisms and providing a source of bioavailable nitrogen to many nitrogen-limited ecosystems. Although current measurements of N2 production and consumption in the oceans indicate that the nitrogen cycle is not balanced, recent findings on the limits of nitrogen fixation suggest that the perceived imbalance is an artifact of an incomplete assessment of marine diazotrophy. One currently poorly studied and potentially underappreciated habitat for diazotrophic organisms is the sediments of the deep-sea. In the present study we investigate the distribution and magnitude of benthic marine diazotrophy at several active deep-sea methane seeps (Mound 12, Costa Rica; Eel River Basin, CA, USA; Hydrate Ridge, OR, USA; and Monterey Canyon, CA, USA). Using 15N2 and 15NH4 sediment incubation experiments followed by single-cell (FISH-NanoSIMS) and bulk isotopic analysis (EA-IRMS), we observed total protein synthesis (15N uptake from 15NH4) and nitrogen fixation (15N update from 15N2). The highest rates of nitrogen fixation observed in the methane seep sediment incubation experiments were over an order of magnitude greater than those previously published from non-seep deep-sea sediments (Hartwig and Stanley, Deep-Sea Research, 1978, 25:411-417). However, methane seep diazotrophy appears to be highly spatially variable, with sediments exhibiting no nitrogen fixation originating only centimeters away from sediments actively incorporating 15N from 15N2. The greatest spatial variability in diazotrophy was observed with depth in the sediment, and corresponded to steep gradients in sulfate and methane. The maximum rates of nitrogen fixation were observed within the methane-sulfate transition zone, where organisms mediating the anaerobic oxidation of methane are typically in high abundance. Additionally, incubation experiments without added methane were observed to have little to no nitrogen fixation activity. In previous work, we demonstrated the capability of uncultured methanotrophic archaea (ANME-2) to fix nitrogen when associated with sulfate reducing bacterial symbionts. These new results suggest that these microbes may be the dominant nitrogen-fixing organisms in methane seep sediment. Intriguingly, characterization of the diversity of nifH genes from our sediment incubations as well as published nifH sequences reported from other seep habitats suggest the potential for other diazotrophic microorganisms in addition to the ANME-2 archaea. To further explore this possibility, FISH-NanoSIMS analyses were conducted on two dominant free-living sulfate-reducing lineages from seep incubations demonstrating nitrogen fixation activity. Preliminary results from this analysis suggest that single cells belonging to the Desulfobulbaceae may also be involved in nitrogen fixation in methane seeps. Despite this demonstrated potential, the extent of methane-independent diazotrophy by non-ANME diazotrophs appears to be low within the methane seep environment. Further studies are necessary to assess the greater diversity and activity of diazotrophs in other deep-sea sedimentary habitats.

The Role of Symbiotic Nitrogen Fixation in Sustainable Production of Biofuels

PubMed Central

Biswas, Bandana; Gresshoff, Peter M.

2014-01-01

With the ever-increasing population of the world (expected to reach 9.6 billion by 2050), and altered life style, comes an increased demand for food, fuel and fiber. However, scarcity of land, water and energy accompanied by climate change means that to produce enough to meet the demands is getting increasingly challenging. Today we must use every avenue from science and technology available to address these challenges. The natural process of symbiotic nitrogen fixation, whereby plants such as legumes fix atmospheric nitrogen gas to ammonia, usable by plants can have a substantial impact as it is found in nature, has low environmental and economic costs and is broadly established. Here we look at the importance of symbiotic nitrogen fixation in the production of biofuel feedstocks; how this process can address major challenges, how improving nitrogen fixation is essential, and what we can do about it. PMID:24786096

State of Climate 2011 - Global Ocean Phytoplankton

NASA Technical Reports Server (NTRS)

Siegel, D. A.; Antoine, D.; Behrenfeld, M. J.; d'Andon, O. H. Fanton; Fields, E.; Franz, B. A.; Goryl, P.; Maritorena, S.; McClain, C. R.; Wang, M.;

Ammonia fixation by humic substances: A nitrogen-15 and carbon-13 NMR study

USGS Publications Warehouse

Thorn, K.A.; Mikita, M.A.

1992-01-01

The process of ammonia fixation has been studied in three well characterized and structurally diverse fulvic and humic acid samples. The Suwannee River fulvic acid, and the IHSS peat and leonardite humic acids, were reacted with 15N-labelled ammonium hydroxide, and analyzed by liquid phase 15N NMR spectrometry. Elemental analyses and liquid phase 13C NMR spectra also were recorded on the samples before and after reaction with ammonium hydroxide. The largest increase in percent nitrogen occurred with the Suwannee River fulvic acid, which had a nitrogen content of 0.88% before fixation and 3.17% after fixation. The 15N NMR spectra revealed that ammonia reacted similarly with all three samples, indicating that the functional groups which react with ammonia exist in structural configurations common to all three samples. The majority of nitrogcn incorporated into the samples appears to be in the form of indole and pyrrole nitrogen, followed by pyridine, pyrazine, amide and aminohydroquinone nitrogen. Chemical changes in the individual samples upon fixation could not be discerned from the 13C NMR spectra.

Regional nitrogen budget of the Lake Victoria Basin, East Africa: syntheses, uncertainties and perspectives

NASA Astrophysics Data System (ADS)

Zhou, Minghua; Brandt, Patric; Pelster, David; Rufino, Mariana C.; Robinson, Timothy; Butterbach-Bahl, Klaus

2014-10-01

Using the net anthropogenic nitrogen input (NANI) approach we estimated the N budget for the Lake Victoria Basin in East Africa. The NANI of the basin ranged from 887 to 3008 kg N km-2 yr-1 (mean: 1827 kg N km-2 yr-1) for the period 1995-2000. The net nitrogen release at basin level is due primarily to livestock and human consumption of feed and foods, contributing between 69% and 85%. Atmospheric oxidized N deposition contributed approximately 14% to the NANI of the Lake Victoria Basin, while either synthetic N fertilizer imports or biological N fixations only contributed less than 6% to the regional NANI. Due to the low N imports of feed and food products (<20 kg N km-2 yr-1), nitrogen release to the watershed must be derived from the mining of soil N stocks. The fraction of riverine N export to Lake Victoria accounted for 16%, which is much lower than for watersheds located in Europe and USA (25%). A significant reduction of the uncertainty of our N budget estimate for Lake Victoria Basin would be possible if better data on livestock systems and riverine N export were available. Our study indicates that at present soil N mining is the main source of nitrogen in the Lake Victoria Basin. Thus, sustainable N management requires increasing agricultural N inputs to guarantee food security and rehabilitation and protection of soils to minimize environmental costs. Moreover, to reduce N pollution of the lake, improving management of human and animal wastes needs to be carefully considered in future.

Spring thaw ionic pulses boost nutrient availability and microbial growth in entombed Antarctic Dry Valley cryoconite holes

PubMed Central

Telling, Jon; Anesio, Alexandre M.; Tranter, Martyn; Fountain, Andrew G.; Nylen, Thomas; Hawkings, Jon; Singh, Virendra B.; Kaur, Preeti; Musilova, Michaela; Wadham, Jemma L.

2014-01-01

The seasonal melting of ice entombed cryoconite holes on McMurdo Dry Valley glaciers provides oases for life in the harsh environmental conditions of the polar desert where surface air temperatures only occasionally exceed 0°C during the Austral summer. Here we follow temporal changes in cryoconite hole biogeochemistry on Canada Glacier from fully frozen conditions through the initial stages of spring thaw toward fully melted holes. The cryoconite holes had a mean isolation age from the glacial drainage system of 3.4 years, with an increasing mass of aqueous nutrients (dissolved organic carbon, total nitrogen, total phosphorus) with longer isolation age. During the initial melt there was a mean nine times enrichment in dissolved chloride relative to mean concentrations of the initial frozen holes indicative of an ionic pulse, with similar mean nine times enrichments in nitrite, ammonium, and dissolved organic matter. Nitrate was enriched twelve times and dissolved organic nitrogen six times, suggesting net nitrification, while lower enrichments for dissolved organic phosphorus and phosphate were consistent with net microbial phosphorus uptake. Rates of bacterial production were significantly elevated during the ionic pulse, likely due to the increased nutrient availability. There was no concomitant increase in photosynthesis rates, with a net depletion of dissolved inorganic carbon suggesting inorganic carbon limitation. Potential nitrogen fixation was detected in fully melted holes where it could be an important source of nitrogen to support microbial growth, but not during the ionic pulse where nitrogen availability was higher. This study demonstrates that ionic pulses significantly alter the timing and magnitude of microbial activity within entombed cryoconite holes, and adds credence to hypotheses that ionic enrichments during freeze-thaw can elevate rates of microbial growth and activity in other icy habitats, such as ice veins and subglacial regelation zones. PMID:25566210

Spring thaw ionic pulses boost nutrient availability and microbial growth in entombed Antarctic Dry Valley cryoconite holes.

PubMed

Telling, Jon; Anesio, Alexandre M; Tranter, Martyn; Fountain, Andrew G; Nylen, Thomas; Hawkings, Jon; Singh, Virendra B; Kaur, Preeti; Musilova, Michaela; Wadham, Jemma L

2014-01-01

The seasonal melting of ice entombed cryoconite holes on McMurdo Dry Valley glaciers provides oases for life in the harsh environmental conditions of the polar desert where surface air temperatures only occasionally exceed 0°C during the Austral summer. Here we follow temporal changes in cryoconite hole biogeochemistry on Canada Glacier from fully frozen conditions through the initial stages of spring thaw toward fully melted holes. The cryoconite holes had a mean isolation age from the glacial drainage system of 3.4 years, with an increasing mass of aqueous nutrients (dissolved organic carbon, total nitrogen, total phosphorus) with longer isolation age. During the initial melt there was a mean nine times enrichment in dissolved chloride relative to mean concentrations of the initial frozen holes indicative of an ionic pulse, with similar mean nine times enrichments in nitrite, ammonium, and dissolved organic matter. Nitrate was enriched twelve times and dissolved organic nitrogen six times, suggesting net nitrification, while lower enrichments for dissolved organic phosphorus and phosphate were consistent with net microbial phosphorus uptake. Rates of bacterial production were significantly elevated during the ionic pulse, likely due to the increased nutrient availability. There was no concomitant increase in photosynthesis rates, with a net depletion of dissolved inorganic carbon suggesting inorganic carbon limitation. Potential nitrogen fixation was detected in fully melted holes where it could be an important source of nitrogen to support microbial growth, but not during the ionic pulse where nitrogen availability was higher. This study demonstrates that ionic pulses significantly alter the timing and magnitude of microbial activity within entombed cryoconite holes, and adds credence to hypotheses that ionic enrichments during freeze-thaw can elevate rates of microbial growth and activity in other icy habitats, such as ice veins and subglacial regelation zones.

Comparison of inhibition of N2 fixation and ureide accumulation under water deficit in four common bean genotypes of contrasting drought tolerance

PubMed Central

Coleto, I.; Pineda, M.; Rodiño, A. P.; De Ron, A. M.; Alamillo, J. M.

2014-01-01

Background and Aims Drought is the principal constraint on world production of legume crops. There is considerable variability among genotypes in sensitivity of nitrogen fixation to drought, which has been related to accumulation of ureides in soybean. The aim of this study was to search for genotypic differences in drought sensitivity and ureide accumulation in common bean (Phaseolus vulgaris) germplasm that may be useful in the improvement of tolerance to water deficit in common bean. Methods Changes in response to water deficit of nitrogen fixation rates, ureide content and the expression and activity of key enzymes for ureide metabolism were measured in four P. vulgaris genotypes differing in drought tolerance. Key Results A variable degree of drought-induced nitrogen fixation inhibition was found among the bean genotypes. In addition to inhibition of nitrogen fixation, there was accumulation of ureides in stems and leaves of sensitive and tolerant genotypes, although this was higher in the leaves of the most sensitive ones. In contrast, there was no accumulation of ureides in the nodules or roots of stressed plants. In addition, the level of ureides in the most sensitive genotype increased after inhibition of nitrogen fixation, suggesting that ureides originate in vegetative tissues as a response to water stress, probably mediated by the induction of allantoinase. Conclusions Variability of drought-induced inhibition of nitrogen fixation among the P. vulgaris genotypes was accompanied by subsequent accumulation of ureides in stems and leaves, but not in nodules. The results indicate that shoot ureide accumulation after prolonged exposure to drought could not be the cause of inhibition of nitrogen fixation, as has been suggested in soybean. Instead, ureides seem to be produced as part of a general response to stress, and therefore higher accumulation might correspond to higher sensitivity to the stressful conditions. PMID:24638821

Comparison of inhibition of N2 fixation and ureide accumulation under water deficit in four common bean genotypes of contrasting drought tolerance.

PubMed

Coleto, I; Pineda, M; Rodiño, A P; De Ron, A M; Alamillo, J M

2014-05-01

Drought is the principal constraint on world production of legume crops. There is considerable variability among genotypes in sensitivity of nitrogen fixation to drought, which has been related to accumulation of ureides in soybean. The aim of this study was to search for genotypic differences in drought sensitivity and ureide accumulation in common bean (Phaseolus vulgaris) germplasm that may be useful in the improvement of tolerance to water deficit in common bean. Changes in response to water deficit of nitrogen fixation rates, ureide content and the expression and activity of key enzymes for ureide metabolism were measured in four P. vulgaris genotypes differing in drought tolerance. A variable degree of drought-induced nitrogen fixation inhibition was found among the bean genotypes. In addition to inhibition of nitrogen fixation, there was accumulation of ureides in stems and leaves of sensitive and tolerant genotypes, although this was higher in the leaves of the most sensitive ones. In contrast, there was no accumulation of ureides in the nodules or roots of stressed plants. In addition, the level of ureides in the most sensitive genotype increased after inhibition of nitrogen fixation, suggesting that ureides originate in vegetative tissues as a response to water stress, probably mediated by the induction of allantoinase. Variability of drought-induced inhibition of nitrogen fixation among the P. vulgaris genotypes was accompanied by subsequent accumulation of ureides in stems and leaves, but not in nodules. The results indicate that shoot ureide accumulation after prolonged exposure to drought could not be the cause of inhibition of nitrogen fixation, as has been suggested in soybean. Instead, ureides seem to be produced as part of a general response to stress, and therefore higher accumulation might correspond to higher sensitivity to the stressful conditions.

Seasonal patterns of climate controls over nitrogen fixation by Alnus viridis subsp

Treesearch

Jennifer S. Mitchell; Roger W. Ruess

2009-01-01

Patterns of and controls over N2 fixation by green alder were studied in post-fire, mid-succession, and white spruce upland forests in interior Alaska, focusing on the hypothesis that ecosystem-level nitrogen (N) inputs decrease with successional development. N2-fixation rates tracked plant phenology during the 1997 (...

Genome-wide transcriptome profiling of nitrogen fixation in Paenibacillus sp. WLY78.

PubMed

Shi, Hao-wen; Wang, Li-ying; Li, Xin-xin; Liu, Xiao-meng; Hao, Tian-yi; He, Xiao-juan; Chen, San-feng

2016-03-01

Diazotrophic (nitrogen-fixing) Gram-positive and endospore-formed Paenibacillus spp. have potential uses as a bacterial fertilizer in agriculture. The transcriptional analysis of nitrogen fixation in Paenibacillus is lacking, although regulation mechanisms of nitrogen fixation have been well studied in Gram-negative diazotrophs. Here we report a global transcriptional profiling analysis of nitrogen fixation in Paenibacillus sp. WLY78 cultured under N2-fixing condition (without O2 and NH4(+)) and non-N2-fixing condition (air and 100 mM NH4(+)). The nif (nitrogen fixation) gene operon composed of 9 genes (nifBHDKENXhesAnifV) in this bacterium was significantly up-regulated in N2-fixing condition compared to non-N2-fixing condition, indicating that nif gene transcription is strictly controlled by NH4(+) and O2. qRT-PCR confirmed that these nif genes were differently expressed. Non-nif genes specifically required in nitrogen fixation, such as mod, feoAB and cys encoding transporters of Mo, Fe and S atoms, were coordinately transcribed with nif genes in N2-fixing condition. The transcript abundance of suf operon specific for synthesis of Fe-S cluster was up-regulated in N2-fixing condition, suggesting that Sul system, which takes place of nifS and nifU, plays important role in the synthesis of nitrogenase. We discover potential specific electron transporters which might provide electron from Fe protein to MoFe protein of nitrogenase. The glnR whose predicted protein might mediate nif transcription regulation by NH4(+) is significantly up-regulated in N2-fixing condition. The transcription levels of nitrogen metabolism and anaerobic respiration were also analyzed. The nif gene operon (nifBHDKENXhesAnifV) in Paenibacillus sp. WLY78 is significantly up-regulated in N2-fixing condition compared to non-N2-fixing condition. Non-nif genes specifically required in nitrogen fixation were also significantly up-regulated in N2-fixing condition. Fur and Fnr which are involved in anaerobic regulation and GlnR which might mediate nif gene transcription regulation by NH4(+) were significantly up-regulated in N2-fixing condition. This study provides valuable insights into nitrogen fixation process and regulation in Gram-positive firmicutes.

Transcriptome analysis of two recombinant inbred lines of common bean contrasting for symbiotic nitrogen fixation

USDA-ARS?s Scientific Manuscript database

Common bean (Phaseolus vulgaris L.) is able to fix atmospheric nitrogen (N2) through symbiotic nitrogen fixation (SNF). Effective utilization of existing variability for SNF in common bean for genetic improvement requires an understanding of underlying genes and molecular mechanisms. The utility of ...

Regional and national significance of biological nitrogen fixation by crops in the United States

EPA Science Inventory

Background/Questions/Methods Biological nitrogen fixation by crops (C-BNF) represents one of the largest anthropogenic inputs of reactive nitrogen (N) to land surfaces around the world. In the United States (US), existing estimates of C-BNF are uncertain because of incomplete o...

Termites create spatial structure and govern ecosystem function by affecting N2 fixation in an East African savanna.

PubMed

Fox-Dobbs, Kena; Doak, Daniel F; Brody, Alison K; Palmer, Todd M

2010-05-01

The mechanisms by which even the clearest of keystone or dominant species exert community-wide effects are only partially understood in most ecosystems. This is especially true when a species or guild influences community-wide interactions via changes in the abiotic landscape. Using stable isotope analyses, we show that subterranean termites in an East African savanna strongly influence a key ecosystem process: atmospheric nitrogen fixation by a monodominant tree species and its bacterial symbionts. Specifically, we applied the 15N natural abundance method in combination with other biogeochemical analyses to assess levels of nitrogen fixation by Acacia drepanolobium and its effects on co-occurring grasses and forbs in areas near and far from mounds and where ungulates were or were not excluded. We find that termites exert far stronger effects than do herbivores on nitrogen fixation. The percentage of nitrogen derived from fixation in Acacia drepanolobium trees is higher (55-80%) away from mounds vs. near mounds (40-50%). Mound soils have higher levels of plant available nitrogen, and Acacia drepanolobium may preferentially utilize soil-based nitrogen sources in lieu of fixed nitrogen when these sources are readily available near termite mounds. At the scale of the landscape, our models predict that termite/soil derived nitrogen sources influence >50% of the Acacia drepanolobium trees in our system. Further, the spatial extent of these effects combine with the spacing of termite mounds to create highly regular patterning in nitrogen fixation rates, resulting in marked habitat heterogeneity in an otherwise uniform landscape. In summary, we show that termite-associated effects on nitrogen processes are not only stronger than those of more apparent large herbivores in the same system, but also occur in a highly regular spatial pattern, potentially adding to their importance as drivers of community and ecosystem structure.

Nutrient budgets in the subtropical ocean gyres dominated by lateral transport

NASA Astrophysics Data System (ADS)

Letscher, Robert T.; Primeau, François; Moore, J. Keith

2016-11-01

Ocean circulation replenishes surface nutrients depleted by biological production and export. Vertical processes are thought to dominate, but estimated vertical nutrient fluxes are insufficient to explain observed net productivity in the subtropical ocean gyres. Lateral inputs help balance the North Atlantic nutrient budget, but their importance for other gyres has not been demonstrated. Here we use an ocean model that couples circulation and ecosystem dynamics to show that lateral transport and biological uptake of inorganic and organic forms of nitrogen and phosphorus from the gyre margins exceeds the vertical delivery of nutrients, supplying 24-36% of the nitrogen and 44-67% of the phosphorus required to close gyre nutrient budgets. At the Bermuda and Hawaii time-series sites, nearly half of the annual lateral supply by lateral transport occurs during the summer-to-fall stratified period, helping explain seasonal patterns of inorganic carbon drawdown and nitrogen fixation. Our study confirms the importance of upper-ocean lateral nutrient transport for understanding the biological cycles of carbon and nutrients in the ocean's largest biome.

The Nitrogen Cycle During the Transition to Euxinia

NASA Astrophysics Data System (ADS)

Meyer, K. M.; Kump, L. R.; Ridgwell, A.

2008-12-01

Nitrogen and phosphorous are essential to life, and their biological availability is hypothesized to regulate marine productivity on short and geologic timescales. The nature of primary production during recurrent intervals of Phanerozoic anoxia is of particular interest because of the redox control of nutrient and trace metal availability. Dissolved phosphate likely increased during transitions from oxic to euxinic marine conditions, while nitrogen availability may have decreased due to extensive denitrification as low-oxygen waters spread. Because nitrogen fixation is both metabolically and trace-metal intensive, a key question in the transition to euxinia is whether nitrogen fixation can "keep pace" with denitrification. If denitrification exceeds nitrogen fixation, diminished export production and oxygen demand in an N-limited ocean would pose a negative feedback that may prevent euxinia altogether or initiate the shift back to oxic conditions. Here we use the GENIE-1 Earth system model to address the biogeochemistry of the oxic-euxinic transition characteristic of some Phanerozoic oceanic anoxic events. As previously demonstrated with box models, phosphate accumulation stimulates both nitrogen fixation and denitrification. While there is an initial transient loss of total fixed nitrogen from the ocean, nitrogen inputs eventually exceed losses, and the marine nitrogen reservoir grows with that of phosphate to significantly exceed its modern value. Nitrogen buildup also corresponds with a shift in ecology of the surface ocean and the unexpected initiation of non-Redfieldian stoichiometry in the chemistry of the deep ocean.

Comparative genomic analysis of carbon and nitrogen assimilation mechanisms in three indigenous bioleaching bacteria: predictions and validations

PubMed Central

Levicán, Gloria; Ugalde, Juan A; Ehrenfeld, Nicole; Maass, Alejandro; Parada, Pilar

2008-01-01

Background Carbon and nitrogen fixation are essential pathways for autotrophic bacteria living in extreme environments. These bacteria can use carbon dioxide directly from the air as their sole carbon source and can use different sources of nitrogen such as ammonia, nitrate, nitrite, or even nitrogen from the air. To have a better understanding of how these processes occur and to determine how we can make them more efficient, a comparative genomic analysis of three bioleaching bacteria isolated from mine sites in Chile was performed. This study demonstrated that there are important differences in the carbon dioxide and nitrogen fixation mechanisms among bioleaching bacteria that coexist in mining environments. Results In this study, we probed that both Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans incorporate CO2 via the Calvin-Benson-Bassham cycle; however, the former bacterium has two copies of the Rubisco type I gene whereas the latter has only one copy. In contrast, we demonstrated that Leptospirillum ferriphilum utilizes the reductive tricarboxylic acid cycle for carbon fixation. Although all the species analyzed in our study can incorporate ammonia by an ammonia transporter, we demonstrated that Acidithiobacillus thiooxidans could also assimilate nitrate and nitrite but only Acidithiobacillus ferrooxidans could fix nitrogen directly from the air. Conclusion The current study utilized genomic and molecular evidence to verify carbon and nitrogen fixation mechanisms for three bioleaching bacteria and provided an analysis of the potential regulatory pathways and functional networks that control carbon and nitrogen fixation in these microorganisms. PMID:19055775

Nitrogen fixation dynamics of two diazotrophic communities in Mono Lake, California

USGS Publications Warehouse

Oremland, R.S.

1990-01-01

Two types of diazotrophic microbial communities were found in the littoral zone of alkaline hypersaline Mono Lake, California. One consisted of anaerobic bacteria inhabiting the flocculent surface layers of sediments. Nitrogen fixation (acetylene reduction) by flocculent surface layers occurred under anaerobic conditions, was not stimulated by light or by additions of organic substrates, and was inhibited by O2, nitrate, and ammonia. The second community consisted of a ball-shaped association of a filamentous chlorophyte (Ctenocladus circinnatus) with diazotrophic, nonheterocystous cyanobacteria, as well as anaerobic bacteria (Ctenocladus balls). Nitrogen fixation by Ctenocladus balls was usually, but not always, stimulated by light. Rates of anaerobic dark fixation equaled those in the light under air. Fixation in the light was stimulated by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and by propanil [N-(3,4-dichlorophenyl)propanamide]. 3-(3,4-Dichlorophenyl)-1,1-dimethyl urea-elicited nitrogenase activity was inhibited by ammonia (96%) and nitrate (65%). Fixation was greatest when Ctenocladus balls were incubated anaerobically in the light with sulfide. Dark anaerobic fixation was not stimulated by organic substrates in short-term (4-h) incubations, but was in long-term (67-h) ones. Areal estimates of benthic N2 fixation were measured seasonally, using chambers. Highest rates (~29.3 ??mol of C2H4 m-2 h-1) occurred under normal diel regimens of light and dark. These estimates indicate that benthic N2 fixation has the potential to be a significant nitrogen source in Mono Lake.

Low temperature delays timing and enhances the cost of nitrogen fixation in the unicellular cyanobacterium Cyanothece

PubMed Central

Brauer, Verena S; Stomp, Maayke; Rosso, Camillo; van Beusekom, Sebastiaan AM; Emmerich, Barbara; Stal, Lucas J; Huisman, Jef

2013-01-01

Marine nitrogen-fixing cyanobacteria are largely confined to the tropical and subtropical ocean. It has been argued that their global biogeographical distribution reflects the physiologically feasible temperature range at which they can perform nitrogen fixation. In this study we refine this line of argumentation for the globally important group of unicellular diazotrophic cyanobacteria, and pose the following two hypotheses: (i) nitrogen fixation is limited by nitrogenase activity at low temperature and by oxygen diffusion at high temperature, which is manifested by a shift from strong to weak temperature dependence of nitrogenase activity, and (ii) high respiration rates are required to maintain very low levels of oxygen for nitrogenase, which results in enhanced respiratory cost per molecule of fixed nitrogen at low temperature. We tested these hypotheses in laboratory experiments with the unicellular cyanobacterium Cyanothece sp. BG043511. In line with the first hypothesis, the specific growth rate increased strongly with temperature from 18 to 30 °C, but leveled off at higher temperature under nitrogen-fixing conditions. As predicted by the second hypothesis, the respiratory cost of nitrogen fixation and also the cellular C:N ratio rose sharply at temperatures below 21 °C. In addition, we found that low temperature caused a strong delay in the onset of the nocturnal nitrogenase activity, which shortened the remaining nighttime available for nitrogen fixation. Together, these results point at a lower temperature limit for unicellular nitrogen-fixing cyanobacteria, which offers an explanation for their (sub)tropical distribution and suggests expansion of their biogeographical range by global warming. PMID:23823493

Projections of Biofuel Growth Patterns Reveal the Potential Importance of Nitrogen Fixation for Miscanthus Productivity

NASA Astrophysics Data System (ADS)

Davis, S. C.; Parton, W. J.; Dohleman, F. G.; Gottel, N. R.; Smith, C. M.; Kent, A. D.; Delucia, E. H.

2008-12-01

Demand for liquid biofuels is increasing because of the disparity between fuel demand and supply. Relative to grain crops, the more intensive harvest required for second generation liquid biofuel production leads to the removal of significantly more carbon and nitrogen from the soil. These elements are conventionally litter products of crops that are returned to the soil and can accumulate over time. This loss of organic matter represents a management challenge because the energy cost associated with fertilizers or external sources of organic matter reduce the net energy value of the biofuel crops. Plants that have exceptional strategies for exploiting nutrients may be the most viable options for sustainable biofuel yields because of low management and energy cost. Miscanthus x giganteus has high N retranslocation rates, maintains high photosynthetic rates over a large temperature range, exploits a longer-than-average growing season, and yields at least twice the biomass of other candidate biofuel grass crops (i.e. switchgrass). We employed the DAYCENT model to project potential productivity of Miscanthus, corn, switchgrass, and mixed prairie communities based on our current knowledge of these species. Ecosystem process descriptions that have been validated for many crop species did not accurately predict Miscanthus yields and lead to new hypotheses about unknown N cycling mechanisms for this species. We tested the hypothesis that Miscanthus hosts N-fixing bacteria in several ways. First, we used enrichment culture and molecular methods to detect N-fixing bacteria in Miscanthus. Then, we demonstrated the plant-growth promoting effect of diazotrophs isolated from Miscanthus rhizomes on a model grass. And finally, we applied 15N2 to the soil and rooting zone of field grown Miscanthus plants to determine if atmospheric N2 was incorporated into plant tissue, a process that requires N-fixation. These experiments are the first tests of N-fixation in Miscanthus x giganteus, and the ecosystem model allowed us to project how much nitrogen may be obtained from N-fixation to support sustainable high biomass yields.

Path of carbon flow during NO/sub 3//sup -/-induced photosynthetic suppression in N-limited Selenastrum minutum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Elrifi, I.R.; Turpin, D.H.

Nitrate addition to nitrate-limited cultures of Selenastrum minutum Naeg. Collins (Chlorophyta) resulted in a 70% suppression of photosynthetic carbon fixation. In /sup 14/CO/sub 2/ pulse/chase experiments nitrate resupply increased radiolabel incorporation into amino and organic acids and decreased radiolabel incorporation into insoluble material. Nitrate resupply increased the concentration of phosphoenolpyruvate and increased the radiolabeling of phosphoenolpyruvate, pyruvate and tricarboxylic acid cycle intermediates, notably citrate, fumarate, and malate. Furthermore, nitrate also increased the pool sizes and radiolabeling of most amino acids, with alanine, aspartate, glutamate, and glutamine showing the largest changes. Nitrate resupply increased the proportion of radiolabel in the C-4more » position of malate and increased the ratios of radiolabel in aspartate to phosphoenolpyruvate and in pyruvate to phosphoenolpyruvate, indicative of increased phosphoenolpyruvate carboxylase and pyruvate kinase activities. Analysis of these data showed that the rate of carbon flow through glutamate (10.6 ..mu..moles glutamate per milligram chlorophyll per hour) and the rate of net glutamate production (7.9 ..mu..moles glutamate per milligram chlorophyll per hour) were both greater than the maximum rate of carbon export from the Calvin cycle which could be maintained during steady state photosynthesis. These results are consistent with the hypothesis that nitrogen resupply to nitrogen-limited microalgae results in a transient suppression of photosynthetic carbon fixation due, in part, to the severity of competition for carbon skeletons between the Calvin cycle and nitrogen assimilation.« less

The Path of Carbon Flow during NO3−-Induced Photosynthetic Suppression in N-Limited Selenastrum minutum1

PubMed Central

Elrifi, Ivor R.; Turpin, David H.

1987-01-01

Nitrate addition to nitrate-limited cultures of Selenastrum minutum Naeg. Collins (Chlorophyta) resulted in a 70% suppression of photosynthetic carbon fixation. In 14CO2 pulse/chase experiments nitrate resupply increased radiolabel incorporation into amino and organic acids and decreased radiolabel incorporation into insoluble material. Nitrate resupply increased the concentration of phosphoenolpyruvate and increased the radiolabeling of phosphoenolpyruvate, pyruvate and tricarboxylic acid cycle intermediates, notably citrate, fumarate, and malate. Furthermore, nitrate also increased the pool sizes and radiolabeling of most amino acids, with alanine, aspartate, glutamate, and glutamine showing the largest changes. Nitrate resupply increased the proportion of radiolabel in the C-4 position of malate and increased the ratios of radiolabel in aspartate to phosphoenolpyruvate and in pyruvate to phosphoenolpyruvate, indicative of increased phosphoenolpyruvate carboxylase and pyruvate kinase activities. Analysis of these data showed that the rate of carbon flow through glutamate (10.6 μmoles glutamate per milligram chlorophyll per hour) and the rate of net glutamate production (7.9 μmoles glutamate per milligram chlorophyll per hour) were both greater than the maximum rate of carbon export from the Calvin cycle which could be maintained during steady state photosynthesis. These results are consistent with the hypothesis that nitrogen resupply to nitrogen-limited microalgae results in a transient suppression of photosynthetic carbon fixation due, in part, to the severity of competition for carbon skeletons between the Calvin cycle and nitrogen assimilation (IR Elrifi, DH Turpin 1986 Plant Physiol 81: 273-279). PMID:16665223

The Path of Carbon Flow during NO(3)-Induced Photosynthetic Suppression in N-Limited Selenastrum minutum.

PubMed

Elrifi, I R; Turpin, D H

1987-01-01

Nitrate addition to nitrate-limited cultures of Selenastrum minutum Naeg. Collins (Chlorophyta) resulted in a 70% suppression of photosynthetic carbon fixation. In (14)CO(2) pulse/chase experiments nitrate resupply increased radiolabel incorporation into amino and organic acids and decreased radiolabel incorporation into insoluble material. Nitrate resupply increased the concentration of phosphoenolpyruvate and increased the radiolabeling of phosphoenolpyruvate, pyruvate and tricarboxylic acid cycle intermediates, notably citrate, fumarate, and malate. Furthermore, nitrate also increased the pool sizes and radiolabeling of most amino acids, with alanine, aspartate, glutamate, and glutamine showing the largest changes. Nitrate resupply increased the proportion of radiolabel in the C-4 position of malate and increased the ratios of radiolabel in aspartate to phosphoenolpyruvate and in pyruvate to phosphoenolpyruvate, indicative of increased phosphoenolpyruvate carboxylase and pyruvate kinase activities. Analysis of these data showed that the rate of carbon flow through glutamate (10.6 mumoles glutamate per milligram chlorophyll per hour) and the rate of net glutamate production (7.9 mumoles glutamate per milligram chlorophyll per hour) were both greater than the maximum rate of carbon export from the Calvin cycle which could be maintained during steady state photosynthesis. These results are consistent with the hypothesis that nitrogen resupply to nitrogen-limited microalgae results in a transient suppression of photosynthetic carbon fixation due, in part, to the severity of competition for carbon skeletons between the Calvin cycle and nitrogen assimilation (IR Elrifi, DH Turpin 1986 Plant Physiol 81: 273-279).

Dynamics of nitrate limitation on gaseous nitrogen exchanges from pristine peatlands

NASA Astrophysics Data System (ADS)

Roobroeck, D.; Brüggemann, N.; Butterbach-Bahl, K.; Boeckx, P.

2009-04-01

The cycling of gaseous nitrogen species in peatland ecosystems and the functioning of driving forces on microbial denitrification rates are poorly accounted. Physico-chemical soil conditions and biotic interactions1 control the availability of nitrate for respiratory denitrification resulting in high spatial variability of gaseous nitrogen exchange rates in nutrient poor peat soils and complicating impact assessment of eutrophication. The responses of nitrous oxide (N2O) and dinitrogen (N2) fluxes to nitrate addition were compared between distinct contrasts in plant growth at a pristine, hummocky peatland. Allowing to determine the dynamics of nitrate limitation on gaseous nitrogen exchanges in accordance to covariance in soil anaerobiosis and resource competition. Two quantification techniques were applied parallel to soil core incubations in order to determine N2O and N2 fluxes. Helium atmosphere incubation was used for direct quantification of net N2O and N2 fluxes. Reducing the background N2 concentration in the soil atmosphere to approx. 20 ppm enabled highly sensitive measurement of N2 fluxes. On the other hand a 15N-N2O tracer technique was explored as a tool to demonstrate and quantify gross consumption rates of atmospheric N2O to N2 and recycling of gaseous N-losses by microbial fixation. The headspace N2O pool was increased with 0.03 ppm 15•15N-N2O rendering an enrichment of ± 9.8 atom% 15•15N-N2O. Triplicate soil core samples were taken from two contrasting soil habitat in a hummocky, Carex dominated fen located in the Biebrza National Park, NE Poland (53 °07′N; 23 °10′E). The hummocks had a gravimetric soil water content of 76.6 ± 2.2% and high root abundance, dissimilar to 83.4 ± 1.0% and little root prevalence in the hollows. Singular nitrate addition, comparable to the atmospheric NOY -deposition, was applied two days in advance of flux measurement. Actual net gaseous nitrogen fluxes and responses to nitrate addition were apparently different for both soil habitat. Hummock soil cores showed to be net sources of N2O sinks (-3.04 ± 0.12 g N2O-N h-1 m-2). Net N2 fluxes measured consistently higher from the hollows than the hummocks (resp. 2622.3 ± 106.3 and 1065.3 ± 139.2 g N2-N h-1 m-2). Nitrate addition to the hummock habitat resulted in a small, non-significant increase of the net N2O flux, while the hollow soil cores showed a drastic shift towards a net N2O source upon nitrate addition (16.27 ± 2.87 g N2O-N h-1 m-2). The N2:N2O ratios and net N2O fluxes clearly illustrated that relatively more bio-available nitrogen is converted to N2O by respiratory denitrification at higher soil nitrate availability. The 15•15N-N2O tracer technique demonstrated consumptive reduction of atmospheric N2O to N2. With NO3-addition the atom percent excess of 15N in N2 decreased for both soil habitat indicating that less atmospheric N2O is reduced to inert N2 when more nitrate is available for microbial denitrification. N2O consumption rates will be discussed on the presentation. Indirect fumigation of soil samples proved that CHCl3-labile nitrogen was significantly enriched in 15N when 15N-N2O was applied in the headspace. Demonstrating that nitrogen lost during respiratory denitrification is recycled to microbial biomass, most likely a result of N2-fixation by soil micro-organisms in this mineral N-depleted ecosystem. Higher root abundance associated with lower soil anaerobiosis and higher resource competition caused net N2O fluxes to be positive, but mitigated the effect of nitrate addition. Lower root abundance associated with higher soil anaerobiosis, caused natural peat soil to be net N2O sinks, but lower resource competition however attributed to higher eutrophication susceptibility. Variance in physico-chemical soil conditions and biotic interactions showed to interfere with the effect of nitrate availability on consumptive reduction of atmospheric N2O to N2. Inverse covariance of soil anaerobiosis and resource competition as a result of variance in plant growth indicated to be a major regulatory dynamic of gaseous nitrogen exchanges from natural peatland, by which the susceptibility to nitrate eutrophication is determined. References 1 Silvan, N., Tuittila, E., Kitunnen, V., Vasander, H., Laine, J., 2005. Nitrate uptake by Eriophorum vaginatum controls N2O production in a restored peatland, Soil Biology & Biochemistry, 37:1519-1526.

Effects of acidic rain and ozone on nitrogen fixation and photosynthesis in the lichen lobaria pulmonaria (L. ) Hoffm

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sigal, L.L.; Johnston, J.W.

1986-01-01

The lichen Lobaria pulmonaria was subjected to ozone fumigations at 118, 235 and 353 mcg/cu m and simulated acidic rain at pH levels of 2.6, 4.2 and 5.6 for 5 days (M,W,F,M,W) during a 10-day period. Acidic rain at pH 2.6 caused significant reduction in nitrogen fixation and gross photosynthesis of 100 and 90%, respectively, and thallus bleaching was apparent. There were no significant differences between the pH 5.6 and 4.2 treatments in either gross photosynthesis or nitrogen fixation, and the color of the lichen thalli was unchanged. The effect of ozone on nitrogen fixation and photosynthesis over the rangemore » of concentrations used was not significant, but there was a trend toward reduced nitrogen fixation with increasing O/sub 3/ concentration. There were no significant ozone-acidic rain interactions. The threshold for response to rain acidity for L. pulmonaria lies between pH 2.6 and 4.2, and the acidity of wet deposition in parts of the United States may fall in the range.« less

Effect of insecticides and phenolics on nitrogen fixation by Nostoc linckia

DOE Office of Scientific and Technical Information (OSTI.GOV)

Megharaj, M.; Venkateswarlu, K.; Rao, A.S.

1988-08-01

The nitrogen-fixing blue-green algae (cyanobacteria) significantly influence the nitrogen economy of temperate and tropical soils. Although the genera Nostoc and Tolypothrix have been particularly implicated in the fixation of significantly large amounts of atmospheric nitrogen, these diazotrophs received little attention in relation to insecticide treatment and the available few reports do not indicate a permanent deleterious effect of insecticides on their nitrogenase activity. As it has been well established that the effect of insecticides on nitrogen fixation by cyanobacteria is independent of that on growth, an attempt was, therefore, made to determine the influence of four insecticides (monocrotophos, quinalphos, cypermethrinmore » and fenvalerate) and four phenolics (p-nitrophenol (PNP), m-nitrophenol (MNP), 2,4-dinitrophenol (DNP) and catechol) on nitrogen-fixing capacity of N.linckia, isolated from a black soil.« less

Methanotrophy induces nitrogen fixation during peatland development

PubMed Central

Larmola, Tuula; Leppänen, Sanna M.; Tuittila, Eeva-Stiina; Aarva, Maija; Merilä, Päivi; Fritze, Hannu; Tiirola, Marja

2014-01-01

Nitrogen (N) accumulation rates in peatland ecosystems indicate significant biological atmospheric N2 fixation associated with Sphagnum mosses. Here, we show that the linkage between methanotrophic carbon cycling and N2 fixation may constitute an important mechanism in the rapid accumulation of N during the primary succession of peatlands. In our experimental stable isotope enrichment study, previously overlooked methane-induced N2 fixation explained more than one-third of the new N input in the younger peatland stages, where the highest N2 fixation rates and highest methane oxidation activities co-occurred in the water-submerged moss vegetation. PMID:24379382

Fixation of nitrogen in the presence of water vapor

DOEpatents

Harteck, Paul

1984-01-01

A process for the fixation of nitrogen is disclosed which comprises combining a mixture of nitrogen, oxygen, metal oxide and water vapor, initially heating the combination to initiate a reaction which forms nitrate, but at a temperature and pressure range below the dissociation pressure of the nitrate. With or without the water component, the yield of fixed nitrogen is increased by the use of a Linde Molecular Sieve Catalyst.

Comparison of Nitrogen Fixation Activity in Tall and Short Spartina alterniflora Salt Marsh Soils 1

PubMed Central

Hanson, Roger B.

1977-01-01

A comparison of the N2 fixers in the tall Spartina alterniflora and short S. alterniflora marsh soils was investigated. Zero-order kinetics and first-order kinetics of acetylene reduction were used to describe the activity of the N2 fixers in marsh soil slurries. It was found that the Vmax values were approximately 10 times greater for the N2 fixers in the tall Spartina than in the short Spartina marsh when raffinose was used as the energy source. In addition, the (Ks + Sn) values were approximately 4 to 15 times lower for the N2 fixers in the tall Spartina than in short Spartina marsh. First-order kinetics of nitrogen fixation for several substrates indicate that the N2 fixers in the tall Spartina marsh were two to seven times more active than those in the short Spartina marsh. Ammonium chloride (25 μg/ml) did not inhibit nitrogen fixation in the tall Spartina marsh, but there was a 50% inhibition in nitrogen fixation in the short Spartina marsh. On the other hand, sodium nitrate inhibited nitrogen fixation almost 100% at 25 μg/ml in both soil environments. Amino nitrogen (25 to 100 μg/ml) had little or no effect on nitrogen fixation. The results indicate that the N2 fixers in the tall Spartina marsh were physiologically more responsive to nutrient addition than those in the short Spartina marsh. This difference in the two populations may be related to the difference in daily tidal influence in the respective areas and thus provide another explanation for the enhanced S. alterniflora production in the creek bank soil system. PMID:16345213

Traffic Lights in Trichodesmium. Regulation of Photosynthesis for Nitrogen Fixation Studied by Chlorophyll Fluorescence Kinetic Microscopy1

PubMed Central

Küpper, Hendrik; Ferimazova, Naila; Šetlík, Ivan; Berman-Frank, Ilana

2004-01-01

We investigated interactions between photosynthesis and nitrogen fixation in the non-heterocystous marine cyanobacterium Trichodesmium IMS101 at the single-cell level by two-dimensional (imaging) microscopic measurements of chlorophyll fluorescence kinetics. Nitrogen fixation was closely associated with the appearance of cells with high basic fluorescence yield (F0), termed bright cells. In cultures aerated with normal air, both nitrogen fixation and bright cells appeared in the middle of the light phase. In cultures aerated with 5% oxygen, both processes occurred at a low level throughout most of the day. Under 50% oxygen, nitrogen fixation commenced at the beginning of the light phase but declined soon afterwards. Rapid reversible switches between fluorescence levels were observed, which indicated that the elevated F0 of the bright cells originates from reversible uncoupling of the photosystem II (PSII) antenna from the PSII reaction center. Two physiologically distinct types of bright cells were observed. Type I had about double F0 compared to the normal F0 in the dark phase and a PSII activity, measured as variable fluorescence (Fv = Fm − F0), similar to normal non-diazotrophic cells. Correlation of type I cells with nitrogen fixation, oxygen concentration, and light suggests that this physiological state is connected to an up-regulation of the Mehler reaction, resulting in oxygen consumption despite functional PSII. Type II cells had more than three times the normal F0 and hardly any PSII activity measurable by variable fluorescence. They did not occur under low-oxygen concentrations, but appeared under high-oxygen levels outside the diazotrophic period, suggesting that this state represents a reaction to oxidative stress not necessarily connected to nitrogen fixation. In addition to the two high-fluorescence states, cells were observed to reversibly enter a low-fluorescence state. This occurred mainly after a cell went through its bright phase and may represent a fluorescence-quenching recovery phase. PMID:15299119

Robust biological nitrogen fixation in a model grass-bacterial association.

PubMed

Pankievicz, Vânia C S; do Amaral, Fernanda P; Santos, Karina F D N; Agtuca, Beverly; Xu, Youwen; Schueller, Michael J; Arisi, Ana Carolina M; Steffens, Maria B R; de Souza, Emanuel M; Pedrosa, Fábio O; Stacey, Gary; Ferrieri, Richard A

2015-03-01

Nitrogen-fixing rhizobacteria can promote plant growth; however, it is controversial whether biological nitrogen fixation (BNF) from associative interaction contributes to growth promotion. The roots of Setaria viridis, a model C4 grass, were effectively colonized by bacterial inoculants resulting in a significant enhancement of growth. Nitrogen-13 tracer studies provided direct evidence for tracer uptake by the host plant and incorporation into protein. Indeed, plants showed robust growth under nitrogen-limiting conditions when inoculated with an ammonium-excreting strain of Azospirillum brasilense. (11)C-labeling experiments showed that patterns in central carbon metabolism and resource allocation exhibited by nitrogen-starved plants were largely reversed by bacterial inoculation, such that they resembled plants grown under nitrogen-sufficient conditions. Adoption of S. viridis as a model should promote research into the mechanisms of associative nitrogen fixation with the ultimate goal of greater adoption of BNF for sustainable crop production. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

Nitrogen fixation activity in biological soil crusts dominated by cyanobacteria in the Subpolar Urals (European North-East Russia).

PubMed

Patova, Elena; Sivkov, Michail; Patova, Anna

2016-09-01

The nitrogen fixation by biological soil crusts with a dominance of cyanobacteria was studied using the acetylene reduction assay in the territory of the Subpolar Urals (65°11' N, 60°18' E), Russia. The field measurements of nitrogen fixation activity were conducted in situ for two different types of soil crusts dominated by Stigonema (V1 type) and Nostoc with Scytonema (V2 type). The nitrogen fixation process had similar dynamics in both crusts but nitrogen fixation rates were different. The crusts of the V2 type showed a significantly higher acetylene reduction activity, with ethylene production rate of 1.76 ± 0.49 g C2H4 m(-2) h(-1) at 15°C, compared with V1-type soil crusts, with a rate of 0.53 ± 0.21 mg C2H4 m(-2) h(-1) at 15°C. The daily value of acetylene reduction activity in V2-type soil crusts was 32.7 ± 6.2 mg C2H4 m(-2) d(-1) and in V1-type crusts, 12.3 ± 1.8 mg C2H4 m(-2) d(-1) After recalculation for N, the daily values of nitrogen fixation were in the range 3.3-22.3 mg N m(-2) d(-1), which is a few times higher than the values of N input from the precipitation to the soil in the studied regions. The dependence of nitrogen-fixation activity on temperature and light intensity of biological soil crusts was investigated. On the basis of temperature models obtained from the dependence, the nitrogen balance was calculated for the growing season (approximately 120 days). The crusts dominated by Stigonema species were fixing 0.3 g N m(-2) (ethylene production rate, 1.10 g C2H4 m(-2)) and crusts dominated by Nostoc and Scytonema were fixing 1.3 g N m(-2) (4.10 g C2H4 m(-2)). © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Medicago truncatula copper transporter 1 (MtCOPT1) delivers copper for symbiotic nitrogen fixation.

PubMed

Senovilla, Marta; Castro-Rodríguez, Rosario; Abreu, Isidro; Escudero, Viviana; Kryvoruchko, Igor; Udvardi, Michael K; Imperial, Juan; González-Guerrero, Manuel

2018-04-01

Copper is an essential nutrient for symbiotic nitrogen fixation. This element is delivered by the host plant to the nodule, where membrane copper (Cu) transporter would introduce it into the cell to synthesize cupro-proteins. COPT family members in the model legume Medicago truncatula were identified and their expression determined. Yeast complementation assays, confocal microscopy and phenotypical characterization of a Tnt1 insertional mutant line were carried out in the nodule-specific M. truncatula COPT family member. Medicago truncatula genome encodes eight COPT transporters. MtCOPT1 (Medtr4g019870) is the only nodule-specific COPT gene. It is located in the plasma membrane of the differentiation, interzone and early fixation zones. Loss of MtCOPT1 function results in a Cu-mitigated reduction of biomass production when the plant obtains its nitrogen exclusively from symbiotic nitrogen fixation. Mutation of MtCOPT1 results in diminished nitrogenase activity in nodules, likely an indirect effect from the loss of a Cu-dependent function, such as cytochrome oxidase activity in copt1-1 bacteroids. These data are consistent with a model in which MtCOPT1 transports Cu from the apoplast into nodule cells to provide Cu for essential metabolic processes associated with symbiotic nitrogen fixation. © 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

Molybdenum and Phosphorus Interact to Constrain Asymbiotic Nitrogen Fixation in Tropical Forests

PubMed Central

Wurzburger, Nina; Bellenger, Jean Philippe; Kraepiel, Anne M. L.; Hedin, Lars O.

2012-01-01

Biological di-nitrogen fixation (N2) is the dominant natural source of new nitrogen to land ecosystems. Phosphorus (P) is thought to limit N2 fixation in many tropical soils, yet both molybdenum (Mo) and P are crucial for the nitrogenase reaction (which catalyzes N2 conversion to ammonia) and cell growth. We have limited understanding of how and when fixation is constrained by these nutrients in nature. Here we show in tropical forests of lowland Panama that the limiting element on asymbiotic N2 fixation shifts along a broad landscape gradient in soil P, where Mo limits fixation in P-rich soils while Mo and P co-limit in P-poor soils. In no circumstance did P alone limit fixation. We provide and experimentally test a mechanism that explains how Mo and P can interact to constrain asymbiotic N2 fixation. Fixation is uniformly favored in surface organic soil horizons - a niche characterized by exceedingly low levels of available Mo relative to P. We show that soil organic matter acts to reduce molybdate over phosphate bioavailability, which, in turn, promotes Mo limitation in sites where P is sufficient. Our findings show that asymbiotic N2 fixation is constrained by the relative availability and dynamics of Mo and P in soils. This conceptual framework can explain shifts in limitation status across broad landscape gradients in soil fertility and implies that fixation depends on Mo and P in ways that are more complex than previously thought. PMID:22470462

Growth, nitrogen fixation and mineral acquisition of Alnus sieboldiana after inoculation of Frankia together with Gigaspora margarita and Pseudomonas putida.

Treesearch

Takashi Yamanaka; Akio Akama; Ching-Yan Li; Hiroaki Okabe

2005-01-01

The role of tetrapartite associations among Frankia, Gigaspora margarita (an arbuscular mycorrhizal fungus), Pseudomonas putida (rhizobacterium), and Alnus sieboldiana in growth, nitrogen fixation, and mineral acquisition of A. sieboldiana was investigated....

Patterns of Nitrogen Fixation and Related Genetic Diversity (nifH) in Microbial Mats and Stromatolites from Different Environments

NASA Astrophysics Data System (ADS)

Beltrán, Y. Y.; Centeno, C.; Falcón, L. I.

2010-04-01

We want to estimate the patterns of nitrogen fixation and the related genetic diversity (nifH) of microbial mats and microbialites on dial and temporal scales along a physicochemical and geographical gradient.

A Medicago truncatula Tobacco Retrotransposon Insertion Mutant Collection with Defects in Nodule Development and Symbiotic Nitrogen Fixation1[W][OA

PubMed Central

Pislariu, Catalina I.; D. Murray, Jeremy; Wen, JiangQi; Cosson, Viviane; Muni, RajaSekhara Reddy Duvvuru; Wang, Mingyi; A. Benedito, Vagner; Andriankaja, Andry; Cheng, Xiaofei; Jerez, Ivone Torres; Mondy, Samuel; Zhang, Shulan; Taylor, Mark E.; Tadege, Million; Ratet, Pascal; Mysore, Kirankumar S.; Chen, Rujin; Udvardi, Michael K.

2012-01-01

A Tnt1-insertion mutant population of Medicago truncatula ecotype R108 was screened for defects in nodulation and symbiotic nitrogen fixation. Primary screening of 9,300 mutant lines yielded 317 lines with putative defects in nodule development and/or nitrogen fixation. Of these, 230 lines were rescreened, and 156 lines were confirmed with defective symbiotic nitrogen fixation. Mutants were sorted into six distinct phenotypic categories: 72 nonnodulating mutants (Nod−), 51 mutants with totally ineffective nodules (Nod+ Fix−), 17 mutants with partially ineffective nodules (Nod+ Fix+/−), 27 mutants defective in nodule emergence, elongation, and nitrogen fixation (Nod+/− Fix−), one mutant with delayed and reduced nodulation but effective in nitrogen fixation (dNod+/− Fix+), and 11 supernodulating mutants (Nod++Fix+/−). A total of 2,801 flanking sequence tags were generated from the 156 symbiotic mutant lines. Analysis of flanking sequence tags revealed 14 insertion alleles of the following known symbiotic genes: NODULE INCEPTION (NIN), DOESN’T MAKE INFECTIONS3 (DMI3/CCaMK), ERF REQUIRED FOR NODULATION, and SUPERNUMERARY NODULES (SUNN). In parallel, a polymerase chain reaction-based strategy was used to identify Tnt1 insertions in known symbiotic genes, which revealed 25 additional insertion alleles in the following genes: DMI1, DMI2, DMI3, NIN, NODULATION SIGNALING PATHWAY1 (NSP1), NSP2, SUNN, and SICKLE. Thirty-nine Nod− lines were also screened for arbuscular mycorrhizal symbiosis phenotypes, and 30 mutants exhibited defects in arbuscular mycorrhizal symbiosis. Morphological and developmental features of several new symbiotic mutants are reported. The collection of mutants described here is a source of novel alleles of known symbiotic genes and a resource for cloning novel symbiotic genes via Tnt1 tagging. PMID:22679222

A Medicago truncatula tobacco retrotransposon insertion mutant collection with defects in nodule development and symbiotic nitrogen fixation.

PubMed

Pislariu, Catalina I; Murray, Jeremy D; Wen, JiangQi; Cosson, Viviane; Muni, RajaSekhara Reddy Duvvuru; Wang, Mingyi; Benedito, Vagner A; Andriankaja, Andry; Cheng, Xiaofei; Jerez, Ivone Torres; Mondy, Samuel; Zhang, Shulan; Taylor, Mark E; Tadege, Million; Ratet, Pascal; Mysore, Kirankumar S; Chen, Rujin; Udvardi, Michael K

2012-08-01

A Tnt1-insertion mutant population of Medicago truncatula ecotype R108 was screened for defects in nodulation and symbiotic nitrogen fixation. Primary screening of 9,300 mutant lines yielded 317 lines with putative defects in nodule development and/or nitrogen fixation. Of these, 230 lines were rescreened, and 156 lines were confirmed with defective symbiotic nitrogen fixation. Mutants were sorted into six distinct phenotypic categories: 72 nonnodulating mutants (Nod-), 51 mutants with totally ineffective nodules (Nod+ Fix-), 17 mutants with partially ineffective nodules (Nod+ Fix+/-), 27 mutants defective in nodule emergence, elongation, and nitrogen fixation (Nod+/- Fix-), one mutant with delayed and reduced nodulation but effective in nitrogen fixation (dNod+/- Fix+), and 11 supernodulating mutants (Nod++Fix+/-). A total of 2,801 flanking sequence tags were generated from the 156 symbiotic mutant lines. Analysis of flanking sequence tags revealed 14 insertion alleles of the following known symbiotic genes: NODULE INCEPTION (NIN), DOESN'T MAKE INFECTIONS3 (DMI3/CCaMK), ERF REQUIRED FOR NODULATION, and SUPERNUMERARY NODULES (SUNN). In parallel, a polymerase chain reaction-based strategy was used to identify Tnt1 insertions in known symbiotic genes, which revealed 25 additional insertion alleles in the following genes: DMI1, DMI2, DMI3, NIN, NODULATION SIGNALING PATHWAY1 (NSP1), NSP2, SUNN, and SICKLE. Thirty-nine Nod- lines were also screened for arbuscular mycorrhizal symbiosis phenotypes, and 30 mutants exhibited defects in arbuscular mycorrhizal symbiosis. Morphological and developmental features of several new symbiotic mutants are reported. The collection of mutants described here is a source of novel alleles of known symbiotic genes and a resource for cloning novel symbiotic genes via Tnt1 tagging.

Assessing nitrogen fixation in mixed- and single-species plantations of Eucalyptus globulus and Acacia mearnsii.

PubMed

Forrester, David I; Schortemeyer, Marcus; Stock, William D; Bauhus, Jürgen; Khanna, Partap K; Cowie, Annette L

2007-09-01

Mixtures of Eucalyptus globulus Labill. and Acacia mearnsii de Wildeman are twice as productive as E. globulus monocultures growing on the same site in East Gippsland, Victoria, Australia, possibly because of increased nitrogen (N) availability owing to N(2) fixation by A. mearnsii. To investigate whether N(2) fixation by A. mearnsii could account for the mixed-species growth responses, we assessed N(2) fixation by the accretion method and the (15)N natural abundance method. Nitrogen gained by E. globulus and A. mearnsii mixtures and monocultures was calculated by the accretion method with plant and soil samples collected 10 years after plantation establishment. Nitrogen in biomass and soil confirmed that A. mearnsii influenced N dynamics. Assuming that the differences in soil, forest floor litter and biomass N of plots containing A. mearnsii compared with E. globulus monocultures were due to N(2) fixation, the 10-year annual mean rates of N(2) fixation were 38 and 86 kg ha(-1) year(-1) in 1:1 mixtures and A. mearnsii monocultures, respectively. Nitrogen fixation by A. mearnsii could not be quantified on the basis of the natural abundance of (15)N because such factors as mycorrhization type and fractionation of N isotopes during N cycling within the plant confounded the effect of the N source on the N isotopic signature of plants. This study shows that A. mearnsii fixed significant quantities of N(2) when mixed with E. globulus. A decline in delta(15)N values of E. globulus and A. mearnsii with time, from 2 to 10 years, is further evidence that N(2) was fixed and cycled through the stands. The increased aboveground biomass production of E. globulus trees in mixtures when compared with monocultures can be attributed to increases in N availability.

Aquatic Plant Control Research Program: The Rhizosphere Microbiology of Rooted Aquatic Plants.

DTIC Science & Technology

1988-04-01

acids. Acetic acid postulated as agent Thalassia Nitrogen fixation in Capone 1983 testudinwn rhizosphere and phyllosphere 13. Fungi living in the...microbial population associated with the rhizome detritus. 26. Durako and Moffler (1987) examined responses of Thalassia testudinum to nitrogen enrichment...economy of seagrasses for communities of Thalassia testudinium and Zostera maina. He concluded that nitrogen fixation is probably more important for

Improvement in nitrogen fixation capacity could be part of the domestication process in soybean

PubMed Central

Muñoz, N; Qi, X; Li, M-W; Xie, M; Gao, Y; Cheung, M-Y; Wong, F-L; Lam, H-M

2016-01-01

Biological nitrogen fixation (BNF) in soybeans is a complex process involving the interplay between the plant host and the symbiotic rhizobia. As nitrogen supply has a crucial role in growth and development, higher nitrogen fixation capacity would be important to achieve bigger plants and larger seeds, which were important selection criteria during plant domestication by humans. To test this hypothesis, we monitored the nitrogen fixation-related performance in 31 cultivated and 17 wild soybeans after inoculation with the slow-growing Bradyrhizobium diazoefficiens sp. nov. USDA110 and the fast-growing Sinorhizobium (Ensifer) fredii CCBAU45436. Our results showed that, in general, cultivated soybeans gave better performance in BNF. Electron microscopic studies indicated that there was an exceptionally high accumulation of poly-β-hydroxybutyrate bodies in bacteroids in the nodules of all wild soybeans tested, suggesting that the C/N balance in wild soybeans may not be optimized for nitrogen fixation. Furthermore, we identified new quantitative trait loci (QTLs) for total ureides and total nodule fresh weight by employing a recombinant inbred population composed of descendants from a cross between a cultivated and a wild parent. Using nucleotide diversity (θπ), divergence index (Fst) and distribution of fixed single-nucleotide polymorphisms as parameters, we found that some regions in the total ureides QTL on chromosome 17 and the total nodule fresh weight QTL on chromosome 12 exhibited very low diversity among cultivated soybeans, suggesting that these were traits specially selected during the domestication and breeding process. PMID:27118154

Biological nitrogen fixation and habitat of running buffalo clover

Treesearch

D.R. Morris; V.S. Baligar; T.M. Schuler; P.J. Harmon

2002-01-01

Running buffalo clover (RBC) [Trifolium stoloniferum (Muhl. ex Eat.)] is an endangered species whose survival is uncertain. An experiment was conducted on extant RBC sites to investigate biological nitrogen (N2) fixation, associated plant species, and soil conditions under natural mountain settings. Isotope (15...

Irreversibly increased nitrogen fixation in Trichodesmium experimentally adapted to elevated carbon dioxide

PubMed Central

Hutchins, David A.; Walworth, Nathan G.; Webb, Eric A.; Saito, Mak A.; Moran, Dawn; McIlvin, Matthew R.; Gale, Jasmine; Fu, Fei-Xue

2015-01-01

Nitrogen fixation rates of the globally distributed, biogeochemically important marine cyanobacterium Trichodesmium increase under high carbon dioxide (CO2) levels in short-term studies due to physiological plasticity. However, its long-term adaptive responses to ongoing anthropogenic CO2 increases are unknown. Here we show that experimental evolution under extended selection at projected future elevated CO2 levels results in irreversible, large increases in nitrogen fixation and growth rates, even after being moved back to lower present day CO2 levels for hundreds of generations. This represents an unprecedented microbial evolutionary response, as reproductive fitness increases acquired in the selection environment are maintained after returning to the ancestral environment. Constitutive rate increases are accompanied by irreversible shifts in diel nitrogen fixation patterns, and increased activity of a potentially regulatory DNA methyltransferase enzyme. High CO2-selected cell lines also exhibit increased phosphorus-limited growth rates, suggesting a potential advantage for this keystone organism in a more nutrient-limited, acidified future ocean. PMID:26327191

Irreversibly increased nitrogen fixation in Trichodesmium experimentally adapted to elevated carbon dioxide

NASA Astrophysics Data System (ADS)

Hutchins, David A.; Walworth, Nathan G.; Webb, Eric A.; Saito, Mak A.; Moran, Dawn; McIlvin, Matthew R.; Gale, Jasmine; Fu, Fei-Xue

2015-09-01

Nitrogen fixation rates of the globally distributed, biogeochemically important marine cyanobacterium Trichodesmium increase under high carbon dioxide (CO2) levels in short-term studies due to physiological plasticity. However, its long-term adaptive responses to ongoing anthropogenic CO2 increases are unknown. Here we show that experimental evolution under extended selection at projected future elevated CO2 levels results in irreversible, large increases in nitrogen fixation and growth rates, even after being moved back to lower present day CO2 levels for hundreds of generations. This represents an unprecedented microbial evolutionary response, as reproductive fitness increases acquired in the selection environment are maintained after returning to the ancestral environment. Constitutive rate increases are accompanied by irreversible shifts in diel nitrogen fixation patterns, and increased activity of a potentially regulatory DNA methyltransferase enzyme. High CO2-selected cell lines also exhibit increased phosphorus-limited growth rates, suggesting a potential advantage for this keystone organism in a more nutrient-limited, acidified future ocean.

Molybdenum Trafficking for Nitrogen Fixation†

PubMed Central

Hernandez, Jose A.; George, Simon J.; Rubio, Luis M.

2009-01-01

The molybdenum nitrogenase is responsible for most biological nitrogen fixation, a prokaryotic metabolic process that determines the global biogeochemical cycles of nitrogen and carbon. Here we describe the trafficking of molybdenum for nitrogen fixation in the model diazotrophic bacterium Azotobacter vinelandii. The genes and proteins involved in molybdenum uptake, homeostasis, storage, regulation, and nitrogenase cofactor biosynthesis are reviewed. Molybdenum biochemistry in A. vinelandii reveals unexpected mechanisms and a new role for iron-sulfur clusters in the sequestration and delivery of molybdenum. PMID:19772354

Biological Hydrogen Production: Simultaneous Saccharification and Fermentation with Nitrogen and Phosphorus Removal from Wastewater Effluent

DTIC Science & Technology

2012-03-01

the Haber - Bosch process, in which hydrogen is first produced from methane (eq. 1), then ammonia is produced from nitrogen and hydrogen: N2 (g...3H2 (g) - 2NH3 (g) (5) Agronomists have calculated that well over one-third of the world’s present population is fed by virtue of the Haber - Bosch ...fixation of nitrogen through the Haber - Bosch process, leading to a potential confluence of energy and fertilizer crises. Biological nitrogen fixation

Nitrogen fixation in a non-heterocystous cyanobacterial mat from a mountain river

NASA Astrophysics Data System (ADS)

Berrendero, Esther; Valiente, Eduardo Fernández; Perona, Elvira; Gómez, Claudia L.; Loza, Virginia; Muñoz-Martín, M. Ángeles; Mateo, Pilar

2016-08-01

In situ nitrogen fixation was investigated in a cyanobacterial mat growing on the bed of rocks of the Muga River, Spain. The filamentous non-heterocystous cyanobacterium Schizothrix dominated the mat, showing nitrogenase activity in the light at similar rates to those found in nearby heterocystous Rivularia colonies. N2 fixation in the light was significantly increased by an inhibitor of PSII and oxygen evolution, DCMU (3-[3,4-dichlorophenyl]-1,1-dimethylurea), and anaerobic conditions. However, no nitrogenase activity was found in the dark. Addition of fructose as a respiratory substrate induced nitrogenase activity in samples incubated under aerobic conditions in the dark but not in anaerobic conditions. Microelectrode oxygen profiles showed internal microaerobic microzones where nitrogen fixation might concentrate. Analyses of the 16S rRNA gene revealed only the presence of sequences belonging to filamentous non-heterocystous cyanobacteria. nifH gene diversity showed that the major phylotypes also belonged to this group. One of the three strains isolated from the Schizothrix mat was capable of fixing N2 and growing in the absence of combined N. This was consistent with the nifH gene analysis. These results suggest a relevant contribution of non-heterocystous cyanobacteria to nitrogen fixation in these mats.

Rising sea level, temperature, and precipitation impact plant and ecosystem responses to elevated CO2 on a Chesapeake Bay wetland: review of a 28-year study.

PubMed

Drake, Bert G

2014-11-01

An ongoing field study of the effects of elevated atmospheric CO2 on a brackish wetland on Chesapeake Bay, started in 1987, is unique as the longest continually running investigation of the effects of elevated CO2 on an ecosystem. Since the beginning of the study, atmospheric CO2 increased 18%, sea level rose 20 cm, and growing season temperature varied with approximately the same range as predicted for global warming in the 21st century. This review looks back at this study for clues about how the effects of rising sea level, temperature, and precipitation interact with high atmospheric CO2 to alter the physiology of C3 and C4 photosynthetic species, carbon assimilation, evapotranspiration, plant and ecosystem nitrogen, and distribution of plant communities in this brackish wetland. Rising sea level caused a shift to higher elevations in the Scirpus olneyi C3 populations on the wetland, displacing the Spartina patens C4 populations. Elevated CO2 stimulated carbon assimilation in the Scirpus C3 species measured by increased shoot and root density and biomass, net ecosystem production, dissolved organic and inorganic carbon, and methane production. But elevated CO2 also decreased biomass of the grass, S. patens C4. The elevated CO2 treatment reduced tissue nitrogen concentration in shoots, roots, and total canopy nitrogen, which was associated with reduced ecosystem respiration. Net ecosystem production was mediated by precipitation through soil salinity: high salinity reduced the CO2 effect on net ecosystem production, which was zero in years of severe drought. The elevated CO2 stimulation of shoot density in the Scirpus C3 species was sustained throughout the 28 years of the study. Results from this study suggest that rising CO2 can add substantial amounts of carbon to ecosystems through stimulation of carbon assimilation, increased root exudates to supply nitrogen fixation, reduced dark respiration, and improved water and nitrogen use efficiency. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

A model using marginal efficiency of investment to analyze carbon and nitrogen interactions in terrestrial ecosystems (ACONITE Version 1)

NASA Astrophysics Data System (ADS)

Thomas, R. Q.; Williams, M.

2014-09-01

Carbon (C) and nitrogen (N) cycles are coupled in terrestrial ecosystems through multiple processes including photosynthesis, tissue allocation, respiration, N fixation, N uptake, and decomposition of litter and soil organic matter. Capturing the constraint of N on terrestrial C uptake and storage has been a focus of the Earth System Modeling community. However, there is little understanding of the trade-offs and sensitivities of allocating C and N to different tissues in order to optimize the productivity of plants. Here we describe a new, simple model of ecosystem C-N cycling and interactions (ACONITE), that builds on theory related to plant economics in order to predict key ecosystem properties (leaf area index, leaf C : N, N fixation, and plant C use efficiency) based on the outcome of assessments of the marginal change in net C or N uptake associated with a change in allocation of C or N to plant tissues. We simulated and evaluated steady-state ecosystem stocks and fluxes in three different forest ecosystems types (tropical evergreen, temperate deciduous, and temperate evergreen). Leaf C : N differed among the three ecosystem types (temperate deciduous < tropical evergreen < temperature evergreen), a result that compared well to observations from a global database describing plant traits. Gross primary productivity (GPP) and net primary productivity (NPP) estimates compared well to observed fluxes at the simulation sites. Simulated N fixation at steady-state, calculated based on relative demand for N and the marginal return on C investment to acquire N, was an order of magnitude higher in the tropical forest than in the temperate forest, consistent with observations. A sensitivity analysis revealed that parameterization of the relationship between leaf N and leaf respiration had the largest influence on leaf area index and leaf C : N. A parameter governing how photosynthesis scales with day length had the largest influence on total vegetation C, GPP, and NPP. Multiple parameters associated with photosynthesis, respiration, and N uptake influenced the rate of N fixation. Overall, our ability to constrain leaf area index and allow spatially and temporally variable leaf C : N can help address challenges simulating these properties in ecosystem and Earth System models. Furthermore, the simple approach with emergent properties based on coupled C-N dynamics has potential for use in research that uses data-assimilation methods to integrate data on both the C and N cycles to improve C flux forecasts.

Nutrient co-limited Trichodesmium as nitrogen source or sink in a future ocean.

PubMed

Walworth, Nathan G; Fu, Fei-Xue; Lee, Michael D; Cai, Xiaoni; Saito, Mak A; Webb, Eric A; Hutchins, David A

2017-11-27

Nitrogen-fixing (N 2 ) cyanobacteria provide bioavailable nitrogen to vast ocean regions but are in turn limited by iron (Fe) and/or phosphorus (P), which may force them to employ alternative nitrogen acquisition strategies. The adaptive responses of nitrogen-fixers to global-change drivers under nutrient-limited conditions could profoundly alter the current ocean nitrogen and carbon cycles. Here, we show that the globally-important N 2 -fixer Trichodesmium fundamentally shifts nitrogen metabolism towards organic-nitrogen scavenging following long-term high-CO 2 adaptation under iron and/or phosphorus (co)-limitation. Global shifts in transcripts and proteins under high CO 2 /Fe-limited and/or P-limited conditions include decreases in the N 2 -fixing nitrogenase enzyme, coupled with major increases in enzymes that oxidize trimethylamine (TMA). TMA is an abundant, biogeochemically-important organic nitrogen compound that supports rapid Trichodesmium growth while inhibiting N 2 fixation. In a future high-CO 2 ocean, this whole-cell energetic reallocation towards organic nitrogen scavenging and away from N 2 -fixation may reduce new-nitrogen inputs by Trichodesmium , while simultaneously depleting the scarce fixed-nitrogen supplies of nitrogen-limited open ocean ecosystems. Importance Trichodesmium is among the most biogeochemically-significant microorganisms in the ocean, since it supplies up to 50% of the new nitrogen supporting open ocean food webs. We used Trichodesmium cultures adapted to high CO 2 for 7 years followed by additional exposure to iron and/or phosphorus (co)-limitation. We show that 'future ocean' conditions of high CO 2 and concurrent nutrient limitation(s) fundamentally shift nitrogen metabolism away from nitrogen fixation, and instead towards upregulation of organic-nitrogen scavenging pathways. We show that Trichodesmium's responses to projected future ocean conditions include decreases in the nitrogen-fixing nitrogenase enzymes, coupled with major increases in enzymes that oxidize the abundant organic nitrogen source trimethylamine (TMA). Such a shift towards organic nitrogen uptake and away from nitrogen fixation may substantially reduce new-nitrogen inputs by Trichodesmium to the rest of the microbial community in the future high-CO 2 ocean, with potential global implications for ocean carbon and nitrogen cycling. Copyright © 2017 American Society for Microbiology.

Application of 15N-enrichment methodologies to estimate nitrogen fixation in Casuarina equisetifolia

Treesearch

John A. Parrotta; Dwight D. Baker; Maurice Fried

1994-01-01

The 15N-enrichment technique for estimating biological nitrogen fixation in Casuarina equisetifolia J.R. & G. Forst. was evaluated under field conditions in single-species and mixed-species plantings (with a nonfixing reference species, Eucalyptus X robusta J.E. Smith) between...

Nitrogen comes down to earth: report from the 5th European Nitrogen Fixation Conference.

PubMed

De Hoff, Peter; Hirsch, Ann M

2003-05-01

For four days and four nights, with almost 50 presentations and more than 175 posters, the 5th European Nitrogen Fixation Conference continued a tradition of excellence, bringing scientists from diverse fields such as microbiology, biochemistry, computational genomics, and plant physiology together to address the complex problems associated with biological nitrogen fixation (BNF). The conference was hosted by the John Innes Center and the University of East Anglia in Norwich, England and took place from September 6 through 10, 2002. A diverse range of topics was presented, from the evolution of rhizobial genomes to the plant genes involved in bacterial and fungal symbiosis, to the structure of nitrogenase, and to the means by which nitrogen is shuttled between the symbiotic bacteria and the plant. Additionally, sessions involving broader issues, such as nitrogen fertilizer use and work being done in developing countries, brought home the importance of the research being carried out in BNF around the world.

Co-inoculation of a Pea Core-Collection with Diverse Rhizobial Strains Shows Competitiveness for Nodulation and Efficiency of Nitrogen Fixation Are Distinct traits in the Interaction

PubMed Central

Bourion, Virginie; Heulin-Gotty, Karine; Aubert, Véronique; Tisseyre, Pierre; Chabert-Martinello, Marianne; Pervent, Marjorie; Delaitre, Catherine; Vile, Denis; Siol, Mathieu; Duc, Gérard; Brunel, Brigitte; Burstin, Judith; Lepetit, Marc

2018-01-01

Pea forms symbiotic nodules with Rhizobium leguminosarum sv. viciae (Rlv). In the field, pea roots can be exposed to multiple compatible Rlv strains. Little is known about the mechanisms underlying the competitiveness for nodulation of Rlv strains and the ability of pea to choose between diverse compatible Rlv strains. The variability of pea-Rlv partner choice was investigated by co-inoculation with a mixture of five diverse Rlv strains of a 104-pea collection representative of the variability encountered in the genus Pisum. The nitrogen fixation efficiency conferred by each strain was determined in additional mono-inoculation experiments on a subset of 18 pea lines displaying contrasted Rlv choice. Differences in Rlv choice were observed within the pea collection according to their genetic or geographical diversities. The competitiveness for nodulation of a given pea-Rlv association evaluated in the multi-inoculated experiment was poorly correlated with its nitrogen fixation efficiency determined in mono-inoculation. Both plant and bacterial genetic determinants contribute to pea-Rlv partner choice. No evidence was found for co-selection of competitiveness for nodulation and nitrogen fixation efficiency. Plant and inoculant for an improved symbiotic association in the field must be selected not only on nitrogen fixation efficiency but also for competitiveness for nodulation. PMID:29367857

Co-inoculation of a Pea Core-Collection with Diverse Rhizobial Strains Shows Competitiveness for Nodulation and Efficiency of Nitrogen Fixation Are Distinct traits in the Interaction.

PubMed

Bourion, Virginie; Heulin-Gotty, Karine; Aubert, Véronique; Tisseyre, Pierre; Chabert-Martinello, Marianne; Pervent, Marjorie; Delaitre, Catherine; Vile, Denis; Siol, Mathieu; Duc, Gérard; Brunel, Brigitte; Burstin, Judith; Lepetit, Marc

2017-01-01

Pea forms symbiotic nodules with Rhizobium leguminosarum sv. viciae (Rlv). In the field, pea roots can be exposed to multiple compatible Rlv strains. Little is known about the mechanisms underlying the competitiveness for nodulation of Rlv strains and the ability of pea to choose between diverse compatible Rlv strains. The variability of pea-Rlv partner choice was investigated by co-inoculation with a mixture of five diverse Rlv strains of a 104-pea collection representative of the variability encountered in the genus Pisum . The nitrogen fixation efficiency conferred by each strain was determined in additional mono-inoculation experiments on a subset of 18 pea lines displaying contrasted Rlv choice. Differences in Rlv choice were observed within the pea collection according to their genetic or geographical diversities. The competitiveness for nodulation of a given pea-Rlv association evaluated in the multi-inoculated experiment was poorly correlated with its nitrogen fixation efficiency determined in mono-inoculation. Both plant and bacterial genetic determinants contribute to pea-Rlv partner choice. No evidence was found for co-selection of competitiveness for nodulation and nitrogen fixation efficiency. Plant and inoculant for an improved symbiotic association in the field must be selected not only on nitrogen fixation efficiency but also for competitiveness for nodulation.

Satellite captures trichodesmium blooms in the southwestern tropical Pacific

NASA Astrophysics Data System (ADS)

Dupouy, Cécile; Neveux, Jacques; Subramaniam, Ajit; Mulholland, Margaret R.; Montoya, Joseph P.; Campbell, Lisa; Carpenter, Edward J.; Capone, Douglas G.

Obtaining a true estimate of nitrogen fixation by cyanobacteria in the oceans, mainly Trichodesmium, is an important step toward understanding the entire nitrogen cycle in the tropical ocean. This strictly anaerobic process, which has a high Fe requirement, could regulate atmospheric CO2 over geological time. For example, during interglacial periods, N2 fixation would be too low (low Fe) to balance denitrification and the ocean would lose its fixed nitrogen [Falkowski, 1997]. Has the level of marine nitrogen fixation been underestimated until now? High N2 fixation rates measured on Trichodesmium spp. communities have led to an upward revision of this marine flux [Capone et al, 1997]. Recent modeling studies and observations predict that N2 fixation could regulate the long-term N:P equilibrium in the oceans and balance denitrification [Tyrell, 1999; J L. Sarmiento and N. Gruber, manuscript in preparation, 1999].The major nitrogen fixer, Trichodesmium spp., which are filamentous, nonheterocystous N2-fixing cyanobacteria, has a nearly ubiquitous distribution in the euphotic zone of tropical and subtropical seas and could play a major role in bringing new N to these oligotrophic systems. Satellite images from Sea-viewing Wide Field-of-view Sensor (SeaWiFs), the recently launched ocean color sensor, and data from a recent cruise, provide further evidence of the importance of Trichodesmium in the southwestern tropical Pacific Ocean.

Screening and Selection of Maize to Enhance Associative Bacterial Nitrogen Fixation 1

PubMed Central

Ela, Stephen W.; Anderson, Mary Ann; Brill, Winston J.

1982-01-01

The ability of maize (corn, Zea mays L.) to support bacterial nitrogen fixation in or on maize roots has been increased, through screening and selection. Isotopic N fixed from 15N2 was found on the roots. The nitrogen-fixing association was found in germplasm from tropical maize, but this activity can be transferred to maize currently used in midwestern United States agriculture. PMID:16662718

Heterotrophic organisms dominate nitrogen fixation in the South Pacific Gyre

PubMed Central

Halm, Hannah; Lam, Phyllis; Ferdelman, Timothy G; Lavik, Gaute; Dittmar, Thorsten; LaRoche, Julie; D'Hondt, Steven; Kuypers, Marcel MM

2012-01-01

Oceanic subtropical gyres are considered biological deserts because of the extremely low availability of nutrients and thus minimum productivities. The major source of nutrient nitrogen in these ecosystems is N2-fixation. The South Pacific Gyre (SPG) is the largest ocean gyre in the world, but measurements of N2-fixation therein, or identification of microorganisms involved, are scarce. In the 2006/2007 austral summer, we investigated nitrogen and carbon assimilation at 11 stations throughout the SPG. In the ultra-oligotrophic waters of the SPG, the chlorophyll maxima reached as deep as 200 m. Surface primary production seemed limited by nitrogen, as dissolved inorganic carbon uptake was stimulated upon additions of 15N-labeled ammonium and leucine in our incubation experiments. N2-fixation was detectable throughout the upper 200 m at most stations, with rates ranging from 0.001 to 0.19 nM N h−1. N2-fixation in the SPG may account for the production of 8–20% of global oceanic new nitrogen. Interestingly, comparable 15N2-fixation rates were measured under light and dark conditions. Meanwhile, phylogenetic analyses for the functional gene biomarker nifH and its transcripts could not detect any common photoautotrophic diazotrophs, such as, Trichodesmium, but a prevalence of γ-proteobacteria and the unicellular photoheterotrophic Group A cyanobacteria. The dominance of these likely heterotrophic diazotrophs was further verified by quantitative PCR. Hence, our combined results show that the ultra-oligotrophic SPG harbors a hitherto unknown heterotrophic diazotrophic community, clearly distinct from other oceanic gyres previously visited. PMID:22170429

Sedimentary denitrification: Isotope fractionation and its impact on water column nitrate isotopes

NASA Astrophysics Data System (ADS)

Dähnke, K.; Thamdrup, B.

2012-04-01

The global marine nitrogen cycle is constrained by one major source and two processes that act as nitrogen sinks: nitrogen fixation on the one side and denitrification or anammox on the other. These processes with their respective isotope effecst set the marine nitrate 15N-isotope value to a relatively constant average of 5 per mil. This value can be used to better assess the magnitude of these source and sink terms, but the underlying assumption at present is that sedimentary denitrification, a process responsible for approximately one third of global nitrogen removal, has little to no isotope effect on the water column. We tested this hypothesis in sediment incubations, measuring net denitrification and nitrogen and oxygen stable isotope fractionation in surface sediments from the coastal Baltic Sea (Boknis Eck, Northern Germany). We found tremendously high denitrification rates, and regardless of current paradigms assuming little fractionation during sediment denitrification, we measured fractionation factors of 19 per mil for nitrogen and 11 per mil for oxygen in nitrate. These results potentially challenge the current view of fractionation during sedimentary denitrification and imply that nitrogen budget calculation may need to consider this variability. Furthermore, the ratio of fractionation factors for nitrogen and oxygen is distinct from the 1 : 1 ratio otherwise found in marine systems, and suggests that isotope kinetics of sedimentary denitrification might be entirely different from water column denitrification. Acknowledgements: This work was funded by the German Research Foundation (DFG) and in parts by the Danish National Research Foundation.

Phylogenetic constraints do not explain the rarity of nitrogen-fixing trees in late-successional temperate forests.

PubMed

Menge, Duncan N L; DeNoyer, Jeanne L; Lichstein, Jeremy W

2010-08-06

Symbiotic nitrogen (N)-fixing trees are rare in late-successional temperate forests, even though these forests are often N limited. Two hypotheses could explain this paradox. The 'phylogenetic constraints hypothesis' states that no late-successional tree taxa in temperate forests belong to clades that are predisposed to N fixation. Conversely, the 'selective constraints hypothesis' states that such taxa are present, but N-fixing symbioses would lower their fitness. Here we test the phylogenetic constraints hypothesis. Using U.S. forest inventory data, we derived successional indices related to shade tolerance and stand age for N-fixing trees, non-fixing trees in the 'potentially N-fixing clade' (smallest angiosperm clade that includes all N fixers), and non-fixing trees outside this clade. We then used phylogenetically independent contrasts (PICs) to test for associations between these successional indices and N fixation. Four results stand out from our analysis of U.S. trees. First, N fixers are less shade-tolerant than non-fixers both inside and outside of the potentially N-fixing clade. Second, N fixers tend to occur in younger stands in a given geographical region than non-fixers both inside and outside of the potentially N-fixing clade. Third, the potentially N-fixing clade contains numerous late-successional non-fixers. Fourth, although the N fixation trait is evolutionarily conserved, the successional traits are relatively labile. These results suggest that selective constraints, not phylogenetic constraints, explain the rarity of late-successional N-fixing trees in temperate forests. Because N-fixing trees could overcome N limitation to net primary production if they were abundant, this study helps to understand the maintenance of N limitation in temperate forests, and therefore the capacity of this biome to sequester carbon.

Phylogenetic Constraints Do Not Explain the Rarity of Nitrogen-Fixing Trees in Late-Successional Temperate Forests

PubMed Central

Menge, Duncan N. L.; DeNoyer, Jeanne L.; Lichstein, Jeremy W.

2010-01-01

Background Symbiotic nitrogen (N)-fixing trees are rare in late-successional temperate forests, even though these forests are often N limited. Two hypotheses could explain this paradox. The ‘phylogenetic constraints hypothesis’ states that no late-successional tree taxa in temperate forests belong to clades that are predisposed to N fixation. Conversely, the ‘selective constraints hypothesis’ states that such taxa are present, but N-fixing symbioses would lower their fitness. Here we test the phylogenetic constraints hypothesis. Methodology/Principal Findings Using U.S. forest inventory data, we derived successional indices related to shade tolerance and stand age for N-fixing trees, non-fixing trees in the ‘potentially N-fixing clade’ (smallest angiosperm clade that includes all N fixers), and non-fixing trees outside this clade. We then used phylogenetically independent contrasts (PICs) to test for associations between these successional indices and N fixation. Four results stand out from our analysis of U.S. trees. First, N fixers are less shade-tolerant than non-fixers both inside and outside of the potentially N-fixing clade. Second, N fixers tend to occur in younger stands in a given geographical region than non-fixers both inside and outside of the potentially N-fixing clade. Third, the potentially N-fixing clade contains numerous late-successional non-fixers. Fourth, although the N fixation trait is evolutionarily conserved, the successional traits are relatively labile. Conclusions/Significance These results suggest that selective constraints, not phylogenetic constraints, explain the rarity of late-successional N-fixing trees in temperate forests. Because N-fixing trees could overcome N limitation to net primary production if they were abundant, this study helps to understand the maintenance of N limitation in temperate forests, and therefore the capacity of this biome to sequester carbon. PMID:20700466

Nitrogen fixation in boreal peatlands: the effects of increased N deposition on N2-fixation

NASA Astrophysics Data System (ADS)

Popma, J. M.; Wieder, R.; Lamers, L.; Vile, M. A.

2013-12-01

Boreal peatlands are of great importance to global carbon and nitrogen cycling. While covering only 3-4 % of the terrestrial surface, they account for 25-30 % of the world's soil C and 9-15 % of the world's soil N. In Western Canada atmospheric dry deposition rates are extremely low: approximately 1 kg N ha-1 yr-1. Though these systems have been functioning as net sinks over the past 11,000 years, natural and anthropogenic disturbances might compromise the historical balance of C and N. Biological N2-fixation has recently been shown to represent a very significant input of N into these systems, contributing to 62% of total N in Western Canada. Interactions between N deposition and biological N2-fixation are as yet, unknown, but the impact of elevated deposition of N-compounds from increased industrial expansion of oil sands mining to peatlands, is concerning. Given that nitrogenase, the enzyme responsible for catalyzing N2-fixation, is energetically costly when active, enhanced inputs of atmospheric N deposition could be a major determinant for enzyme activity and rates of biological N input to these bogs. Understanding interactions between N deposition and N2 fixation in boreal peatlands can aid in predicting the consequences of increased N deposition and setting critical loads. We conducted a field-fertilization experiment in a poor fen in Alberta, Canada, to determine the effects of enhanced N deposition on a dominant fen species Sphagnum angustifolium. The experiment consisted of seven N treatments: Control, 0, 5, 10, 15, 20 and 25 kg N ha-1 y1, n=3. N2-fixation was measured during summer 2012 and 2013 using the acetylene reduction assay (ARA). ARA rates were converted to rates of N2-fixation by calibrating ARA with paired 15N2-incubations. In both 2012 and 2013, with increasing N deposition from 0 kg N ha-1 yr-1 to 25 kg N ha-1 yr-1, rates of N2 fixation decreased, with highest rates in the 0 kg N ha-1 yr-1 treatment mosses (54.2 × 1.40; 48.58 × 7.12 kg N ha-1 yr-1, mean × std err for 2012 and 2013, respectively) followed by progressively lower rates with a low of 5.02 × 0.87 in 2012 and 8.94 × 3.09 in 2013 (mean × std err). As biological N2-fixation is an energetically costly process, up-regulating enzyme activity when N availability is low and down-regulating activity when N deposition is enhanced makes thermodynamic and evolutionary sense. N2-fixation shows to be one of the most early-warning indicators to the early response of boreal peatlands to increased N deposition, and can aid in setting critical loads to protect these historically pristine ecosystems.

Proteomic Phenotyping of Novosphingobium nitrogenifigens Reveals a Robust Capacity for Simultaneous Nitrogen Fixation, Polyhydroxyalkanoate Production, and Resistance to Reactive Oxygen Species

PubMed Central

Strabala, Timothy J.; Peng, Lifeng; Rawson, Pisana; Lloyd-Jones, Gareth; Jordan, T. William

2012-01-01

Novosphingobium nitrogenifigens Y88T (Y88) is a free-living, diazotrophic Alphaproteobacterium, capable of producing 80% of its biomass as the biopolymer polyhydroxybutyrate (PHB). We explored the potential utility of this species as a polyhydroxybutyrate production strain, correlating the effects of glucose, nitrogen availability, dissolved oxygen concentration, and extracellular pH with polyhydroxybutyrate production and changes in the Y88 proteomic profile. Using two-dimensional differential in-gel electrophoresis and tandem mass spectrometry, we identified 217 unique proteins from six growth conditions. We observed reproducible, characteristic proteomic signatures for each of the physiological states we examined. We identified proteins that changed in abundance in correlation with either nitrogen fixation, dissolved oxygen concentration, or acidification of the growth medium. The proteins that correlated with nitrogen fixation were identified either as known nitrogen fixation proteins or as novel proteins that we predict play roles in aspects of nitrogen fixation based on their proteomic profiles. In contrast, the proteins involved in central carbon and polyhydroxybutyrate metabolism were constitutively abundant, consistent with the constitutive polyhydroxybutyrate production that we observed in this species. Three proteins with roles in detoxification of reactive oxygen species were identified in this obligate aerobe. The most abundant protein in all experiments was a polyhydroxyalkanoate granule-associated protein, phasin. The full-length isoform of this protein has a long, intrinsically disordered Ala/Pro/Lys-rich N-terminal segment, a feature that appears to be unique to sphingomonad phasins. The data suggest that Y88 has potential as a PHB production strain due to its aerobic tolerance and metabolic orientation toward polyhydroxybutyrate accumulation, even in low-nitrogen growth medium. PMID:22582058

Increasing subtropical North Pacific Ocean nitrogen fixation since the Little Ice Age.

PubMed

Sherwood, Owen A; Guilderson, Thomas P; Batista, Fabian C; Schiff, John T; McCarthy, Matthew D

2014-01-02

The North Pacific subtropical gyre (NPSG) plays a major part in the export of carbon and other nutrients to the deep ocean. Primary production in the NPSG has increased in recent decades despite a reduction in nutrient supply to surface waters. It is thought that this apparent paradox can be explained by a shift in plankton community structure from mostly eukaryotes to mostly nitrogen-fixing prokaryotes. It remains uncertain, however, whether the plankton community domain shift can be linked to cyclical climate variability or a long-term global warming trend. Here we analyse records of bulk and amino-acid-specific (15)N/(14)N isotopic ratios (δ(15)N) preserved in the skeletons of long-lived deep-sea proteinaceous corals collected from the Hawaiian archipelago; these isotopic records serve as a proxy for the source of nitrogen-supported export production through time. We find that the recent increase in nitrogen fixation is the continuation of a much larger, centennial-scale trend. After a millennium of relatively minor fluctuation, δ(15)N decreases between 1850 and the present. The total shift in δ(15)N of -2 per mil over this period is comparable to the total change in global mean sedimentary δ(15)N across the Pleistocene-Holocene transition, but it is happening an order of magnitude faster. We use a steady-state model and find that the isotopic mass balance between nitrate and nitrogen fixation implies a 17 to 27 per cent increase in nitrogen fixation over this time period. A comparison with independent records suggests that the increase in nitrogen fixation might be linked to Northern Hemisphere climate change since the end of the Little Ice Age.

Estimation of nitrogen fixation in Leucaena leucocephala using 15N-enrichment methodologies

Treesearch

John A. Parrotta; Dwight D. Baker; Maurice Fried

1994-01-01

An estimation of biological nitrogen fixation by Leucaena leucocephala (Lam.) de Wit in monoculture and mixed-species plantations (with Casuarina equisetifolia L. ex J.R. & G. Forst., and Eucalyptus robusta Sm.) was undertaken over a two-year period in Puerto Rico using the 15N-enrichment...

Genome-wide association analysis of symbiotic nitrogen fixation in common bean

USDA-ARS?s Scientific Manuscript database

A genome-wide association study (GWAS) was conducted to explore the genetic basis of variation for symbiotic nitrogen fixation (SNF) and related traits in the Andean diversity panel (ADP) comprised of 259 common bean (Phaseolus vulgaris) genotypes. The ADP was evaluated for SNF and related traits in...

The role of nitrogen fixation in neotropical dry forests: insights from ecosystem modeling and field data

NASA Astrophysics Data System (ADS)

Trierweiler, A.; Xu, X.; Gei, M. G.; Powers, J. S.; Medvigy, D.

2016-12-01

Tropical dry forests (TDFs) have immense functional diversity and face multiple resource constraints (both water and nutrients). Legumes are abundant and exhibit a wide diversity of N2-fixing strategies in TDFs. The abundance and diversity of legumes and their interaction with N2-fixing bacteria may strongly control the coupled carbon-nitrogen cycle in the biome and influence whether TDFs will be particularly vulnerable or uniquely adapted to projected global change. However, the importance of N2-fixation in TDFs and the carbon cost of acquiring N through symbiotic relationships are not fully understood. Here, we use models along with field measurements to examine the role of legumes, nitrogen fixation, and plant-symbiont nutrient exchanges in TDFs. We use a new version of the Ecosystem Demography (ED2) model that has been recently parameterized for TDFs. The new version incorporates plant-mycorrhizae interactions and multiple resource constraints (carbon, nitrogen, phosphorus, and water). We represent legumes and other functional groups found in TDFs with a range of resource acquisition strategies. In the model, plants then can dynamically adjust their carbon allocation and nutrient acquisition strategies (e.g. N2-fixing bacteria and mycorrhizal fungi) according to the nutrient limitation status. We test (i) the model's performance against a nutrient gradient of field sites in Costa Rica and (ii) the model's sensitivity to the carbon cost to acquire N through fixation and mycorrhizal relationships. We also report on simulated tree community responses to ongoing field nutrient fertilization experiments. We found that the inclusion of the N2-fixation legume plant functional traits were critical to reproducing community dynamics of Costa Rican field TDF sites and have a large impact on forest biomass. Simulated ecosystem fixation rates matched the magnitude and temporal patterns of field measured fixation. Our results show that symbiotic nitrogen fixation plays an important role in tropical dry forests and biomass accumulation. Also, we suggest that fixation's tight link to the rainy season could result in potential nutrient cycling vulnerabilities with projected rainfall changes.

Two MicroRNAs Linked to Nodule Infection and Nitrogen-Fixing Ability in the Legume Lotus japonicus1[W

PubMed Central

De Luis, Ana; Markmann, Katharina; Cognat, Valérie; Holt, Dennis B.; Charpentier, Myriam; Parniske, Martin; Stougaard, Jens; Voinnet, Olivier

2012-01-01

Legumes overcome nitrogen shortage by developing root nodules in which symbiotic bacteria fix atmospheric nitrogen in exchange for host-derived carbohydrates and mineral nutrients. Nodule development involves the distinct processes of nodule organogenesis, bacterial infection, and the onset of nitrogen fixation. These entail profound, dynamic gene expression changes, notably contributed to by microRNAs (miRNAs). Here, we used deep-sequencing, candidate-based expression studies and a selection of Lotus japonicus mutants uncoupling different symbiosis stages to identify miRNAs involved in symbiotic nitrogen fixation. Induction of a noncanonical miR171 isoform, which targets the key nodulation transcription factor Nodulation Signaling Pathway2, correlates with bacterial infection in nodules. A second candidate, miR397, is systemically induced in the presence of active, nitrogen-fixing nodules but not in that of noninfected or inactive nodule organs. It is involved in nitrogen fixation-related copper homeostasis and targets a member of the laccase copper protein family. These findings thus identify two miRNAs specifically responding to symbiotic infection and nodule function in legumes. PMID:23071252

Biological nitrogen fixation in non-legume plants.

PubMed

Santi, Carole; Bogusz, Didier; Franche, Claudine

2013-05-01

Nitrogen is an essential nutrient in plant growth. The ability of a plant to supply all or part of its requirements from biological nitrogen fixation (BNF) thanks to interactions with endosymbiotic, associative and endophytic symbionts, confers a great competitive advantage over non-nitrogen-fixing plants. Because BNF in legumes is well documented, this review focuses on BNF in non-legume plants. Despite the phylogenic and ecological diversity among diazotrophic bacteria and their hosts, tightly regulated communication is always necessary between the microorganisms and the host plant to achieve a successful interaction. Ongoing research efforts to improve knowledge of the molecular mechanisms underlying these original relationships and some common strategies leading to a successful relationship between the nitrogen-fixing microorganisms and their hosts are presented. Understanding the molecular mechanism of BNF outside the legume-rhizobium symbiosis could have important agronomic implications and enable the use of N-fertilizers to be reduced or even avoided. Indeed, in the short term, improved understanding could lead to more sustainable exploitation of the biodiversity of nitrogen-fixing organisms and, in the longer term, to the transfer of endosymbiotic nitrogen-fixation capacities to major non-legume crops.

Distribution and rates of nitrogen fixation in the western tropical South Pacific Ocean constrained by nitrogen isotope budgets

NASA Astrophysics Data System (ADS)

Knapp, Angela N.; McCabe, Kelly M.; Grosso, Olivier; Leblond, Nathalie; Moutin, Thierry; Bonnet, Sophie

2018-05-01

Constraining the rates and spatial distribution of dinitrogen (N2) fixation fluxes to the ocean informs our understanding of the environmental sensitivities of N2 fixation as well as the timescale over which the fluxes of nitrogen (N) to and from the ocean may respond to each other. Here we quantify rates of N2 fixation as well as its contribution to export production along a zonal transect in the western tropical South Pacific (WTSP) Ocean using N isotope (δ15N) budgets. Comparing measurements of water column nitrate + nitrite δ15N with the δ15N of sinking particulate N at a western, central, and eastern station, these δ15N budgets indicate high, modest, and low rates of N2 fixation at the respective stations. The results also imply that N2 fixation supports exceptionally high, i.e. ≥ 50 %, of export production at the western and central stations, which are also proximal to the largest iron sources. These geochemically based rates of N2 fixation are equal to or greater than those previously reported in the tropical North Atlantic, indicating that the WTSP Ocean has the capacity to support globally significant rates of N2 fixation, which may compensate for N removal in the oxygen-deficient zones of the eastern tropical Pacific.

Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions.

PubMed

Setten, Lorena; Soto, Gabriela; Mozzicafreddo, Matteo; Fox, Ana Romina; Lisi, Christian; Cuccioloni, Massimiliano; Angeletti, Mauro; Pagano, Elba; Díaz-Paleo, Antonio; Ayub, Nicolás Daniel

2013-01-01

Nitrogen is the second most critical factor for crop production after water. In this study, the beneficial rhizobacterium Pseudomonas protegens Pf-5 was genetically modified to fix nitrogen using the genes encoding the nitrogenase of Pseudomonas stutzeri A1501 via the X940 cosmid. Pf-5 X940 was able to grow in L medium without nitrogen, displayed high nitrogenase activity and released significant quantities of ammonium to the medium. Pf-5 X940 also showed constitutive expression and enzymatic activity of nitrogenase in ammonium medium or in nitrogen-free medium, suggesting a constitutive nitrogen fixation. Similar to Pseudomonas protegens Pf-5, Pseudomonas putida, Pseudomonas veronii and Pseudomonas taetrolens but not Pseudomonas balearica and Pseudomonas stutzeri transformed with cosmid X940 showed constitutive nitrogenase activity and high ammonium production, suggesting that this phenotype depends on the genome context and that this technology to obtain nitrogen-fixing bacteria is not restricted to Pf-5. Interestingly, inoculation of Arabidopsis, alfalfa, tall fescue and maize with Pf-5 X940 increased the ammonium concentration in soil and plant productivity under nitrogen-deficient conditions. In conclusion, these results open the way to the production of effective recombinant inoculants for nitrogen fixation on a wide range of crops.

Engineering Pseudomonas protegens Pf-5 for Nitrogen Fixation and its Application to Improve Plant Growth under Nitrogen-Deficient Conditions

PubMed Central

Setten, Lorena; Soto, Gabriela; Mozzicafreddo, Matteo; Fox, Ana Romina; Lisi, Christian; Cuccioloni, Massimiliano; Angeletti, Mauro; Pagano, Elba; Díaz-Paleo, Antonio; Ayub, Nicolás Daniel

2013-01-01

Nitrogen is the second most critical factor for crop production after water. In this study, the beneficial rhizobacterium Pseudomonas protegens Pf-5 was genetically modified to fix nitrogen using the genes encoding the nitrogenase of Pseudomonas stutzeri A1501 via the X940 cosmid. Pf-5 X940 was able to grow in L medium without nitrogen, displayed high nitrogenase activity and released significant quantities of ammonium to the medium. Pf-5 X940 also showed constitutive expression and enzymatic activity of nitrogenase in ammonium medium or in nitrogen-free medium, suggesting a constitutive nitrogen fixation. Similar to Pseudomonas protegens Pf-5, Pseudomonas putida, Pseudomonas veronii and Pseudomonas taetrolens but not Pseudomonas balearica and Pseudomonas stutzeri transformed with cosmid X940 showed constitutive nitrogenase activity and high ammonium production, suggesting that this phenotype depends on the genome context and that this technology to obtain nitrogen-fixing bacteria is not restricted to Pf-5. Interestingly, inoculation of Arabidopsis, alfalfa, tall fescue and maize with Pf-5 X940 increased the ammonium concentration in soil and plant productivity under nitrogen-deficient conditions. In conclusion, these results open the way to the production of effective recombinant inoculants for nitrogen fixation on a wide range of crops. PMID:23675499

Nitrogen fixation and nifH diversity in human gut microbiota

PubMed Central

Igai, Katsura; Itakura, Manabu; Nishijima, Suguru; Tsurumaru, Hirohito; Suda, Wataru; Tsutaya, Takumi; Tomitsuka, Eriko; Tadokoro, Kiyoshi; Baba, Jun; Odani, Shingo; Natsuhara, Kazumi; Morita, Ayako; Yoneda, Minoru; Greenhill, Andrew R.; Horwood, Paul F.; Inoue, Jun-ichi; Ohkuma, Moriya; Hongoh, Yuichi; Yamamoto, Taro; Siba, Peter M.; Hattori, Masahira; Minamisawa, Kiwamu; Umezaki, Masahiro

2016-01-01

It has been hypothesized that nitrogen fixation occurs in the human gut. However, whether the gut microbiota truly has this potential remains unclear. We investigated the nitrogen-fixing activity and diversity of the nitrogenase reductase (NifH) genes in the faecal microbiota of humans, focusing on Papua New Guinean and Japanese individuals with low to high habitual nitrogen intake. A 15N2 incorporation assay showed significant enrichment of 15N in all faecal samples, irrespective of the host nitrogen intake, which was also supported by an acetylene reduction assay. The fixed nitrogen corresponded to 0.01% of the standard nitrogen requirement for humans, although our data implied that the contribution in the gut in vivo might be higher than this value. The nifH genes recovered in cloning and metagenomic analyses were classified in two clusters: one comprising sequences almost identical to Klebsiella sequences and the other related to sequences of Clostridiales members. These results are consistent with an analysis of databases of faecal metagenomes from other human populations. Collectively, the human gut microbiota has a potential for nitrogen fixation, which may be attributable to Klebsiella and Clostridiales strains, although no evidence was found that the nitrogen-fixing activity substantially contributes to the host nitrogen balance. PMID:27554344

Influence of pulsed-light irradiation on the productivity and nitrogen-fixing ability of blue-green algae nostoc muscorum Ag

DOE Office of Scientific and Technical Information (OSTI.GOV)

Umarov, G.Ya.; Kuchkarova, M.A.; Maksudov, T.U.

1975-01-01

The utilization of pulsed concentrated sunlight to improve the productivity of nostoc muscorum Ag. algae was investigated. In laboratory experiments the greatest accumulation of biomass was found after 5-min irradiation; there was a 10 percent increase in nitrogen fixation. For cultivation under the open sky productivity and nitrogen fixation rose after 10- and 20-min irradiation by pulsed concentrated sunlight.

Microgravity Effects on the Early Events of Biological Nitrogen Fixation in Medicago Truncatula: Results from the SyNRGE Experiment

NASA Technical Reports Server (NTRS)

Stutte, Gary W.; Roberts, Michael

2012-01-01

SyNRGE (Symbiotic Nodulation in a Reduced Gravity Environment) was a sortie mission on STS-135 in the Biological Research in Canisters (BRIC) hardware to study the effect of microgravity on a plant-microbe symbiosis resulting in biological nitrogen fixation. Medicago truncatula, a model species for th legume family, was inoculated with its bacterial symbiont, Sinorhizobium meliloti, to observe early biomolecular events associated with infection and nodulation in Petri Dish Fixation Units (PDFU's).

Quantifying nitrogen-fixation in feather moss carpets of boreal forests.

PubMed

DeLuca, Thomas H; Zackrisson, Olle; Nilsson, Marie-Charlotte; Sellstedt, Anita

2002-10-31

Biological nitrogen (N) fixation is the primary source of N within natural ecosystems, yet the origin of boreal forest N has remained elusive. The boreal forests of Eurasia and North America lack any significant, widespread symbiotic N-fixing plants. With the exception of scattered stands of alder in early primary successional forests, N-fixation in boreal forests is considered to be extremely limited. Nitrogen-fixation in northern European boreal forests has been estimated at only 0.5 kg N ha(-1) yr(-1); however, organic N is accumulated in these ecosystems at a rate of 3 kg N ha(-1) yr(-1) (ref. 8). Our limited understanding of the origin of boreal N is unacceptable given the extent of the boreal forest region, but predictable given our imperfect knowledge of N-fixation. Herein we report on a N-fixing symbiosis between a cyanobacterium (Nostoc sp.) and the ubiquitous feather moss, Pleurozium schreberi (Bird) Mitt. that alone fixes between 1.5 and 2.0 kg N ha(-1) yr(-1) in mid- to late-successional forests of northern Scandinavia and Finland. Previous efforts have probably underestimated N-fixation potential in boreal forests.

Coordinated regulation of growth, activity and transcription in natural populations of the unicellular nitrogen-fixing cyanobacterium Crocosphaera.

PubMed

Wilson, Samuel T; Aylward, Frank O; Ribalet, Francois; Barone, Benedetto; Casey, John R; Connell, Paige E; Eppley, John M; Ferrón, Sara; Fitzsimmons, Jessica N; Hayes, Christopher T; Romano, Anna E; Turk-Kubo, Kendra A; Vislova, Alice; Armbrust, E Virginia; Caron, David A; Church, Matthew J; Zehr, Jonathan P; Karl, David M; DeLong, Edward F

2017-07-31

The temporal dynamics of phytoplankton growth and activity have large impacts on fluxes of matter and energy, yet obtaining in situ metabolic measurements of sufficient resolution for even dominant microorganisms remains a considerable challenge. We performed Lagrangian diel sampling with synoptic measurements of population abundances, dinitrogen (N 2 ) fixation, mortality, productivity, export and transcription in a bloom of Crocosphaera over eight days in the North Pacific Subtropical Gyre (NPSG). Quantitative transcriptomic analyses revealed clear diel oscillations in transcript abundances for 34% of Crocosphaera genes identified, reflecting a systematic progression of gene expression in diverse metabolic pathways. Significant time-lagged correspondence was evident between nifH transcript abundance and maximal N 2 fixation, as well as sepF transcript abundance and cell division, demonstrating the utility of transcriptomics to predict the occurrence and timing of physiological and biogeochemical processes in natural populations. Indirect estimates of carbon fixation by Crocosphaera were equivalent to 11% of net community production, suggesting that under bloom conditions this diazotroph has a considerable impact on the wider carbon cycle. Our cross-scale synthesis of molecular, population and community-wide data underscores the tightly coordinated in situ metabolism of the keystone N 2 -fixing cyanobacterium Crocosphaera, as well as the broader ecosystem-wide implications of its activities.

CO2 Fixation, Lipid Production, and Power Generation by a Novel Air-Lift-Type Microbial Carbon Capture Cell System.

PubMed

Hu, Xia; Liu, Baojun; Zhou, Jiti; Jin, Ruofei; Qiao, Sen; Liu, Guangfei

2015-09-01

An air-lift-type microbial carbon capture cell (ALMCC) was constructed for the first time by using an air-lift-type photobioreactor as the cathode chamber. The performance of ALMCC in fixing high concentration of CO2, producing energy (power and biodiesel), and removing COD together with nutrients was investigated and compared with the traditional microbial carbon capture cell (MCC) and air-lift-type photobioreactor (ALP). The ALMCC system produced a maximum power density of 972.5 mW·m(-3) and removed 86.69% of COD, 70.52% of ammonium nitrogen, and 69.24% of phosphorus, which indicate that ALMCC performed better than MCC in terms of power generation and wastewater treatment efficiency. Besides, ALMCC demonstrated 9.98- and 1.88-fold increases over ALP and MCC in the CO2 fixation rate, respectively. Similarly, the ALMCC significantly presented a higher lipid productivity compared to those control reactors. More importantly, the preliminary analysis of energy balance suggested that the net energy of the ALMCC system was significantly superior to other systems and could theoretically produce enough energy to cover its consumption. In this work, the established ALMCC system simultaneously achieved the high level of CO2 fixation, energy recycle, and municipal wastewater treatment effectively and efficiently.

The relationship between dissolved hydrogen and nitrogen fixation in ocean waters

NASA Astrophysics Data System (ADS)

Moore, Robert M.; Punshon, Stephen; Mahaffey, Claire; Karl, David

2009-09-01

Fixed nitrogen is a key nutrient involved in regulating global marine productivity and hence the global oceanic carbon cycle. Oceanic nitrogen (N 2) fixation is estimated to supply 8×10 12 moles N y -1 to the ocean, approximately equal to current riverine and the atmospheric inputs of fixed N, and between 50 and 100% of current estimates of oceanic denitrification. However, the spatial and temporal variability of N 2 fixation remains uncertain, mostly because of the normal low resolution sampling for diazotroph distribution and fixation rates. It is well established that N 2 fixation, mediated by the enzyme nitrogenase, is a source of hydrogen (H 2), but the extent to which it leads to supersaturation of H 2 in oceanic waters is unresolved. Here, we present simultaneous measurements of upper ocean dissolved H 2 concentration (nmol L -1), and rates of N 2 fixation (μmol N m -3 d -1), determined using 15N 2 tracer techniques (at 7 or 15 m), on a transect from Fiji to Hawaii. We find a significant correlation ( r=0.98) between dissolved H 2 and rates of N 2 fixation, with the greatest supersaturation of H 2 and highest rates of N 2 fixation being observed in the subtropical gyres at the southern (˜18°S) and northern (18°N) reaches of the transect. The lowest H 2 saturation and N 2 fixation were observed in the equatorial region between 8°S and 14°N. We propose that an empirical relationship between H 2 supersaturations and N 2 fixation measurements could be used to guide sampling for 15N fixation measurements or to aid the spatial interpolation of such measurements.

Genome-wide association analysis of diverse soybean genotypes reveals novel markers for nitrogen traits

USDA-ARS?s Scientific Manuscript database

Nitrogen is a primary plant nutrient that plays a major role in achieving maximum economic yield. Insufficient availability most often limits soybean crop growth. Symbiotic N2 fixation in soybean is highly sensitive to limited water availability, and breeding for reduced N2 fixation sensitivity to ...

CADMIUM EFFECTS ON THE NITROGEN FIXATION SYSTEM OF RED ALDER

EPA Science Inventory

Red alder (Alnus rubra) was grown in sand culture in the greenhouse to obtain data on the effects of cadmium (Cd) on a symbiotic nitrogen (N) fixation system which contributes to the fertility of forest soils. Treatment of red alder seedlings for 11 weeks with 0.545 to 136 microm...

Biological N2 fixation mainly controlled by Sphagnum tissue N:P ratio in ombrotrophic bogs

NASA Astrophysics Data System (ADS)

Zivkovic, Tatjana; Moore, Tim R.

2017-04-01

Most of the 18 Pg nitrogen (N) accumulated in northern nutrient-poor and Sphagnum-dominated peatlands (bogs and fens) can be attributed to N2-fixation by diazotrophs either associated with the live Sphagnum or non-symbiotically in the deeper peat such as through methane consumption close to the water table. Where atmospheric N deposition is low (< 0.2 g m-2 y-1), ombrotrophic bogs rely on N2-fixation as the primary source of N that sustains primary production. Due to high energetic requirements, N2-fixation depends on the available phosphorus (P). Anthropogenic impacts in the last 200 years increased atmospheric N deposition, resulting in a switch from N to P limitation in Sphagnum, suggested by the increase in tissue N:P to >16. It is unclear how Sphagnum-hosted diazotrophic activity may be affected by N deposition and thus changes in N:P ratio. First, we investigated the effects of long-term addition of different sources of nitrogen (0, 1.6, 3.2 and 6.4 g N m-2 y-1as NH4Cl and NaNO3), and phosphorus (5 g P m-2 y-1as KH2PO4) on Sphagnum nutrient status (N, P and N:P ratio), net primary productivity (NPP) and Sphagnum-associated N2fixation at Mer Bleue, a temperate ombrotrophic bog. We show that N concentration in Sphagnum tissue increased with larger rates of N addition, with a stronger effect on Sphagnum from NH4 than NO3. The addition of P created a 3.5 fold increase in Sphagnum P content compared to controls. Sphagnum NPP decreased linearly with the rise in N:P ratio, while linear growth declined exponentially with increase in Sphagnum N content. Rates of N2-fixation determined in the laboratory significantly decreased in response to even the smallest addition of both N species. In contrast, the addition of P increased N2 fixation by up to 100 times compared to N treatments and up to 5-30 times compared to controls. The change in N2-fixation was best modeled by the N:P ratio, across all experimental treatments. Secondly, to test the role of N:P ratio on N2-fixation across a range of bogs, eight study sites along the latitudinal gradient from temperate, boreal to subarctic zone in eastern Canada were selected. From each bog, two predominant microptopographies, hummocks and hollows, were tested for both N2-fixation activity in the laboratory and Sphagnum tissue concentrations of N, P and N:P ratio. We found that 65% of the variance in the increase of N2-fixation activity was explained by the decrease in N:P ratio in hollows (n = 73) but only about 20% in hummocks (n = 78). Changes in neither N or P concentration alone explained the increase in N2-fixation better than N:P ratio. We interpret that the difference between hollows and hummocks results from the availability of moisture that further limits N2-fixation. When moisture is not a limiting factor, i.e. in hollows, N:P ratio will be the best predictor of N2-fixation in bogs. Both studies suggest that the relative P availability to Sphagnum-associated diazotrophs, measured as a tissue N:P ratio, best describes N2-fixation activity in bogs, especially ones exposed to larger N deposition.

A Minimal Nitrogen Fixation Gene Cluster from Paenibacillus sp. WLY78 Enables Expression of Active Nitrogenase in Escherichia coli

PubMed Central

Zhao, Dehua; Liu, Xiaomeng; Zhang, Bo; Xie, Jianbo; Hong, Yuanyuan; Li, Pengfei; Chen, Sanfeng; Dixon, Ray; Li, Jilun

2013-01-01

Most biological nitrogen fixation is catalyzed by molybdenum-dependent nitrogenase, an enzyme complex comprising two component proteins that contains three different metalloclusters. Diazotrophs contain a common core of nitrogen fixation nif genes that encode the structural subunits of the enzyme and components required to synthesize the metalloclusters. However, the complement of nif genes required to enable diazotrophic growth varies significantly amongst nitrogen fixing bacteria and archaea. In this study, we identified a minimal nif gene cluster consisting of nine nif genes in the genome of Paenibacillus sp. WLY78, a gram-positive, facultative anaerobe isolated from the rhizosphere of bamboo. We demonstrate that the nif genes in this organism are organized as an operon comprising nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV and that the nif cluster is under the control of a σ70 (σA)-dependent promoter located upstream of nifB. To investigate genetic requirements for diazotrophy, we transferred the Paenibacillus nif cluster to Escherichia coli. The minimal nif gene cluster enables synthesis of catalytically active nitrogenase in this host, when expressed either from the native nifB promoter or from the T7 promoter. Deletion analysis indicates that in addition to the core nif genes, hesA plays an important role in nitrogen fixation and is responsive to the availability of molybdenum. Whereas nif transcription in Paenibacillus is regulated in response to nitrogen availability and by the external oxygen concentration, transcription from the nifB promoter is constitutive in E. coli, indicating that negative regulation of nif transcription is bypassed in the heterologous host. This study demonstrates the potential for engineering nitrogen fixation in a non-nitrogen fixing organism with a minimum set of nine nif genes. PMID:24146630

Drivers of the dynamics of diazotrophs and denitrifiers in North Sea bottom waters and sediments

PubMed Central

Fan, Haoxin; Bolhuis, Henk; Stal, Lucas J.

2015-01-01

The fixation of dinitrogen (N2) and denitrification are two opposite processes in the nitrogen cycle. The former transfers atmospheric dinitrogen gas into bound nitrogen in the biosphere, while the latter returns this bound nitrogen back to atmospheric dinitrogen. It is unclear whether or not these processes are intimately connected in any microbial ecosystem or that they are spatially and/or temporally separated. Here, we measured seafloor nitrogen fixation and denitrification as well as pelagic nitrogen fixation by using the stable isotope technique. Alongside, we measured the diversity, abundance, and activity of nitrogen-fixing and denitrifying microorganisms at three stations in the southern North Sea. Nitrogen fixation ranged from undetectable to 2.4 nmol N L−1 d−1 and from undetectable to 8.2 nmol N g−1 d−1 in the water column and seafloor, respectively. The highest rates were measured in August at Doggersbank, both for the water column and for the seafloor. Denitrification ranged from 1.7 to 208.8 μmol m−2 d−1 and the highest rates were measured in May at the Oyster Grounds. DNA sequence analysis showed sequences of nifH, a structural gene for nitrogenase, related to sequences from anaerobic sulfur/iron reducers and sulfate reducers. Sequences of the structural gene for nitrite reductase, nirS, were related to environmental clones from marine sediments. Quantitative polymerase chain reaction (qPCR) data revealed the highest abundance of nifH and nirS genes at the Oyster Grounds. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) data revealed the highest nifH expression at Doggersbank and the highest nirS expression at the Oyster Grounds. The distribution of the diazotrophic and denitrifying communities seems to be subject to different selecting factors, leading to spatial and temporal separation of nitrogen fixation and denitrification. These selecting factors include temperature, organic matter availability, and oxygen concentration. PMID:26257718

Composition and diversity of nifH genes of nitrogen-fixing cyanobacteria associated with boreal forest feather mosses.

PubMed

Ininbergs, Karolina; Bay, Guillaume; Rasmussen, Ulla; Wardle, David A; Nilsson, Marie-Charlotte

2011-10-01

Recent studies have revealed that nitrogen fixation by cyanobacteria living in association with feather mosses is a major input of nitrogen to boreal forests. We characterized the community composition and diversity of cyanobacterial nifH phylotypes associated with each of two feather moss species (Pleurozium schreberi and Hylocomium splendens) on each of 30 lake islands varying in ecosystem properties in northern Sweden. Nitrogen fixation was measured using acetylene reduction, and nifH sequences were amplified using general and cyanobacterial selective primers, separated and analyzed using density gradient gel electrophoresis (DGGE) or cloning, and further sequenced for phylogenetic analyses. Analyses of DGGE fingerprinting patterns revealed two host-specific clusters (one for each moss species), and sequence analysis showed five clusters of nifH phylotypes originating from heterocystous cyanobacteria. For H. splendens only, N(2) fixation was related to both nifH composition and diversity among islands. We demonstrated that the cyanobacterial communities associated with feather mosses show a high degree of host specificity. However, phylotype composition and diversity, and nitrogen fixation, did not differ among groups of islands that varied greatly in their availability of resources. These results suggest that moss species identity, but not extrinsic environmental conditions, serves as the primary determinant of nitrogen-fixing cyanobacterial communities that inhabit mosses. © 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.

The physiological cost of diazotrophy for Trichodesmium erythraeum IMS101

PubMed Central

Davey, Phillip A.; Lawson, Tracy; Geider, Richard J.

2018-01-01

Trichodesmium plays a significant role in the oligotrophic oceans, fixing nitrogen in an area corresponding to half of the Earth’s surface, representing up to 50% of new production in some oligotrophic tropical and subtropical oceans. Whilst Trichodesmium blooms at the surface exhibit a strong dependence on diazotrophy, colonies at depth or at the surface after a mixing event could be utilising additional N-sources. We conducted experiments to establish how acclimation to varying N-sources affects the growth, elemental composition, light absorption coefficient, N2 fixation, PSII electron transport rate and the relationship between net and gross photosynthetic O2 exchange in T. erythraeum IMS101. To do this, cultures were acclimated to growth medium containing NH4+ and NO3- (replete concentrations) or N2 only (diazotrophic control). The light dependencies of O2 evolution and O2 uptake were measured using membrane inlet mass spectrometry (MIMS), while PSII electron transport rates were measured from fluorescence light curves (FLCs). We found that at a saturating light intensity, Trichodesmium growth was ~ 10% and 13% lower when grown on N2 than with NH4+ and NO3-, respectively. Oxygen uptake increased linearly with net photosynthesis across all light intensities ranging from darkness to 1100 μmol photons m-2 s-1. The maximum rates and initial slopes of light response curves for C-specific gross and net photosynthesis and the slope of the relationship between gross and net photosynthesis increased significantly under non-diazotrophic conditions. We attribute these observations to a reduced expenditure of reductant and ATP for nitrogenase activity under non-diazotrophic conditions which allows NADPH and ATP to be re-directed to CO2 fixation and/or biosynthesis. The energy and reductant conserved through utilising additional N-sources could enhance Trichodesmium’s productivity and growth and have major implications for its role in ocean C and N cycles. PMID:29641568

Solar Water Splitting and Nitrogen Fixation with Layered Bismuth Oxyhalides.

PubMed

Li, Jie; Li, Hao; Zhan, Guangming; Zhang, Lizhi

2017-01-17

Hydrogen and ammonia are the chemical molecules that are vital to Earth's energy, environmental, and biological processes. Hydrogen with renewable, carbon-free, and high combustion-enthalpy hallmarks lays the foundation of next-generation energy source, while ammonia furnishes the building blocks of fertilizers and proteins to sustain the lives of plants and organisms. Such merits fascinate worldwide scientists in developing viable strategies to produce hydrogen and ammonia. Currently, at the forefronts of hydrogen and ammonia syntheses are solar water splitting and nitrogen fixation, because they go beyond the high temperature and pressure requirements of methane stream reforming and Haber-Bosch reaction, respectively, as the commercialized hydrogen and ammonia production routes, and inherit the natural photosynthesis virtues that are green and sustainable and operate at room temperature and atmospheric pressure. The key to propelling such photochemical reactions lies in searching photocatalysts that enable water splitting into hydrogen and nitrogen fixation to make ammonia efficiently. Although the past 40 years have witnessed significant breakthroughs using the most widely studied TiO 2 , SrTiO 3 , (Ga 1-x Zn x )(N 1-x O x ), CdS, and g-C 3 N 4 for solar chemical synthesis, two crucial yet still unsolved issues challenge their further progress toward robust solar water splitting and nitrogen fixation, including the inefficient steering of electron transportation from the bulk to the surface and the difficulty of activating the N≡N triple bond of N 2 . This Account details our endeavors that leverage layered bismuth oxyhalides as photocatalysts for efficient solar water splitting and nitrogen fixation, with a focus on addressing the above two problems. We first demonstrate that the layered structures of bismuth oxyhalides can stimulate an internal electric field (IEF) that is capable of efficiently separating electrons and holes after their formation and of precisely channeling their migration from the bulk to the surface along the different directions, thus enabling more electrons to reach the surface for water splitting and nitrogen fixation. Simultaneously, their oxygen termination feature and the strain differences between interlayers and intralayers render the easy generation of surface oxygen vacancies (OVs) that afford Lewis-base and unsaturated-unsaturated sites for nitrogen activation. With these rationales as the guideline, we can obtain striking visible-light hydrogen- and ammonia-evolving rates without using any noble-metal cocatalysts. Then we show how to utilize IEF and OV based strategies to improve the solar water splitting and nitrogen fixation performances of bismuth oxyhalide photocatalysts. Finally, we highlight the challenges remaining in using bismuth oxyhalides for solar hydrogen and ammonia syntheses, and the prospect of further development of this research field. We believe that our mechanistic insights could serve as a blueprint for the design of more efficient solar water splitting and nitrogen fixation systems, and layered bismuth oxyhalides might open up new photocatalyst paradigm for such two solar chemical syntheses.

Incorporating a prognostic representation of marine nitrogen fixers into the global ocean biogeochemical model HAMOCC

NASA Astrophysics Data System (ADS)

Paulsen, Hanna; Ilyina, Tatiana; Six, Katharina D.; Stemmler, Irene

2017-03-01

Nitrogen (N2) fixation is a major source of bioavailable nitrogen to the euphotic zone, thereby exerting an important control on ocean biogeochemical cycling. This paper presents the incorporation of prognostic N2 fixers into the HAMburg Ocean Carbon Cycle model (HAMOCC), a component of the Max Planck Institute Earth System Model (MPI-ESM). Growth dynamics of N2 fixers in the model are based on physiological characteristics of the cyanobacterium Trichodesmium. The applied temperature dependency confines diazotrophic growth and N2 fixation to the tropical and subtropical ocean roughly between 40°S and 40°N. Simulated large-scale spatial patterns compare well with observations, and the global N2 fixation rate of 135.6 Tg N yr-1 is within the range of current estimates. The vertical distribution of N2 fixation also matches well the observations, with a major fraction of about 85% occurring in the upper 20 m. The observed seasonal variability at the stations BATS and ALOHA is reasonably reproduced, with highest fixation rates in northern summer/fall. Iron limitation was found to be an important factor in controlling the simulated distribution of N2 fixation, especially in the Pacific Ocean. The new model component considerably improves the representation of present-day N2 fixation in HAMOCC. It provides the basis for further studies on the role of diazotrophs in global biogeochemical cycles, as well as on the response of N2 fixation to changing environmental conditions.

Biological nitrogen fixation in non-legume plants

PubMed Central

Santi, Carole; Bogusz, Didier; Franche, Claudine

2013-01-01

Background Nitrogen is an essential nutrient in plant growth. The ability of a plant to supply all or part of its requirements from biological nitrogen fixation (BNF) thanks to interactions with endosymbiotic, associative and endophytic symbionts, confers a great competitive advantage over non-nitrogen-fixing plants. Scope Because BNF in legumes is well documented, this review focuses on BNF in non-legume plants. Despite the phylogenic and ecological diversity among diazotrophic bacteria and their hosts, tightly regulated communication is always necessary between the microorganisms and the host plant to achieve a successful interaction. Ongoing research efforts to improve knowledge of the molecular mechanisms underlying these original relationships and some common strategies leading to a successful relationship between the nitrogen-fixing microorganisms and their hosts are presented. Conclusions Understanding the molecular mechanism of BNF outside the legume–rhizobium symbiosis could have important agronomic implications and enable the use of N-fertilizers to be reduced or even avoided. Indeed, in the short term, improved understanding could lead to more sustainable exploitation of the biodiversity of nitrogen-fixing organisms and, in the longer term, to the transfer of endosymbiotic nitrogen-fixation capacities to major non-legume crops. PMID:23478942

Biological nitrogen fixation under primordial Martian partial pressures of dinitrogen

NASA Technical Reports Server (NTRS)

Klingler, J. M.; Mancinelli, R. L.; White, M. R.

1989-01-01

One of the most striking differences between the conditions on early Mars and earth was a low (18 mb) partial pressure of N2 (pN2) on early Mars, as opposed to 780 mb N2 on earth. To investigate the possibility of biological nitrogen fixation under conditions of early Mars, experiments were carried out on the growth of Azotobacter vinelandii and Azomonas agilis in nitrogen-free synthetic medium under various partial pressures of N2 (ranging from 780 to 0 mb). It was found that, although the biomass, cell number, and growth rate of these bacteria decreased with decreasing pN2 values below pN2 of 400 mb, both microorganisms were capable of growing at pN2 as low as 5 mb (but not at of below 1 mb), indicating that biological fixation of nitrogen could have occurred on primordial Mars.

Moss-cyanobacteria associations as biogenic sources of nitrogen in boreal forest ecosystems.

PubMed

Rousk, Kathrin; Jones, Davey L; Deluca, Thomas H

2013-01-01

The biological fixation of atmospheric nitrogen (N) is a major pathway for available N entering ecosystems. In N-limited boreal forests, a significant amount of N2 is fixed by cyanobacteria living in association with mosses, contributing up to 50% to the total N input. In this review, we synthesize reports on the drivers of N2 fixation in feather moss-cyanobacteria associations to gain a deeper understanding of their role for ecosystem-N-cycling. Nitrogen fixation in moss-cyanobacteria associations is inhibited by N inputs and therefore, significant fixation occurs only in low N-deposition areas. While it has been shown that artificial N additions in the laboratory as well as in the field inhibit N2 fixation in moss-cyanobacteria associations, the type, as well as the amounts of N that enters the system, affect N2 fixation differently. Another major driver of N2 fixation is the moisture status of the cyanobacteria-hosting moss, wherein moist conditions promote N2 fixation. Mosses experience large fluctuations in their hydrological status, undergoing significant natural drying and rewetting cycles over the course of only a few hours, especially in summer, which likely compromises the N input to the system via N2 fixation. Perhaps the most central question, however, that remains unanswered is the fate of the fixed N2 in mosses. The cyanobacteria are likely to leak N, but whether this N is transferred to the soil and if so, at which rates and timescales, is unknown. Despite our increasing understanding of the drivers of N2 fixation, the role moss-cyanobacteria associations play in ecosystem-N-cycling remains unresolved. Further, the relationship mosses and cyanobacteria share is unknown to date and warrants further investigation.

Annual Removal of Aboveground Plant Biomass Alters Soil Microbial Responses to Warming

PubMed Central

Xue, Kai; Yuan, Mengting M.; Xie, Jianping; Li, Dejun; Qin, Yujia; Wu, Liyou; Deng, Ye; He, Zhili; Van Nostrand, Joy D.; Luo, Yiqi; Tiedje, James M.

2016-01-01

ABSTRACT Clipping (i.e., harvesting aboveground plant biomass) is common in agriculture and for bioenergy production. However, microbial responses to clipping in the context of climate warming are poorly understood. We investigated the interactive effects of grassland warming and clipping on soil properties and plant and microbial communities, in particular, on microbial functional genes. Clipping alone did not change the plant biomass production, but warming and clipping combined increased the C4 peak biomass by 47% and belowground net primary production by 110%. Clipping alone and in combination with warming decreased the soil carbon input from litter by 81% and 75%, respectively. With less carbon input, the abundances of genes involved in degrading relatively recalcitrant carbon increased by 38% to 137% in response to either clipping or the combined treatment, which could weaken long-term soil carbon stability and trigger positive feedback with respect to warming. Clipping alone also increased the abundance of genes for nitrogen fixation, mineralization, and denitrification by 32% to 39%. Such potentially stimulated nitrogen fixation could help compensate for the 20% decline in soil ammonium levels caused by clipping alone and could contribute to unchanged plant biomass levels. Moreover, clipping tended to interact antagonistically with warming, especially with respect to effects on nitrogen cycling genes, demonstrating that single-factor studies cannot predict multifactorial changes. These results revealed that clipping alone or in combination with warming altered soil and plant properties as well as the abundance and structure of soil microbial functional genes. Aboveground biomass removal for biofuel production needs to be reconsidered, as the long-term soil carbon stability may be weakened. PMID:27677789

Bacterial nitrogen fixation in sand bioreactors treating winery wastewater with a high carbon to nitrogen ratio.

PubMed

Welz, Pamela J; Ramond, Jean-Baptiste; Braun, Lorenz; Vikram, Surendra; Le Roes-Hill, Marilize

2018-02-01

Heterotrophic bacteria proliferate in organic-rich environments and systems containing sufficient essential nutrients. Nitrogen, phosphorus and potassium are the nutrients required in the highest concentrations. The ratio of carbon to nitrogen is an important consideration for wastewater bioremediation because insufficient nitrogen may result in decreased treatment efficiency. It has been shown that during the treatment of effluent from the pulp and paper industry, bacterial nitrogen fixation can supplement the nitrogen requirements of suspended growth systems. This study was conducted using physicochemical analyses and culture-dependent and -independent techniques to ascertain whether nitrogen-fixing bacteria were selected in biological sand filters used to treat synthetic winery wastewater with a high carbon to nitrogen ratio (193:1). The systems performed well, with the influent COD of 1351 mg/L being reduced by 84-89%. It was shown that the nitrogen fixing bacterial population was influenced by the presence of synthetic winery effluent in the surface layers of the biological sand filters, but not in the deeper layers. It was hypothesised that this was due to the greater availability of atmospheric nitrogen at the surface. The numbers of culture-able nitrogen-fixing bacteria, including presumptive Azotobacter spp. exhibited 1-2 log increases at the surface. The results of this study confirm that nitrogen fixation is an important mechanism to be considered during treatment of high carbon to nitrogen wastewater. If biological treatment systems can be operated to stimulate this phenomenon, it may obviate the need for nitrogen addition. Copyright © 2017 Elsevier Ltd. All rights reserved.

Light and spectral properties as determinants of C:N:P-ratios in phytoplankton

NASA Astrophysics Data System (ADS)

Hessen, Dag O.; Leu, Eva; Færøvig, Per J.; Falk Petersen, Stig

2008-10-01

Light is a major determinant not only for carbon (C)-fixation in autotrophs, but also for the cellular proportions of major elements like C, nitrogen (N) and phosphorus (P). High intensities of photosynthetically active radiation (PAR) increase C:P-ratios in experiments with arctic marine and freshwater phytoplankton species. While high levels of PAR promote high autotrophic productivity, the increased C:P may invoke a "paradox of enrichment" effect since this means lower stoichiometric food quality for herbivores. In contrast, exposure to ultraviolet radiation (UVR) gave reduced cellular C:P-ratios (and N:P) in phytoplankton. This was partly owing to a strong reduction in C-fixation under UVR, but also due to enhanced uptake of P, presumably in response to increased demands for nucleotide repair under UVR stress. The net outcome of these opposing effects will depend on optical properties and mixing depth in the water column. These stoichiometric responses could cause deviations from Redfield ratio in phytoplankton as well as affecting biogeochemical cycling and trophic transfer efficiency in aquatic food-webs.

Net anthropogenic nitrogen accumulation in the Beijing metropolitan region.

PubMed

Han, Yuguo; Li, Xuyong; Nan, Zhe

2011-03-01

A rapid increase in anthropogenic nitrogen inputs has a strong impact on terrestrial and aquatic ecosystems. We have estimated net anthropogenic nitrogen accumulation (NANA) as an index of nitrogen (N) pollution potential in the Beijing metropolitan region, China. Our research provides a basis for understanding the potential impact of anthropogenic N inputs on environmental problems, such as nation-wide water quality degradation under the current rapid urban expansion in modern China. The NANA estimation is based on an inventory of atmospheric N deposition, N fertilizer use, consumption of human food and animal feed, N fixation, and riverine N import and export. We calculated N accumulation values for the years 1991, 1997, 2003, and 2007. The average NANA values for the urban and suburban areas from 1991 to 2007 were 24,038 and 13,090 kg N km(-2) year(-1), respectively. NANA is higher in eastern and southern areas than in northern and western areas, and higher in the urban area than in the suburban area. The overall average NANA in Beijing has a downward trend from 15,187 kg N km(-2) year(-1) in 1991 to 11,606 kg N km(-2) year(-1) in 2007, but is still two to five times as that of developed countries. N input from nitrogenous fertilizer is the largest source of NANA, accounting for 44.4% (6,764 kg N km(-2) year(-1)) of the total N input, followed by atmospheric N deposition and N in human food and animal feed. NANA is closely related to land use, on average 23,140 kg N km(-2) year(-1) in densely populated developed land, 17,904 kg N km(-2) year(-1) in agricultural land, and 10,445 kg N km(-2) year(-1) in forest land. Human population density is the best single predictor of NANA.

Regulation of Carbon Flow by Nitrogen and Light in the Red Alga, Gelidium coulteri.

PubMed

Macler, B A

1986-09-01

The red alga Gelidium coulteri Harv. photosynthetically fixed [(14)C] bicarbonate at high rates under defined conditions in unialgal laboratory culture. The fixation rate and flow of photosynthate into various end products were dependent on the nitrogen status of the tissue. Plants fed luxury levels of nitrogen (approximately 340 micromolar) showed fixation rates several-fold higher than those seen for plants starved for nitrogen. The addition of NO(3) (-) or NH(4) (+) to such starved plants further inhibited fixation over at least the first several hours after addition. The majority of (14)C after incubations of 30 minutes to 8 hours was found in the compounds floridoside, agar and floridean starch. In addition, amino acids and intermediate compounds of the reductive pentose phosphate pathway, glycolytic pathway and tricarboxylic acid cycle were detected. Nitrogen affected the partitioning of labeled carbon into these compounds. Plants under luxury nitrogen conditions had higher floridoside levels and markedly lower amounts of agar and starch than found in plants limited for nitrogen. Amino acid, phycobiliprotein and chlorophyll levels were also significantly higher in nitrogen-enriched plants. Addition of NO(3) (-) to starved plants led to an increase in floridoside, tricarboxylic acid cycle intermediates and amino acids within 1 hour and inhibited carbon flow into agar and starch. Carbon fixation in the dark was only 1 to 7% of that seen in the light. Dark fixation of [(14)C]bicarbonate yielded label primarily in tricarboxylic acid cycle intermediates, amino acids and polysaccharides. Nitrogen stimulated amino acid synthesis at the expense of agar and starch. Floridoside was only a minor component in the dark. Pulse-chase experiments, designed to show carbon turnover, indicated a 2-fold increase in labeling of agar over 96 hours of chase in the light. No increases were seen in the dark. Low molecular weight pools, including floridoside, decreased 2- to 5-fold over this period under both light and dark conditions. Nitrogen status did not influence turnover. There was little or no organic carbon released into the culture medium over this period. The results are consistent with biosynthetic pathways to floridoside and agar that share the common intermediate UDP-d-galactose. It is hypothesized that synthesis of floridoside is regulated by nitrogen and light at the enzymic level.

Regulation of Carbon Flow by Nitrogen and Light in the Red Alga, Gelidium coulteri1

PubMed Central

Macler, Bruce A.

1986-01-01

The red alga Gelidium coulteri Harv. photosynthetically fixed [14C] bicarbonate at high rates under defined conditions in unialgal laboratory culture. The fixation rate and flow of photosynthate into various end products were dependent on the nitrogen status of the tissue. Plants fed luxury levels of nitrogen (approximately 340 micromolar) showed fixation rates several-fold higher than those seen for plants starved for nitrogen. The addition of NO3− or NH4+ to such starved plants further inhibited fixation over at least the first several hours after addition. The majority of 14C after incubations of 30 minutes to 8 hours was found in the compounds floridoside, agar and floridean starch. In addition, amino acids and intermediate compounds of the reductive pentose phosphate pathway, glycolytic pathway and tricarboxylic acid cycle were detected. Nitrogen affected the partitioning of labeled carbon into these compounds. Plants under luxury nitrogen conditions had higher floridoside levels and markedly lower amounts of agar and starch than found in plants limited for nitrogen. Amino acid, phycobiliprotein and chlorophyll levels were also significantly higher in nitrogen-enriched plants. Addition of NO3− to starved plants led to an increase in floridoside, tricarboxylic acid cycle intermediates and amino acids within 1 hour and inhibited carbon flow into agar and starch. Carbon fixation in the dark was only 1 to 7% of that seen in the light. Dark fixation of [14C]bicarbonate yielded label primarily in tricarboxylic acid cycle intermediates, amino acids and polysaccharides. Nitrogen stimulated amino acid synthesis at the expense of agar and starch. Floridoside was only a minor component in the dark. Pulse-chase experiments, designed to show carbon turnover, indicated a 2-fold increase in labeling of agar over 96 hours of chase in the light. No increases were seen in the dark. Low molecular weight pools, including floridoside, decreased 2- to 5-fold over this period under both light and dark conditions. Nitrogen status did not influence turnover. There was little or no organic carbon released into the culture medium over this period. The results are consistent with biosynthetic pathways to floridoside and agar that share the common intermediate UDP-d-galactose. It is hypothesized that synthesis of floridoside is regulated by nitrogen and light at the enzymic level. PMID:16664980

Methanotrophy Induces Nitrogen Fixation in Boreal Mosses

NASA Astrophysics Data System (ADS)

Tiirola, M. A.

2014-12-01

Many methanotrophic bacterial groups fix nitrogen in laboratory conditions. Furthermore, nitrogen (N) is a limiting nutrient in many environments where methane concentrations are highest. Despite these facts, methane-induced N fixation has previously been overlooked, possibly due to methodological problems. To study the possible link between methanotrophy and diazotrophy in terrestrial and aquatic habitats, we measured the co-occurrence of these two processes in boreal forest, peatland and stream mosses using a stable isotope labeling approach (15 N2 and 13 CH4 double labeling) and sequencing of the nifH gene marker. N fixation associated with forest mosses was dependent on the annual N deposition, whereas methane stimulate N fixation neither in high (>3 kg N ha -1 yr -1) nor low deposition areas, which was in accordance with the nifH gene sequencing showing that forest mosses (Pleurozium schreberi and Hylocomium splendens ) carried mainly cyanobacterial N fixers. On the other extreme, in stream mosses (Fontinalis sp.) methane was actively oxidized throughout the year, whereas N fixation showed seasonal fluctuation. The co-occurrence of the two processes in single cell level was proven by co-localizing both N and methane-carbon fixation with the secondary ion mass spectrometry (SIMS) approach. Methanotrophy and diazotrophy was also studied in peatlands of different primary successional stages in the land-uplift coast of Bothnian Bay, in the Siikajoki chronosequence, where N accumulation rates in peat profiles indicate significant N fixation. Based on experimental evidence it was counted that methane-induced N fixation explained over one-third of the new N input in the younger peatland successional stages, where the highest N fixation rates and highest methane oxidation activities co-occurred in the water-submerged Sphagnum moss vegetation. The linkage between methanotrophic carbon cycling and N fixation may therefore constitute an important mechanism in the rapid accumulation of N during the primary succession of peatlands. It is still an open issue whether methanotrophy induces N fixation directly or by enhancing phototrophic or heterotrophic N fixation.

Diurnal variation in the functioning of cowpea nodules.

PubMed

Rainbird, R M; Atkins, C A; Pate, J S

1983-06-01

Nitrogenase (EC 1.7.99.2) activity of nodules of cowpea (Vigna unguiculata [L.] Walp), maintained under conditions of a 12-hour day at 30 degrees C and 800 to 1,000 microeinsteins per square meter per second (photosynthetically active radiation) and a 12-hour night at 20 degrees C, showed a marked diurnal variation with the total electron flux through the enzyme at night being 60% of that in the photoperiod. This diurnal pattern was, however, due to changes in hydrogen evolution. The rate of nitrogen fixation, measured by short-term (15)N(2) assimilation or estimated from the difference in hydrogen evolution in air or Ar:O(2) (80:20; v/v), showed no diurnal variation. Carbon dioxide released from nodules showed a diurnal variation synchronized with that of nitrogenase functioning and, as a consequence, the apparent ;respiratory cost' of nitrogen fixation in the photoperiod was almost double that at night (9.74 +/- 0.38 versus 5.70 +/- 0.90 moles CO(2) evolved per mole N(2) fixed). Separate carbon and nitrogen balances constructed for nodules during the photoperiod and dark period showed that, at night, nodule functioning required up to 40% less carbohydrate to achieve the same level of nitrogen fixation as during the photoperiod (2.4 versus 1.4 moles hexose per mole N(2) fixed).Stored reserves of nonstructural carbohydrate of the nodule only partly satisfied the requirement for carbon at night, and fixation was dependent on continued import of translocated assimilates at all times. Measurements of the soluble nitrogen pools of the nodule together with (15)N studies indicated that, both during the day and night, nitrogenous products of fixation were effectively translocated to all organs of the host plant despite low rates of transpiration at night. Reduced fluxes of water through the plant at night were apparently counteracted by increased concentration of nitrogen, especially as ureides, in the xylem stream.

Crassulacean acid metabolism in the shade. Studies on an epiphytic fern, Pyrrosia longifolia, and other rainforest species from Australia.

PubMed

Winter, K; Osmond, C B; Hubick, K T

1986-01-01

Crassulacean acid metabolism (CAM) was studied in a tropical epiphytic fern, Pyrrosia longifolia, from a fully sun-exposed and from a very shaded site in Northern Queensland, Australia. Measurements of instantaneous net CO 2 exchange showed carbon gain via CO 2 dark fixation with some net CO 2 uptake also occuring during late afternoon, in both sun and shade fronds. Maximum rates of net CO 2 uptake and the nocturnal increase in titratable acidity were lower in shade than in sun fronds. δ 13 C values of sun and shade fronds were not significantly different, and ranged between-14 and-15‰ suggesting that, in the long term, carbon gain was mainly via CO 2 dark fixation. Sun fronds had a higher light compensation point of photosynthesis than shade fronds but the same quantum yield. Yet there was no acclimation of photosynthetic O 2 evolution, (measured at 5% CO 2 ) in sun and shade fronds and photosynthesis saturated at between 200 and 400 μmol quanta m -2 s -1 . Use of higher light intensities for photosynthesis of sun fronds was probably precluded by low nutrient availability. Total nitrogen was less than 1% of dry weight in fully expanded sun and shade fronds. Exposure of shade fronds to full sunlight for 6 h led to a 60% decline in the quantum yield of photosynthesis and to a decline in variable fluorescence measured at room temperature. Photoinhibition by high light was also observed in Hoya nicholsoniae, a rainforest climber growing in deep shade. This species also exhibited CAM as demonstrated by nocturnal net CO 2 uptake, nocturnal acidification and a δ 13 C value of-14‰. Photosynthetic O 2 evolution in this species was saturated at 2.5% of full sunlight. Two species of Dendrobium (Orchidaceae) from sun-exposed sites, one species exhibiting CAM and the other one exhibiting net CO 2 uptake exclusively during daytime via conventional C 3 photosynthesis, showed similar light response curves and the same quantum yield for photosynthetic O 2 evolution.

Bulk vs. amino acid stable N isotope estimations of metabolic status and contributions of nitrogen fixation to size-fractionated zooplankton biomass in the subtropical N Atlantic

NASA Astrophysics Data System (ADS)

Mompeán, Carmen; Bode, Antonio; Gier, Elizabeth; McCarthy, Matthew D.

2016-08-01

A comparative analysis of natural abundance of stable N isotopes (δ15N) in individual amino acids and bulk organic matter of size-fractionated plankton revealed the differential impact of nitrogen fixation through the food web in a transect across the subtropical North Atlantic. All δ15N measurements showed low values in the central region, followed by the western zone, while maximum δ15N values were found in the eastern zone. These results were consistent with the prevalence of nitrogen fixation in the central and western zones, and the influence of the west Africa upwelling in the eastern zone. Use of compound-specific amino acid isotope data (CSI-AA) revealed relatively low variability in the impact of diazotrophic nitrogen within the different plankton size fractions, while δ15N of bulk organic matter showed high variability with size. Explicit CSI-AA trophic position estimates showed a small increase with mean plankton size class and varied in a relatively narrow range 1.8-2.5), with the lowest values in the central zone. High correlations between bulk plankton δ15N and individual amino acids (in particular Phe and Thr), as well as reconstructed total protein δ15N values, suggest a set of new relationships that may be important to tracing direct plankton contributions to nitrogen recycling in the ocean, including detrital organic nitrogen pools. Overall, these new results represent the most detailed investigation of CSI-AA data in plankton size classes to date, and indicated a greater importance of diazotrophic N than suggested by concurrent measurements of bulk δ15N, abundance of large nitrogen fixing organisms or nitrogen fixation rates.

The anthropogenic perturbation of the marine nitrogen cycle by atmospheric deposition: Nitrogen cycle feedbacks and the 15N Haber-Bosch effect

NASA Astrophysics Data System (ADS)

Yang, Simon; Gruber, Nicolas

2016-10-01

Over the last 100 years, anthropogenic emissions have led to a strong increase of atmospheric nitrogen deposition over the ocean, yet the resulting impacts and feedbacks are neither well understood nor quantified. To this end, we run a suite of simulations with the ocean component of the Community Earth System Model v1.2 forced with five scenarios of nitrogen deposition over the period from 1850 through 2100, while keeping all other forcings unchanged. Even though global oceanic net primary production increases little in response to this fertilization, the higher export and the resulting expansion of the oxygen minimum zones cause an increase in pelagic and benthic denitrification and burial by about 5%. In addition, the enhanced availability of fixed nitrogen in the surface ocean reduces global ocean N2 fixation by more than 10%. Despite the compensating effects through these negative feedbacks that eliminate by the year 2000 about 60% of the deposited nitrogen, the anthropogenic nitrogen input forced the upper ocean N budget into an imbalance of between 9 and 22 Tg N yr-1 depending on the deposition scenario. The excess nitrogen accumulates to highly detectable levels and causes in most areas a distinct negative trend in the δ15N of the oceanic fixed nitrogen pools—a trend we refer to as the 15N Haber-Bosch effect. Changes in surface nitrate utilization and the nitrogen feedbacks induce further changes in the δ15N of NO3-, making it a good but complex recorder of the overall impact of the changes in atmospheric deposition.

Accounting for nitrogen fixation in simple models of lake nitrogen loading/export.

PubMed

Ruan, Xiaodan; Schellenger, Frank; Hellweger, Ferdi L

2014-05-20

Coastal eutrophication, an important global environmental problem, is primarily caused by excess nitrogen and management efforts consequently focus on lowering watershed N export (e.g., by reducing fertilizer use). Simple quantitative models are needed to evaluate alternative scenarios at the watershed scale. Existing models generally assume that, for a specific lake/reservoir, a constant fraction of N loading is exported downstream. However, N fixation by cyanobacteria may increase when the N loading is reduced, which may change the (effective) fraction of N exported. Here we present a model that incorporates this process. The model (Fixation and Export of Nitrogen from Lakes, FENL) is based on a steady-state mass balance with loading, output, loss/retention, and N fixation, where the amount fixed is a function of the N/P ratio of the loading (i.e., when N/P is less than a threshold value, N is fixed). Three approaches are used to parametrize and evaluate the model, including microcosm lab experiments, lake field observations/budgets and lake ecosystem model applications. Our results suggest that N export will not be reduced proportionally with N loading, which needs to be considered when evaluating management scenarios.

Nodule and Leaf Nitrate Reductases and Nitrogen Fixation in Medicago sativa L. under Water Stress

PubMed Central

Aparicio-Tejo, P.; Sánchez-Díaz, Manuel

1982-01-01

The effect of water stress on patterns of nitrate reductase activity in the leaves and nodules and on nitrogen fixation were investigated in Medicago sativa L. plants watered 1 week before drought with or without NO3−. Nitrogen fixation was decreased by water stress and also inhibited strongly by the presence of NO3−. During drought, leaf nitrate reductase activity (NRA) decreased significantly particularly in plants watered with NO3−, while with rewatering, leaf NRA recovery was quite important especially in the NO3−-watered plants. As water stress progressed, the nodular NRA increased both in plants watered with NO3− and in those without NO3− contrary to the behavior of the leaves. Beyond −15.105 pascal, nodular NRA began to decrease in plants watered with NO3−. This phenomenon was not observed in nodules of plants given water only. Upon rewatering, it was observed that in plants watered with NO3− the nodular NRA increased again, while in plants watered but not given NO3−, such activity began to decrease. Nitrogen fixation increased only in plants without NO3−. PMID:16662233

Nitrogen fixation in the mucus of Red Sea corals.

PubMed

Grover, Renaud; Ferrier-Pagès, Christine; Maguer, Jean-François; Ezzat, Leila; Fine, Maoz

2014-11-15

Scleractinian corals are essential constituents of tropical reef ecological diversity. They live in close association with diazotrophs [dinitrogen (N2)-fixing microbes], which can fix high rates of N2. Whether corals benefit from this extrinsic nitrogen source is still under debate. Until now, N2 fixation rates have been indirectly estimated using the acetylene reduction assay, which does not permit assessment of the amount of nitrogen incorporated into the different compartments of the coral holobiont. In the present study, the (15)N2 technique was applied for the first time on three Red Sea coral species. Significant (15)N enrichment was measured in particles released by corals to the surrounding seawater. N2 fixation rates were species specific and as high as 1.6-2 ng N day(-1) l(-1). However, no significant enrichment was measured in the symbiotic dinoflagellates or the coral host tissues, suggesting that corals do not benefit from diazotrophic N2 fixation. © 2014. Published by The Company of Biologists Ltd.

Expression of drought-tolerant N2 fixation in heterogeneous inbred families derived from PI 471938 and Hutcheson soybean

USDA-ARS?s Scientific Manuscript database

Nitrogen fixation of soybean is particularly vulnerable to drought, since, in most genotypes, N2 fixation activity decreases very early in the soil drying cycle. Although a few soybean genotypes, including ‘PI 471938’, have been identified that express N2 fixation tolerance of drought, it is unknown...

Nutrient feedbacks to soil heterotrophic nitrogen fixation in forests

USGS Publications Warehouse

Perakis, Steven; Pett-Ridge, Julie C.; Catricala, Christina E.

2017-01-01

Multiple nutrient cycles regulate biological nitrogen (N) fixation in forests, yet long-term feedbacks between N-fixation and coupled element cycles remain largely unexplored. We examined soil nutrients and heterotrophic N-fixation across a gradient of 24 temperate conifer forests shaped by legacies of symbiotic N-fixing trees. We observed positive relationships among mineral soil pools of N, carbon (C), organic molybdenum (Mo), and organic phosphorus (P) across sites, evidence that legacies of symbiotic N-fixing trees can increase the abundance of multiple elements important to heterotrophic N-fixation. Soil N accumulation lowered rates of heterotrophic N-fixation in organic horizons due to both N inhibition of nitrogenase enzymes and declines in soil organic matter quality. Experimental fertilization of organic horizon soil revealed widespread Mo limitation of heterotrophic N-fixation, especially at sites where soil Mo was scarce relative to C. Fertilization also revealed widespread absence of P limitation, consistent with high soil P:Mo ratios. Responses of heterotrophic N-fixation to added Mo (positive) and N (negative) were correlated across sites, evidence that multiple nutrient controls of heterotrophic N-fixation were more common than single-nutrient effects. We propose a conceptual model where symbiotic N-fixation promotes coupled N, C, P, and Mo accumulation in soil, leading to positive feedback that relaxes nutrient limitation of overall N-fixation, though heterotrophic N-fixation is primarily suppressed by strong negative feedback from long-term soil N accumulation.

Quantifying the contribution of single microbial cells to nitrogen assimilation in aquatic environments

NASA Astrophysics Data System (ADS)

Musat, N.; Kuypers, M. M. M.

2009-04-01

Nitrogen is a primary productivity-limiting nutrient in the ocean. The nitrogen limitation of productivity may be overcome by organisms capable of converting dissolved N2 into fixed nitrogen available to the ecosystem. In many oceanic regions, growth of phytoplankton is nitrogen limited because fixation of N2 cannot make up for the removal of fixed inorganic nitrogen (NH4+, NO2-, NO3-) by anaerobic microbial processes. The amount of available fixed nitrogen in the ocean can be changed by the biological processes of heterotrophic denitrification, anaerobic ammonium oxidation and nitrogen fixation. For a complete understanding of nitrogen cycling in the ocean a link between the microbial and biogeochemical processes at the single cell level and their role in global biogeochemical cycles is essential. Here we report a recently developed method, Halogen In Situ Hybridization-Secondary Ion Mass Spectroscopy (HISH-SIMS) and its potential application to study the nitrogen-cycle processes in the ocean. The method allows simultaneous phylogenetic identification and quantitation of metabolic activities of single microbial cells in the environment. It uses horseradish-peroxidase-labeled oligonucleotide probes and fluorine-containing tyramides for the identification of microorganisms in combination with stable-isotope-labeling experiments for analyzing the metabolic function of single microbial cells. HISH-SIMS was successfully used to study nitrogen assimilation and nitrogen fixation by anaerobic phototrophs in a meromictic alpine lake. The HISH-SIMS method enables studies of the ecophysiology of individual, phylogenetically identified microorganisms involved in the N-cycle and allows us to track the flow of nitrogen within microbial communities.

Catalytic transformation of dinitrogen into ammonia and hydrazine by iron-dinitrogen complexes bearing pincer ligand

PubMed Central

Kuriyama, Shogo; Arashiba, Kazuya; Nakajima, Kazunari; Matsuo, Yuki; Tanaka, Hiromasa; Ishii, Kazuyuki; Yoshizawa, Kazunari; Nishibayashi, Yoshiaki

2016-01-01

Synthesis and reactivity of iron-dinitrogen complexes have been extensively studied, because the iron atom plays an important role in the industrial and biological nitrogen fixation. As a result, iron-catalyzed reduction of molecular dinitrogen into ammonia has recently been achieved. Here we show that an iron-dinitrogen complex bearing an anionic PNP-pincer ligand works as an effective catalyst towards the catalytic nitrogen fixation, where a mixture of ammonia and hydrazine is produced. In the present reaction system, molecular dinitrogen is catalytically and directly converted into hydrazine by using transition metal-dinitrogen complexes as catalysts. Because hydrazine is considered as a key intermediate in the nitrogen fixation in nitrogenase, the findings described in this paper provide an opportunity to elucidate the reaction mechanism in nitrogenase. PMID:27435503

The contribution of nitrogen fixation by cyanobacteria to particulate organic nitrogen in a constructed wetland

NASA Astrophysics Data System (ADS)

Zhang, X.; PAN, X.; MA, M.; Li, W.; Cui, L.

2016-12-01

N-fixing cyanobacteria can create extra nitrogen for aquatic ecosystems. Previous studies reported inconsistence patterns of the contribution of biological nitrogen fixation to the nitrogen pools in aquatic ecosystems. However, there were few studies concerning the effect of fixed nitrogen by cyanobacteria on the nitrogen removal efficiency in constructed wetlands. This study was performed at the Beijing Wildlife Rescue and Rehabilitation Centre, where a constructed lake for the habitation of waterfowls and a constructed wetland for purifying sewage from the lake are located. The composition of phytoplankton communities, the concentrations of particulate organic nitrogen (PON) and nitrogen fixation rates (Rn) in the constructed lake and the constructed wetland were compared throughout a growing season. We counted the densities of genus Anabaena and Microcystis cells, and explored their relationships with PON and Rn in water. The proportions of PON from various sources, including the ambient N2, waterfowl faeces, wetland sediments and the nitrates, were calculated by the natural abundance of 15N with the IsoSource software. The result revealed that the constructed lake was alternately dominated by Anabaena and Microcystis throughout the growing season, and the Rn was positively correlated with PON and the cell density of Anabaena (P < 0.05). This implied that the fixed nitrogen by N-fixing Anabaena might be utilized by non-N-fixing Microcystis, maintaining the fixed nitrogen with PON form. The ambient N2 composed 0.5 82% and 50.0 84.7% to the PON in the constructed lake and wetland respectively during the growing season. The proportions of PON from N2 increased to more than 80% when the Rn reached the highest in September. The result demonstrated that the nitrogen fixed by Anabaena might be utilized by non-N-fixing Microcystis which formed water blooms in summer. Therefore, the decline of the removal efficiency of PON in the constructed wetland in summer might indirectly result from the nitrogen fixation, since the proliferated algal were difficult to sediment in surface flow wetlands.

Nitrogen Fixation (Acetylene Reduction) Associated with Duckweed (Lemnaceae) Mats

PubMed Central

Zuberer, D. A.

1982-01-01

Duckweed (Lemnaceae) mats in Texas and Florida were investigated, using the acetylene reduction assay, to determine whether nitrogen fixation occurred in these floating aquatic macrophyte communities. N2-fixing microorganisms were enumerated by plating or most-probable-number techniques, using appropriate N-free media. Results of the investigations indicated that substantial N2-fixation (C2H2) was associated with duckweed mats in Texas and Florida. Acetylene reduction values ranged from 1 to 18 μmol of C2H4 g (dry weight)−1 day−1 for samples incubated aerobically in light. Dark N2 fixation was always two- to fivefold lower. 3-(3,4-Dichlorophenyl)-1,1-dimethylurea (7 to 10 μM) reduced acetylene reduction to levels intermediate between light and dark incubation. Acetylene reduction was generally greatest for samples incubated anaerobically in the light. It was estimated that 15 to 20% of the N requirement of the duckweed could be supplied through biological nitrogen fixation. N2-fixing heterotrophic bacteria (105 cells g [wet weight]−1 and cyanobacteria (105 propagules g [wet weight]−1 were associated with the duckweed mats. Azotobacter sp. was not detected in these investigations. One diazotrophic isolate was classified as Klebsiella. PMID:16345992

Prebiotic Chemistry and Atmospheric Warming of Early Earth by an Active Young Sun

NASA Technical Reports Server (NTRS)

Airapetian, V. S.; Glocer, A.; Gronoff, G.; Hebrard, E.; Danchi, W.

2016-01-01

Nitrogen is a critical ingredient of complex biological molecules. Molecular nitrogen, however, which was outgassed Into the Earth's early atmosphere, is relatively chemically inert and nitrogen fixation into more chemically reactive compounds requires high temperatures. Possible mechanisms of nitrogen fixation include lightning, atmospheric shock heating by meteorites, and solar ultraviolet radiation. Here we show that nitrogen fixation in the early terrestrial atmosphere can be explained by frequent and powerful coronal mass ejection events from the young Sun -- so-called superflares. Using magnetohydrodynamic simulations constrained by Kepler Space Telescope observations, we find that successive superflare ejections produce shocks that accelerate energetic particles, which would have compressed the early Earth's magnetosphere. The resulting extended polar cap openings provide pathways for energetic particles to penetrate into the atmosphere and, according to our atmospheric chemistry simulations, initiate reactions converting molecular nitrogen, carbon dioxide and methane to the potent greenhouse gas nitrous oxide as well as hydrogen cyanide, an essential compound for life. Furthermore, the destruction of N2, C02 and CH, suggests that these greenhouse gases cannot explain the stability of liquid water on the early Earth. Instead, we propose that the efficient formation of nitrous oxide could explain a warm early Earth.

Ca2+ Requirement for Aerobic Nitrogen Fixation by Heterocystous Blue-Green Algae 1

PubMed Central

Rodríguez, Herminia; Rivas, Joaquín; Guerrero, Miguel G.; Losada, Manuel

1990-01-01

The requirement of Ca2+ for growth and nitrogen fixation has been investigated in two strains of heterocystous blue-green algae (Anabaena sp. and Anabaena ATCC 33047). With combined nitrogen (nitrate or ammonium) or with N2 under microaerobic conditions, Ca2+ was not required for growth, at least in concentrations greater than traces. In contrast, Ca2+ was required as a macronutrient for growth and nitrogen fixation with air as the nitrogen source. Addition of Ca2+ to an aerobic culture without Ca2+ promoted, after a lag of several hours, development of nitrogenase activity and cell growth. Provision of air to a microaerobic culture in the absence of Ca2+ promoted a drastic drop in nitrogenase activity, which rapidly recovered its initial level upon restoration of microaerobic conditions. Development of nitrogenase activity in response to either Ca2+ or low oxygen tension was dependent on de novo protein synthesis. The role of Ca2+ seems to be related to protection of nitrogenase from inactivation, by conferring heterocysts resistance to oxygen. PMID:16667401

Environmental forcing of nitrogen fixation in the eastern tropical and sub-tropical North Atlantic Ocean.

PubMed

Rijkenberg, Micha J A; Langlois, Rebecca J; Mills, Matthew M; Patey, Matthew D; Hill, Polly G; Nielsdóttir, Maria C; Compton, Tanya J; Laroche, Julie; Achterberg, Eric P

2011-01-01

During the winter of 2006 we measured nifH gene abundances, dinitrogen (N(2)) fixation rates and carbon fixation rates in the eastern tropical and sub-tropical North Atlantic Ocean. The dominant diazotrophic phylotypes were filamentous cyanobacteria, which may include Trichodesmium and Katagnymene, with up to 10(6) L(-1)nifH gene copies, unicellular group A cyanobacteria with up to 10(5) L(-1)nifH gene copies and gamma A proteobacteria with up to 10(4) L(-1)nifH gene copies. N(2) fixation rates were low and ranged between 0.032-1.28 nmol N L(-1) d(-1) with a mean of 0.30 ± 0.29 nmol N L(-1) d(-1) (1σ, n = 65). CO(2)-fixation rates, representing primary production, appeared to be nitrogen limited as suggested by low dissolved inorganic nitrogen to phosphate ratios (DIN:DIP) of about 2 ± 3.2 in surface waters. Nevertheless, N(2) fixation rates contributed only 0.55 ± 0.87% (range 0.03-5.24%) of the N required for primary production. Boosted regression trees analysis (BRT) showed that the distribution of the gamma A proteobacteria and filamentous cyanobacteria nifH genes was mainly predicted by the distribution of Prochlorococcus, Synechococcus, picoeukaryotes and heterotrophic bacteria. In addition, BRT indicated that multiple a-biotic environmental variables including nutrients DIN, dissolved organic nitrogen (DON) and DIP, trace metals like dissolved aluminum (DAl), as a proxy of dust inputs, dissolved iron (DFe) and Fe-binding ligands as well as oxygen and temperature influenced N(2) fixation rates and the distribution of the dominant diazotrophic phylotypes. Our results suggest that lower predicted oxygen concentrations and higher temperatures due to climate warming may increase N(2) fixation rates. However, the balance between a decreased supply of DIP and DFe from deep waters as a result of more pronounced stratification and an enhanced supply of these nutrients with a predicted increase in deposition of Saharan dust may ultimately determine the consequences of climate warming for N(2) fixation in the North Atlantic.

Nitrogen Fixation (Acetylene Reduction) by Epiphytes of Freshwater Macrophytes

PubMed Central

Finke, Linda R.; Seeley, H. W.

1978-01-01

The involvement of epiphytic microorganisms in nitrogen fixation was investigated in a shallow freshwater pond near Ithaca, N.Y. The acetylene reduction technique was used to follow diel and seasonal cycles of nitrogen fixation by epiphytes of Myriophyllum spicatum. Acetylene-reducing activity was maximal between noon and 6 p.m., but substantial levels of activity relative to daytime rates continued through the night. Experiments with the seasonal course of activity showed a gradual decline during the autumn months and no activity in January or February. Activity commenced in May, with an abrupt increase to levels between 0.45 and 0.95 nmol of ethylene formed per mg (dry weight) of plant per h. Through most of the summer months, mean rates of acetylene reduction remained between 0.15 and 0.60 nmol/mg (dry weight) per h. It was calculated from diel and seasonal cycles that, in the pond areas studied, epiphytes were capable of adding from 7.5 to 12.5 μg of N per mg of plant per year to the pond. This amount is significant relative to the total amount of nitrogen incorporated into the plant. Blue-green algae (cyanobacteria), particularly Gloeotrichia, appeared to bear prime responsibility for nitrogen fixation, but photosynthetic bacteria of the genus Rhodopseudomonas were isolated from M. spicatum and shown to support high rates of acetylene reduction. PMID:16345301

Transcriptional Profiling of Nitrogen Fixation in Azotobacter vinelandii▿†

PubMed Central

Hamilton, Trinity L.; Ludwig, Marcus; Dixon, Ray; Boyd, Eric S.; Dos Santos, Patricia C.; Setubal, João C.; Bryant, Donald A.; Dean, Dennis R.; Peters, John W.

2011-01-01

Most biological nitrogen (N2) fixation results from the activity of a molybdenum-dependent nitrogenase, a complex iron-sulfur enzyme found associated with a diversity of bacteria and some methanogenic archaea. Azotobacter vinelandii, an obligate aerobe, fixes nitrogen via the oxygen-sensitive Mo nitrogenase but is also able to fix nitrogen through the activities of genetically distinct alternative forms of nitrogenase designated the Vnf and Anf systems when Mo is limiting. The Vnf system appears to replace Mo with V, and the Anf system is thought to contain Fe as the only transition metal within the respective active site metallocofactors. Prior genetic analyses suggest that a number of nif-encoded components are involved in the Vnf and Anf systems. Genome-wide transcription profiling of A. vinelandiicultured under nitrogen-fixing conditions under various metal amendments (e.g., Mo or V) revealed the discrete complement of genes associated with each nitrogenase system and the extent of cross talk between the systems. In addition, changes in transcript levels of genes not directly involved in N2fixation provided insight into the integration of central metabolic processes and the oxygen-sensitive process of N2fixation in this obligate aerobe. The results underscored significant differences between Mo-dependent and Mo-independent diazotrophic growth that highlight the significant advantages of diazotrophic growth in the presence of Mo. PMID:21724999

Anaerobic Nitrogen Fixers on Mars

NASA Astrophysics Data System (ADS)

Lewis, B. G.

2000-07-01

The conversion of atmospheric nitrogen gas to the protein of living systems is an amazing process of nature. The first step in the process is biological nitrogen fixation, the transformation of N2 to NH3. The phenomenon is crucial for feeding the billions of our species on Earth. On Mars, the same process may allow us to discover how life can adapt to a hostile environment, and render it habitable. Hostile environments also exist on Earth. For example, nothing grows in coal refuse piles due to the oxidation of pyrite and marcasite to sulfuric acid. Yet, when the acidity is neutralized, alfalfa and soybean plants develop root nodules typical of symbiotic nitrogen fixation with Rhizobium species possibly living in the pyritic material. When split open, these nodules exhibited the pinkish color of leghemoglobin, a protein in the nodule protecting the active nitrogen-fixing enzyme nitrogenase against the toxic effects of oxygen. Although we have not yet obtained direct evidence of nitrogenase activity in these nodules (reduction of acetylene to ethylene, for example), these findings suggested the possibility that nitrogen fixation was taking place in this hostile, non-soil material. This immediately raises the possibility that freeliving anaerobic bacteria which fix atmospheric nitrogen on Earth, could do the same on Mars.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Capone, D.G.; Penhale, P.A.; Oremland, R.S.

N/sub 2/ (C/sub 2/H/sub 2/) fixation and primary production were measured in communities of Thalassia testudinum at two sites in Bimini Harbor (Bahamas). Production was determined by uptake of (/sup 14/C)NaHCO/sub 3/, by leaf growth measurements, and by applying an empirical formula based on leaf dimensions. The last two methods gave similar results but the /sup 14/C method gave higher values. Anaerobic sediment N/sub 2/ fixation supplied about 1/4 to 1/2 of the nitrogen demand for leaf production (by leaf growth method) and there was a significant correlation between N/sub 2/ fixation and CO/sub 2/ fixation rates when all componentsmore » of the communities were considered (macrophyte, phyllosphere epiphytes, and detrital leaves). N/sub 2/ fixation is important to production in Thalassia communities and the plant and its leaf epiphytes may be distinct entities in terms of nitrogen and carbon metabolism.« less

Evaluation of the tepary bean (Phaseolus acutifolius) diversity panel for response to the NL 3 strain of Bean Common Mosaic Necrosis Virus (BCMNV) and for biological nitrogen fixation with Bradyrhizobium strains

USDA-ARS?s Scientific Manuscript database

Aphid-transmitted Bean Common Mosaic Necrosis Virus (BCMNV) and Bean Common Mosaic Virus (BCMV) are potyviruses that are seed transmitted in tepary bean. Developing resistance to these viruses will be critical for expanding production in areas where they are endemic. Biological nitrogen fixation (BN...

Seasonal patterns of periphyton nitrogen fixation in calcareous wetlands

NASA Astrophysics Data System (ADS)

Liao, X.; Inglett, P.

2011-12-01

Periphyton mats are an ecologically important component of the Everglades ecosystem and plays various vital ecological functions. However, nitrogen fixation of periphyton, has received little attention throughout much of the Everglades system. The objective of this study was to characterize the seasonal pattern of periphyton N2 fixation in the Hole-in-the-Donut (HID) of Florida Everglades, where farmed marl prairie wetlands have been restored through complete soil removal (CSR) to reduce nutrient levels. Two restored areas (i.e., cleared in 2000 and 2003) and a reference (natural and unfarmed) marl prairie wetland sites were selected in the HID. Seven times of sampling were performed across the wet and dry season during the 2010 and 2011. The annual fixed nitrogen was approximately 0.4gN m-2 yr-1 in the restored sites which was higher in the reference site (~0.2gN m-2 yr-1). All the three sites showed similar seasonal patterns of N2 fixation that is higher values were observed in the wet season; but the peak value was one month later in reference sits (i.e., September) comparing to the restored areas (i.e., July). The peak of periphyton AR rates in the 2000- and 2003-restored areas appeared in July (i.e., wet season) within the range of 20-79 nmols g-1dw h-1 and 31-53nmols g-1dw h-1, respectively. In contrast, the peak of reference site was observed in September with the range of 2-5 nmols g-1dw h-1. Stable N isotopic ratios (i.e., δ15N) also varied with time but didn't show consistent seasonal pattern as nitrogen fixation. N2 fixation positively correlated with periphyton total phosphorus (TP) and negatively correlated with total nitrogen and phosphorus molar ratios (TN:TP), indicating that N2 fixation would be a indicator of nutrient limitation. In general, δ15N was negatively correlated with nitrogenase activity but the correlation became weakened in the wet season, especially in the flooded July and September, which would be explained by other environmental factors and higher denitrification activities in that time.

Oxygenation of Earth's atmosphere and its impact on the evolution of nitrogen-based metabolisms

NASA Astrophysics Data System (ADS)

Papineau, D.; Mojzsis, S. J.

2002-12-01

The evolution of metabolic pathways is closely linked to the evolution of the redox state of the terrestrial atmosphere. Nitrogen has been an essential biological element since the emergence of life when reduced nitrogen compounds (e.g. ammonia) were utilized in the prebiotic synthesis of proteins and nucleic acids. The nitrogen isotopic composition of sediments has been used to trace the origin of sedimentary organic matter in the rock record. Nitrogen is therefore suitable as a biosignature to trace the emergence of life on Earth or other planetary bodies as well as to follow the subsequent evolution of the biosphere in response to global redox changes. Evidence is strong that biological nitrogen fixation evolved very early in the history of life. The Last Common Ancestor (LCA) on Earth was most likely capable of nitrogen fixation as seen from the phylogenetic distribution of nitrogen-fixing organisms in both the domains of Bacteria and Archaea. Phylogenetic trees plotted with nitrogen-fixing gene (Nif) sequences from lineages of Bacteria and Archaea suggest that the Nif genes originated in a common ancestor of the two domains. Other phylogenetic analyses have also demonstrated that the paralogous duplication of the nifDK and nifEN operons, central to nitrogen fixation, predated the divergence of Archaea from Bacteria and therefore occurred prior to the emergence of the LCA. Although the same may be true for denitrification, this metabolic pathway probably did not become dominant until atmospheric pO2 increased between ~2.4 to 1.9 Ga during the Great Oxygenation Event (GOE). Recent work has shown a general depletion in 15N content of Archean (pre-2.5 Ga) relative to Phanerozoic (<540 Ma) kerogens. Studies have shown that the distribution of the δ15N values in kerogens shift from negative values in the Early Archean (from -6 to +6‰ with an average near 0‰ ) to approximately contemporary positive values (from +2 to +10‰ with an average at +6‰ ) by the Early Proterozoic. In the anoxic Archean atmosphere, the nitrogen cycle must have been different and the instability of oxidized nitrogen species such as NO3- under low pO2 must have limited its availability. Therefore nitrogen fixation probably was the dominant nitrogen-based metabolic pathway during the Archean until the rise of oxygen in the atmosphere in the Early Proterozoic, which resulted in more favorable conditions for denitrification to become the dominant nitrogen-based metabolism. Stable isotopic ratios can be used to detect denitrification (which positively fractionates δ15N) and nitrogen fixation (which negatively fractionates δ15N) in K+-containing minerals such as biotite and muscovite. In an effort to characterize the transition from the essentially anoxic Archean atmosphere to the modern oxidizing atmosphere, we are investigating sedimentary rocks spanning 2.4 to 1.9 Ga from Rovaniemi, Finland. Structural NH4+ in minerals is characterized by μFTIR spectroscopy. Our new μFTIR measurements of Early Proterozoic metasediments spanning the 2.4 - 1.9 Ga time interval (and focused on the well-represented 2.2 - 1.9 Ga time interval from Rovaniemi) provide a high-resolution record of ammonium content during the GOE as a prelude to future detailed δ15N measurements by laser mass spectrometry. This work traces the evolution of nitrogen fixation in the biosphere, its response to changes in global redox and provides a tool for the investigation of biosignatures in samples returned from other ancient planetary surfaces.

Controls on the distributions of organic carbon and nitrogen in the eastern Pacific Ocean

NASA Astrophysics Data System (ADS)

Hansell, Dennis A.; Waterhouse, Tye Y.

1997-05-01

Measurements of total organic carbon (TOC) and nitrogen (TON) were made on the WOCE P18 line (from 67°S to 23°N along 103°/110°W). There was an accumulation of TOC on the equator and in the oligotrophic waters north and south of the equator. The concentrations of TOC were well correlated with temperature, indicating an important physical control on its distribution. The boundary separating shallow, TOC-rich water from deep, TOC-poor water overlaid the main thermocline. This observation suggests that water column stability or residence time imparted by the main thermocline is a primary determinant of TOC accumulation. Elevated TON concentrations were found in all surface waters, with the lowest values found in the region of 20-35°S. Net TON drawdown in the South Pacific subtropical gyre, likely due to biological utilization and vertical export of the nitrogen, was initiated with depletion of equatorially upwelled nitrate. The degree to which inorganic nitrogen was limiting in the surface layer south of the equator served to control the concentrations of TON. Such controls were not exerted on organic carbon, as reflected by increasing C:N ratios of organic matter as TON was removed. Unlike the findings in the South Pacific, TON concentrations in oligotrophic waters north of the equator were frequently higher than on the equator. Such accumulations are hypothesized to be maintained from nitrogen fixation, nitrogen input due to vertical migration of autotrophs or diffusive flux of inorganic nitrogen into the euphotic zone across the relatively shallow nitracline.

Effect of mineral phosphates on growth and nitrogen fixation of diazotrophic cyanobacteria Anabaena variabilis and Westiellopsis prolifica.

PubMed

Yandigeri, Mahesh S; Yadav, Arvind K; Meena, Kamlesh Kumar; Pabbi, Sunil

2010-03-01

The nitrogen fixing cyanobacterial strains namely Anabaena variabilis (Nostocales, Nostocaceae) and Westiellopsis prolifica (Nostocales, Hapalosiphonaceae) were evaluated for their nitrogen fixation and growth potential in response to different concentrations (10, 20 and 30 mg P) of the alternate insoluble P-sources Mussorie Rock Phosphate and Tricalcium Phosphate. Distinct and significant intergeneric differences were observed with respect to nitrogen fixation measured as Acetylene Reduction Activity (ARA) and growth potential as soluble proteins, total carbohydrate content, dry weight and total chlorophyll content in response to different concentrations of Mussorie Rock Phosphate and Tricalcium Phosphate. Both the strains showed higher soluble protein content at 20 mg P (Mussorie Rock Phosphate) that increased with time of incubation in A. variabilis. Both cyanobacteria recorded maximum Acetylene Reduction Activity at 20 mg P (Tricalcium Phosphate) followed by activity in presence of soluble phosphate (K2HPO4). The mean activity at all concentrations of insoluble phosphate (Mussorie Rock Phosphate and Tricalcium Phosphate) was more than in the presence of soluble phosphate.

Endophytic colonization and in planta nitrogen fixation by a diazotrophic Serratia sp. in rice.

PubMed

Sandhiya, G S; Sugitha, T C K; Balachandar, D; Kumar, K

2005-09-01

Nitrogen fixing endophytic Serratia sp. was isolated from rice and characterized. Re-colonization ability of Serratia sp. in the rice seedlings as endophyte was studied under laboratory condition. For detecting the re-colonization potential in the rice seedlings, Serratia sp. was marked with reporter genes (egfp and Kmr) using transposon mutagenesis. The conjugants were screened for re-colonization ability and presence of nif genes using PCR. Further, the influence of flavonoids and growth hormones on the endophytic colonization and in planta nitrogen fixation of Serratia was also investigated. The flavonoids, quercetin (3 microg/ml) and diadzein (2 microg/ml) significantly increased the re-colonization ability of the endophytic Serratia, whereas the growth hormones like IAA and NAA (5 microg/ml) reduced the endophytic colonization ability of Serratia sp. Similarly, the in planta nitrogen fixation by Serratia sp. in rice was significantly increased due to flavonoids. The inoculation of endophytic diazotrophs increased the plant biomass and biochemical constituents.

Biological nitrogen fixation and biomass accumulation within poplar clones as a result of inoculations with diazotrophic endophyte consortia.

PubMed

Knoth, Jenny L; Kim, Soo-Hyung; Ettl, Gregory J; Doty, Sharon L

2014-01-01

Sustainable production of biomass for bioenergy relies on low-input crop production. Inoculation of bioenergy crops with plant growth-promoting endophytes has the potential to reduce fertilizer inputs through the enhancement of biological nitrogen fixation (BNF). Endophytes isolated from native poplar growing in nutrient-poor conditions were selected for a series of glasshouse and field trials designed to test the overall hypothesis that naturally occurring diazotrophic endophytes impart growth promotion of the host plants. Endophyte inoculations contributed to increased biomass over uninoculated control plants. This growth promotion was more pronounced with multi-strain consortia than with single-strain inocula. Biological nitrogen fixation was estimated through (15)N isotope dilution to be 65% nitrogen derived from air (Ndfa). Phenotypic plasticity in biomass allocation and branch production observed as a result of endophyte inoculations may be useful in bioenergy crop breeding and engineering programs. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

Impact of Trichodesmium Sp. on Pacific Primary production

NASA Astrophysics Data System (ADS)

Dutheil, C.; Menkes, C.; Aumont, O.; Shiozaki, T.; Bonnet, S.; Rodier, M.; Bopp, L.; Lorrain, A.

2016-12-01

Recent sea-experiments have suggested that the South Pacific is one of the world's hot spot for nitrogen fixation. In that region, diazotrophs Trichodesmium Sp. have been shown to be one of its major contributors. Here we assess the climatological impact of these diazotrophs in the Pacific by using a 1°x1° coupled model dynamical-biogeochemical model ROMS-PISCES in which an explicit a Trichodesmium compartment is implemented. Firstly, we validate our model on the main limiting components (phosphate, iron, and temperature) of Trichodesmium growth. Phosphate patterns show modelled values and structures in qualitatively good agreement with observations. Iron concentrations are in good agreement with the observations. We also validate our model on nitrogen fixation rates. The regional spatial patterns of strong fixation are coherent with the observations. In the South Pacific, the model is able to reproduce the strong east-west gradient. Secondly, we evaluate the climatological effects of Trichodesmium on the biogeochemical conditions of the Tropical Pacific by adding with the explicit Trichodesmium compartment. The implementation of this compartment improves the model ability to reproduce the observed chlorophyll fields in the South West Pacific and the northern hemisphere, especially around Hawaii. In regions where there are strong nitrogen fixation rates, we observe an increase in the primary production by more than 100%, and an increase by more than 60 % in the production due to nanophytoplankton and diatoms, between the simulation with trichodesmium and without nitrogen fixation.

Connecting biodiversity and potential functional role in modern euxinic environments by microbial metagenomics

PubMed Central

Llorens-Marès, Tomàs; Yooseph, Shibu; Goll, Johannes; Hoffman, Jeff; Vila-Costa, Maria; Borrego, Carles M; Dupont, Chris L; Casamayor, Emilio O

2015-01-01

Stratified sulfurous lakes are appropriate environments for studying the links between composition and functionality in microbial communities and are potentially modern analogs of anoxic conditions prevailing in the ancient ocean. We explored these aspects in the Lake Banyoles karstic area (NE Spain) through metagenomics and in silico reconstruction of carbon, nitrogen and sulfur metabolic pathways that were tightly coupled through a few bacterial groups. The potential for nitrogen fixation and denitrification was detected in both autotrophs and heterotrophs, with a major role for nitrogen and carbon fixations in Chlorobiaceae. Campylobacterales accounted for a large percentage of denitrification genes, while Gallionellales were putatively involved in denitrification, iron oxidation and carbon fixation and may have a major role in the biogeochemistry of the iron cycle. Bacteroidales were also abundant and showed potential for dissimilatory nitrate reduction to ammonium. The very low abundance of genes for nitrification, the minor presence of anammox genes, the high potential for nitrogen fixation and mineralization and the potential for chemotrophic CO2 fixation and CO oxidation all provide potential clues on the anoxic zones functioning. We observed higher gene abundance of ammonia-oxidizing bacteria than ammonia-oxidizing archaea that may have a geochemical and evolutionary link related to the dominance of Fe in these environments. Overall, these results offer a more detailed perspective on the microbial ecology of anoxic environments and may help to develop new geochemical proxies to infer biology and chemistry interactions in ancient ecosystems. PMID:25575307

USE OF 15N IN THE STUDY OF FIXATION OF ATMOSPHERIC NITROGEN BY NON- NODULATED SEED PLANTS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stevenson, G.

1959-10-31

Both from observation of non-leguminous plants growing under natural conditions and also from measurements made of plot experiments with grasses it has been found that large amounts of nitrogen, of the order of 50-lb N/acre/year, accumulate both in the soil and in plant material. Measurements of the contribution made by nonsymbiotic nitrogen-fixing bacteria are only of the order of 2 to 3 lb N/acre/year, so that it appears likely that some other mechanism operates which leads to fixation of nitrogen with the growth of many nonleguminous plants. Experiments were carried out with the following species which grow well in Newmore » Zealand under poor nutrient conditions, especially as regards nitrogen: Pinus radiata, Coprosma robusta, Epilobium erectum and Dactylis glomerata. Plants have been grown in sand watered with a nitrogen-free nutrient solution when they have shown signs of nitrogen starvation, but, nevertheless, they have made considerable growth. Some plants have been exposed to an isotopically enriched atmosphere for periods of 7 to 14 days, and significant amounts of nitrogen-15 have been recovered from the combined nitrogen in the plants indicating that fixation of molecular nitrogen has occurred. The effect is not due to any of the known nonsymbiotic nitrogen-fixing bacteria which were shown to be absent from the sand cultures. Two possible explanations considered are that the effect may be due to microorganisms present in or on the plants, and that the effect may be due to some activity of the plants themselves. (auth)« less

Abiotic nitrogen fixation on terrestrial planets: reduction of NO to ammonia by FeS.

PubMed

Summers, David P; Basa, Ranor C B; Khare, Bishun; Rodoni, David

2012-02-01

Understanding the abiotic fixation of nitrogen and how such fixation can be a supply of prebiotic nitrogen is critical for understanding both the planetary evolution of, and the potential origin of life on, terrestrial planets. As nitrogen is a biochemically essential element, sources of biochemically accessible nitrogen, especially reduced nitrogen, are critical to prebiotic chemistry and the origin of life. Loss of atmospheric nitrogen can result in loss of the ability to sustain liquid water on a planetary surface, which would impact planetary habitability and hydrological processes that shape the surface. It is known that NO can be photochemically converted through a chain of reactions to form nitrate and nitrite, which can be subsequently reduced to ammonia. Here, we show that NO can also be directly reduced, by FeS, to ammonia. In addition to removing nitrogen from the atmosphere, this reaction is particularly important as a source of reduced nitrogen on an early terrestrial planet. By converting NO directly to ammonia in a single step, ammonia is formed with a higher product yield (~50%) than would be possible through the formation of nitrate/nitrite and subsequent conversion to ammonia. In conjunction with the reduction of NO, there is also a catalytic disproportionation at the mineral surface that converts NO to NO₂ and N₂O. The NO₂ is then converted to ammonia, while the N₂O is released back in the gas phase, which provides an abiotic source of nitrous oxide.

New, national bottom-up estimate for tree-based biological nitrogen fixation in the US

EPA Science Inventory

Nitrogen is a limiting nutrient in many ecosystems, but is also a chief pollutant from human activity. Quantifying human impacts on the nitrogen cycle and investigating natural ecosystem nitrogen cycling both require an understanding of the magnitude of nitrogen inputs from biolo...

Nitrogen and Martian Habitability: Insights from Five Years of Curiosity Measurements

NASA Astrophysics Data System (ADS)

Stern, J. C.; Sutter, B.; Navarro-Gonzalez, R.; McKay, C.; Ming, D. W.; Mahaffy, P. R.; Archer, D., Jr.; Franz, H. B.; Freissinet, C.; Jackson, W. A.; Conrad, P. G.; Glavin, D. P.; Trainer, M. G.; Malespin, C.; McAdam, A.; Eigenbrode, J. L.; Teinturier, S.; Manning, C.

2017-12-01

The detection of "fixed" N on Mars in the form of nitrate by the Sample Analysis at Mars (SAM) instrument suite on the Mars Science Laboratory (MSL) Curiosity Rover [1] has major implications for martian habitability. "Follow the nitrogen" has been proposed as a strategy in the search for both extant and extinct life on Mars [e.g., 2]. Nitrogen is so crucial to life on Earth that life developed metabolic pathways to break the triple bond of N2 and "fix" atmospheric nitrogen to more biologically available molecules for use in proteins and informational polymers. Sequestration of nitrate in regolith has long been predicted to contribute to the removal of N from the martian atmosphere [e.g., 3], and our detections confirm that nitrogen fixation was occurring on ancient Mars. Detections of fixed nitrogen, particularly within the context of the habitable environment in Yellowknife Bay characterized by the MSL payload, are an important tool to assess whether life ever could have existed on ancient Mars. We present 5 years of analyses and interpretation of nitrate in solid martian drilled and scooped samples by SAM on MSL. Nitrate abundance reported by SAM in situ measurements ranges from non-detection to 681 ± 304 mg/kg [1,4] in the samples examined to date. The measured abundances are consistent with nitrogen fixation via impact generated thermal shock on ancient Mars and/or dry deposition from photochemistry of thermospheric NO. We review the integration of SAM data with terrestrial Mars analog work in order to better understand the timing of nitrogen fixation and mobility of nitrogen on Mars, and thus its availability to putative biology. In particular, the relationship between nitrate and other soluble salts, such as perchlorate, may help reveal the timing of nitrogen fixation and post-depositional behavior of nitrate on Mars [4]. Finally, we present a comparison of isotopic composition (δ15N) of nitrate with δ15N of atmospheric nitrogen (δ15N ≈ 574‰, [5]), which can be used to constrain atmospheric loss of N2 and model the evolution of the atmosphere on Mars.

Iron availability influences 15N-isotope fractionation during nitrogen fixation by aerobic chemoheterotroph Azotobacter vinelandii

NASA Astrophysics Data System (ADS)

Zhang, X.; Kopf, S.; Lee, A. C.

2016-12-01

The N stable isotope composition (δ15N) of biomass provides a powerful tool for reconstructing present and past N cycling, but its interpretation hinges on a complete understanding of the isotopic signature of biological nitrogen fixation, which sets the δ15N of newly fixed N. All biological nitrogen fixation is catalyzed by the metalloenzyme nitrogenase in a complex reaction that reduces inert atmospheric N2 gas into bioavailable ammonium. Recent investigations into the metal cofactor variants of nitrogenase revealed that the canonical Mo-, and alternative V-, and Fe-only isoforms of nitrogenase impart different isotope fractionations during N2 fixation in vivo, challenging the traditional view that N2 fixation only imparts small, invariable isotope effects of 0-2‰. However, the mechanistic basis for the fractionation of N2 fixation remains largely unknown. To better understand mechanisms underlying fractionation, we varied Fe availability and measured in vivo fractionations for the aerobic chemoheterotroph Azotobacter vinelandii utilizing Mo- or V-nitrogenase under batch culture conditions. Under all iron conditions, N2 fixation based on Mo-nitrogenase yielded lower fractionations (heavier biomasss δ15N) compared to V-nitrogenase. For fractionations associated with a single metalloenzyme, higher Fe concentrations, which correlated with faster growth rates, yielded small but systematically larger fractionations ( 1 ‰ increase for Mo- and V- nitrogenases). To directly determine the effect of growth rate on fractionation, we grew Mo-nitrogenase expressing A. vinelandii in Fe-replete medium at different growth rates using chemostats and found that growth rate alone does not alter fractionation. The results indicate that Fe availability, in addition to the type of nitrogenase metalloenzyme, controls 15N fractionation during N2 fixation by A. vinelandii.

Effect of salinity on diazotrophic activity and microbial composition of phototrophic communities from Bitter-1 soda lake (Kulunda Steppe, Russia).

PubMed

Namsaraev, Zorigto; Samylina, Olga; Sukhacheva, Marina; Borisenko, Gennadii; Sorokin, Dimitry Y; Tourova, Tatiana

2018-04-16

Bitter-1 is a shallow hypersaline soda lake in Kulunda Steppe (Altai region, Russia). During a study period between 2005 and 2016, the salinity in the littoral area of the lake fluctuated within the range from 85 to 400 g/L (in July of each year). Light-dependent nitrogen fixation occurred in this lake up to the salt-saturating conditions. The rates increased with a decrease in salinity, both under environmental conditions and in laboratory simulations. The salinities below 100 g/L were favorable for light-dependent nitrogen fixation, while the process was dramatically inhibited above 200 g/L salts. The analysis of nifH genes in environmental samples and in enrichment cultures of diazotrophic phototrophs suggested that anaerobic fermenting and sulfate-reducing bacteria could participate in the dark nitrogen fixation process up to soda-saturating conditions. However, we cannot exclude the possibility that haloalkaliphilic nonheterocystous cyanobacteria (Euhalothece sp. and Geitlerinema sp.) and anoxygenic purple sulfur bacteria (Ectothiorhodospira sp.) might also play a role in the process at light conditions. The heterocystous cyanobacterium Nodularia sp. develops at low salinity (below 80 g/L) that is not characteristic for Bitter-1 Lake and thus does not make a significant contribution to the nitrogen fixation in this lake.

Novel European free-living, non-diazotrophic Bradyrhizobium isolates from contrasting soils that lack nodulation and nitrogen fixation genes - a genome comparison

NASA Astrophysics Data System (ADS)

Jones, Frances Patricia; Clark, Ian M.; King, Robert; Shaw, Liz J.; Woodward, Martin J.; Hirsch, Penny R.

2016-05-01

The slow-growing genus Bradyrhizobium is biologically important in soils, with different representatives found to perform a range of biochemical functions including photosynthesis, induction of root nodules and symbiotic nitrogen fixation and denitrification. Consequently, the role of the genus in soil ecology and biogeochemical transformations is of agricultural and environmental significance. Some isolates of Bradyrhizobium have been shown to be non-symbiotic and do not possess the ability to form nodules. Here we present the genome and gene annotations of two such free-living Bradyrhizobium isolates, named G22 and BF49, from soils with differing long-term management regimes (grassland and bare fallow respectively) in addition to carbon metabolism analysis. These Bradyrhizobium isolates are the first to be isolated and sequenced from European soil and are the first free-living Bradyrhizobium isolates, lacking both nodulation and nitrogen fixation genes, to have their genomes sequenced and assembled from cultured samples. The G22 and BF49 genomes are distinctly different with respect to size and number of genes; the grassland isolate also contains a plasmid. There are also a number of functional differences between these isolates and other published genomes, suggesting that this ubiquitous genus is extremely heterogeneous and has roles within the community not including symbiotic nitrogen fixation.

Molybdate in Rhizobial Seed-Coat Formulations Improves the Production and Nodulation of Alfalfa

PubMed Central

Zhou, Jiqiong; Deng, Bo; Zhang, Yingjun; Cobb, Adam B.; Zhang, Zhao

2017-01-01

Rhizobia-legume symbiosis is the most well researched biological nitrogen fixation system. Coating legume seeds with rhizobia is now a recognized practical measure for improving the production of legume corp. However, the efficacy of some commercial rhizobia inoculants cannot be guaranteed in China due to the low rate of live rhizobia in these products. A greenhouse experiment was conducted to assess the effects of different rhizobial inoculant formulations on alfalfa productivity and nitrogen fixation. Two rhizobia strains, (ACCC17631 and ACCC17676), that are effective partners with alfalfa variety Zhongmu No. 1 were assessed with different concentrations of ammonium molybdate in seed-coat formulations with two different coating adhesives. Our study showed that the growth, nodulation, and nitrogen fixation ability of the plants inoculated with the ACCC17631 rhizobial strain were greatest when the ammonium molybdate application was0.2% of the formulation. An ammonium molybdate concentration of 0.1% was most beneficial to the growth of the plants inoculated with the ACCC17676 rhizobial strain. The sodium carboxymethyl cellulose and sodium alginate, used as coating adhesives, did not have a significant effect on alfalfa biomass and nitrogen fixation. However, the addition of skimmed milk to the adhesive improved nitrogenase activity. These results demonstrate that a new rhizobial seed-coat formulation benefitted alfalfa nodulation and yield. PMID:28099471

Molybdate in Rhizobial Seed-Coat Formulations Improves the Production and Nodulation of Alfalfa.

PubMed

Zhou, Jiqiong; Deng, Bo; Zhang, Yingjun; Cobb, Adam B; Zhang, Zhao

2017-01-01

Rhizobia-legume symbiosis is the most well researched biological nitrogen fixation system. Coating legume seeds with rhizobia is now a recognized practical measure for improving the production of legume corp. However, the efficacy of some commercial rhizobia inoculants cannot be guaranteed in China due to the low rate of live rhizobia in these products. A greenhouse experiment was conducted to assess the effects of different rhizobial inoculant formulations on alfalfa productivity and nitrogen fixation. Two rhizobia strains, (ACCC17631 and ACCC17676), that are effective partners with alfalfa variety Zhongmu No. 1 were assessed with different concentrations of ammonium molybdate in seed-coat formulations with two different coating adhesives. Our study showed that the growth, nodulation, and nitrogen fixation ability of the plants inoculated with the ACCC17631 rhizobial strain were greatest when the ammonium molybdate application was0.2% of the formulation. An ammonium molybdate concentration of 0.1% was most beneficial to the growth of the plants inoculated with the ACCC17676 rhizobial strain. The sodium carboxymethyl cellulose and sodium alginate, used as coating adhesives, did not have a significant effect on alfalfa biomass and nitrogen fixation. However, the addition of skimmed milk to the adhesive improved nitrogenase activity. These results demonstrate that a new rhizobial seed-coat formulation benefitted alfalfa nodulation and yield.

Cyclic variations in nitrogen uptake rate of soybean plants: ammonium as a nitrogen source

NASA Technical Reports Server (NTRS)

Henry, L. T.; Raper, C. D. Jr

1989-01-01

When NO3- is the sole nitrogen source in flowing solution culture, the net rate of nitrogen uptake by nonnodulated soybean (Glycine max L. Merr. cv Ransom) plants cycles between maxima and minima with a periodicity of oscillation that corresponds with the interval of leaf emergence. Since soybean plants accumulate similar quantities of nitrogen when either NH4+ or NO3- is the sole source in solution culture controlled at pH 6.0, an experiment was conducted to determine if the oscillations in net rate of nitrogen uptake also occur when NH4+ is the nitrogen source. During a 21-day period of vegetative development, net uptake of NH4+ was measured daily by ion chromatography as depletion of NH4+ from a replenished nutrient solution containing 1.0 millimolar NH4+. The net rate of NH4+ uptake oscillated with a periodicity that was similar to the interval of leaf emergence. Instances of negative net rates of uptake indicate that the transition between maxima and minima involved changes in influx and efflux components of net NH4+ uptake.

Genome evolution and nitrogen fixation in bacterial ectosymbionts of a protist inhabiting wood-feeding cockroaches

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tai, Vera; Carpenter, Kevin J.; Weber, Peter K.

By combining genomics and isotope imaging analysis using high-resolution secondary ion mass spectrometry (NanoSIMS), we examined the function and evolution of Bacteroidales ectosymbionts of the protistBarbulanymphafrom the hindguts of the wood-eating cockroachCryptocercus punctulatus. In particular, we investigated the structure of ectosymbiont genomes, which, in contrast to those of endosymbionts, has been little studied to date, and tested the hypothesis that these ectosymbionts fix nitrogen. Unlike with most obligate endosymbionts, genome reduction has not played a major role in the evolution of the Barbulanympha ectosymbionts. Instead, interaction with the external environment has remained important for this symbiont as genes for synthesismore » of transporters, outer membrane proteins, lipopolysaccharides, and lipoproteins have been retained. The ectosymbiont genome carried two complete operons for nitrogen fixation, a urea transporter, and a urease, indicating the availability of nitrogen as a driving force behind the symbiosis. NanoSIMS analysis ofC. punctulatushindgut symbionts exposedin vivoto 15N 2 supports the hypothesis thatBarbulanymphaectosymbionts are capable of nitrogen fixation. This genomic andin vivofunctional investigation of protist ectosymbionts highlights the diversity of evolutionary forces and trajectories that shape symbiotic interactions. The ecological and evolutionary importance of symbioses is increasingly clear, but the overall diversity of symbiotic interactions remains poorly explored. Here in this study, we investigated the evolution and nitrogen fixation capabilities of ectosymbionts attached to the protist Barbulanympha from the hindgut of the wood-eating cockroach Cryptocercus punctulatus. In addressing genome evolution of protist ectosymbionts, our data suggest that the ecological pressures influencing the evolution of extracellular symbionts clearly differ from intracellular symbionts and organelles. Using NanoSIMS analysis, we also obtained direct imaging evidence of a specific hindgut microbe playing a role in nitrogen fixation. These results demonstrate the power of combining NanoSIMS and genomics tools for investigating the biology of uncultivable microbes. This investigation paves the way for a more precise understanding of microbial interactions in the hindguts of wood-eating insects and further exploration of the diversity and ecological significance of symbiosis between microbes.« less

Genome evolution and nitrogen fixation in bacterial ectosymbionts of a protist inhabiting wood-feeding cockroaches

DOE PAGES

Tai, Vera; Carpenter, Kevin J.; Weber, Peter K.; ...

2016-05-27

By combining genomics and isotope imaging analysis using high-resolution secondary ion mass spectrometry (NanoSIMS), we examined the function and evolution of Bacteroidales ectosymbionts of the protistBarbulanymphafrom the hindguts of the wood-eating cockroachCryptocercus punctulatus. In particular, we investigated the structure of ectosymbiont genomes, which, in contrast to those of endosymbionts, has been little studied to date, and tested the hypothesis that these ectosymbionts fix nitrogen. Unlike with most obligate endosymbionts, genome reduction has not played a major role in the evolution of the Barbulanympha ectosymbionts. Instead, interaction with the external environment has remained important for this symbiont as genes for synthesismore » of transporters, outer membrane proteins, lipopolysaccharides, and lipoproteins have been retained. The ectosymbiont genome carried two complete operons for nitrogen fixation, a urea transporter, and a urease, indicating the availability of nitrogen as a driving force behind the symbiosis. NanoSIMS analysis ofC. punctulatushindgut symbionts exposedin vivoto 15N 2 supports the hypothesis thatBarbulanymphaectosymbionts are capable of nitrogen fixation. This genomic andin vivofunctional investigation of protist ectosymbionts highlights the diversity of evolutionary forces and trajectories that shape symbiotic interactions. The ecological and evolutionary importance of symbioses is increasingly clear, but the overall diversity of symbiotic interactions remains poorly explored. Here in this study, we investigated the evolution and nitrogen fixation capabilities of ectosymbionts attached to the protist Barbulanympha from the hindgut of the wood-eating cockroach Cryptocercus punctulatus. In addressing genome evolution of protist ectosymbionts, our data suggest that the ecological pressures influencing the evolution of extracellular symbionts clearly differ from intracellular symbionts and organelles. Using NanoSIMS analysis, we also obtained direct imaging evidence of a specific hindgut microbe playing a role in nitrogen fixation. These results demonstrate the power of combining NanoSIMS and genomics tools for investigating the biology of uncultivable microbes. This investigation paves the way for a more precise understanding of microbial interactions in the hindguts of wood-eating insects and further exploration of the diversity and ecological significance of symbiosis between microbes.« less

Prebiotic chemistry and atmospheric warming of early Earth by an active young Sun

NASA Astrophysics Data System (ADS)

Airapetian, V. S.; Glocer, A.; Gronoff, G.; Hébrard, E.; Danchi, W.

2016-06-01

Nitrogen is a critical ingredient of complex biological molecules. Molecular nitrogen, however, which was outgassed into the Earth’s early atmosphere, is relatively chemically inert and nitrogen fixation into more chemically reactive compounds requires high temperatures. Possible mechanisms of nitrogen fixation include lightning, atmospheric shock heating by meteorites, and solar ultraviolet radiation. Here we show that nitrogen fixation in the early terrestrial atmosphere can be explained by frequent and powerful coronal mass ejection events from the young Sun--so-called superflares. Using magnetohydrodynamic simulations constrained by Kepler Space Telescope observations, we find that successive superflare ejections produce shocks that accelerate energetic particles, which would have compressed the early Earth’s magnetosphere. The resulting extended polar cap openings provide pathways for energetic particles to penetrate into the atmosphere and, according to our atmospheric chemistry simulations, initiate reactions converting molecular nitrogen, carbon dioxide and methane to the potent greenhouse gas nitrous oxide as well as hydrogen cyanide, an essential compound for life. Furthermore, the destruction of N2, CO2 and CH4 suggests that these greenhouse gases cannot explain the stability of liquid water on the early Earth. Instead, we propose that the efficient formation of nitrous oxide could explain a warm early Earth.

Response of enzymes involved in the processes of antioxidation towards benthiocarb and methylparathion in cyanobacteria Nostoc muscorum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bhunia, A.K.; Roy, D.; Basu, N.K.

1991-08-01

Recently, it has been observed in the authors' laboratory that growth, nitrogen fixation, protein content of cyanobacteria Nostoc muscorum were reduced by methylparathion and benthiocarb treatment. Though many works on toxicity of pesticides on cyanobacteria, specially on growth, photosynthesis and nitrogen fixation are available, the effects of pesticides on antioxidant enzyme levels is still unclear. In this communication, studies have been presented on the effects of organophosphate insecticide methyl-parathione and carbamate herbicide benthiocarb, on glutathione content, glutathione reductase (GR) and superoxide dismutase (SOD) activities of filamentous, nitrogen-fixing cyanobacteria Nostoc muscorum.

The impact of simulated chronic nitrogen deposition on the biomass and N₂-fixation activity of two boreal feather moss-cyanobacteria associations.

PubMed

Gundale, Michael J; Bach, Lisbet H; Nordin, Annika

2013-01-01

Bryophytes achieve substantial biomass and play several key functional roles in boreal forests that can influence how carbon (C) and nitrogen (N) cycling respond to atmospheric deposition of reactive nitrogen (Nr). They associate with cyanobacteria that fix atmospheric N₂, and downregulation of this process may offset anthropogenic Nr inputs to boreal systems. Bryophytes also promote soil C accumulation by thermally insulating soils, and changes in their biomass influence soil C dynamics. Using a unique large-scale (0.1 ha forested plots), long-term experiment (16 years) in northern Sweden where we simulated anthropogenic Nr deposition, we measured the biomass and N₂-fixation response of two bryophyte species, the feather mosses Hylocomium splendens and Pleurozium schreberi. Our data show that the biomass declined for both species; however, N₂-fixation rates per unit mass and per unit area declined only for H. splendens. The low and high treatments resulted in a 29% and 54% reduction in total feather moss biomass, and a 58% and 97% reduction in total N₂-fixation rate per unit area, respectively. These results help to quantify the sensitivity of feather moss biomass and N₂ fixation to chronic Nr deposition, which is relevant for modelling ecosystem C and N balances in boreal ecosystems.

Influence of arbuscular mycorrhizae on biomass production and nitrogen fixation of berseem clover plants subjected to water stress.

PubMed

Saia, Sergio; Amato, Gaetano; Frenda, Alfonso Salvatore; Giambalvo, Dario; Ruisi, Paolo

2014-01-01

Several studies, performed mainly in pots, have shown that arbuscular mycorrhizal symbiosis can mitigate the negative effects of water stress on plant growth. No information is available about the effects of arbuscular mycorrhizal symbiosis on berseem clover growth and nitrogen (N) fixation under conditions of water shortage. A field experiment was conducted in a hilly area of inner Sicily, Italy, to determine whether symbiosis with AM fungi can mitigate the detrimental effects of drought stress (which in the Mediterranean often occurs during the late period of the growing season) on forage yield and symbiotic N2 fixation of berseem clover. Soil was either left under water stress (i.e., rain-fed conditions) or the crop was well-watered. Mycorrhization treatments consisted of inoculation of berseem clover seeds with arbuscular mycorrhizal spores or suppression of arbuscular mycorrhizal symbiosis by means of fungicide treatments. Nitrogen biological fixation was assessed using the 15N-isotope dilution technique. Arbuscular mycorrhizal symbiosis was able to mitigate the negative effect of water stress on berseem clover grown in a typical semiarid Mediterranean environment. In fact, under water stress conditions, arbuscular mycorrhizal symbiosis resulted in increases in total biomass, N content, and N fixation, whereas no effect of crop mycorrhization was observed in the well-watered treatment.

Interaction of root nodule size and oxygen pressure on the rate of nitrogen fixation by cowpea and peanut

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sen, D.; Weaver, R.W.

1987-04-01

Size and anatomical features of nodules influence the rate of O/sub 2/ diffusion into nodules. Availability of oxygen can be a limiting factor in nitrogen fixation. Larger nodules have thicker cortices and low surface to volume ratio leading to lower rates of gaseous diffusion. Increased oxygen pressure in the environment alters the rate of nitrogen fixation but the rate of change may depend on the nodule size. This was investigated by measuring /sup 15/N/sub 2/ incorporation into nodules. Root nodules from 38 day old cowpea and peanut plants were collected and sorted into size groups having diameters of >3 mm,more » 2-3 mm, and just below 2 mm. Samples of each size group were enclosed in tubes and exposed to various combination of oxygen (8-28%) and /sup 15/N/sub 2/. With higher O/sub 2/ pressure all nodules showed increased N/sub 2/ fixation but the largest nodules showed the maximum increase. Specific activity of larger nodules was higher for N/sub 2/ fixation. For the sizes of nodules examined the largest nodules did not reflect any of the disadvantages of the large size but the benefits of higher rates of O/sub 2/ entry was evident.« less

Role of PII proteins in nitrogen fixation control of Herbaspirillum seropedicae strain SmR1

PubMed Central

2011-01-01

Background The PII protein family comprises homotrimeric proteins which act as transducers of the cellular nitrogen and carbon status in prokaryotes and plants. In Herbaspirillum seropedicae, two PII-like proteins (GlnB and GlnK), encoded by the genes glnB and glnK, were identified. The glnB gene is monocistronic and its expression is constitutive, while glnK is located in the nlmAglnKamtB operon and is expressed under nitrogen-limiting conditions. Results In order to determine the involvement of the H. seropedicae glnB and glnK gene products in nitrogen fixation, a series of mutant strains were constructed and characterized. The glnK- mutants were deficient in nitrogen fixation and they were complemented by plasmids expressing the GlnK protein or an N-truncated form of NifA. The nitrogenase post-translational control by ammonium was studied and the results showed that the glnK mutant is partially defective in nitrogenase inactivation upon addition of ammonium while the glnB mutant has a wild-type phenotype. Conclusions Our results indicate that GlnK is mainly responsible for NifA activity regulation and ammonium-dependent post-translational regulation of nitrogenase in H. seropedicae. PMID:21223584

Role of PII proteins in nitrogen fixation control of Herbaspirillum seropedicae strain SmR1.

PubMed

Noindorf, Lilian; Bonatto, Ana C; Monteiro, Rose A; Souza, Emanuel M; Rigo, Liu U; Pedrosa, Fabio O; Steffens, Maria B R; Chubatsu, Leda S

2011-01-11

The PII protein family comprises homotrimeric proteins which act as transducers of the cellular nitrogen and carbon status in prokaryotes and plants. In Herbaspirillum seropedicae, two PII-like proteins (GlnB and GlnK), encoded by the genes glnB and glnK, were identified. The glnB gene is monocistronic and its expression is constitutive, while glnK is located in the nlmAglnKamtB operon and is expressed under nitrogen-limiting conditions. In order to determine the involvement of the H. seropedicae glnB and glnK gene products in nitrogen fixation, a series of mutant strains were constructed and characterized. The glnK- mutants were deficient in nitrogen fixation and they were complemented by plasmids expressing the GlnK protein or an N-truncated form of NifA. The nitrogenase post-translational control by ammonium was studied and the results showed that the glnK mutant is partially defective in nitrogenase inactivation upon addition of ammonium while the glnB mutant has a wild-type phenotype. Our results indicate that GlnK is mainly responsible for NifA activity regulation and ammonium-dependent post-translational regulation of nitrogenase in H. seropedicae.

Understanding the Red Sea nutrient cycle - a first look into nitrogen fixation in the Red Sea

NASA Astrophysics Data System (ADS)

Mohamed, Roslinda; Arrieta, Jesus; Alam, Intikhab; Duarte, Carlos

2016-04-01

The Red Sea is an elongated and semi-enclosed system bordered by Africa and Saudi Arabia. Positioned in an arid, tropical zone, the system receives high solar irradiance and heat flux, extensive evaporation, low rainfall and therefore high salinity. These harsh environmental conditions has set the Red Sea to be one of the fastest warming and saltiest ecosystem in the world. Although nutrients are known to be at very low concentrations, the ultimately limiting nutrient in the system is still undefined. Therefore, like most other oligotrophic systems, we regard the Red Sea as being nitrogen-limited and we foresee nitrogen fixation as the most probable bottleneck in the Red Sea nitrogen budget. On the basis of metagenomes from pelagic microbial communities along the Red Sea, we looked into the distribution of nitrogenase, an enzyme involved in nitrogen fixation, in this system and provide a first insight into the microbial community that is involved in the process. The implications of this study will not only help improve our understanding of the Red Sea nutrient regime, but may also hint on future ocean responses to rising climates.

Iron availability limits the ocean nitrogen inventory stabilizing feedbacks between marine denitrification and nitrogen fixation

NASA Astrophysics Data System (ADS)

Moore, J. Keith; Doney, Scott C.

2007-06-01

Recent upward revisions in key sink/source terms for fixed nitrogen (N) in the oceans imply a short residence time and strong negative feedbacks involving denitrification and N fixation to prevent large swings in the ocean N inventory over timescales of a few centuries. We tested the strength of these feedbacks in a global biogeochemical elemental cycling (BEC) ocean model that includes water column denitrification and an explicit N fixing phytoplankton group. In the northern Indian Ocean and over longer timescales in the tropical Atlantic, we find strong stabilizing feedbacks that minimize changes in marine N inventory over timescales of ˜30-200 years. In these regions high atmospheric dust/iron inputs lead to phosphorus limitation of diazotrophs, and thus a tight link between N fixation and surface water N/P ratios. Maintenance of the oxygen minimum zones in these basins depends on N fixation driven export. The stabilizing feedbacks in other regions are significant but weaker owing to iron limitation of the diazotrophs. Thus Fe limitation appears to restrict the ability of N fixation to compensate for changes in denitrification in the current climate, perhaps leading the oceans to lose fixed N. We suggest that iron is the ultimate limiting nutrient leading to nitrogen being the proximate limiting nutrient over wide regions today. Iron stress was at least partially alleviated during more dusty, glacial times, leading to a higher marine N inventory, increased export production, and perhaps widespread phosphorus limitation of the phytoplankton community. The increased efficiency of the biological pump would have contributed to the glacial drawdown in atmospheric CO2.

Microbial photosynthesis in coral reef sediments (Heron Reef, Australia)

NASA Astrophysics Data System (ADS)

Werner, Ursula; Blazejak, Anna; Bird, Paul; Eickert, Gabriele; Schoon, Raphaela; Abed, Raeid M. M.; Bissett, Andrew; de Beer, Dirk

2008-03-01

We investigated microphytobenthic photosynthesis at four stations in the coral reef sediments at Heron Reef, Australia. The microphytobenthos was dominated by diatoms, dinoflagellates and cyanobacteria, as indicated by biomarker pigment analysis. Conspicuous algae firmly attached to the sand grains (ca. 100 μm in diameter, surrounded by a hard transparent wall) were rich in peridinin, a marker pigment for dinoflagellates, but also showed a high diversity based on cyanobacterial 16S rDNA gene sequence analysis. Specimens of these algae that were buried below the photic zone exhibited an unexpected stimulation of respiration by light, resulting in an increase of local oxygen concentrations upon darkening. Net photosynthesis of the sediments varied between 1.9 and 8.5 mmol O 2 m -2 h -1 and was strongly correlated with Chl a content, which lay between 31 and 84 mg m -2. An estimate based on our spatially limited dataset indicates that the microphytobenthic production for the entire reef is in the order of magnitude of the production estimated for corals. Photosynthesis stimulated calcification at all investigated sites (0.2-1.0 mmol Ca 2+ m -2 h -1). The sediments of at least three stations were net calcifying. Sedimentary N 2-fixation rates (measured by acetylene reduction assays at two sites) ranged between 0.9 to 3.9 mmol N 2 m -2 h -1 and were highest in the light, indicating the importance of heterocystous cyanobacteria. In coral fingers no N 2-fixation was measurable, which stresses the importance of the sediment compartment for reef nitrogen cycling.

Global terrestrial carbon and nitrogen cycling insensitive to estimates of biological N fixation

NASA Astrophysics Data System (ADS)

Steinkamp, J.; Weber, B.; Werner, C.; Hickler, T.

2015-12-01

Dinitrogen (N2) is the most abundant molecule in the atmosphere and incorporated in other molecules an essential nutrient for life on earth. However, only few natural processes can initiate a reaction of N2. These natural processes are fire, lightning and biological nitrogen fixation (BNF) with BNF being the largest source. In the course of the last century humans have outperformed the natural processes of nitrogen fixation by the production of fertilizer. Industrial and other human emission of reactive nitrogen, as well as fire and lightning lead to a deposition of 63 Tg (N) per year. This is twice the amount of BNF estimated by the default setup of the dynamic global vegetation model LPJ-GUESS (30 Tg), which is a conservative approach. We use different methods and parameterizations for BNF in LPJ-GUESS: 1.) varying total annual amount; 2.) annual evenly distributed and daily calculated fixation rates; 3.) an improved dataset of BNF by cryptogamic covers (free-living N-fixers). With this setup BNF is ranging from 30 Tg to 60 Tg. We assess the impact of BNF on carbon storage and grand primary production (GPP) of the natural vegetation. These results are compared to and evaluated against available independent datasets. We do not see major differences in the productivity and carbon stocks with these BNF estimates, suggesting that natural vegetation is insensitive to BNF on a global scale and the vegetation can compensate for the different nitrogen availabilities. Current deposition of nitrogen compounds and internal cycling through mineralization and uptake is sufficient for natural vegetation productivity. However, due to the coarse model grid and spatial heterogeneity in the real world this conclusion does not exclude the existence of habitats constrained by BNF.

Synergistic Effects of Nitrogen and Potassium on Quantitative Limitations to Photosynthesis in Rice ( Oryza sativa L.).

PubMed

Hou, Wenfeng; Yan, Jinyao; Jákli, Bálint; Lu, Jianwei; Ren, Tao; Cong, Rihuan; Li, Xiaokun

2018-05-23

The inhibition of the net CO 2 assimilation ( A) during photosynthesis is one of the major physiological effects of both nitrogen (N) and potassium (K) deficiencies on rice growth. Whether the reduction in A arises from a limitation in either the diffusion and biochemical fixation of CO 2 or photochemical energy conversion is still debated in relation to N and K deficiencies. In this study, the gas exchange parameters of rice under different N and K levels were evaluated and limitations within the photosynthetic carbon capture process were quantified. A was increased by 17.3 and 12.1% for the supply of N and K, respectively. The suitable N/K ratio should be maintained from 1.42 to 1.50. The limitation results indicated that A is primarily limited by the biochemical process. The stomatal conductance ( L S ), mesophyll conductance ( L M ), and biochemical ( L B ) limitations were regulated by 26.6-79.9, 24.4-54.1, and 44.1-75.2%, respectively, with the N and K supply.

Bacteria and the Nitrogen Economy.

ERIC Educational Resources Information Center

Ayanaba, A.

1982-01-01

Biological nitrogen fixation accounts for almost 70 percent of nitrogen for plant growth. If food is to keep abreast of population growth, even more nitrogen must be fixed. For this international research institutes continue the search for natural variants in the bacterial population while also pursuing novel genetic engineering methods. (Author)

Hydrologic controls on nitrogen availability in a high-latitude, semi-arid floodplain.

Treesearch

Nicholas J. Lisuzzo; Knut Kielland; Jeremy B. Jones

2008-01-01

Past research shows a discrepancy between apparent nitrogen supply and the annual growth requirements for early successional plant communities along the Tanana River floodplain in interior Alaska. Because previous measurements of nitrogen fixation, mineralization, and deposition can only account for approximately 26% of these communities' nitrogen requirements,...

Effects of Water and Nitrogen Availability on Nitrogen Contribution by the Legume, Lupinus argenteus Pursh.

USDA-ARS?s Scientific Manuscript database

Nitrogen-fixing species contribute to ecosystem nitrogen budgets, but background resource levels influence nodulation, fixation, and plant growth. We conducted a greenhouse experiment to examine the separate and interacting effects of water and N availability on biomass production, tissue N concentr...

Aerobic and anaerobic nitrogen transformation processes in N2-fixing cyanobacterial aggregates

PubMed Central

Klawonn, Isabell; Bonaglia, Stefano; Brüchert, Volker; Ploug, Helle

2015-01-01

Colonies of N2-fixing cyanobacteria are key players in supplying new nitrogen to the ocean, but the biological fate of this fixed nitrogen remains poorly constrained. Here, we report on aerobic and anaerobic microbial nitrogen transformation processes that co-occur within millimetre-sized cyanobacterial aggregates (Nodularia spumigena) collected in aerated surface waters in the Baltic Sea. Microelectrode profiles showed steep oxygen gradients inside the aggregates and the potential for nitrous oxide production in the aggregates' anoxic centres. 15N-isotope labelling experiments and nutrient analyses revealed that N2 fixation, ammonification, nitrification, nitrate reduction to ammonium, denitrification and possibly anaerobic ammonium oxidation (anammox) can co-occur within these consortia. Thus, N. spumigena aggregates are potential sites of nitrogen gain, recycling and loss. Rates of nitrate reduction to ammonium and N2 were limited by low internal nitrification rates and low concentrations of nitrate in the ambient water. Presumably, patterns of N-transformation processes similar to those observed in this study arise also in other phytoplankton colonies, marine snow and fecal pellets. Anoxic microniches, as a pre-condition for anaerobic nitrogen transformations, may occur within large aggregates (⩾1 mm) even when suspended in fully oxygenated waters, whereas anoxia in small aggregates (<1 to ⩾0.1 mm) may only arise in low-oxygenated waters (⩽25 μM). We propose that the net effect of aggregates on nitrogen loss is negligible in NO3−-depleted, fully oxygenated (surface) waters. In NO3−-enriched (>1.5 μM), O2-depleted water layers, for example, in the chemocline of the Baltic Sea or the oceanic mesopelagic zone, aggregates may promote N-recycling and -loss processes. PMID:25575306

Aerobic and anaerobic nitrogen transformation processes in N2-fixing cyanobacterial aggregates.

PubMed

Klawonn, Isabell; Bonaglia, Stefano; Brüchert, Volker; Ploug, Helle

2015-06-01

Colonies of N(2)-fixing cyanobacteria are key players in supplying new nitrogen to the ocean, but the biological fate of this fixed nitrogen remains poorly constrained. Here, we report on aerobic and anaerobic microbial nitrogen transformation processes that co-occur within millimetre-sized cyanobacterial aggregates (Nodularia spumigena) collected in aerated surface waters in the Baltic Sea. Microelectrode profiles showed steep oxygen gradients inside the aggregates and the potential for nitrous oxide production in the aggregates' anoxic centres. (15)N-isotope labelling experiments and nutrient analyses revealed that N(2) fixation, ammonification, nitrification, nitrate reduction to ammonium, denitrification and possibly anaerobic ammonium oxidation (anammox) can co-occur within these consortia. Thus, N. spumigena aggregates are potential sites of nitrogen gain, recycling and loss. Rates of nitrate reduction to ammonium and N(2) were limited by low internal nitrification rates and low concentrations of nitrate in the ambient water. Presumably, patterns of N-transformation processes similar to those observed in this study arise also in other phytoplankton colonies, marine snow and fecal pellets. Anoxic microniches, as a pre-condition for anaerobic nitrogen transformations, may occur within large aggregates (⩾1 mm) even when suspended in fully oxygenated waters, whereas anoxia in small aggregates (<1 to ⩾0.1 mm) may only arise in low-oxygenated waters (⩽25 μM). We propose that the net effect of aggregates on nitrogen loss is negligible in NO(3)(-)-depleted, fully oxygenated (surface) waters. In NO(3)(-)-enriched (>1.5 μM), O(2)-depleted water layers, for example, in the chemocline of the Baltic Sea or the oceanic mesopelagic zone, aggregates may promote N-recycling and -loss processes.

Nitrogen mass balance in the Brazilian Amazon: an update.

PubMed

Martinelli, L A; Pinto, A S; Nardoto, G B; Ometto, J P H B; Filoso, S; Coletta, L D; Ravagnani, E C

2012-08-01

The main purpose of this study is to perform a nitrogen budget survey for the entire Brazilian Amazon region. The main inputs of nitrogen to the region are biological nitrogen fixation occurring in tropical forests (7.7 Tg.yr(-1)), and biological nitrogen fixation in agricultural lands mainly due to the cultivation of a large area with soybean, which is an important nitrogen-fixing crop (1.68 Tg.yr(-1)). The input due to the use of N fertilizers (0.48 Tg.yr(-1)) is still incipient compared to the other two inputs mentioned above. The major output flux is the riverine flux, equal to 2.80 Tg.yr(-1) and export related to foodstuff, mainly the transport of soybean and beef to other parts of the country. The continuous population growth and high rate of urbanization may pose new threats to the nitrogen cycle of the region through the burning of fossil fuel and dumping of raw domestic sewage in rivers and streams of the region.

ATMOSPHERIC NITROGEN FIXATION BY METHANE-OXIDIZING BACTERIA

PubMed Central

Davis, J. B.; Coty, V. F.; Stanley, J. P.

1964-01-01

Davis, J. B. (Socony Mobil Oil Co., Inc., Dallas, Tex.), V. F. Coty, and J. P. Stanley. Atmospheric nitrogen fixation by methane-oxidizing bacteria. J. Bacteriol. 88:468–472. 1964.—Methane-oxidizing bacteria capable of fixing atmospheric nitrogen were isolated from garden soil, pond mud, oil field soil, and soil exposed to natural gas, indicating a rather wide prevalence in nature. This may explain the high concentration of organic nitrogen commonly found in soils exposed to gas leakage from pipelines or natural-gas seeps. Added molybdenum was a requirement for growth in a nitrogen-free mineral salts medium. All nitrogen-fixing, methane-oxidizing bacteria isolated were gram-negative, nonsporeforming, usually motile rods. Colonies were light yellow, yellow, or white. The most common isolate, which formed light-yellow colonies, is referred to as Pseudomonas methanitrificans sp. n., and is distinguished from Pseudomonas (Methanomonas) methanica by nitrogen-fixing ability and a preponderance of poly-β-hydroxybutyrate in the cellular lipid fraction. Images PMID:14203365

Assessment of free-living nitrogen fixing microorganisms for commercial nitrogen fixation. [economic analysis of ammonia production

NASA Technical Reports Server (NTRS)

Stokes, B. O.; Wallace, C. J.

1978-01-01

Ammonia production by Klebsiella pneumoniae is not economical with present strains and improving nitrogen fixation to its theoretical limits in this organism is not sufficient to achieve economic viability. Because the value of both the hydrogen produced by this organism and the methane value of the carbon source required greatly exceed the value of the ammonia formed, ammonia (fixed nitrogen) should be considered the by-product. The production of hydrogen by KLEBSIELLA or other anaerobic nitrogen fixers should receive additional study, because the activity of nitrogenase offers a significant improvement in hydrogen production. The production of fixed nitrogen in the form of cell mass by Azotobacter is also uneconomical and the methane value of the carbon substrate exceeds the value of the nitrogen fixed. Parametric studies indicate that as efficiencies approach the theoretical limits the economics may become competitive. The use of nif-derepressed microorganisms, particularly blue-green algae, may have significant potential for in situ fertilization in the environment.

Kinetics of nif Gene Expression in a Nitrogen-Fixing Bacterium

PubMed Central

Poza-Carrión, César; Jiménez-Vicente, Emilio; Navarro-Rodríguez, Mónica; Echavarri-Erasun, Carlos

2014-01-01

Nitrogen fixation is a tightly regulated trait. Switching from N2 fixation-repressing conditions to the N2-fixing state is carefully controlled in diazotrophic bacteria mainly because of the high energy demand that it imposes. By using quantitative real-time PCR and quantitative immunoblotting, we show here how nitrogen fixation (nif) gene expression develops in Azotobacter vinelandii upon derepression. Transient expression of the transcriptional activator-encoding gene, nifA, was followed by subsequent, longer-duration waves of expression of the nitrogenase biosynthetic and structural genes. Importantly, expression timing, expression levels, and NifA dependence varied greatly among the nif operons. Moreover, the exact concentrations of Nif proteins and their changes over time were determined for the first time. Nif protein concentrations were exquisitely balanced, with FeMo cofactor biosynthetic proteins accumulating at levels 50- to 100-fold lower than those of the structural proteins. Mutants lacking nitrogenase structural genes or impaired in FeMo cofactor biosynthesis showed overenhanced responses to derepression that were proportional to the degree of nitrogenase activity impairment, consistent with the existence of at least two negative-feedback regulatory mechanisms. The first such mechanism responded to the levels of fixed nitrogen, whereas the second mechanism appeared to respond to the levels of the mature NifDK component. Altogether, these findings provide a framework to engineer N2 fixation in nondiazotrophs. PMID:24244007

Kinetics of Nif gene expression in a nitrogen-fixing bacterium.

PubMed

Poza-Carrión, César; Jiménez-Vicente, Emilio; Navarro-Rodríguez, Mónica; Echavarri-Erasun, Carlos; Rubio, Luis M

2014-02-01

Nitrogen fixation is a tightly regulated trait. Switching from N2 fixation-repressing conditions to the N2-fixing state is carefully controlled in diazotrophic bacteria mainly because of the high energy demand that it imposes. By using quantitative real-time PCR and quantitative immunoblotting, we show here how nitrogen fixation (nif) gene expression develops in Azotobacter vinelandii upon derepression. Transient expression of the transcriptional activator-encoding gene, nifA, was followed by subsequent, longer-duration waves of expression of the nitrogenase biosynthetic and structural genes. Importantly, expression timing, expression levels, and NifA dependence varied greatly among the nif operons. Moreover, the exact concentrations of Nif proteins and their changes over time were determined for the first time. Nif protein concentrations were exquisitely balanced, with FeMo cofactor biosynthetic proteins accumulating at levels 50- to 100-fold lower than those of the structural proteins. Mutants lacking nitrogenase structural genes or impaired in FeMo cofactor biosynthesis showed overenhanced responses to derepression that were proportional to the degree of nitrogenase activity impairment, consistent with the existence of at least two negative-feedback regulatory mechanisms. The first such mechanism responded to the levels of fixed nitrogen, whereas the second mechanism appeared to respond to the levels of the mature NifDK component. Altogether, these findings provide a framework to engineer N2 fixation in nondiazotrophs.

The Activity of Nodules of the Supernodulating Mutant Mtsunn Is not Limited by Photosynthesis under Optimal Growth Conditions

PubMed Central

Cabeza, Ricardo A.; Lingner, Annika; Liese, Rebecca; Sulieman, Saad; Senbayram, Mehmet; Tränkner, Merle; Dittert, Klaus; Schulze, Joachim

2014-01-01

Legumes match the nodule number to the N demand of the plant. When a mutation in the regulatory mechanism deprives the plant of that ability, an excessive number of nodules are formed. These mutants show low productivity in the fields, mainly due to the high carbon burden caused through the necessity to supply numerous nodules. The objective of this study was to clarify whether through optimal conditions for growth and CO2 assimilation a higher nodule activity of a supernodulating mutant of Medicago truncatula (M. truncatula) can be induced. Several experimental approaches reveal that under the conditions of our experiments, the nitrogen fixation of the supernodulating mutant, designated as sunn (super numeric nodules), was not limited by photosynthesis. Higher specific nitrogen fixation activity could not be induced through short- or long-term increases in CO2 assimilation around shoots. Furthermore, a whole plant P depletion induced a decline in nitrogen fixation, however this decline did not occur significantly earlier in sunn plants, nor was it more intense compared to the wild-type. However, a distinctly different pattern of nitrogen fixation during the day/night cycles of the experiment indicates that the control of N2 fixing activity of the large number of nodules is an additional problem for the productivity of supernodulating mutants. PMID:24727372

Relevance of ammonium oxidation within biological soil crust communities

USGS Publications Warehouse

Johnson, S.L.; Budinoff, C.R.; Belnap, J.; Garcia-Pichel, F.

2005-01-01

Thin, vertically structured topsoil communities that become ecologically important in arid regions (biological soil crusts or BSCs) are responsible for much of the nitrogen inputs into pristine arid lands. We studied N2 fixation and ammonium oxidation (AO) at subcentimetre resolution within BSCs from the Colorado Plateau. Pools of dissolved porewater nitrate/ nitrite, ammonium and organic nitrogen in wetted BSCs were high in comparison with those typical of aridosoils. They remained stable during incubations, indicating that input and output processes were of similar magnitude. Areal N2 fixation rates (6.5-48 ??mol C2H2 m-2 h -1) were high, the vertical distribution of N2 fixation peaking close to the surface if populations of heterocystous cyanobacteria were present, but in the subsurface if they were absent. Areal AO rates (19-46 ??mol N m-2 h-1) were commensurate with N2 fixation inputs. When considering oxygen availability, AO activity invariably peaked 2-3 mm deep and was limited by oxygen (not ammonium) supply. Most probable number (MPN)-enumerated ammonia-oxidizing bacteria (6.7-7.9 ?? 103 cells g-1 on average) clearly peaked at 2-3 mm depth. Thus, AO (hence nitrification) is a spatially restricted but important process in the nitrogen cycling of BSC, turning much of the biologically fixed nitrogen into oxidized forms, the fate of which remains to be determined.

Nitrogen Fixation in Thermophilic Chemosynthetic Microbial Communities Depending on Hydrogen, Sulfate, and Carbon Dioxide

PubMed Central

Nishihara, Arisa; Haruta, Shin; McGlynn, Shawn E.; Thiel, Vera; Matsuura, Katsumi

2018-01-01

The activity of nitrogen fixation measured by acetylene reduction was examined in chemosynthetic microbial mats at 72–75°C in slightly-alkaline sulfidic hot springs in Nakabusa, Japan. Nitrogenase activity markedly varied from sampling to sampling. Nitrogenase activity did not correlate with methane production, but was detected in samples showing methane production levels less than the maximum amount, indicating a possible redox dependency of nitrogenase activity. Nitrogenase activity was not affected by 2-bromo-ethane sulfonate, an inhibitor of methanogenesis. However, it was inhibited by the addition of molybdate, an inhibitor of sulfate reduction and sulfur disproportionation, suggesting the involvement of sulfate-reducing or sulfur-disproportionating organisms. Nitrogenase activity was affected by different O2 concentrations in the gas phase, again supporting the hypothesis of a redox potential dependency, and was decreased by the dispersion of mats with a homogenizer. The loss of activity that occurred from dispersion was partially recovered by the addition of H2, sulfate, and carbon dioxide. These results suggested that the observed activity of nitrogen fixation was related to chemoautotrophic sulfate reducers, and fixation may be active in a limited range of ambient redox potential. Since thermophilic chemosynthetic communities may resemble ancient microbial communities before the appearance of photosynthesis, the present results may be useful when considering the ancient nitrogen cycle on earth. PMID:29367473

A model using marginal efficiency of investment to analyse carbon and nitrogen interactions in forested ecosystems

NASA Astrophysics Data System (ADS)

Thomas, R. Q.; Williams, M.

2014-12-01

Carbon (C) and nitrogen (N) cycles are coupled in terrestrial ecosystems through multiple processes including photosynthesis, tissue allocation, respiration, N fixation, N uptake, and decomposition of litter and soil organic matter. Capturing the constraint of N on terrestrial C uptake and storage has been a focus of the Earth System modelling community. Here we explore the trade-offs and sensitivities of allocating C and N to different tissues in order to optimize the productivity of plants using a new, simple model of ecosystem C-N cycling and interactions (ACONITE). ACONITE builds on theory related to plant economics in order to predict key ecosystem properties (leaf area index, leaf C:N, N fixation, and plant C use efficiency) based on the optimization of the marginal change in net C or N uptake associated with a change in allocation of C or N to plant tissues. We simulated and evaluated steady-state and transient ecosystem stocks and fluxes in three different forest ecosystems types (tropical evergreen, temperate deciduous, and temperate evergreen). Leaf C:N differed among the three ecosystem types (temperate deciduous < tropical evergreen < temperature evergreen), a result that compared well to observations from a global database describing plant traits. Gross primary productivity (GPP) and net primary productivity (NPP) estimates compared well to observed fluxes at the simulation sites. A sensitivity analysis revealed that parameterization of the relationship between leaf N and leaf respiration had the largest influence on leaf area index and leaf C:N. Also, a widely used linear leaf N-respiration relationship did not yield a realistic leaf C:N, while a more recently reported non-linear relationship simulated leaf C:N that compared better to the global trait database than the linear relationship. Overall, our ability to constrain leaf area index and allow spatially and temporally variable leaf C:N can help address challenges simulating these properties in ecosystem and Earth System models. Furthermore, the simple approach with emergent properties based on coupled C-N dynamics has potential for use in research that uses data-assimilation methods to integrate data on both the C and N cycles to improve C flux forecasts.

Resolution of Conflicting Signals at the Single-Cell Level in the Regulation of Cyanobacterial Photosynthesis and Nitrogen Fixation.

PubMed

Mohr, Wiebke; Vagner, Tomas; Kuypers, Marcel M M; Ackermann, Martin; Laroche, Julie

2013-01-01

Unicellular, diazotrophic cyanobacteria temporally separate dinitrogen (N2) fixation and photosynthesis to prevent inactivation of the nitrogenase by oxygen. This temporal segregation is regulated by a circadian clock with oscillating activities of N2 fixation in the dark and photosynthesis in the light. On the population level, this separation is not always complete, since the two processes can overlap during transitions from dark to light. How do single cells avoid inactivation of nitrogenase during these periods? One possibility is that phenotypic heterogeneity in populations leads to segregation of the two processes. Here, we measured N2 fixation and photosynthesis of individual cells using nanometer-scale secondary ion mass spectrometry (nanoSIMS) to assess both processes in a culture of the unicellular, diazotrophic cyanobacterium Crocosphaera watsonii during a dark-light and a continuous light phase. We compared single-cell rates with bulk rates and gene expression profiles. During the regular dark and light phases, C. watsonii exhibited the temporal segregation of N2 fixation and photosynthesis commonly observed. However, N2 fixation and photosynthesis were concurrently measurable at the population level during the subjective dark phase in which cells were kept in the light rather than returned to the expected dark phase. At the single-cell level, though, cells discriminated against either one of the two processes. Cells that showed high levels of photosynthesis had low nitrogen fixing activities, and vice versa. These results suggest that, under ambiguous environmental signals, single cells discriminate against either photosynthesis or nitrogen fixation, and thereby might reduce costs associated with running incompatible processes in the same cell.

Resolution of Conflicting Signals at the Single-Cell Level in the Regulation of Cyanobacterial Photosynthesis and Nitrogen Fixation

PubMed Central

Mohr, Wiebke; Vagner, Tomas; Kuypers, Marcel M. M.; Ackermann, Martin; LaRoche, Julie

2013-01-01

Unicellular, diazotrophic cyanobacteria temporally separate dinitrogen (N2) fixation and photosynthesis to prevent inactivation of the nitrogenase by oxygen. This temporal segregation is regulated by a circadian clock with oscillating activities of N2 fixation in the dark and photosynthesis in the light. On the population level, this separation is not always complete, since the two processes can overlap during transitions from dark to light. How do single cells avoid inactivation of nitrogenase during these periods? One possibility is that phenotypic heterogeneity in populations leads to segregation of the two processes. Here, we measured N2 fixation and photosynthesis of individual cells using nanometer-scale secondary ion mass spectrometry (nanoSIMS) to assess both processes in a culture of the unicellular, diazotrophic cyanobacterium Crocosphaera watsonii during a dark-light and a continuous light phase. We compared single-cell rates with bulk rates and gene expression profiles. During the regular dark and light phases, C. watsonii exhibited the temporal segregation of N2 fixation and photosynthesis commonly observed. However, N2 fixation and photosynthesis were concurrently measurable at the population level during the subjective dark phase in which cells were kept in the light rather than returned to the expected dark phase. At the single-cell level, though, cells discriminated against either one of the two processes. Cells that showed high levels of photosynthesis had low nitrogen fixing activities, and vice versa. These results suggest that, under ambiguous environmental signals, single cells discriminate against either photosynthesis or nitrogen fixation, and thereby might reduce costs associated with running incompatible processes in the same cell. PMID:23805199

Exchange of nitrogen and phosphorus between a shallow lagoon and coastal waters

USGS Publications Warehouse

Hayn, Melanie; Howarth, Robert W.; Ganju, Neil K.; Berg, Peter; Foreman, Kenneth H.; Giblin, Anne E.; McGlathery, Karen

2014-01-01

West Falmouth Harbor, a shallow lagoon on Cape Cod, has experienced a threefold increase in nitrogen load since the mid- to late 1990s due to input from a groundwater plume contaminated by a municipal wastewater treatment plant. We measured the exchange of nitrogen and phosphorus between the harbor and the coastal waters of Buzzards Bay over several years when the harbor was experiencing this elevated nitrogen load. During summer months, the harbor not only retained the entire watershed nitrogen load but also had a net import of nitrogen from Buzzards Bay. During the spring and fall, the harbor had a net export of nitrogen to Buzzards Bay. We did not measure the export in winter, but assuming the winter net export was less than 112 % of the load, the harbor exported less than half of the watershed nitrogen load on an annual basis. For phosphorus, the harbor had a net import from coastal waters in the spring and summer months and a net export in the fall. Despite the large increase in nitrogen load to the harbor, the summertime import of phosphorus from Buzzards Bay was sufficient to maintain nitrogen limitation of primary productivity during the summer. Our findings illustrate that shallow systems dominated by benthic producers have the potential to retain large terrestrial nitrogen loads when there is sufficient supply of phosphorus from exchange with coastal waters.

Response of an Alpine Tundra in the Southern Rocky Mountains to Climate Change by 2100: Projections of Water, Carbon, and Nitrogen Cycling under RCP 4.5 and RCP 8.5 Scenarios

NASA Astrophysics Data System (ADS)

Dong, Z.; Driscoll, C. T.; Hayhoe, K.; Pourmokhtarian, A.; Stoner, A. M. K.

2016-12-01

Biogeochemical cycling of water, carbon, and nitrogen in alpine tundra ecosystems are closely related to the water and nutrient supply and ecosystem function of watersheds. While studies on the response of alpine tundra to climate change have largely focused on ecosystem structure, research on response of ecosystem function and element cycling are less well established. Using downscaled future climate scenarios under Representative Concentration Pathways (RCP) and revised algorithm of the ecosystem model, PnET-BGC, we investigated water, carbon, and nitrogen cycling of an alpine tundra ecosystem under different projections of future climate change at Saddle site of Niwot Ridge, Colorado. Simulations from this study suggest that future water supply from the alpine tundra was well predicted by the Budyko curve, which contrasts with findings from several previous studies. Although foliar display is projected to decrease due to summer water stress, an extend growing season and increasing atmospheric CO2 concentrations reverse its effects on carbon fixation by allowing longer period of photosynthesis and greater photosynthetic rate per leaf area. As a result of the increasing carbon sequestration, large increases in carbon storage are projected in living and dead biomass. Decomposition of soil organic carbon and mineralization of soil organic nitrogen increase with temperature and soil moisture, but also related to the period of snow cover which likely enhances microbial activity and associated soil decomposition and N immobilization. Future increase in winter precipitation leads to increasing snow water content which increases spring soil moisture and decomposition. Shorter future snow cover period and decreased summer soil moisture caused lower decomposition in both seasons, therefore negligible long-term pattern is projected. Future net N mineralization generally followed the pattern of organic carbon decomposition, but slightly increased because of decreasing winter immobilization due to projected shorter snow cover period. Nitrogen uptake is projected to be higher under radiative forcing scenarios of higher primary production and greater net N mineralization.

Study on the effect of magnetic field treatment of newly isolated Paenibacillus sp.

PubMed

Li, Jie; Yi, Yanli; Cheng, Xilei; Zhang, Dageng; Irfan, Muhammad

2015-12-01

Symbiotic nitrogen fixation in plants occurs in roots with the help of some bacteria which help in soil nitrogen fertility management. Isolation of significant environment friendly bacteria for nitrogen fixation is very important to enhance yield in plants. In this study effect of different magnetic field intensity and treatment time was studied on the morphology, physiology and nitrogen fixing capacity of newly isolated Paenibaccilus sp. from brown soil. The bacterium was identified by 16S rDNA sequence having highest similarity (99%) with Paenibacillus sp as revealed by BLAST. Different magnetic intensities such as 100mT, 300mT and 500mT were applied with processing time of 0, 5, 10, 20 and 30 minutes. Of all these treatment 300mT with processing time of 10 minutes was found to be most suitable treatment. Results revealed that magnetic treatment improve the growth rate with shorter generation time leading to increased enzyme activities (catalase, peroxidase and superoxide dismutase) and nitrogen fixing efficiencies. High magnetic field intensity (500mT) caused ruptured cell morphology and decreased enzyme activities which lead to less nitrogen fixation. It is concluded that appropriate magnetic field intensity and treatment time play a vital role in the growth of soil bacteria which increases the nitrogen fixing ability which affects the yield of plant. These results were very helpful in future breading programs to enhance the yield of soybean.

Light-dark (12:12) cycle of carbon and nitrogen metabolism in Crocosphaera watsonii WH8501: relation to the cell cycle.

PubMed

Dron, Anthony; Rabouille, Sophie; Claquin, Pascal; Le Roy, Bertrand; Talec, Amélie; Sciandra, Antoine

2012-04-01

This study provides with original data sets on the physiology of the unicellular diazotrophic cyanobacterium Crocosphaera watsonii WH8501, maintained in continuous culture in conditions of obligate diazotrophy. Cultures were exposed to a 12:12 light-dark regime, representative of what they experience in nature and where growth is expected to be balanced. Nitrogen and carbon metabolism were monitored at high frequency and their dynamics was compared with the cell cycle. Results reveal a daily cycle in the physiological and biochemical parameters, tightly constrained by the timely decoupled processes of N(2) fixation and carbon acquisition. The cell division rate increased concomitantly to carbon accumulation and peaked 6 h into the light. The carbon content reached a maximum at the end of the light phase. N(2) fixation occurred mostly during the dark period and peaked between 9 and 10 h into the night, while DNA synthesis, reflected by DNA fluorescence, increased until the end of the night. Consequently, cells in G1- and S-phases present a marked decrease in their C:N ratio. Nitrogen acquisition through N(2) fixation exceeded 1.3- to 3-fold the nitrogen requirements for growth, suggesting that important amounts of nitrogen are excreted even under conditions supposed to favour balanced, carbon and nitrogen acquisitions. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.

Dinitrogen fixation associated with shoots of aquatic carnivorous plants: is it ecologically important?

PubMed Central

Sirová, Dagmara; Šantrůček, Jiří; Adamec, Lubomír; Bárta, Jiří; Borovec, Jakub; Pech, Jiří; Owens, Sarah M.; Šantrůčková, Hana; Schäufele, Rudi; Štorchová, Helena; Vrba, Jaroslav

2014-01-01

Background and Aims Rootless carnivorous plants of the genus Utricularia are important components of many standing waters worldwide, as well as suitable model organisms for studying plant–microbe interactions. In this study, an investigation was made of the importance of microbial dinitrogen (N2) fixation in the N acquisition of four aquatic Utricularia species and another aquatic carnivorous plant, Aldrovanda vesiculosa. Methods 16S rRNA amplicon sequencing was used to assess the presence of micro-organisms with known ability to fix N2. Next-generation sequencing provided information on the expression of N2 fixation-associated genes. N2 fixation rates were measured following 15N2-labelling and were used to calculate the plant assimilation rate of microbially fixed N2. Key Results Utricularia traps were confirmed as primary sites of N2 fixation, with up to 16 % of the plant-associated microbial community consisting of bacteria capable of fixing N2. Of these, rhizobia were the most abundant group. Nitrogen fixation rates increased with increasing shoot age, but never exceeded 1·3 μmol N g–1 d. mass d–1. Plant assimilation rates of fixed N2 were detectable and significant, but this fraction formed less than 1 % of daily plant N gain. Although trap fluid provides conditions favourable for microbial N2 fixation, levels of nif gene transcription comprised <0·01 % of the total prokaryotic transcripts. Conclusions It is hypothesized that the reason for limited N2 fixation in aquatic Utricularia, despite the large potential capacity, is the high concentration of NH4-N (2·0–4·3 mg L–1) in the trap fluid. Resulting from fast turnover of organic detritus, it probably inhibits N2 fixation in most of the microorganisms present. Nitrogen fixation is not expected to contribute significantly to N nutrition of aquatic carnivorous plants under their typical growth conditions; however, on an annual basis the plant–microbe system can supply nitrogen in the order of hundreds of mg m–2 into the nutrient-limited littoral zone, where it may thus represent an important N source. PMID:24817095

The impact of simulated chronic nitrogen deposition on the biomass and N2-fixation activity of two boreal feather moss–cyanobacteria associations

PubMed Central

Gundale, Michael J.; Bach, Lisbet H.; Nordin, Annika

2013-01-01

Bryophytes achieve substantial biomass and play several key functional roles in boreal forests that can influence how carbon (C) and nitrogen (N) cycling respond to atmospheric deposition of reactive nitrogen (Nr). They associate with cyanobacteria that fix atmospheric N2, and downregulation of this process may offset anthropogenic Nr inputs to boreal systems. Bryophytes also promote soil C accumulation by thermally insulating soils, and changes in their biomass influence soil C dynamics. Using a unique large-scale (0.1 ha forested plots), long-term experiment (16 years) in northern Sweden where we simulated anthropogenic Nr deposition, we measured the biomass and N2-fixation response of two bryophyte species, the feather mosses Hylocomium splendens and Pleurozium schreberi. Our data show that the biomass declined for both species; however, N2-fixation rates per unit mass and per unit area declined only for H. splendens. The low and high treatments resulted in a 29% and 54% reduction in total feather moss biomass, and a 58% and 97% reduction in total N2-fixation rate per unit area, respectively. These results help to quantify the sensitivity of feather moss biomass and N2 fixation to chronic Nr deposition, which is relevant for modelling ecosystem C and N balances in boreal ecosystems. PMID:24196519

Interactions between nitrogen cycling and methane oxidation in the pelagic waters of the Gulf of Mexico.

NASA Astrophysics Data System (ADS)

Joye, S. B.; Weber, S.; Battles, J.; Montoya, J. P.

2014-12-01

Methane is an important greenhouse gas that plays a critical role in climate variation. Although a variety of marine methane sources and sinks have been identified, key aspects of the fate of methane in the ocean remain poorly constrained. At cold seeps in the Gulf of Mexico and elsewhere, methane is introduced into the overlying water column via fluid escape from the seabed. We quantified the fate of methane in the water column overlying seafloor cold seeps, in a brine basin, and at several control sites. Our goals were to determine the factors that regulated methane consumption and assimilation and to explore how these controlling factors varied among and between sites. In particular, we examined the impact of nitrogen availability on methane oxidation and studied the ability of methane oxidizing bacteria to fix molecular nitrogen. Methane oxidation rates were highest in the methane rich bottom waters of natural hydrocabron seeps. At these sites, inorganic nitrogen addition stimulated methane oxidation in laboratory experiments. In vitro shipboard experiments revealed that rates of methane oxidation and nitrogen fixation were correlated strongly, suggesting that nitrogen fixation may have been mediated by methanotrophic bacteria. The highest rates of methane oxidation and nitrogen fixation were observed in the deepwater above at natural hydrocarbon seeps. Rates of methane oxidation were substantial along the chemocline of a brine basin but in these ammonium-rich brines, addition of inorganic nitrogen had little impact on methane oxidation suggesting that methanotrophy in these waters were not nitrogen limited. Control sites exhibited the lowest methane concentrations and methane oxidation rates but even these waters exhibited substantial potential for methane oxidation when methane and inorganic nitrogen concentrations were increased. Together, these data suggest that the availability of inorganic nitrogen plays a critical role in regulating methane oxidation in pelagic ocean waters. Some methanotrophs may obtain a competitive advantage in nitrogen-limited oceanic environments by fixing molecular nitrogen. The importance of such "methano-diazotrophy" on a global scale warrants further investigation.

The importance of nodule CO2 fixation for the efficiency of symbiotic nitrogen fixation in pea at vegetative growth and during pod formation.

PubMed

Fischinger, Stephanie Anastasia; Schulze, Joachim

2010-05-01

Nodule CO2 fixation is of pivotal importance for N2 fixation. The process provides malate for bacteroids and oxaloacetate for nitrogen assimilation. The hypothesis of the present paper was that grain legume nodules would adapt to higher plant N demand and more restricted carbon availability at pod formation through increased nodule CO2 fixation and a more efficient N2 fixation. Growth, N2 fixation, and nodule composition during vegetative growth and at pod formation were studied in pea plants (Pisum sativum L.). In parallel experiments, 15N2 and 13CO2 uptake, as well as nodule hydrogen and CO2 release, was measured. Plants at pod formation showed higher growth rates and N2 fixation per plant when compared with vegetative growth. The specific activity of active nodules was about 25% higher at pod formation. The higher nodule activity was accompanied by higher amino acid concentration in nodules and xylem sap with a higher share of asparagine. Nodule 13CO2 fixation was increased at pod formation, both per plant and per 15N2 fixed unit. However, malate concentration in nodules was only 40% of that during vegetative growth and succinate was no longer detectable. The data indicate that increased N2 fixation at pod formation is connected with strongly increased nodule CO2 fixation. While the sugar concentration in nodules at pod formation was not altered, the concentration of organic acids, namely malate and succinate, was significantly lower. It is concluded that strategies to improve the capability of nodules to fix CO2 and form organic acids might prolong intensive N2 fixation into the later stages of pod formation and pod filling in grain legumes.

GROWTH AND METABOLISM OF INDIVIDUAL BACTERIAL CELLS UTILIZING NANOSIMS

DOE Office of Scientific and Technical Information (OSTI.GOV)

NEALSON, H. K.

This work involved the use of the Nano-SIMS Instrument at Lawrence Livermore Laboratory, in an effort to utilize this unique tool for experiments in Biology. The work consisted primarily of experiments to measure in real time, C and N fixation in cyanobacteria. The work revealed a number of the difficulties in using the nano-SIMS approach with biological material, but with collaboration from a number of individuals at USC and LLNL, major progress was made. The collaborators from LLNL were from the Chemistry Group (Dr. Peter Weber), and the Biology Group (Dr. Jennifer Pett-Ridge). In addition, there were a number ofmore » other scientists involved from LLNL. The USC group consisted of Dr. K.H. Nealson, the PI on the grant, Dr. R. Popa, a postdoctoral fellow and research associate at USC, Professor Douglas Capone, and Juliet Finze, a graduate student in biology. Two major experiments were done, both of which yielded new and exciting data. (1) We studied nitrogen and carbon fixation in Anabaena, demonstrating that fixation ofN occurred rapidly in the heterocysts, and that the fixed N was transported rapidly and completely to the vegetative cells. C fixation occurred in the vegetative cells, with labeled C remaining in these cells in support of their growth and metabolism. This work was accepted in the ISME Journal (Nature Publication), and published last month. (2) We studied nitrogen and carbon fixation in Trichodesmium, a non-heterocystous cyanobacterium that also fixes nitrogen. Interestingly, the nitrogen fixation was confined to regions within the filaments that seem to be identical to the so-called cyanophycaen granules. The fixed N is then transported to other parts of the cyanobacterium, as judged by movement of the heavy N throughout the filaments. On the basis of these very exciting results, we have applied for funding from the NSF to continue the collaboration with LLNL. The results of both studies were presented in the summer of 2007 at the Gordon Research Conference (Applied Environmental Microbiol.).« less

Acetylene reduction (nitrogen fixation) associated with corn inoculated with Spirillum.

PubMed

Barber, L E; Tjepkema, J D; Russell, S A; Evans, H J

1976-07-01

Sorghum and corn breeding lines were grown in soil in field and greenhouse experiments with and without an inoculum of N2-fixing in Spirillum strains from Brazil. Estimated rates of N2 fixation associated with field-grown corn and sorghum plants were less than 4 g of N2/ha per day. The mean estimated N2-fixation rates determined on segments of roots from corn inoculated with Spirillum and grown in the greenhouse at 24 to 27 degrees C were 15 g of N2/ha per day (16 inbreds), 25 g of N2/ha per day (six hybrids), and 165 g of N2/ha per day for one hybird which was heavily inoculated. The corresponding mean rates determined from measurements of in situ cultures of the same series of corn plants (i.e., 16 inbreds, six hybrids, and one heavily inoculated hybrid) were 0.4, 2.3, and 1.1 g of N2/ha per day, respectively. Lower rates of C2H2 reduction were associated with control corn cultures which had been treated with autoclaved Spirillum than with cultures inoculated with live Spirillum. No C2H2 reduction was detected in plant cultures treated with ammonium nitrate. Numbers of nitrogen-fixing bacteria on excised roots of corn plants increased an average of about 30-fold during an overnight preincubation period, and as a result acetylene reduction assays of root samples after preincubation failed to serve as a valid basis for estimating N2 fixation by corn in pot cultures. Plants grown without added nitrogen either with or without inoculum exhibited severe symptoms of nitrogen deficiency and in most cases produced significantly less dry weight than those supplied with fixed nitrogen. Although substantial rates of C2H2 reduction by excised corn roots were observed after preincubation under limited oxygen, the yield and nitrogen content of inoculated plants and the C2H2-reduction rates by inoculated pot cultures of corn, in situ, provided no evidence of appreciable N2 fixation.

The interactive effects of temperature and light on biological nitrogen fixation in boreal forests.

PubMed

Gundale, Michael J; Nilsson, Madeleine; Bansal, Sheel; Jäderlund, Anders

2012-04-01

Plant productivity is predicted to increase in northern latitudes as a result of climate warming; however, this may depend on whether biological nitrogen (N)-fixation also increases. We evaluated how the variation in temperature and light affects N-fixation by two boreal feather mosses, Pleurozium schreberi and Hylocomium splendens, which are the primary source of N-fixation in most boreal environments. We measured N-fixation rates 2 and 4 wk after exposure to a factorial combination of environments of normal, intermediate and high temperature (16.3, 22.0 and 30.3°C) and light (148.0, 295.7 and 517.3 μmol m(-2) s(-1)). Our results showed that P. schreberi achieved higher N-fixation rates relative to H. splendens in response to warming treatments, but that the highest warming treatment eventually caused N-fixation to decline for both species. Light strongly interacted with warming treatments, having positive effects at low or intermediate temperatures and damaging effects at high temperatures. These results suggest that climate warming may increase N-fixation in boreal forests, but that increased shading by the forest canopy or the occurrence of extreme temperature events could limit increases. They also suggest that P. schreberi may become a larger source of N in boreal forests relative to H. splendens as climate warming progresses. © 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.

Contrasting seasonal responses in dinitrogen fixation between shallow and deep-water colonies of the model coral Stylophora pistillata in the northern Red Sea.

PubMed

Bednarz, Vanessa N; Naumann, Malik S; Cardini, Ulisse; van Hoytema, Nanne; Rix, Laura; Al-Rshaidat, Mamoon M D; Wild, Christian

2018-01-01

Tropical corals are often associated with dinitrogen (N2)-fixing bacteria (diazotrophs), and seasonal changes in key environmental parameters, such as dissolved inorganic nitrogen (DIN) availability and seawater temperature, are known to affect N2 fixation in coral-microbial holobionts. Despite, then, such potential for seasonal and depth-related changes in N2 fixation in reef corals, such variation has not yet been investigated. Therefore, this study quantified seasonal (winter vs. summer) N2 fixation rates associated with the reef-building coral Stylophora pistillata collected from depths of 5, 10 and 20 m in the northern Gulf of Aqaba (Red Sea). Findings revealed that corals from all depths exhibited the highest N2 fixation rates during the oligotrophic summer season, when up to 11% of their photo-metabolic nitrogen demand (CPND) could be met by N2 fixation. While N2 fixation remained seasonally stable for deep corals (20 m), it significantly decreased for the shallow corals (5 and 10 m) during the DIN-enriched winter season, accounting for less than 2% of the corals' CPND. This contrasting seasonal response in N2 fixation across corals of different depths could be driven by 1) release rates of coral-derived organic matter, 2) the community composition of the associated diazotrophs, and/or 3) nutrient acquisition by the Symbiodinium community.

Effects of water and nitrogen availability on nitrogen contribution by the legume, Lupinus argenteus Pursh

Treesearch

Erin Goergen; Jeanne C. Chambers; Robert Blank

2009-01-01

Nitrogen-fixing species contribute to ecosystem nitrogen budgets, but background resource levels influence nodulation, fixation, and plant growth. We conducted a greenhouse experiment to examine the separate and interacting effects of water and N availability on biomass production, tissue N concentration, nodulation, nodule activity, and rhizodeposition of ...

Crops, Nitrogen, Water: Are Legumes Friend, Foe, or Misunderstood Ally?

PubMed

Adams, Mark A; Buchmann, Nina; Sprent, Janet; Buckley, Thomas N; Turnbull, Tarryn L

2018-06-01

Biological nitrogen fixation (BNF) by crop legumes reduces demand for industrial nitrogen fixation (INF). Nonetheless, rates of BNF in agriculture remain low, with strong negative feedback to BNF from reactive soil nitrogen (N) and drought. We show that breeding for yield has resulted in strong relationships between photosynthesis and leaf N in non-leguminous crops, whereas grain legumes show strong relations between leaf N and water use efficiency (WUE). We contrast these understandings with other studies that draw attention to the water costs of grain legume crops, and their potential for polluting the biosphere with N. We propose that breeding grain legumes for reduced stomatal conductance can increase WUE without compromising production or BNF. Legume crops remain a better bet than relying on INF. Copyright © 2018 Elsevier Ltd. All rights reserved.

The Role of Nitrogen Fixation in Cyanobacterial Bloom Toxicity in a Temperate, Eutrophic Lake

PubMed Central

Beversdorf, Lucas J.; Miller, Todd R.; McMahon, Katherine D.

2013-01-01

Toxic cyanobacterial blooms threaten freshwaters worldwide but have proven difficult to predict because the mechanisms of bloom formation and toxin production are unknown, especially on weekly time scales. Water quality management continues to focus on aggregated metrics, such as chlorophyll and total nutrients, which may not be sufficient to explain complex community changes and functions such as toxin production. For example, nitrogen (N) speciation and cycling play an important role, on daily time scales, in shaping cyanobacterial communities because declining N has been shown to select for N fixers. In addition, subsequent N pulses from N2 fixation may stimulate and sustain toxic cyanobacterial growth. Herein, we describe how rapid early summer declines in N followed by bursts of N fixation have shaped cyanobacterial communities in a eutrophic lake (Lake Mendota, Wisconsin, USA), possibly driving toxic Microcystis blooms throughout the growing season. On weekly time scales in 2010 and 2011, we monitored the cyanobacterial community in a eutrophic lake using the phycocyanin intergenic spacer (PC-IGS) region to determine population dynamics. In parallel, we measured microcystin concentrations, N2 fixation rates, and potential environmental drivers that contribute to structuring the community. In both years, cyanobacterial community change was strongly correlated with dissolved inorganic nitrogen (DIN) concentrations, and Aphanizomenon and Microcystis alternated dominance throughout the pre-toxic, toxic, and post-toxic phases of the lake. Microcystin concentrations increased a few days after the first significant N2 fixation rates were observed. Then, following large early summer N2 fixation events, Microcystis increased and became most abundant. Maximum microcystin concentrations coincided with Microcystis dominance. In both years, DIN concentrations dropped again in late summer, and N2 fixation rates and Aphanizomenon abundance increased before the lake mixed in the fall. Estimated N inputs from N2 fixation were large enough to supplement, or even support, the toxic Microcystis blooms. PMID:23405255

Evaluating the role of Actinobacteria in the gut of wood-feeding termites (Reticulitermes spp.)

Treesearch

Rachel A. Arango; Frederick Green III; Vina W. Yang; Joliene R. Lindholm; Nathaniel P. Chotlos; Kenneth F. Raffa

2017-01-01

Nitrogen has been shown to be a limiting nutrient across a range of xylophagous insects. These insects often rely on symbiotic microorganisms in the gut for nitrogen acquisition, via fixation of atmospheric nitrogen or break down of other available nitrogenous substances. In phylogenetically lower, wood-feeding termites, the role of nitrogen fixing bacteria has been...

[Characteristics of nitrogen fixation and denitrification in Termites Neotermes castaneus, Zootermopsis angusticollis, and Reticulitermes lucifugus].

PubMed

Golichenkov, M V; Kostina, N V; Ul'ianova, T A; Dobrovol'skaia, T G; Umarov, M M

2002-01-01

We studied specific features of microbial nitrogen fixation and denitrification in laboratory cultures of the termites Neotermes castaneus, Zootermopsis angusticollis, and Reticulitermes lucifugus, as well as in their nest materials. The nitrogenase activity in the termites was much higher than in the materials of termitarium. Denitrification was found only in the nest materials of termitarium. Studies of the bacterial community of gut nitrogen fixers Neotermes castaneus have shown the predominance of anaerobic and facultatively anaerobic bacteria that amount to up to 60% of the total number of "gut" bacteria. In the materials of termitarium, aerobic cellulose-destroying myxobacteria predominated, which are typical inhabitants of plant substrates, a food for the termite.

Growth-promoting Sphingomonas paucimobilis ZJSH1 associated with Dendrobium officinale through phytohormone production and nitrogen fixation

PubMed Central

Yang, Suijuan; Zhang, Xinghai; Cao, Zhaoyun; Zhao, Kaipeng; Wang, Sai; Chen, Mingxue; Hu, Xiufang

2014-01-01

Growth-promoting Sphingomonas paucimobilis ZJSH1, associated with Dendrobium officinale, a traditional Chinese medicinal plant, was characterized. At 90 days post-inoculation, strain ZJSH1 significantly promoted the growth of D. officinale seedlings, with increases of stems by 8.6% and fresh weight by 7.5%. Interestingly, the polysaccharide content extracted from the inoculated seedlings was 0.6% higher than that of the control. Similar growth promotion was observed with the transplants inoculated with strain ZJSH1. The mechanism of growth promotion was attributed to a combination of phytohormones and nitrogen fixation. Strain ZJSH1 was found using the Kjeldahl method to have a nitrogen fixation activity of 1.15 mg l−1, which was confirmed by sequencing of the nifH gene. Using high-performance liquid chromatography-mass spectrometry, strain ZJSH1 was found to produce various phytohormones, including salicylic acid (SA), indole-3-acetic acid (IAA), Zeatin and abscisic acid (ABA). The growth curve showed that strain ZJSH1 grew well in the seedlings, especially in the roots. Accordingly, much higher contents of SA, ABA, IAA and c-ZR were detected in the inoculated seedlings, which may play roles as both phytohormones and ‘Systemic Acquired Resistance’ drivers. Nitrogen fixation and secretion of plant growth regulators (SA, IAA, Zeatin and ABA) endow S. paucimobilis ZJSH1 with growth-promoting properties, which provides a potential for application in the commercial growth of D. officinale. PMID:25142808

ORCHIDEE-CNP: Site-Scale Evaluation against Observations from a Soil Formation Chronosequence in Hawaii

NASA Astrophysics Data System (ADS)

Goll, D. S.; Vuichard, N.; Maignan, F.; Jornet-Puig, A.; Sardans, J.; Peng, S.; Sun, Y.; Kvakić, M.; Guimberteau, M.; Guenet, B.; Zaehle, S.; Penuelas, J.; Jannssens, I.; Ciais, P.

2017-12-01

Land surface models rarely incorporate the terrestrial phosphorus cycle and its interactions with the carbon cycle, despite the extensive scientific debate about the importance of nitrogen and phosphorus supply for future land carbon uptake. We describe a representation of the terrestrial phosphorus cycle for the land surface model ORCHIDEE, and evaluate it with data from nutrient manipulation experiments along a soil formation chronosequence in Hawaii. ORCHIDEE accounts for influence of nutritional state of vegetation on tissue nutrient concentrations, photosynthesis, plant growth, biomass allocation, biochemical (phosphatase-mediated) mineralization and biological nitrogen fixation. Changes in nutrient content (quality) of litter affect the carbon use efficiency of decomposition and in return the nutrient availability to vegetation. The model explicitly accounts for root zone depletion of phosphorus as a function of root phosphorus uptake and phosphorus transport from soil to the root surface. The model captures the observed differences in the foliage stoichiometry of vegetation between an early (300yr) and a late stage (4.1 Myr) of soil development. The contrasting sensitivities of net primary productivity to the addition of either nitrogen, phosphorus or both among sites are in general reproduced by the model. As observed, the model simulates a preferential stimulation of leaf level productivity when nitrogen stress is alleviated, while leaf level productivity and leaf area index are stimulated equally when phosphorus stress is alleviated. The nutrient use efficiencies in the model are lower as observed primarily due to biases in the nutrient content and turnover of woody biomass.

Genomic features separating ten strains of Neorhizobium galegae with different symbiotic phenotypes.

PubMed

Österman, Janina; Mousavi, Seyed Abdollah; Koskinen, Patrik; Paulin, Lars; Lindström, Kristina

2015-05-02

The symbiotic phenotype of Neorhizobium galegae, with strains specifically fixing nitrogen with either Galega orientalis or G. officinalis, has made it a target in research on determinants of host specificity in nitrogen fixation. The genomic differences between representative strains of the two symbiovars are, however, relatively small. This introduced a need for a dataset representing a larger bacterial population in order to make better conclusions on characteristics typical for a subset of the species. In this study, we produced draft genomes of eight strains of N. galegae having different symbiotic phenotypes, both with regard to host specificity and nitrogen fixation efficiency. These genomes were analysed together with the previously published complete genomes of N. galegae strains HAMBI 540T and HAMBI 1141. The results showed that the presence of an additional rpoN sigma factor gene in the symbiosis gene region is a characteristic specific to symbiovar orientalis, required for nitrogen fixation. Also the nifQ gene was shown to be crucial for functional symbiosis in both symbiovars. Genome-wide analyses identified additional genes characteristic of strains of the same symbiovar and of strains having similar plant growth promoting properties on Galega orientalis. Many of these genes are involved in transcriptional regulation or in metabolic functions. The results of this study confirm that the only symbiosis-related gene that is present in one symbiovar of N. galegae but not in the other is an rpoN gene. The specific function of this gene remains to be determined, however. New genes that were identified as specific for strains of one symbiovar may be involved in determining host specificity, while others are defined as potential determinant genes for differences in efficiency of nitrogen fixation.

Alnus peptides modify membrane porosity and induce the release of nitrogen-rich metabolites from nitrogen-fixing Frankia.

PubMed

Carro, Lorena; Pujic, Petar; Alloisio, Nicole; Fournier, Pascale; Boubakri, Hasna; Hay, Anne E; Poly, Franck; François, Philippe; Hocher, Valerie; Mergaert, Peter; Balmand, Severine; Rey, Marjolaine; Heddi, Abdelaziz; Normand, Philippe

2015-08-01

Actinorhizal plant growth in pioneer ecosystems depends on the symbiosis with the nitrogen-fixing actinobacterium Frankia cells that are housed in special root organs called nodules. Nitrogen fixation occurs in differentiated Frankia cells known as vesicles. Vesicles lack a pathway for assimilating ammonia beyond the glutamine stage and are supposed to transfer reduced nitrogen to the plant host cells. However, a mechanism for the transfer of nitrogen-fixation products to the plant cells remains elusive. Here, new elements for this metabolic exchange are described. We show that Alnus glutinosa nodules express defensin-like peptides, and one of these, Ag5, was found to target Frankia vesicles. In vitro and in vivo analyses showed that Ag5 induces drastic physiological changes in Frankia, including an increased permeability of vesicle membranes. A significant release of nitrogen-containing metabolites, mainly glutamine and glutamate, was found in N2-fixing cultures treated with Ag5. This work demonstrates that the Ag5 peptide is central for Frankia physiology in nodules and uncovers a novel cellular function for this large and widespread defensin peptide family.

Acetogenesis from H2 plus CO2 and nitrogen fixation by an endosymbiotic spirochete of a termite-gut cellulolytic protist

PubMed Central

Ohkuma, Moriya; Noda, Satoko; Hattori, Satoshi; Iida, Toshiya; Yuki, Masahiro; Starns, David; Inoue, Jun-ichi; Darby, Alistair C.; Hongoh, Yuichi

2015-01-01

Symbiotic associations of cellulolytic eukaryotic protists and diverse bacteria are common in the gut microbial communities of termites. Besides cellulose degradation by the gut protists, reductive acetogenesis from H2 plus CO2 and nitrogen fixation by gut bacteria play crucial roles in the host termites’ nutrition by contributing to the energy demand of termites and supplying nitrogen poor in their diet, respectively. Fractionation of these activities and the identification of key genes from the gut community of the wood-feeding termite Hodotermopsis sjoestedti revealed that substantial activities in the gut—nearly 60% of reductive acetogenesis and almost exclusively for nitrogen fixation—were uniquely attributed to the endosymbiotic bacteria of the cellulolytic protist in the genus Eucomonympha. The rod-shaped endosymbionts were surprisingly identified as a spirochete species in the genus Treponema, which usually exhibits a characteristic spiral morphology. The endosymbionts likely use H2 produced by the protist for these dual functions. Although H2 is known to inhibit nitrogen fixation in some bacteria, it seemed to rather stimulate this important mutualistic process. In addition, the single-cell genome analyses revealed the endosymbiont's potentials of the utilization of sugars for its energy requirement, and of the biosynthesis of valuable nutrients such as amino acids from the fixed nitrogen. These metabolic interactions are suitable for the dual functions of the endosymbiont and reconcile its substantial contributions in the gut. PMID:25979941

Metaproteomics reveals differential modes of metabolic coupling among ubiquitous oxygen minimum zone microbes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hawley, Alyse K.; Brewer, Heather M.; Norbeck, Angela D.

2014-08-05

Oxygen minimum zones (OMZs) are intrinsic water column features arising from respiratory oxygen demand during organic matter degradation in stratified marine waters. Currently OMZs are expanding due to global climate change. This expansion alters marine ecosystem function and the productivity of fisheries due to habitat compression and changes in biogeochemical cycling leading to fixed nitrogen loss and greenhouse gas production. Here we use metaproteomics to chart spatial and temporal patterns of gene expression along defined redox gradients in a seasonally anoxic fjord, Saanich Inlet to better understand microbial community responses to OMZ expansion. The expression of metabolic pathway components formore » nitrification, anaerobic ammonium oxidation (anammox), denitrification and inorganic carbon fixation predominantly co-varied with abundance and distribution patterns of Thaumarchaeota, Nitrospira, Planctomycetes and SUP05/ARCTIC96BD-19 Gammaproteobacteria. Within these groups, pathways mediating inorganic carbon fixation and nitrogen and sulfur transformations were differentially expressed across the redoxcline. Nitrification and inorganic carbon fixation pathways affiliated with Thaumarchaeota dominated dysoxic waters and denitrification, sulfur-oxidation and inorganic carbon fixation pathways affiliated with SUP05 dominated suboxic and anoxic waters. Nitrite-oxidation and anammox pathways affiliated with Nitrospina and Planctomycetes respectively, also exhibited redox partitioning between dysoxic and suboxic waters. The differential expression of these pathways under changing water column redox conditions has quantitative implications for coupled biogeochemical cycling linking different modes of inorganic carbon fixation with distributed nitrogen and sulfur-based energy metabolism extensible to coastal and open ocean OMZs.« less

[Effects of reduced nitrogen application and soybean intercropping on nitrogen balance of sugarcane field].

PubMed

Liu, Yu; Zhang, Ying; Yang, Wen-ting; Li, Zhi-xian; Guan, Ao-mei

2015-03-01

A four-year (2010-2013) field experiment was carried out to explore the effects of three planting patterns (sugarcane, soybean monoculture and sugarcane-soybean 1:2 intercropping) with two nitrogen input levels (300 and 525 kg . hm-2) on soybean nitrogen fixation, sugarcane and soybean nitrogen accumulation, and ammonia volatilization and nitrogen leaching in sugarcane field. The results showed that the soybean nitrogen fixation efficiency (NFE) of sugarcane-soybean inter-cropping was lower than that of soybean monoculture. There was no significant difference in NFE among the treatments with the two nitrogen application rates. The nitrogen application rate and inter-cropping did not remarkably affect nitrogen accumulation of sugarcane and soybean. The ammonia volatilization of the reduced nitrogen input treatment was significantly lower than that of the conventional nitrogen input treatment. Furthermore, there was no significant difference in nitrogen leaching at different nitrogen input levels and among different planting patterns. The sugarcane field nitrogen balance analysis indicated that the nitrogen application rate dominated the nitrogen budget of sugarcane field. During the four-year experiment, all treatments leaved a nitrogen surplus (from 73.10 to 400.03 kg . hm-2) , except a nitrogen deficit of 66.22 kg . hm-2 in 2011 in the treatment of sugarcane monoculture with the reduced nitrogen application. The excessive nitrogen surplus might increase the risk of nitrogen pollution in the field. In conclusion, sugarcane-soybean intercropping with reduced nitrogen application is feasible to practice in consideration of enriching the soil fertility, reducing nitrogen pollution and saving production cost in sugarcane field.

The importance of potassium in resistance to crown rot disease in alfalfa

USDA-ARS?s Scientific Manuscript database

Nitrogen, phosphate, and potassium (potash, K2O) are the three most important nutrients in alfalfa growth and development. Nitrogen fertilization is not required because alfalfa has a high rate of biological nitrogen fixation. Phosphorus and potassium are frequently applied as fertilizer, but potass...

Variable Nitrogen Fixation in Wild Populus

PubMed Central

Doty, Sharon L.; Sher, Andrew W.; Fleck, Neil D.; Khorasani, Mahsa; Bumgarner, Roger E.; Khan, Zareen; Ko, Andrew W. K.; Kim, Soo-Hyung; DeLuca, Thomas H.

2016-01-01

The microbiome of plants is diverse, and like that of animals, is important for overall health and nutrient acquisition. In legumes and actinorhizal plants, a portion of essential nitrogen (N) is obtained through symbiosis with nodule-inhabiting, N2-fixing microorganisms. However, a variety of non-nodulating plant species can also thrive in natural, low-N settings. Some of these species may rely on endophytes, microorganisms that live within plants, to fix N2 gas into usable forms. Here we report the first direct evidence of N2 fixation in the early successional wild tree, Populus trichocarpa, a non-leguminous tree, from its native riparian habitat. In order to measure N2 fixation, surface-sterilized cuttings of wild poplar were assayed using both 15N2 incorporation and the commonly used acetylene reduction assay. The 15N label was incorporated at high levels in a subset of cuttings, suggesting a high level of N-fixation. Similarly, acetylene was reduced to ethylene in some samples. The microbiota of the cuttings was highly variable, both in numbers of cultured bacteria and in genetic diversity. Our results indicated that associative N2-fixation occurred within wild poplar and that a non-uniformity in the distribution of endophytic bacteria may explain the variability in N-fixation activity. These results point to the need for molecular studies to decipher the required microbial consortia and conditions for effective endophytic N2-fixation in trees. PMID:27196608

Assessing planetary and regional nitrogen boundaries related to food security and adverse environmental impacts

NASA Astrophysics Data System (ADS)

de Vries, Wim; Kros, Hans; Kroeze, Carolien; Seitzinger, Sybil

2014-05-01

In this presentation, we first discuss the concept of -, governance interest in- and criticism on planetary boundaries, specifically with respect to the nitrogen (N) cycle. We then systematically evaluate the criticism and argue that planetary N boundaries need to include both the benefits and adverse impacts of reactive N (Nr) and the spatial variability of Nr impacts, in terms of shortage and surplus, being main arguments for not deriving such boundaries. Next, we present an holistic approach for an updated planetary N boundary by considering the need to: (i) avoid adverse impacts of elevated Nr emissions to water, air and soils, and (ii) feed the world population in an adequate way. The derivation of a planetary N boundary, in terms of anthropogenic fixation of di-nitrogen (N2) by growing legumes and production of N fertilizer, is illustrated by (i) identification of multiple threat N indicators and setting critical limits for them, (ii) back calculating critical N losses from critical limits for N indicators, while accounting for the spatial variability of indicators and their exceedance and (iii) back calculating critical N fixation rates from critical N losses. The derivation of the needed planetary N fixation is assessed from the global population, the recommended dietary N consumption per capita and the N use efficiency in the complete chain from N fixation to N consumption. Results of example applications show that the previously suggested planetary N boundary of 25% of the current value is too low in view of needed N fixation and also unnecessary in view of most environmental impacts. We also illustrate the impacts of changes in the N use efficiency on planetary boundaries in terms of critical N fixation rates.

Nitrogen fixation apparatus

DOEpatents

Chen, Hao-Lin

1984-01-01

A method and apparatus for achieving nitrogen fixation includes a volumetric electric discharge chamber. The volumetric discharge chamber provides an even distribution of an electron beam, and enables the chamber to be maintained at a controlled energy to pressure (E/p) ratio. An E/p ratio of from 5 to 15 kV/atm of O.sub.2 /cm promotes the formation of vibrationally excited N.sub.2. Atomic oxygen interacts with vibrationally excited N.sub.2 at a much quicker rate than unexcited N.sub.2, greatly improving the rate at which NO is formed.

Nitrogen fixation method and apparatus

DOEpatents

Chen, Hao-Lin

1983-01-01

A method and apparatus for achieving nitrogen fixation includes a volumetric electric discharge chamber. The volumetric discharge chamber provides an even distribution of an electron beam, and enables the chamber to be maintained at a controlled energy to pressure (E/p) ratio. An E/p ratio of from 5 to 15 kV/atm of O.sub.2 /cm promotes the formation of vibrationally excited N.sub.2. Atomic oxygen interacts with vibrationally excited N.sub.2 at a much quicker rate than unexcited N.sub.2, greatly improving the rate at which NO is formed.

Nitrogen fixation method and apparatus

DOEpatents

Chen, H.L.

1983-08-16

A method and apparatus for achieving nitrogen fixation includes a volumetric electric discharge chamber. The volumetric discharge chamber provides an even distribution of an electron beam, and enables the chamber to be maintained at a controlled energy to pressure (E/p) ratio. An E/p ratio of from 5 to 15 kV/atm of O[sub 2]/cm promotes the formation of vibrationally excited N[sub 2]. Atomic oxygen interacts with vibrationally excited N[sub 2] at a much quicker rate than unexcited N[sub 2], greatly improving the rate at which NO is formed. 1 fig.

Net nitrogen mineralization in natural ecosystems across the conterminous US

Treesearch

LeeAnna Y. Chapman; Steven G. McNulty; Ge Sun; Yang Zhang

2013-01-01

Nitrogen is the primary nutrient limiting ecosystem productivity over most of the US. Although soil nitrogen content is important, knowledge about its spatial extent at the continental scale is limited. The objective of this study was to estimate net nitrogen mineralization for the conterminous US (CONUS) using an empirical modeling approach by scaling up site level...

AUTORADIOGRAPHIC LOCALIZATION OF 13N AFTER FIXATION OF 13N-LABELED NITROGEN GAS BY A HETEROCYST-FORMING BLUE-GREEN ALGA

PubMed Central

Wolk, C. Peter; Austin, Sam M.; Bortins, John; Galonsky, Aaron

1974-01-01

13N, generated by proton bombardment of 13C powder, is rapidly and easily converted to 13N-N2, 0.01 atm pressure, ca. 10 mCi/ml, by automated Dumas combustion. 13N fixed (as 13N-N2) by algal filaments was localized by an autoradiographic technique which permits track autoradiography with isotopes having short half-lives. Our findings show directly that a minimum of about 25% of the N2 fixation by intact, aerobically grown filaments of Anabaena cylindrica is carried out by the heterocysts. If all of the N2 fixation takes place in the heterocysts, then the movement of nitrogen along the filaments can be characterized by a constant τ < ca. 5 s (cell-2). PMID:4208073

The role of non-rainfall water on physiological activation in desert biological soil crusts

NASA Astrophysics Data System (ADS)

Zheng, Jiaoli; Peng, Chengrong; Li, Hua; Li, Shuangshuang; Huang, Shun; Hu, Yao; Zhang, Jinli; Li, Dunhai

2018-01-01

Non-rainfall water (NRW, e.g. fog and dew), in addition to rainfall and snowfall, are considered important water inputs to drylands. At the same time, biological soil crusts (BSCs) are important components of drylands. However, little information is available regarding the effect of NRW inputs on BSC activation. In this study, the effects of NRW on physiological activation in three BSC successional stages, including the cyanobacteria crust stage (Crust-C), moss colonization stage (Crust-CM), and moss crust stage (Crust-M), were studied in situ. Results suggest NRW inputs hydrated and activated physiological activity (Fv/Fm, carbon exchange, and nitrogen fixation) in BSCs but led to a negative carbon balance and low rates of nitrogen fixation in BSCs. One effective NRW event could hydrate BSCs for 7 h. Following simulated rainfall, the physiological activities recovered within 3 h, and net carbon gain occurred until 3 h after hydration, whereas NRW-induced physiological recovery processes were slower and exhibited lower activities, leading to a negative carbon balance. There were significant positive correlations between NRW amounts and the recovered values of Fv/Fm in all the three BSC stages (p < .001). The thresholds for Fv/Fm activation decreased with BSC succession, and the annual effective NRW events increased with BSC succession, with values of 29.8, 89.2, and 110.7 in Crust-C, Crust-CM and Crust-M, respectively. The results suggest that moss crust and moss-cyanobacteria crust use NRW to prolong metabolic activity and reduce drought stress more efficiently than cyanobacteria crusts. Therefore, these results suggest that BSCs utilize NRW to sustain life while growth and biomass accumulation require precipitation (rainfall) events over a certain threshold.

Nitrate and Ammonium Induced Photosynthetic Suppression in N-Limited Selenastrum minutum1

PubMed Central

Birch, Douglas G.; Elrifi, Ivor R.; Turpin, David H.

1986-01-01

The effects of nitrate and ammonium addition on net and gross photosynthesis, CO2 efflux and the dissolved inorganic carbon compensation point of nitrogen-limited Selenastrum minutum Naeg. Collins (Chlorophyta) were studied. Cultures pulsed with nitrate or ammonium exhibited a marked decrease in both net and gross photosynthetic carbon fixation. During this period of suppression the specific activity of exogenous dissolved inorganic carbon decreased rapidly in comparison to control cells indicating an increase in the rate of CO2 efflux in the light. The nitrate and ammmonium induced rates of CO2 efflux were 31.0 and 33.8 micromoles CO2 per milligram chlorophyll per hour, respectively, and represented 49 and 48% of the rate of gross photosynthesis. Nitrate addition to cells at dissolved inorganic carbon compensation point caused an increase in compensation point while ammonium had no effect. In the presence of the tricarboxylic acid cycle inhibitor fluoroacetate, the nitrate-induced change in compensation point was greatly reduced suggesting the source of this CO2 was the tricarboxylic acid cycle. These results are consistent with the mechanism of N-induced photosynthetic suppression outlined by Elrifi and Turpin (1986 Plant Physiol 81: 273-279). PMID:16665097

The marine diatom and diazotroph under future climate: Role of Iron

NASA Astrophysics Data System (ADS)

Li, Xuefeng; Fonseca-batista, Debany; Brouwers, Julie; Roevros, Nathalie; Dehairs, Frank; Chou, Lei

2016-04-01

Diatoms constitute a major group of phytoplankton, accounting for one quarter of the world's net primary productivity. Diazotrophs provide the largest input of new nitrogen (N) to the ocean and control the marine N budgets. It has been shown that iron (Fe) can be the limiting factor for diatom growth, in particular, in the HNLC (High Nutrient Low Chlorophyll) regions. This trace element can also govern the development of marine diazotrophs due to the high Fe demand necessary for biological N2 fixation. Iron plays thus an essential role in governing the marine primary productivity and the efficiency of biological carbon pump. Ocean systems are undergoing continuous modifications at varying rates and magnitudes as a result of changing climate. The objectives of our research is to evaluate 1) how climate change (dust deposition, ocean warming and acidification) can affect Fe biogeochemistry and the growth of diatoms and diazotrophs, and 2) the role of Fe in the control of biological N2 fixation under future climate scenarios. Laboratory culture experiments using Chaetoceros socialis were examined at two temperatures (13°C and 18°C) and two CO2 conditions (400 μatm and 800 μatm). The present study demonstrates clearly the influence of ocean acidification on the release of Fe upon dust deposition. It also shows that dust particles could provide a readily utilizable source of Fe and other macronutrients (dissolved phosphate and silicate) for phytoplankton growth. Elevated pCO2 concentrations may have adverse impact on the diatom growth; seawater warming may cause poleward shifts in the biogeographic distribution of diatoms. The impact of Fe on the natural N2 fixation was tested via field incubation experiments using natureal phytoplankton assemblage in the Bay of Biscay and along the Iberian Margin. N2 fixation rates in oligotrophic waters were greatly stimulated through the addition of dissolved Fe compared to the control, demonstrating the limitation of N2 fixation by Fe. Numerous factors can affect the extent of N2 fixation, but a better understanding of the major controlling factors is highly required. Semi-continuous dilution culture experiments were conducted on Trichodesmium IMS-101 under future high pCO2 and warming seawater conditions. Additionally, special attention has been given to studying the effects of mineral dust deposition which is believed to promote N2 fixation through increasing Fe availability.

Climatic and Grazing Controls on Biological Soil Crust Nitrogen Fixation in Semi-arid Ecosystems

NASA Astrophysics Data System (ADS)

Schwabedissen, S. G.; Reed, S.; Lohse, K. A.; Magnuson, T. S.

2014-12-01

Nitrogen, next to water, is believed to be the main limiting resource in arid and semi-arid ecosystems. Biological soil crusts (biocrusts) -a surface community of mosses, lichens and cyanobacteria-have been found to be the main influx of "new" nitrogen (N) into many dryland ecosystems. Controls on biocrust N fixation rates include climate (temperature and moisture), phosphorus availability, and disturbance factors such as trampling, yet a systematic examination of climatic and disturbance controls on biocrusts communities is lacking. Biocrust samples were collected along an elevation gradient in the Reynolds Creek Experimental Watershed near Murphy, Idaho. Four sites were selected from a sagebrush steppe ecosystem with precipitation ranging from ≤250mm/yr to ≥1100mm/yr. Each site included 5 grazed plots and one historic exclosure plot that has been free from grazing for more than 40 years. Five samples each were collected from under plants and from interplant spaces from the grazed plots and exclosures and analyzed for potential N fixation using an acetylene reduction assay. We hypothesized that N fixation rates would be the highest in the exclosures of the two middle sites along the elevation gradient, due to the lack of disturbance and optimal temperature and moisture, respectively. As predicted, results showed higher rates of potential N fixation in exclosures than non-exclosures at a mid-elevation 8.4 ± 3.1 kg N/ha/yr in the exclosures compared to 1.8 ± 1.5 kg N/ha/yr indicating that grazing may reduce N fixation activity. Interestingly, rates were 2-5 times lower under plant canopies compared to interplant spaces at all but the highest elevation site. Findings from our study suggest that biocrust N fixation may be a dominant input of N into theses dryland systems and, in line with our hypotheses, that climate, location within the landscape, and disturbance may interact to regulate the rates of this fundamental ecosystem process.

Nitrogen isotope fractionation by alternative nitrogenases and past ocean anoxia

PubMed Central

Zhang, Xinning; Sigman, Daniel M.; Morel, François M. M.; Kraepiel, Anne M. L.

2014-01-01

Biological nitrogen fixation constitutes the main input of fixed nitrogen to Earth’s ecosystems, and its isotope effect is a key parameter in isotope-based interpretations of the N cycle. The nitrogen isotopic composition (δ15N) of newly fixed N is currently believed to be ∼–1‰, based on measurements of organic matter from diazotrophs using molybdenum (Mo)-nitrogenases. We show that the vanadium (V)- and iron (Fe)-only “alternative” nitrogenases produce fixed N with significantly lower δ15N (–6 to –7‰). An important contribution of alternative nitrogenases to N2 fixation provides a simple explanation for the anomalously low δ15N (<–2‰) in sediments from the Cretaceous Oceanic Anoxic Events and the Archean Eon. A significant role for the alternative nitrogenases over Mo-nitrogenase is also consistent with evidence of Mo scarcity during these geologic periods, suggesting an additional dimension to the coupling between the global cycles of trace elements and nitrogen. PMID:24639508

Energy filtering transmission electron microscopy immunocytochemistry and antigen retrieval of surface layer proteins from Tannerella forsythensis using microwave or autoclave heating with citraconic anhydride

PubMed Central

2012-01-01

Tannerella forsythensis (Bacteroides forsythus), an anaerobic Gram-negative species of bacteria that plays a role in the progression of periodontal disease, has a unique bacterial protein profile. It is characterized by two unique protein bands with molecular weights of more than 200 kDa. It also is known to have a typical surface layer (S-layer) consisting of regularly arrayed subunits outside the outer membrane. We examined the relationship between high molecular weight proteins and the S-layer using electron microscopic immunolabeling with chemical fixation and an antigen retrieval procedure consisting of heating in a microwave oven or autoclave with citraconic anhydride. Immunogold particles were localized clearly at the outermost cell surface. We also used energy-filtering transmission electron microscopy (EFTEM) to visualize 3, 3′-diaminobenzidine tetrahydrochloride (DAB) reaction products after microwave antigen retrieval with 1% citraconic anhydride. The three-window method for electron spectroscopic images (ESI) of nitrogen by the EFTEM reflected the presence of moieties demonstrated by the DAB reaction with horseradish peroxidase (HRP)-conjugated secondary antibodies instead of immunogold particles. The mapping patterns of net nitrogen were restricted to the outermost cell surface. PMID:22984898

Energy filtering transmission electron microscopy immunocytochemistry and antigen retrieval of surface layer proteins from Tannerella forsythensis using microwave or autoclave heating with citraconic anhydride.

PubMed

Moriguchi, K; Mitamura, Y; Iwami, J; Hasegawa, Y; Higuchi, N; Murakami, Y; Maeda, H; Yoshimura, F; Nakamura, H; Ohno, N

2012-11-01

Tannerella forsythensis (Bacteroides forsythus), an anaerobic Gram-negative species of bacteria that plays a role in the progression of periodontal disease, has a unique bacterial protein profile. It is characterized by two unique protein bands with molecular weights of more than 200 kDa. It also is known to have a typical surface layer (S-layer) consisting of regularly arrayed subunits outside the outer membrane. We examined the relationship between high molecular weight proteins and the S-layer using electron microscopic immunolabeling with chemical fixation and an antigen retrieval procedure consisting of heating in a microwave oven or autoclave with citraconic anhydride. Immunogold particles were localized clearly at the outermost cell surface. We also used energy-filtering transmission electron microscopy (EFTEM) to visualize 3, 3'-diaminobenzidine tetrahydrochloride (DAB) reaction products after microwave antigen retrieval with 1% citraconic anhydride. The three-window method for electron spectroscopic images (ESI) of nitrogen by the EFTEM reflected the presence of moieties demonstrated by the DAB reaction with horseradish peroxidase (HRP)-conjugated secondary antibodies instead of immunogold particles. The mapping patterns of net nitrogen were restricted to the outermost cell surface.

BMAA Inhibits Nitrogen Fixation in the Cyanobacterium Nostoc sp. PCC 7120

PubMed Central

Berntzon, Lotta; Erasmie, Sven; Celepli, Narin; Eriksson, Johan; Rasmussen, Ulla; Bergman, Birgitta

2013-01-01

Cyanobacteria produce a range of secondary metabolites, one being the neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA), proposed to be a causative agent of human neurodegeneration. As for most cyanotoxins, the function of BMAA in cyanobacteria is unknown. Here, we examined the effects of BMAA on the physiology of the filamentous nitrogen-fixing cyanobacterium Nostoc sp. PCC 7120. Our data show that exogenously applied BMAA rapidly inhibits nitrogenase activity (acetylene reduction assay), even at micromolar concentrations, and that the inhibition was considerably more severe than that induced by combined nitrogen sources and most other amino acids. BMAA also caused growth arrest and massive cellular glycogen accumulation, as observed by electron microscopy. With nitrogen fixation being a process highly sensitive to oxygen species we propose that the BMAA effects found here may be related to the production of reactive oxygen species, as reported for other organisms. PMID:23966039

Black Sea nitrogen cycling and the preservation of phytoplankton δ15N signals during the Holocene

NASA Astrophysics Data System (ADS)

Fulton, James M.; Arthur, Michael A.; Freeman, Katherine H.

2012-06-01

The stable isotopic compositions of bulk, clay-bound, organic, and compound-specific nitrogen were determined for mid to late Holocene Black Sea sediments from a set of box and gravity cores. The data demonstrate that cyanobacterial N2fixation provided ˜55% of phytoplankton-derived N preserved in the top 1-2 cm of the sediments. Prior to widespread agricultural and industrial development in the catchment, N2fixation was more prominent, providing 70-80% of phytoplankton N. Organic and clay-bound nitrogen fractions record different down-coreδ15N trends that reflect phytoplankton and detrital sources, respectively, and in samples with low organic matter content, the clay-bound fraction comprises up to 38% of bulk nitrogen. Compared with bulk samples, pyropheophytina (Pphe a), which is a chlorophyll a (Chl a) degradation product, provides a more accurate record of changing phytoplankton δ15N values during the Holocene. An examination of the δ15NPphe a values in light of published and new estimates of the isotopic difference between biomass and Chl a suggests that most of the preserved Pphe a was derived from eukaryotic algae, not cyanobacteria. We infer from these data that cyanobacterial biomass is rapidly recycled in the photic zone, with 15N-depleted NH4+ released during heterotrophy and assimilated by other phytoplankton. A conceptual model for N2 fixation in the Black Sea is presented, drawing upon water column nutrient and hydrographic data as well as regional climate variability to explain the proposed temporal variability in N2 fixation.

Decoupling of soil carbon and nitrogen turnover partly explains increased net ecosystem production in response to nitrogen fertilization

NASA Astrophysics Data System (ADS)

Ehtesham, Emad; Bengtson, Per

2017-04-01

During the last decade there has been an ongoing controversy regarding the extent to which nitrogen fertilization can increase carbon sequestration and net ecosystem production in forest ecosystems. The debate is complicated by the fact that increased nitrogen availability caused by nitrogen deposition has coincided with increasing atmospheric carbon dioxide concentrations. The latter could further stimulate primary production but also result in increased allocation of carbon to root exudates, which could potentially ‘prime’ the decomposition of soil organic matter. Here we show that increased input of labile carbon to forest soil caused a decoupling of soil carbon and nitrogen cycling, which was manifested as a reduction in respiration of soil organic matter that coincided with a substantial increase in gross nitrogen mineralization. An estimate of the magnitude of the effect demonstrates that the decoupling could potentially result in an increase in net ecosystem production by up to 51 kg C ha-1 day-1 in nitrogen fertilized stands during peak summer. Even if the effect is several times lower on an annual basis, the results still suggest that nitrogen fertilization can have a much stronger influence on net ecosystem production than can be expected from a direct stimulation of primary production alone.

Influence of elevated CO2 concentrations on cell division and nitrogen fixation rates in the bloom-forming cyanobacterium Nodularia spumigena

NASA Astrophysics Data System (ADS)

Czerny, J.; Ramos, J. Barcelos E.; Riebesell, U.

2009-09-01

The surface ocean absorbs large quantities of the CO2 emitted to the atmosphere from human activities. As this CO2 dissolves in seawater, it reacts to form carbonic acid. While this phenomenon, called ocean acidification, has been found to adversely affect many calcifying organisms, some photosynthetic organisms appear to benefit from increasing [CO2]. Among these is the cyanobacterium Trichodesmium, a predominant diazotroph (nitrogen-fixing) in large parts of the oligotrophic oceans, which responded with increased carbon and nitrogen fixation at elevated pCO2. With the mechanism underlying this CO2 stimulation still unknown, the question arises whether this is a common response of diazotrophic cyanobacteria. In this study we therefore investigate the physiological response of Nodularia spumigena, a heterocystous bloom-forming diazotroph of the Baltic Sea, to CO2-induced changes in seawater carbonate chemistry. N. spumigena reacted to seawater acidification/carbonation with reduced cell division rates and nitrogen fixation rates, accompanied by significant changes in carbon and phosphorus quota and elemental composition of the formed biomass. Possible explanations for the contrasting physiological responses of Nodularia compared to Trichodesmium may be found in the different ecological strategies of non-heterocystous (Trichodesmium) and heterocystous (Nodularia) cyanobacteria.

Rhizobacterial population density and nitrogen fixation in seagrass community of Gulf of Mannar, India.

PubMed

Raja, S; Thangaradjou, T; Sivakumar, K; Kannan, L

2012-11-01

Seagrass rhizosphere generally supports high bacterial population density which plays a major role in determining the nutrient cycles of the sea. Higher densities of total heterotrphic bacteria (26.3 x 10(6) CFU g(-1)), nitrogen fixing (27.3 x 10(3) CFUg(-1), ammonifying (44.66 x 10(6) MPN g(-1)) and nitrifying bacteria (42.33 X 10(6) MPN g(-1)) have been registered in the seagrass areas than the non seagrass area. In particular, all these rhizosphere microbial population was higher in Thalassia hemprichii. The rates of nitrogen fixation was recorded in the different species of seagrasses such as Enhalus acoroides (1.166 n mol g(-1) d(-1)), Halophila ovalis (0.166 n mol g(-1) d(-1)), Thalassia hemprichii(18.5 n mol g(-1) d(-1)), Cymodocea serrulata (10.5 n mol g(-1) d(-1)), Halodule uninervis (5.375 n mol g(-1) d(-1)) and Syringodium isoetifolium (0.666 n mol g(-1) d(-1)) using gas chromatography. The average nitrogen fixation by the seagrasses of Gulf of Mannar alone was estimated to be 7640.58 n mol m(-2) d(-1) and the contributions from the rhizosphere microbes will increase the quantity to many fold.

Abundances of iron-binding photosynthetic and nitrogen-fixing proteins of Trichodesmium both in culture and in situ from the North Atlantic.

PubMed

Richier, Sophie; Macey, Anna I; Pratt, Nicola J; Honey, David J; Moore, C Mark; Bibby, Thomas S

2012-01-01

Marine cyanobacteria of the genus Trichodesmium occur throughout the oligotrophic tropical and subtropical oceans, where they can dominate the diazotrophic community in regions with high inputs of the trace metal iron (Fe). Iron is necessary for the functionality of enzymes involved in the processes of both photosynthesis and nitrogen fixation. We combined laboratory and field-based quantifications of the absolute concentrations of key enzymes involved in both photosynthesis and nitrogen fixation to determine how Trichodesmium allocates resources to these processes. We determined that protein level responses of Trichodesmium to iron-starvation involve down-regulation of the nitrogen fixation apparatus. In contrast, the photosynthetic apparatus is largely maintained, although re-arrangements do occur, including accumulation of the iron-stress-induced chlorophyll-binding protein IsiA. Data from natural populations of Trichodesmium spp. collected in the North Atlantic demonstrated a protein profile similar to iron-starved Trichodesmium in culture, suggestive of acclimation towards a minimal iron requirement even within an oceanic region receiving a high iron-flux. Estimates of cellular metabolic iron requirements are consistent with the availability of this trace metal playing a major role in restricting the biomass and activity of Trichodesmium throughout much of the subtropical ocean.

Construction and simulation of the Bradyrhizobium diazoefficiens USDA110 metabolic network: a comparison between free-living and symbiotic states.

PubMed

Yang, Yi; Hu, Xiao-Pan; Ma, Bin-Guang

2017-02-28

Bradyrhizobium diazoefficiens is a rhizobium able to convert atmospheric nitrogen into ammonium by establishing mutualistic symbiosis with soybean. It has been recognized as an important parent strain for microbial agents and is widely applied in agricultural and environmental fields. In order to study the metabolic properties of symbiotic nitrogen fixation and the differences between a free-living cell and a symbiotic bacteroid, a genome-scale metabolic network of B. diazoefficiens USDA110 was constructed and analyzed. The metabolic network, iYY1101, contains 1031 reactions, 661 metabolites, and 1101 genes in total. Metabolic models reflecting free-living and symbiotic states were determined by defining the corresponding objective functions and substrate input sets, and were further constrained by high-throughput transcriptomic and proteomic data. Constraint-based flux analysis was used to compare the metabolic capacities and the effects on the metabolic targets of genes and reactions between the two physiological states. The results showed that a free-living rhizobium possesses a steady state flux distribution for sustaining a complex supply of biomass precursors while a symbiotic bacteroid maintains a relatively condensed one adapted to nitrogen-fixation. Our metabolic models may serve as a promising platform for better understanding the symbiotic nitrogen fixation of this species.

Modeling long-term changes in tundra carbon balance following wildfire, climate change, and potential nutrient addition.

PubMed

Jiang, Yueyang; Rastetter, Edward B; Shaver, Gaius R; Rocha, Adrian V; Zhuang, Qianlai; Kwiatkowski, Bonnie L

2017-01-01

To investigate the underlying mechanisms that control long-term recovery of tundra carbon (C) and nutrients after fire, we employed the Multiple Element Limitation (MEL) model to simulate 200-yr post-fire changes in the biogeochemistry of three sites along a burn severity gradient in response to increases in air temperature, CO 2 concentration, nitrogen (N) deposition, and phosphorus (P) weathering rates. The simulations were conducted for severely burned, moderately burned, and unburned arctic tundra. Our simulations indicated that recovery of C balance after fire was mainly determined by the internal redistribution of nutrients among ecosystem components (controlled by air temperature), rather than the supply of nutrients from external sources (e.g., nitrogen deposition and fixation, phosphorus weathering). Increases in air temperature and atmospheric CO 2 concentration resulted in (1) a net transfer of nutrient from soil organic matter to vegetation and (2) higher C : nutrient ratios in vegetation and soil organic matter. These changes led to gains in vegetation biomass C but net losses in soil organic C stocks. Under a warming climate, nutrients lost in wildfire were difficult to recover because the warming-induced acceleration in nutrient cycles caused further net nutrient loss from the system through leaching. In both burned and unburned tundra, the warming-caused acceleration in nutrient cycles and increases in ecosystem C stocks were eventually constrained by increases in soil C : nutrient ratios, which increased microbial retention of plant-available nutrients in the soil. Accelerated nutrient turnover, loss of C, and increasing soil temperatures will likely result in vegetation changes, which further regulate the long-term biogeochemical succession. Our analysis should help in the assessment of tundra C budgets and of the recovery of biogeochemical function following fire, which is in turn necessary for the maintenance of wildlife habitat and tundra vegetation. © 2016 by the Ecological Society of America.

Coupling of microbial nitrogen transformations and climate in sclerophyll forest soils from the Mediterranean Region of central Chile.

PubMed

Pérez, Cecilia A; Armesto, Juan J

2018-06-01

The Mediterranean region of central Chile is experiencing extensive "mega-droughts" with detrimental effects for the environment and economy of the region. In the northern hemisphere, nitrogen (N) limitation of Mediterranean ecosystems has been explained by the decoupling between N inputs and plant uptake during the dormant season. In central Chile, soils have often been considered N-rich in comparison to other Mediterranean ecosystems of the world, yet the impacts of expected intensification of seasonal drought remain unknown. In this work, we seek to disentangle patterns of microbial N transformations and their seasonal coupling with climate in the Chilean sclerophyll forest-type. We aim to assess how water limitation affects microbial N transformations, thus addressing the impact of ongoing regional climate trends on soil N status. We studied four stands of the sclerophyll forest-type in Chile. Field measurements in surface soils showed a 67% decline of free-living diazotrophic activity (DA) and 59% decrease of net N mineralization rates during the summer rainless and dormant season, accompanied by a stimulation of in-situ denitrification rates to values 70% higher than in wetter winter. Higher rates of both free-living DA and net N mineralization found during spring, provided evidence for strong coupling of these two processes during the growing season. Overall, the experimental addition of water in the field to litter samples almost doubled DA but had no effect on denitrification rates. We conclude that coupling of microbial mediated soil N transformations during the wetter growing season explains the N enrichment of sclerophyll forest soils. Expected increases in the length and intensity of the dry period, according to climate change models, reflected in the current mega-droughts may drastically reduce biological N fixation and net N mineralization, increasing at the same time denitrification rates, thereby potentially reducing long-term soil N capital. Copyright © 2017 Elsevier B.V. All rights reserved.

Carbon sequestration in soybean crop soils: the role of hydrogen-coupled CO2 fixation

NASA Astrophysics Data System (ADS)

Graham, A.; Layzell, D. B.; Scott, N. A.; Cen, Y.; Kyser, T. K.

2011-12-01

Conversion of native vegetation to agricultural land in order to support the world's growing population is a key factor contributing to global climate change. However, the extent to which agricultural activities contribute to greenhouse gas emissions compared to carbon storage is difficult to ascertain, especially for legume crops, such as soybeans. Soybean establishment often leads to an increase in N2O emissions because N-fixation leads to increased soil available N during decomposition of the low C:N legume biomass. However, soybean establishment may also reduce net greenhouse gas emissions by increasing soil fertility, plant growth, and soil carbon storage. The mechanism behind increased carbon storage, however, remains unclear. One explanation points to hydrogen coupled CO2 fixation; the process by which nitrogen fixation releases H2 into the soil system, thereby promoting chemoautotrophic carbon fixation by soil microbes. We used 13CO2 as a tracer to track the amount and fate of carbon fixed by hydrogen coupled CO2 fixation during one-year field and laboratory incubations. The objectives of the research are to 1) quantify rates of 13CO2 fixation in soil collected from a field used for long-term soybean production 2) examine the impact of H2 gas concentration on rates of 13CO2 fixation, and 3) measure changes in δ13C signature over time in 3 soil fractions: microbial biomass, light fraction, and acid stable fraction. If this newly-fixed carbon is incorporated into the acid-stable soil C fraction, it has a good chance of contributing to long-term soil C sequestration under soybean production. Soil was collected in the field both adjacent to root nodules (nodule soil) and >3cm away (root soil) and labelled with 13CO2 (1% v/v) in the presence and absence of H2 gas. After a two week labelling period, δ13C signatures already revealed differences in the four treatments of bulk soil: -17.1 for root, -17.6 for nodule, -14.2 for root + H2, and -6.1 for nodule + H2. Labelled soil was then placed in nylon mesh bags and buried in the field at a depth of 15cm in a soybean field at the Central Experiment Farm in Ottawa, Ontario. Samples will be removed at intervals of 1,2,3,6,9,12, and 15 months, and the δ13C of three soil fractions will be examined to reveal changes in carbon storage over time. Our results will provide insights into the fate of carbon fixed during hydrogen coupled CO2 fixation, and demonstrate whether this CO2 fixation can contribute to the long-term greenhouse gas balance of soybean production systems.

Constraints on nitrogen cycling at the subtropical North Pacific Station ALOHA from isotopic measurements of nitrate and particulate nitrogen

NASA Astrophysics Data System (ADS)

Casciotti, K. L.; Trull, T. W.; Glover, D. M.; Davies, D.

2008-07-01

Nitrogen supply to surface waters can play an important role in the productivity and ecology of subtropical ecosystems. As part of the Vertical Transport in the Global Ocean (VERTIGO) program, we examined the fluxes of nitrogen into and out of the euphotic zone at station ALOHA in the North Pacific Subtropical Gyre using natural abundance stable isotopic measurements of nitrate ( δN and δO), as well as sinking and suspended particulate nitrogen (δ 15N PN). Paralleling the steep gradient in nitrate concentration in the upper thermocline at ALOHA, we observed a steep gradient in δN, decreasing from a maximum of +7.1‰ at 500 meters (m) to +1.5-2.4‰ at 150 m. δO values also decreased from +3.0‰ at 300 m to +0.7-0.9‰ at 150 m. The decreases in both δN and δO require inputs of isotopically "light" nitrate to balance the upward flux of nitrate with high δN (and δO). We conclude that both nitrogen fixation and diagenetic alteration of the sinking flux contribute to the decrease in δN and δO in the upper thermocline at station ALOHA. While nitrogen fixation is required to explain the nitrogen isotope patterns, the rates of nitrogen fixation may be lower than previously estimated. By including high-resolution nitrate isotope measurements in the nitrogen isotope budget for the euphotic zone at ALOHA, we estimate that approximately 25%, rather than 50%, of export production was fueled by N 2 fixation during our study. On the other hand, this input of N 2-derived production accumulates in the upper thermocline over time, playing a significant role in subtropical nutrient cycling through maintenance of the subsurface nitrate pool. An increase in sinking δ 15N PN between 150 and 300 m, also suggests that fractionation during remineralization contributed to the low δN values observed in this depth range by introducing a subsurface nitrate source that is 0.5‰ lower in δ 15N than the particle flux exported from the euphotic zone. While the time scale of these observations are currently limited, they highlight the need for inclusion of δN measurements in a time series program to allow a broader assessment of the variations in subsurface δN values and the links between subsurface nitrate and export flux at station ALOHA.

Inorganic nitrogen availability after severe stand-replacing fire in the Greater Yellowstone ecosystem

PubMed Central

Turner, Monica G.; Smithwick, Erica A. H.; Metzger, Kristine L.; Tinker, Daniel B.; Romme, William H.

2007-01-01

Understanding ecosystem processes as they relate to wildfire and vegetation dynamics is of growing importance as fire frequency and extent increase throughout the western United States. However, the effects of severe, stand-replacing wildfires are poorly understood. We studied inorganic nitrogen pools and mineralization rates after stand-replacing wildfires in the Greater Yellowstone Ecosystem, Wyoming. After fires that burned in summer 2000, soil ammonium concentration peaked in 2001 (33 mg NH4-N· kgsoil−1); soil nitrate increased subsequently (2.7 mg NO3-N·kgsoil−1 in 2003) but was still low. However, annual net ammonification rates were largely negative from 2001 to 2004, indicating ammonium depletion. Thus, although net nitrification rates were positive, annual net nitrogen mineralization (net ammonification plus net nitrification) remained low. Aboveground net primary production (ANPP) increased from 0.25 to 1.6 Mg·ha−1·yr−1 from 2001 to 2004, but variation in ANPP among stands was not related to net nitrogen mineralization rates. Across a broader temporal gradient (stand age zero to >250 yr), negative rates of net annual ammonification were especially pronounced in the first postfire year. Laboratory incubations using 15N isotope pool dilution revealed that gross production of ammonium was reduced and ammonium consumption greatly exceeded gross production during the initial postfire years. Our results suggest a microbial nitrogen sink for several years after severe, stand-replacing fire, confirming earlier hypotheses about postdisturbance succession and nutrient cycling in cold, fire-dominated coniferous forests. Postfire forests in Yellowstone seem to be highly conservative for nitrogen, and microbial immobilization of ammonium plays a key role during early succession. PMID:17360349

Root exudates drive interspecific facilitation by enhancing nodulation and N2 fixation

PubMed Central

Li, Bai; Li, Yu-Ying; Wu, Hua-Mao; Zhang, Fang-Fang; Li, Chun-Jie; Li, Xue-Xian; Lambers, Hans; Li, Long

2016-01-01

Plant diversity in experimental systems often enhances ecosystem productivity, but the mechanisms causing this overyielding are only partly understood. Intercropping faba beans (Vicia faba L.) and maize (Zea mays L.) result in overyielding and also, enhanced nodulation by faba beans. By using permeable and impermeable root barriers in a 2-y field experiment, we show that root–root interactions between faba bean and maize significantly increase both nodulation and symbiotic N2 fixation in intercropped faba bean. Furthermore, root exudates from maize promote faba bean nodulation, whereas root exudates from wheat and barley do not. Thus, a decline of soil nitrate concentrations caused by intercropped cereals is not the sole mechanism for maize promoting faba bean nodulation. Intercropped maize also caused a twofold increase in exudation of flavonoids (signaling compounds for rhizobia) in the systems. Roots of faba bean treated with maize root exudates exhibited an immediate 11-fold increase in the expression of chalcone–flavanone isomerase (involved in flavonoid synthesis) gene together with a significantly increased expression of genes mediating nodulation and auxin response. After 35 d, faba beans treated with maize root exudate continued to show up-regulation of key nodulation genes, such as early nodulin 93 (ENOD93), and promoted nitrogen fixation. Our results reveal a mechanism for how intercropped maize promotes nitrogen fixation of faba bean, where maize root exudates promote flavonoid synthesis in faba bean, increase nodulation, and stimulate nitrogen fixation after enhanced gene expression. These results indicate facilitative root–root interactions and provide a mechanism for a positive relationship between species diversity and ecosystem productivity. PMID:27217575

Organic forms dominate hydrologic nitrogen export from a lowland tropical watershed.

PubMed

Taylor, Philip G; Wieder, William R; Weintraub, Samantha; Cohen, Sagy; Cleveland, Cory C; Townsend, Alan R

2015-05-01

Observations of high dissolved inorganic nitrogen (DIN) concentrations in stream water have reinforced the notion that primary tropical rain forests cycle nitrogen (N) in relative excess compared to phosphorus. Here we test this notion by evaluating hydrologic N export from a small watershed on the Osa Peninsula, Costa Rica, where prior research has shown multiple indicators of conservative N cycling throughout the ecosystem. We repeatedly measured a host of factors known to influence N export for one year, including stream water chemistry and upslope litterfall, soil N availability and net N processing rates, and soil solution chemistry at the surface, 15- and 50-cm depths. Contrary to prevailing assumptions about the lowland N cycle, we find that dissolved organic nitrogen (DON) averaged 85% of dissolved N export for 48 of 52 consecutive weeks. For most of the year stream water nitrate (NO3-) export was very low, which reflected minimal net N processing and DIN leaching from upslope soils. Yet, for one month in the dry season, NO3- was the major component of N export due to a combination of low flows and upslope nitrification that concentrated NO3- in stream water. Particulate organic N (PON) export was much larger than dissolved forms at 14.6 kg N x ha(-1) x yr(-1), driven by soil erosion during storms. At this rate, PON export was slightly greater than estimated inputs from free-living N fixation and atmospheric N deposition, which suggests that erosion-driven PON export could constrain ecosystem level N stocks over longer timescales. This phenomenon is complimentary to the "DON leak" hypothesis, which postulates that the long-term accumulation of ecosystem N in unpolluted ecosystems is constrained by the export of organic N independently of biological N demand. Using an established global sediment generation model, we illustrate that PON erosion may be an important vector for N loss in tropical landscapes that are geomorphically active. This study supports an emerging view that landscape geomorphology influences nutrient biogeochemistry and limitation, though more research is needed to understand the mechanisms and spatial significance of erosional N loss from terrestrial ecosystems.

Phytoplankton Productivity in an Arctic Fjord (West Greenland): Estimating Electron Requirements for Carbon Fixation and Oxygen Production

PubMed Central

Hancke, Kasper; Dalsgaard, Tage; Sejr, Mikael Kristian; Markager, Stiig; Glud, Ronnie Nøhr

2015-01-01

Accurate quantification of pelagic primary production is essential for quantifying the marine carbon turnover and the energy supply to the food web. Knowing the electron requirement (Κ) for carbon (C) fixation (Κ C) and oxygen (O2) production (Κ O2), variable fluorescence has the potential to quantify primary production in microalgae, and hereby increasing spatial and temporal resolution of measurements compared to traditional methods. Here we quantify Κ C and Κ O2 through measures of Pulse Amplitude Modulated (PAM) fluorometry, C fixation and O2 production in an Arctic fjord (Godthåbsfjorden, W Greenland). Through short- (2h) and long-term (24h) experiments, rates of electron transfer (ETRPSII), C fixation and/or O2 production were quantified and compared. Absolute rates of ETR were derived by accounting for Photosystem II light absorption and spectral light composition. Two-hour incubations revealed a linear relationship between ETRPSII and gross 14C fixation (R2 = 0.81) during light-limited photosynthesis, giving a Κ C of 7.6 ± 0.6 (mean ± S.E.) mol é (mol C)−1. Diel net rates also demonstrated a linear relationship between ETRPSII and C fixation giving a Κ C of 11.2 ± 1.3 mol é (mol C)−1 (R2 = 0.86). For net O2 production the electron requirement was lower than for net C fixation giving 6.5 ± 0.9 mol é (mol O2)−1 (R2 = 0.94). This, however, still is an electron requirement 1.6 times higher than the theoretical minimum for O2 production [i.e. 4 mol é (mol O2)−1]. The discrepancy is explained by respiratory activity and non-photochemical electron requirements and the variability is discussed. In conclusion, the bio-optical method and derived electron requirement support conversion of ETR to units of C or O2, paving the road for improved spatial and temporal resolution of primary production estimates. PMID:26218096

Contrasted Reactivity to Oxygen Tensions in Frankia sp. Strain CcI3 throughout Nitrogen Fixation and Assimilation

PubMed Central

Ghodhbane-Gtari, Faten; Hezbri, Karima; Ktari, Amir; Sbissi, Imed; Beauchemin, Nicholas; Gtari, Maher; Tisa, Louis S.

2014-01-01

Reconciling the irreconcilable is a primary struggle in aerobic nitrogen-fixing bacteria. Although nitrogenase is oxygen and reactive oxygen species-labile, oxygen tension is required to sustain respiration. In the nitrogen-fixing Frankia, various strategies have been developed through evolution to control the respiration and nitrogen-fixation balance. Here, we assessed the effect of different oxygen tensions on Frankia sp. strain CcI3 growth, vesicle production, and gene expression under different oxygen tensions. Both biomass and vesicle production were correlated with elevated oxygen levels under both nitrogen-replete and nitrogen-deficient conditions. The mRNA levels for the nitrogenase structural genes (nifHDK) were high under hypoxic and hyperoxic conditions compared to oxic conditions. The mRNA level for the hopanoid biosynthesis genes (sqhC and hpnC) was also elevated under hyperoxic conditions suggesting an increase in the vesicle envelope. Under nitrogen-deficient conditions, the hup2 mRNA levels increased with hyperoxic environment, while hup1 mRNA levels remained relatively constant. Taken together, these results indicate that Frankia protects nitrogenase by the use of multiple mechanisms including the vesicle-hopanoid barrier and increased respiratory protection. PMID:24987692

Insights into mechanisms governing forest carbon response to nitrogen deposition: a model-data comparison using observed responses to nitrogen addition

NASA Astrophysics Data System (ADS)

Thomas, R. Q.; Bonan, G. B.; Goodale, C. L.

2013-01-01

In many forest ecosystems, nitrogen (N) deposition enhances plant uptake of carbon dioxide, thus reducing climate warming from fossil fuel emissions. Therefore, accurately modeling how forest carbon (C) sequestration responds to N deposition is critical for understanding how future changes in N availability will influence climate. Here, we use observations of forest C response to N inputs along N deposition gradients and at five temperate forest sites with fertilization experiments to test and improve a~global biogeochemical model (CLM-CN 4.0). We show that the CLM-CN plant C growth response to N deposition was smaller than observed and the modeled response to N fertilization was larger than observed. A set of modifications to the CLM-CN improved the correspondence between model predictions and observational data (1) by increasing the aboveground C storage in response to historical N deposition (1850-2004) from 14 to 34 kg C per additional kg N added through deposition and (2) by decreasing the aboveground net primary productivity response to N fertilization experiments from 91 to 57 g C m-2 yr-1. Modeled growth response to N deposition was most sensitive to altering the processes that control plant N uptake and the pathways of N loss. The response to N deposition also increased with a more closed N cycle (reduced N fixation and N gas loss) and decreased when prioritizing microbial over plant uptake of soil inorganic N. The net effect of all the modifications to the CLM-CN resulted in greater retention of N deposition and a greater role of synergy between N deposition and rising atmospheric CO2 as a mechanism governing increases in temperate forest primary production over the 20th century. Overall, testing models with both the response to gradual increases in N inputs over decades (N deposition) and N pulse additions of N over multiple years (N fertilization) allows for greater understanding of the mechanisms governing C-N coupling.

Insights into mechanisms governing forest carbon response to nitrogen deposition: a model-data comparison using observed responses to nitrogen addition

NASA Astrophysics Data System (ADS)

Thomas, R. Q.; Bonan, G. B.; Goodale, C. L.

2013-06-01

In many forest ecosystems, nitrogen (N) deposition enhances plant uptake of carbon dioxide, thus reducing climate warming from fossil fuel emissions. Therefore, accurately modeling how forest carbon (C) sequestration responds to N deposition is critical for understanding how future changes in N availability will influence climate. Here, we use observations of forest C response to N inputs along N deposition gradients and at five temperate forest sites with fertilization experiments to test and improve a global biogeochemical model (CLM-CN 4.0). We show that the CLM-CN plant C growth response to N deposition was smaller than observed and the modeled response to N fertilization was larger than observed. A set of modifications to the CLM-CN improved the correspondence between model predictions and observational data (1) by increasing the aboveground C storage in response to historical N deposition (1850-2004) from 14 to 34 kg C per additional kg N added through deposition and (2) by decreasing the aboveground net primary productivity response to N fertilization experiments from 91 to 57 g C m-2 yr-1. Modeled growth response to N deposition was most sensitive to altering the processes that control plant N uptake and the pathways of N loss. The response to N deposition also increased with a more closed N cycle (reduced N fixation and N gas loss) and decreased when prioritizing microbial over plant uptake of soil inorganic N. The net effect of all the modifications to the CLM-CN resulted in greater retention of N deposition and a greater role of synergy between N deposition and rising atmospheric CO2 as a mechanism governing increases in temperate forest primary production over the 20th century. Overall, testing models with both the response to gradual increases in N inputs over decades (N deposition) and N pulse additions of N over multiple years (N fertilization) allows for greater understanding of the mechanisms governing C-N coupling.

Experimental investigation of anaerobic nitrogen fixation rates with varying pressure, temperature and metal concentration with application to the atmospheric evolution of early Earth and Mars.

NASA Astrophysics Data System (ADS)

Gupta, Prateek

2012-07-01

The atmosphere of the early Earth is thought to have been significantly different than the modern composition of 21% O2 and 78% N2, yet the planet has been clearly established as hosting microbial life as far back as 3.8 billion years ago. As such, constraining the atmospheric composition of the early Earth is fundamental to establishing a database of habitable atmospheric compositions. A similar argument can be made for the planet Mars, where nitrates have been hypothesized to exist in the subsurface. During the early period on Mars when liquid water was likely more abundant, life may have developed to take advantage of available nitrates and a biologically-driven Martian nitrogen cycle could have evolved. Early Earth atmospheric composition has been investigated numerically, but only recently has the common assumption of a pN2 different than modern been investigated. Nonetheless, these latest attempts fail to take into account a key atmospheric parameter: life. On modern Earth, nitrogen is cycled vigorously by biology. The nitrogen cycle likely operated on the early Earth, but probably differed in the metabolic processes responsible, dominantly due to the lack of abundant oxygen which stabilizes oxidized forms of N that drive de-nitrification today. Recent advances in evolutionary genomics suggest that microbial pathways that are relatively uncommon today (i.e. vanadium and iron-based nitrogen fixation) probably played important roles in the early N cycle. We quantitatively investigate in the laboratory the effects of variable pressure, temperature and metal concentration on the rates of anoxic nitrogen fixation, as possible inputs for future models investigating atmospheric evolution, and better understand the evolution of the nitrogen cycle on Earth. A common anaerobic methanogenic archaeal species with i) a fully sequenced genome, ii) all three nitrogenases (molybdenum, vanadium and iron-based) and iii) the ability to be genetically manipulated will be used as a model species. This species will be genetically modified to create knock-out mutants lacking one or more nitrogenase genes. These mutants will be used in variable pressure, temperature and metal-concentration experiments. Nitrogen fixation rate and nitrogenase gene expression will be measured using isotope dilution and quantitative polymerase chain reaction, respectively.

Tracing nitrogen accumulation in decaying wood and examining its impact on wood decomposition rate

NASA Astrophysics Data System (ADS)

Rinne, Katja T.; Rajala, Tiina; Peltoniemi, Krista; Chen, Janet; Smolander, Aino; Mäkipää, Raisa

2016-04-01

Decomposition of dead wood, which is controlled primarily by fungi is important for ecosystem carbon cycle and has potentially a significant role in nitrogen fixation via diazotrophs. Nitrogen content has been found to increase with advancing wood decay in several studies; however, the importance of this increase to decay rate and the sources of external nitrogen remain unclear. Improved knowledge of the temporal dynamics of wood decomposition rate and nitrogen accumulation in wood as well as the drivers of the two processes would be important for carbon and nitrogen models dealing with ecosystem responses to climate change. To tackle these questions we applied several analytical methods on Norway spruce logs from Lapinjärvi, Finland. We incubated wood samples (density classes from I to V, n=49) in different temperatures (from 8.5oC to 41oC, n=7). After a common seven day pre-incubation period at 14.5oC, the bottles were incubated six days in their designated temperature prior to CO2 flux measurements with GC to determine the decomposition rate. N2 fixation was measured with acetylene reduction assay after further 48 hour incubation. In addition, fungal DNA, (MiSeq Illumina) δ15N and N% composition of wood for samples incubated at 14.5oC were determined. Radiocarbon method was applied to obtain age distribution for the density classes. The asymbiotic N2 fixation rate was clearly dependent on the stage of wood decay and increased from stage I to stage IV but was substantially reduced in stage V. CO2 production was highest in the intermediate decay stage (classes II-IV). Both N2 fixation and CO2 production were highly temperature sensitive having optima in temperature 25oC and 31oC, respectively. We calculated the variation of annual levels of respiration and N2 fixation per hectare for the study site, and used the latter data together with the 14C results to determine the amount of N2 accumulated in wood in time. The proportion of total nitrogen in wood originating from N2 increased from 0.4% (class I) to 22% (V). Despite significant N inputs, N2 fixation explained only 34%-57% of the increase in wood N content of classes III-V. The DNA results indicated that mycorrhizal colonization of wood could only partially explain the remaining increase in N content. However, majority of the samples contained one or more wood decomposing fungal species that have been reported to have the capability to produce rhizomorphs or mycelial cords used for scavenging nutrients from outside sources. Assuming that the remaining increase in N content was due to fungal activity, we modelled the δ15N variation of wood from class I to V and compared the modelled and measured δ15N values (r = 0.95, p<0.05). The increase in wood nitrogen content in time was observed to have a significant, positive impact on the respiration rate (I-IV: r = 0.57, p<0.01).

Fundamental organometallic reactions: Applications on the CYBER 205

NASA Technical Reports Server (NTRS)

Rappe, A. K.

1984-01-01

Two of the most challenging problems of Organometallic chemistry (loosely defined) are pollution control with the large space velocities needed and nitrogen fixation, a process so capably done by nature and so relatively poorly done by man (industry). For a computational chemist these problems are on the fringe of what is possible with conventional computers (large models needed and accurate energetics required). A summary of the algorithmic modification needed to address these problems on a vector processor such as the CYBER 205 and a sketch of findings to date on deNOx catalysis and nitrogen fixation are presented.

Quantifying the Stress Responses of Brassica Rapa Genotypes, With Experimental Drought in Two Nitrogen Treatments

NASA Astrophysics Data System (ADS)

Hickerson, J. L.; Pleban, J. R.; Mackay, D. S.; Aston, T.; Ewers, B. E.; Weinig, C.

2014-12-01

In a greenhouse study designed to quantify and compare stress responses of four genotypes of Brassica rapa, broccolette (bro), cabbage (cab), turnip (tur), and oil, leaf water potential and net CO2 assimilations were measured. Individuals from each genotype, grown either with high or low nitrogen, were exposed to experimental drought of the same duration. One hypothesis was that the genotypes would differ significantly in their responses to periodic drought. The other hypothesis was that the nitrogen treatment versus no nitrogen treatment would play a significant role in the stress responses during drought. It would be expected that the nitrogen treated would have greater dry leaf mass. A LI-6400 XT portable photosynthesis system was used to obtain A/Ci curves (net CO2 assimilation rate versus substomatal CO2) for each treatment group. Predawn and midday water potentials were obtained throughout the hydrated and drought periods using a Model 670 pressure chamber. The dry leaf mass was significantly greater among the high nitrogen group versus the low nitrogen group for each genotype. Nitrogen and genotype were both determinants in variation of water potentials and net CO2 assimilation. Bro and cab genotypes with high nitrogen showed the highest net CO2 assimilation rates during hydration, but the assimilation rates dropped to the lowest during droughts. The water potentials for bro and cab were lower than values for tur and oil. Nitrogen treated genotypes had lower water potentials, but higher net CO2 assimilation rates. Bayesian ecophysiological modeling with the TREES model showed significant differences in trait expression, quantified in terms of differences in model parameter posteriors, among the four genotypes.

Nutrient availability and the ultimate control of the biological carbon pump in the western tropical South Pacific Ocean

NASA Astrophysics Data System (ADS)

Moutin, Thierry; Wagener, Thibaut; Caffin, Mathieu; Fumenia, Alain; Gimenez, Audrey; Baklouti, Melika; Bouruet-Aubertot, Pascale; Pujo-Pay, Mireille; Leblanc, Karine; Lefevre, Dominique; Helias Nunige, Sandra; Leblond, Nathalie; Grosso, Olivier; de Verneil, Alain

2018-05-01

Surface waters (0-200 m) of the western tropical South Pacific (WTSP) were sampled along a longitudinal 4000 km transect (OUTPACE cruise, DOI: 10.17600/15000900) during the austral summer (stratified) period (18 February to 3 April 2015) between the Melanesian Archipelago (MA) and the western part of the SP gyre (WGY). Two distinct areas were considered for the MA, the western MA (WMA), and the eastern MA (EMA). The main carbon (C), nitrogen (N), and phosphorus (P) pools and fluxes provide a basis for the characterization of the expected trend from oligotrophy to ultra-oligotrophy, and the building of first-order budgets at the daily and seasonal timescales (using climatology). Sea surface chlorophyll a well reflected the expected oligotrophic gradient with higher values obtained at WMA, lower values at WGY, and intermediate values at EMA. As expected, the euphotic zone depth, the deep chlorophyll maximum, and nitracline depth deepen from west to east. Nevertheless, phosphaclines and nitraclines did not match. The decoupling between phosphacline and nitracline depths in the MA allows for excess P to be locally provided in the upper water by winter mixing. We found a significant biological soft tissue carbon pump in the MA sustained almost exclusively by dinitrogen (N2) fixation and essentially controlled by phosphate availability in this iron-rich environment. The MA appears to be a net sink for atmospheric CO2, while the WGY is in quasi-steady state. We suggest that the necessary excess P, allowing the success of nitrogen fixers and subsequent carbon production and export, is mainly brought to the upper surface by local deep winter convection at an annual timescale rather than by surface circulation. While the origin of the decoupling between phosphacline and nitracline remains uncertain, the direct link between local P upper water enrichment, N2 fixation, and organic carbon production and export, offers a possible shorter timescale than previously thought between N input by N2 fixation and carbon export. The low iron availability in the SP gyre and P availability in the MA during the stratified period may appear as the ultimate control of N input by N2 fixation. Because of the huge volume of water to consider, and because the SP Ocean is the place of intense denitrification in the east (N sink) and N2 fixation in the west (N source), precise seasonal C, N, P, and iron (Fe) budgets would be of prime interest to understand the efficiency, at the present time and in the future, of the oceanic biological carbon pump.

Isotopic Biomarkers of Nitrogenase Metalloenzymes: Forging Links Between the Cycles of Nitrogen and Trace Metals

NASA Astrophysics Data System (ADS)

Zhang, X.; McRose, D. L.; Darnajoux, R.; Bellenger, J. P.; Kraepiel, A. M. L.

2015-12-01

Biological N2 fixation, catalyzed by the metalloenzyme nitrogenase, is a critical process that makes life possible on Earth. Environmental N2 fixation has been automatically attributed to canonical Mo-based nitrogenases despite over two decades of knowledge that two other metalloenzyme forms of nitrogenase exist: those containing catalytic V or Fe-only. A key area of missing information is the contribution of the "alternative" V and Fe-only nitrogenases, as the interpretation of field data to construct budgets and assess N availability depends on the type of nitrogenase metalloenzyme used to fix N2. Additionally, substantial changes in metal speciation over geological time may have favored the use of different metalloenzymes, with implications for evolution of the biosphere. Despite the potential importance of alternative nitrogenases in modern and ancient N cycling, few methods can determine their contributions to environmental N2 fixation. Here, we present new isotopic methods to distinguish between the activities of Mo, V, and Fe-only nitrogenases. We show evidence for alternative N2 fixation in diverse environments (cyanolichens, microbial mats, sediments, leaf litter), thereby linking a key process in the nitrogen cycle to specific metalloenzyme forms of nitrogenase. The results invite a reexamination of the conditions under which the different nitrogenase metalloenzymes are active and may lead to new insights into the coupling of the cycles of nitrogen and trace metals.

Ontogenetic resource-use strategies in a rare long-lived cycad along environmental gradients

PubMed Central

Álvarez-Yépiz, Juan C.; Cueva, Alejandro; Dovčiak, Martin; Teece, Mark; Yepez, Enrico A.

2014-01-01

Functional traits can drive plant responses to short- and long-term stressful conditions, with potential effects on species persistence in local habitats, changes in population size and structure, and potential species range shifts in changing environments. We investigated whether ecophysiological traits in a rare cycad vary along environmental gradients and with ontogeny to understand intra-specific resource-use variation (e.g. symbiotic nitrogen fixation, nitrogen- and water-use efficiency) and possible species adaptations for different environments. Environmental gradients were characterized with 14 soil and topographic variables. Nitrogen- and water-use efficiency improved with ontogeny (from seedling to juvenile and adult stages) but declined as soil fertility decreased with increasing elevation. Conversely, reliance on symbiotic nitrogen fixation increased with elevation and varied slightly with ontogeny. Improved water-use efficiency at lower elevation and nitrogen fixation at higher elevation may represent key functional strategies for maintaining the lower and upper altitudinal species range limits, especially in arid environments where stressful conditions are intensifying due to climatic and land-use changes. In addition to facilitation linked to the regeneration niche, improved resource-use efficiency linked to the adult niche may strongly influence cycad distribution and persistence in contemporary environments. A functional approach to conservation of rare or endangered plant species may be needed in order to target the most sensitive stages to changing environmental conditions and to better understand potential range shifts and adaptive responses to global land-use and climate changes. PMID:27293655

Estimation of biological nitrogen fixation by black locust in short-rotation forests using natural 15N abundance method

NASA Astrophysics Data System (ADS)

Veste, M.; Böhm, C.; Quinckenstein, A.; Freese, D.

2012-04-01

The importance of short rotation forests and agroforestry systems for woody biomass production for bioenergy will increase in Central Europe within the next decades. In this context, black locust (Robinia pseudoacacia) has a high growth potential especially at marginal, drought-susceptible sites such as occur in Brandenburg State (Eastern Germany). As a pioneer tree species black locust grows under a wide range of site conditions. The native range of black locust in Northern America is classified by a humid to sub-humid climate with a mean annual precipitation of 1020 to 1830 mm. In Central and Eastern Europe, this species is cultivated in a more continental climate with an annual precipitation often below 600 mm. Therefore, black locust is known to be relatively drought tolerant compared to other temperate, deciduous tree species. Because of its N2-fixation ability black locust plays generally an important role for the improvement of soil fertility. This effect is of particular interest at marginal sites in the post-mining landscapes. In order to estimate the N2-fixation potential of black locust at marginal sites leaf samples were taken from black locust trees in short rotation plantations planted between 1995 and 2007 in post-mining sites south of Cottbus (Brandenburg, NE Germany). The variation of the natural 15N abundance was measured to evaluate the biological nitrogen fixation. The nitrogen derived from the atmosphere can be calculated using a two-pool model from the quotient of the natural 15N abundances of the N2-fixing plant and the plant available soil N. Because representatively determining the plant available soil N is difficult, a non-N2-fixing reference plant growing at the same site with a similar root system and temporal N uptake pattern to the N2-fixing plant is often used. In our case we used red oak (Quercus rubra) as a reference. The average nitrogen content in the leaves of black locust ranged from 3.1% (C/N 14.8) in 15 years old trees to 3.4% (C/N 14.4) in 3 year-old trees, respectively. A higher content of nitrogen was found in leaves of re-sprouted trees with 4.3% (C/N 11.5). The estimated percentage of nitrogen derived from the atmosphere (% NdfA) in black locust was 63% - 83% compared to 56% in seabuckthorn (Hippophaë rhamnoides) and 79% in common broom (Genista scuparia). The annual leaf biomass production of black locust varied between 1325 (2 years old trees) and 2576 kg/ha a (4 years old trees). The estimated leaf nitrogen fixed by Robinia was approx. 30.5 - 59.2 kg/ha a. From the results, we can conclude that the biological nitrogen fixation by Robina is an important factor for the nitrogen balance of short-rotation plantations on nutrient poor-soils.

The role of symbiotic nitrogen fixation in nitrogen availability, competition and plant invasion into the sagebrush steppe

Treesearch

Erin M. Goergen

2009-01-01

In the semi-arid sagebrush steppe of the Northeastern Sierra Nevada, resources are both spatially and temporally variable, arguably making resource availability a primary factor determining invasion success. N fixing plant species, primarily native legumes, are often relatively abundant in sagebrush steppe and can contribute to ecosystem nitrogen budgets. ...

Nitrogen fixation and metabolism by groundwater-dependent perennial plants in a hyperarid desert.

PubMed

Arndt, Stefan K; Kahmen, Ansgar; Arampatsis, Christina; Popp, Marianne; Adams, Mark

2004-11-01

The Central Asian Taklamakan desert is characterized by a hyperarid climate with less than 50 mm annual precipitation but a permanent shallow groundwater table. The perched groundwater (2-16 m) could present a reliable and constant source of nitrogen throughout the growing season and help overcome temporal nitrogen limitations that are common in arid environments. We investigated the importance of groundwater and nitrogen fixation in the nitrogen metabolism of desert plants by assessing the possible forms and availability of soil N and atmospheric N and the seasonal variation in concentration as well as isotopic composition of plant N. Water availability was experimentally modified in the desert foreland through simulated flooding to estimate the contribution of surface water and temporally increased soil moisture for nutrient uptake and plant-water relations. The natural vegetation of the Taklamakan desert is dominated by plants with high foliar nitrogen concentrations (2-3% DM) and leaf nitrate reductase activity (NRA) (0.2-1 micromol NO2- g(-1) FW h(-1)). There is little evidence that nitrogen is a limiting resource as all perennial plants exhibited fast rates of growth. The extremely dry soil conditions preclude all but minor contributions of soil N to total plant N so that groundwater is suggested as the dominant source of N with concentrations of 100 microM NO3-. Flood irrigation had little beneficial effect on nitrogen metabolism and growth, further confirming the dependence on groundwater. Nitrogen fixation was determined by the 15N natural abundance method and was a significant component of the N-requirement of the legume Alhagi, the average contribution of biologically fixed nitrogen in Alhagi was 54.8%. But nitrogen fixing plants had little ecological advantage owing to the more or less constant supply of N available from groundwater. From our data we conclude that the perennial species investigated have adapted to the environmental conditions through development of root systems that access groundwater to satisfy demands for both water and nutrients. This is an ecologically favourable strategy since only groundwater is a predictable and stable resource.

Carbon and nitrogen fixation differ between successional stages of biological soil crusts in the Colorado Plateau and Chihuahuan Desert

USGS Publications Warehouse

Housman, D.C.; Powers, H.H.; Collins, A.D.; Belnap, J.

2006-01-01

Biological soil crusts (cyanobacteria, mosses and lichens collectively) perform essential ecosystem services, including carbon (C) and nitrogen (N) fixation. Climate and land-use change are converting later successional soil crusts to early successional soil crusts with lower C and N fixation rates. To quantify the effect of such conversions on C and N dynamics in desert ecosystems we seasonally measured diurnal fixation rates in different biological soil crusts. We classified plots on the Colorado Plateau (Canyonlands) and Chihuahuan Desert (Jornada) as early (Microcoleus) or later successional (Nostoc/Scytonema or Placidium/Collema) and measured photosynthesis (Pn), nitrogenase activity (NA), and chlorophyll fluorescence (Fv/Fm) on metabolically active (moist) soil crusts. Later successional crusts typically had greater Pn, averaging 1.2-1.3-fold higher daily C fixation in Canyonlands and 2.4-2.8-fold higher in the Jornada. Later successional crusts also had greater NA, averaging 1.3-7.5-fold higher daily N fixation in Canyonlands and 1.3-25.0-fold higher in the Jornada. Mean daily Fv/Fm was also greater in later successional Canyonlands crusts during winter, and Jornada crusts during all seasons except summer. Together these findings indicate conversion of soil crusts back to early successional stages results in large reductions of C and N inputs into these ecosystems.

Effect of air pollution on nitrogen fixation in lichens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kallio, S.; Varheenmaa, T.

1974-01-01

Clear decrease (80-90%) of acetylene reduction in Stereocaulon paschale and Nephroma arcticum was observed in the polluted city area of Turku (SW Finland) within a period of three-four weeks, while nitrogenase activity remained unchanged in the specimens outside the city area. Carbon dioxide fixation in these lichens lowered 20-50%.

Comparative Genomic Analysis of N2-Fixing and Non-N2-Fixing Paenibacillus spp.: Organization, Evolution and Expression of the Nitrogen Fixation Genes

PubMed Central

Xie, Jian-Bo; Du, Zhenglin; Bai, Lanqing; Tian, Changfu; Zhang, Yunzhi; Xie, Jiu-Yan; Wang, Tianshu; Liu, Xiaomeng; Chen, Xi; Cheng, Qi; Chen, Sanfeng; Li, Jilun

2014-01-01

We provide here a comparative genome analysis of 31 strains within the genus Paenibacillus including 11 new genomic sequences of N2-fixing strains. The heterogeneity of the 31 genomes (15 N2-fixing and 16 non-N2-fixing Paenibacillus strains) was reflected in the large size of the shell genome, which makes up approximately 65.2% of the genes in pan genome. Large numbers of transposable elements might be related to the heterogeneity. We discovered that a minimal and compact nif cluster comprising nine genes nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV encoding Mo-nitrogenase is conserved in the 15 N2-fixing strains. The nif cluster is under control of a σ70-depedent promoter and possesses a GlnR/TnrA-binding site in the promoter. Suf system encoding [Fe–S] cluster is highly conserved in N2-fixing and non-N2-fixing strains. Furthermore, we demonstrate that the nif cluster enabled Escherichia coli JM109 to fix nitrogen. Phylogeny of the concatenated NifHDK sequences indicates that Paenibacillus and Frankia are sister groups. Phylogeny of the concatenated 275 single-copy core genes suggests that the ancestral Paenibacillus did not fix nitrogen. The N2-fixing Paenibacillus strains were generated by acquiring the nif cluster via horizontal gene transfer (HGT) from a source related to Frankia. During the history of evolution, the nif cluster was lost, producing some non-N2-fixing strains, and vnf encoding V-nitrogenase or anf encoding Fe-nitrogenase was acquired, causing further diversification of some strains. In addition, some N2-fixing strains have additional nif and nif-like genes which may result from gene duplications. The evolution of nitrogen fixation in Paenibacillus involves a mix of gain, loss, HGT and duplication of nif/anf/vnf genes. This study not only reveals the organization and distribution of nitrogen fixation genes in Paenibacillus, but also provides insight into the complex evolutionary history of nitrogen fixation. PMID:24651173

Evidence for foliar endophytic nitrogen fixation in a widely distributed subalpine conifer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moyes, Andrew B.; Kueppers, Lara M.; Pett-Ridge, Jennifer

Coniferous forest nitrogen (N) budgets indicate unknown sources of N. A consistent association between limber pine ( Pinus flexilis) and potential N 2-fixing acetic acid bacteria (AAB) indicates that native foliar endophytes may supply subalpine forests with N.

Evidence for foliar endophytic nitrogen fixation in a widely distributed subalpine conifer

DOE PAGES

Moyes, Andrew B.; Kueppers, Lara M.; Pett-Ridge, Jennifer; ...

2016-02-01

Coniferous forest nitrogen (N) budgets indicate unknown sources of N. A consistent association between limber pine ( Pinus flexilis) and potential N 2-fixing acetic acid bacteria (AAB) indicates that native foliar endophytes may supply subalpine forests with N.

Widespread occurrence of Sinorhizobium meliloti strains with a type IV secretion system

USDA-ARS?s Scientific Manuscript database

Improvement in symbiotic nitrogen fixation would reduce reliance on nitrogen fertilizers but is hampered by poor establishment of introduced strains due to competition by indigenous symbiotic strains. This study investigated the origins, diversity, and competitiveness of Sinorhizobium meliloti (now ...

The small unicellular diazotrophic symbiont, UCYN-A, is a key player in the marine nitrogen cycle.

PubMed

Martínez-Pérez, Clara; Mohr, Wiebke; Löscher, Carolin R; Dekaezemacker, Julien; Littmann, Sten; Yilmaz, Pelin; Lehnen, Nadine; Fuchs, Bernhard M; Lavik, Gaute; Schmitz, Ruth A; LaRoche, Julie; Kuypers, Marcel M M

2016-09-12

Microbial dinitrogen (N 2 ) fixation, the nitrogenase enzyme-catalysed reduction of N 2 gas into biologically available ammonia, is the main source of new nitrogen (N) in the ocean. For more than 50 years, oceanic N 2 fixation has mainly been attributed to the activity of the colonial cyanobacterium Trichodesmium 1,2 . Other smaller N 2 -fixing microorganisms (diazotrophs)-in particular the unicellular cyanobacteria group A (UCYN-A)-are, however, abundant enough to potentially contribute significantly to N 2 fixation in the surface waters of the oceans 3-6 . Despite their abundance, the contribution of UCYN-A to oceanic N 2 fixation has so far not been directly quantified. Here, we show that in one of the main areas of oceanic N 2 fixation, the tropical North Atlantic 7 , the symbiotic cyanobacterium UCYN-A contributed to N 2 fixation similarly to Trichodesmium. Two types of UCYN-A, UCYN-A1 and -A2, were observed to live in symbioses with specific eukaryotic algae. Single-cell analyses showed that both algae-UCYN-A symbioses actively fixed N 2 , contributing ∼20% to N 2 fixation in the tropical North Atlantic, revealing their significance in this region. These symbioses had growth rates five to ten times higher than Trichodesmium, implying a rapid transfer of UCYN-A-fixed N into the food web that might significantly raise their actual contribution to N 2 fixation. Our analysis of global 16S rRNA gene databases showed that UCYN-A occurs in surface waters from the Arctic to the Antarctic Circle and thus probably contributes to N 2 fixation in a much larger oceanic area than previously thought. Based on their high rates of N 2 fixation and cosmopolitan distribution, we hypothesize that UCYN-A plays a major, but currently overlooked role in the oceanic N cycle.

Phytoplankton plasticity drives large variability in carbon fixation efficiency

NASA Astrophysics Data System (ADS)

Ayata, Sakina-Dorothée.; Lévy, Marina; Aumont, Olivier; Resplandy, Laure; Tagliabue, Alessandro; Sciandra, Antoine; Bernard, Olivier

2014-12-01

Phytoplankton C:N stoichiometry is highly flexible due to physiological plasticity, which could lead to high variations in carbon fixation efficiency (carbon consumption relative to nitrogen). However, the magnitude, as well as the spatial and temporal scales of variability, remains poorly constrained. We used a high-resolution biogeochemical model resolving various scales from small to high, spatially and temporally, in order to quantify and better understand this variability. We find that phytoplankton C:N ratio is highly variable at all spatial and temporal scales (5-12 molC/molN), from mesoscale to regional scale, and is mainly driven by nitrogen supply. Carbon fixation efficiency varies accordingly at all scales (±30%), with higher values under oligotrophic conditions and lower values under eutrophic conditions. Hence, phytoplankton plasticity may act as a buffer by attenuating carbon sequestration variability. Our results have implications for in situ estimations of C:N ratios and for future predictions under high CO2 world.

A model using marginal efficiency of investment to analyse carbon and nitrogen interactions in terrestrial ecosystems (ACONITE Version 1)

NASA Astrophysics Data System (ADS)

Thomas, R. Q.; Williams, M.

2014-04-01

Carbon (C) and nitrogen (N) cycles are coupled in terrestrial ecosystems through multiple processes including photosynthesis, tissue allocation, respiration, N fixation, N uptake, and decomposition of litter and soil organic matter. Capturing the constraint of N on terrestrial C uptake and storage has been a focus of the Earth System modelling community. However there is little understanding of the trade-offs and sensitivities of allocating C and N to different tissues in order to optimize the productivity of plants. Here we describe a new, simple model of ecosystem C-N cycling and interactions (ACONITE), that builds on theory related to plant economics in order to predict key ecosystem properties (leaf area index, leaf C : N, N fixation, and plant C use efficiency) using emergent constraints provided by marginal returns on investment for C and/or N allocation. We simulated and evaluated steady-state ecosystem stocks and fluxes in three different forest ecosystems types (tropical evergreen, temperate deciduous, and temperate evergreen). Leaf C : N differed among the three ecosystem types (temperate deciduous < tropical evergreen < temperature evergreen), a result that compared well to observations from a global database describing plant traits. Gross primary productivity (GPP) and net primary productivity (NPP) estimates compared well to observed fluxes at the simulation sites. Simulated N fixation at steady-state, calculated based on relative demand for N and the marginal return on C investment to acquire N, was an order of magnitude higher in the tropical forest than in the temperate forest, consistent with observations. A sensitivity analysis revealed that parameterization of the relationship between leaf N and leaf respiration had the largest influence on leaf area index and leaf C : N. Also, a widely used linear leaf N-respiration relationship did not yield a realistic leaf C : N, while a more recently reported non-linear relationship performed better. A parameter governing how photosynthesis scales with day length had the largest influence on total vegetation C, GPP, and NPP. Multiple parameters associated with photosynthesis, respiration, and N uptake influenced the rate of N fixation. Overall, our ability to constrain leaf area index and have spatially and temporally variable leaf C : N helps address challenges for ecosystem and Earth System models. Furthermore, the simple approach with emergent properties based on coupled C-N dynamics has potential for use in research that uses data-assimilation methods to integrate data on both the C and N cycles to improve C flux forecasts.

Root-Zone-Specific Oxygen Tolerance of Azospirillum spp. and Diazotrophic Rods Closely Associated with Kallar Grass.

PubMed

Hurek, T; Reinhold, B; Fendrik, I; Niemann, E G

1987-01-01

The effect of oxygen on N(2)-dependent growth of two Azospirillum strains and two diazotrophic rods closely associated with roots of Kallar grass (Leptochloa fusca) was studied. To enable precise comparison, bacteria were grown in dissolved-oxygen-controlled batch and continuous cultures. Steady states were obtained from about 1 to 30 muM O(2), some of them being carbon limited. All strains needed a minimum amount of oxygen for N(2)-dependent growth. Nitrogen contents between 10 and 13% of cell dry weight were observed. The response of steady-state cultures to increasing O(2) concentrations suggested that carbon limitation shifted to internal nitrogen limitation when N(2) fixation became so low that the bacteria could no longer meet their requirements for fixed nitrogen. For Azospirillum lipoferum Rp5, increase of the dilution rate resulted in decreased N(2) fixation in steady-state cultures with internal nitrogen limitation. Oxygen tolerance was found to be strain specific in A. lipoferum with strain Sp59b as a reference organism. Oxygen tolerance of strains from Kallar grass was found to be root zone specific. A. halopraeferens Au 4 and A. lipoferum Rp5, predominating on the rhizoplane of Kallar grass, and strains H6a2 and BH72, predominating in the endorhizosphere, differed in their oxygen tolerance profiles. Strains H6a2 and BH72 still grew and fixed nitrogen in steady-state cultures at O(2) concentrations exceeding those which absolutely inhibited nitrogen fixation of both Azospirillum strains. It is proposed that root-zone-specific oxygen tolerance reflects an adaptation of the isolates to the microenvironments provided by the host plant.

The Genetics of Symbiotic Nitrogen Fixation: Comparative Genomics of 14 Rhizobia Strains by Resolution of Protein Clusters

PubMed Central

Black, Michael; Moolhuijzen, Paula; Chapman, Brett; Barrero, Roberto; Howieson, John; Hungria, Mariangela; Bellgard, Matthew

2012-01-01

The symbiotic relationship between legumes and nitrogen fixing bacteria is critical for agriculture, as it may have profound impacts on lowering costs for farmers, on land sustainability, on soil quality, and on mitigation of greenhouse gas emissions. However, despite the importance of the symbioses to the global nitrogen cycling balance, very few rhizobial genomes have been sequenced so far, although there are some ongoing efforts in sequencing elite strains. In this study, the genomes of fourteen selected strains of the order Rhizobiales, all previously fully sequenced and annotated, were compared to assess differences between the strains and to investigate the feasibility of defining a core ‘symbiome’—the essential genes required by all rhizobia for nodulation and nitrogen fixation. Comparison of these whole genomes has revealed valuable information, such as several events of lateral gene transfer, particularly in the symbiotic plasmids and genomic islands that have contributed to a better understanding of the evolution of contrasting symbioses. Unique genes were also identified, as well as omissions of symbiotic genes that were expected to be found. Protein comparisons have also allowed the identification of a variety of similarities and differences in several groups of genes, including those involved in nodulation, nitrogen fixation, production of exopolysaccharides, Type I to Type VI secretion systems, among others, and identifying some key genes that could be related to host specificity and/or a better saprophytic ability. However, while several significant differences in the type and number of proteins were observed, the evidence presented suggests no simple core symbiome exists. A more abstract systems biology concept of nitrogen fixing symbiosis may be required. The results have also highlighted that comparative genomics represents a valuable tool for capturing specificities and generalities of each genome. PMID:24704847

Nitrogen Fixation by Gliding Arc Plasma: Better Insight by Chemical Kinetics Modelling.

PubMed

Wang, Weizong; Patil, Bhaskar; Heijkers, Stjin; Hessel, Volker; Bogaerts, Annemie

2017-05-22

The conversion of atmospheric nitrogen into valuable compounds, that is, so-called nitrogen fixation, is gaining increased interest, owing to the essential role in the nitrogen cycle of the biosphere. Plasma technology, and more specifically gliding arc plasma, has great potential in this area, but little is known about the underlying mechanisms. Therefore, we developed a detailed chemical kinetics model for a pulsed-power gliding-arc reactor operating at atmospheric pressure for nitrogen oxide synthesis. Experiments are performed to validate the model and reasonable agreement is reached between the calculated and measured NO and NO 2 yields and the corresponding energy efficiency for NO x formation for different N 2 /O 2 ratios, indicating that the model can provide a realistic picture of the plasma chemistry. Therefore, we can use the model to investigate the reaction pathways for the formation and loss of NO x . The results indicate that vibrational excitation of N 2 in the gliding arc contributes significantly to activating the N 2 molecules, and leads to an energy efficient way of NO x production, compared to the thermal process. Based on the underlying chemistry, the model allows us to propose solutions on how to further improve the NO x formation by gliding arc technology. Although the energy efficiency of the gliding-arc-based nitrogen fixation process at the present stage is not comparable to the world-scale Haber-Bosch process, we believe our study helps us to come up with more realistic scenarios of entering a cutting-edge innovation in new business cases for the decentralised production of fertilisers for agriculture, in which low-temperature plasma technology might play an important role. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Isolation and molecular identification of endophytic diazotrophs from seeds and stems of three cereal crops.

PubMed

Liu, Huawei; Zhang, Lei; Meng, Aihua; Zhang, Junbiao; Xie, Miaomiao; Qin, Yaohong; Faulk, Dylan Chase; Zhang, Baohong; Yang, Shushen; Qiu, Li

2017-01-01

Ten strains of endophytic diazotroph were isolated and identified from the plants collected from three different agricultural crop species, wheat, rice and maize, using the nitrogen-free selective isolation conditions. The nitrogen-fixing ability of endophytic diazotroph was verified by the nifH-PCR assay that showed positive nitrogen fixation ability. These identified strains were classified by 879F-RAPD and 16S rRNA sequence analysis. RAPD analyses revealed that the 10 strains were clustered into seven 879F-RAPD groups, suggesting a clonal origin. 16S rRNA sequencing analyses allowed the assignment of the 10 strains to known groups of nitrogen-fixing bacteria, including organisms from the genera Paenibacillus, Enterobacter, Klebsiella and Pantoea. These representative genus are not endophytic diazotrophs in the conventional sense. They may have obtained nitrogen fixation ability through lateral gene transfer, however, the evolutionary forces of lateral gene transfer are not well known. Molecular identification results from 16S rRNA analyses were also confirmed by morphological and biochemical data. The test strains SH6A and MZB showed positive effect on the growth of plants.

Soil Fertility Map for Food Legumes Production Areas in China

NASA Astrophysics Data System (ADS)

Li, Ling; Yang, Tao; Redden, Robert; He, Weifeng; Zong, Xuxiao

2016-05-01

Given the limited resources of fossil energy, and the environmental risks of excess fertilizer on crops, it is time to reappraise the potential role of food legume biological nitrogen fixation (BNF) as sources of nitrogen for cropping systems in China. 150 soil samples across 17 provinces and 2 municipalities of China were collected and analyzed. A distribution map of the soil fertilities and their patterns of distribution was constructed. The pH results indicated that soils were neutral to slightly alkaline overall. The soil organic matter (SOM) and the available nitrogen (AN) content were relatively low, while the available phosphorus (AP) and available potassium (AK) contents were from moderate to high. Production areas of food legumes (faba bean, pea, adzuki bean, mung bean and common bean) were clearly separated into 4 soil fertility type clusters. In addition, regions with SOM, AN, AP and AK deficiency, high acidity and high alkalinity were listed as target areas for further soil improvement. The potential was considered for biological nitrogen fixation to substitute for the application of mineral nitrogen fertiliser.

Importance of salt fingering for new nitrogen supply in the oligotrophic ocean.

PubMed

Fernández-Castro, B; Mouriño-Carballido, B; Marañón, E; Chouciño, P; Gago, J; Ramírez, T; Vidal, M; Bode, A; Blasco, D; Royer, S-J; Estrada, M; Simó, R

2015-09-09

The input of new nitrogen into the euphotic zone constrains the export of organic carbon to the deep ocean and thereby the biologically mediated long-term CO2 exchange between the ocean and atmosphere. In low-latitude open-ocean regions, turbulence-driven nitrate diffusion from the ocean's interior and biological fixation of atmospheric N2 are the main sources of new nitrogen for phytoplankton productivity. With measurements across the tropical and subtropical Atlantic, Pacific and Indian oceans, we show that nitrate diffusion (171±190 μmol m(-2) d(-1)) dominates over N2 fixation (9.0±9.4 μmol m(-2) d(-1)) at the time of sampling. Nitrate diffusion mediated by salt fingers is responsible for ca. 20% of the new nitrogen supply in several provinces of the Atlantic and Indian Oceans. Our results indicate that salt finger diffusion should be considered in present and future ocean nitrogen budgets, as it could supply globally 0.23-1.00 Tmol N yr(-1) to the euphotic zone.

Soil Fertility Map for Food Legumes Production Areas in China

PubMed Central

Li, Ling; Yang, Tao; Redden, Robert; He, Weifeng; Zong, Xuxiao

2016-01-01

Given the limited resources of fossil energy, and the environmental risks of excess fertilizer on crops, it is time to reappraise the potential role of food legume biological nitrogen fixation (BNF) as sources of nitrogen for cropping systems in China. 150 soil samples across 17 provinces and 2 municipalities of China were collected and analyzed. A distribution map of the soil fertilities and their patterns of distribution was constructed. The pH results indicated that soils were neutral to slightly alkaline overall. The soil organic matter (SOM) and the available nitrogen (AN) content were relatively low, while the available phosphorus (AP) and available potassium (AK) contents were from moderate to high. Production areas of food legumes (faba bean, pea, adzuki bean, mung bean and common bean) were clearly separated into 4 soil fertility type clusters. In addition, regions with SOM, AN, AP and AK deficiency, high acidity and high alkalinity were listed as target areas for further soil improvement. The potential was considered for biological nitrogen fixation to substitute for the application of mineral nitrogen fertiliser. PMID:27212262

Nitrogen Fixation By Sulfate-Reducing Bacteria in Coastal and Deep-Sea Sediments

NASA Astrophysics Data System (ADS)

Bertics, V. J.; Löscher, C.; Salonen, I.; Schmitz-Streit, R.; Lavik, G.; Kuypers, M. M.; Treude, T.

2011-12-01

Sulfate-reducing bacteria (SRB) can greatly impact benthic nitrogen (N) cycling, by for instance inhibiting coupled denitrification-nitrification through the production of sulfide or by increasing the availability of fixed N in the sediment via dinitrogen (N2)-fixation. Here, we explored several coastal and deep-sea benthic habitats within the Atlantic Ocean and Baltic Sea, for the occurrence of N2-fixation mediated by SRB. A combination of different methods including microbial rate measurements of N2-fixation and sulfate reduction, geochemical analyses (porewater nutrient profiles, mass spectrometry), and molecular analyses (CARD-FISH, HISH-SIMS, "nested" PCR, and QPCR) were applied to quantify and identify the responsible processes and organisms, respectively. Furthermore, we looked deeper into the question of whether the observed nitrogenase activity was associated with the final incorporation of N into microbial biomass or whether the enzyme activity served another purpose. At the AGU Fall Meeting, we will present and compare data from numerous stations with different water depths, temperatures, and latitudes, as well as differences in key geochemical parameters, such as organic carbon content and oxygen availability. Current metabolic and molecular data indicate that N2-fixation is occurring in many of these benthic environments and that a large part of this activity may linked to SRB.

Namib Desert primary productivity is driven by cryptic microbial community N-fixation.

PubMed

Ramond, Jean-Baptiste; Woodborne, Stephan; Hall, Grant; Seely, Mary; Cowan, Don A

2018-05-02

Carbon exchange in drylands is typically low, but during significant rainfall events (wet anomalies) drylands act as a C sink. During these anomalies the limitation on C uptake switches from water to nitrogen. In the Namib Desert of southern Africa, the N inventory in soil organic matter available for mineralisation is insufficient to support the observed increase in primary productivity. The C4 grasses that flourish after rainfall events are not capable of N fixation, and so there is no clear mechanism for adequate N fixation in dryland ecosystems to support rapid C uptake. Here we demonstrate that N fixation by photoautotrophic hypolithic communities forms the basis for the N budget for plant productivity events in the Namib Desert. Stable N isotope (δ 15 N) values of Namib Desert hypolithic biomass, and surface and subsurface soils were measured over 3 years across dune and gravel plain biotopes. Hypoliths showed significantly higher biomass and lower δ 15 N values than soil organic matter. The δ 15 N values of hypoliths approach the theoretical values for nitrogen fixation. Our results are strongly indicative that hypolithic communities are the foundation of productivity after rain events in the Namib Desert and are likely to play similar roles in other arid environments.

The genome of Paenibacillus sabinae T27 provides insight into evolution, organization and functional elucidation of nif and nif-like genes.

PubMed

Li, Xinxin; Deng, Zhiping; Liu, Zhanzhi; Yan, Yongliang; Wang, Tianshu; Xie, Jianbo; Lin, Min; Cheng, Qi; Chen, Sanfeng

2014-08-27

Most biological nitrogen fixation is catalyzed by the molybdenum nitrogenase. This enzyme is a complex which contains the MoFe protein encoded by nifDK and the Fe protein encoded by nifH. In addition to nifHDK, nifHDK-like genes were found in some Archaea and Firmicutes, but their function is unclear. We sequenced the genome of Paenibacillus sabinae T27. A total of 4,793 open reading frames were predicted from its 5.27 Mb genome. The genome of P. sabinae T27 contains fifteen nitrogen fixation (nif) genes, including three nifH, one nifD, one nifK, four nifB, two nifE, two nifN, one nifX and one nifV. Of the 15 nif genes, eight nif genes (nifB, nifH, nifD, nifK, nifE, nifN, nifX and nifV) and two non-nif genes (orf1 and hesA) form a complete nif gene cluster. In addition to the nif genes, there are nitrogenase-like genes, including two nifH-like genes and five pairs of nifDK-like genes. IS elements on the flanking regions of nif and nif-like genes imply that these genes might have been obtained by horizontal gene transfer. Phylogenies of the concatenated 8 nif gene (nifB, nifH, nifD, nifK, nifE, nifN, nifX and nifV) products suggest that P. sabinae T27 is closely related to Frankia. RT-PCR analysis showed that the complete nif gene cluster is organized as an operon. We demonstrated that the complete nif gene cluster under the control of σ70-dependent promoter enabled Escherichia coli JM109 to fix nitrogen. Also, here for the first time we demonstrated that unlike nif genes, the transcriptions of nifHDK-like genes were not regulated by ammonium and oxygen, and nifH-like or nifD-like gene could not restore the nitrogenase activity of Klebsiella pneumonia nifH- and nifD- mutant strains, respectively, suggesting that nifHDK-like genes were not involved in nitrogen fixation. Our data and analysis reveal the contents and distribution of nif and nif-like genes and contribute to the study of evolutionary history of nitrogen fixation in Paenibacillus. For the first time we demonstrated that the transcriptions of nifHDK-like genes were not regulated by ammonium and oxygen and nifHDK-like genes were not involved in nitrogen fixation.

Ecosystem nitrogen fixation throughout the snow-free period in subarctic tundra: effects of willow and birch litter addition and warming.

PubMed

Rousk, Kathrin; Michelsen, Anders

2017-04-01

Nitrogen (N) fixation in moss-associated cyanobacteria is one of the main sources of available N for N-limited ecosystems such as subarctic tundra. Yet, N 2 fixation in mosses is strongly influenced by soil moisture and temperature. Thus, temporal scaling up of low-frequency in situ measurements to several weeks, months or even the entire growing season without taking into account changes in abiotic conditions cannot capture the variation in moss-associated N 2 fixation. We therefore aimed to estimate moss-associated N 2 fixation throughout the snow-free period in subarctic tundra in field experiments simulating climate change: willow (Salix myrsinifolia) and birch (Betula pubescens spp. tortuosa) litter addition, and warming. To achieve this, we established relationships between measured in situ N 2 fixation rates and soil moisture and soil temperature and used high-resolution measurements of soil moisture and soil temperature (hourly from May to October) to model N 2 fixation. The modelled N 2 fixation rates were highest in the warmed (2.8 ± 0.3 kg N ha -1 ) and birch litter addition plots (2.8 ± 0.2 kg N ha -1 ), and lowest in the plots receiving willow litter (1.6 ± 0.2 kg N ha -1 ). The control plots had intermediate rates (2.2 ± 0.2 kg N ha -1 ). Further, N 2 fixation was highest during the summer in the warmed plots, but was lowest in the litter addition plots during the same period. The temperature and moisture dependence of N 2 fixation was different between the climate change treatments, indicating a shift in the N 2 fixer community. Our findings, using a combined empirical and modelling approach, suggest that a longer snow-free period and increased temperatures in a future climate will likely lead to higher N 2 fixation rates in mosses. Yet, the consequences of increased litter fall on moss-associated N 2 fixation due to shrub expansion in the Arctic will depend on the shrub species' litter traits. © 2016 John Wiley & Sons Ltd.

Cost of nitrogen use in the US

EPA Science Inventory

Growing human demands for food, fuel and fiber have accelerated the human-driven fixation of reactive nitrogen (N) by at least 10-fold over the last century. This acceleration is one of the most dramatic changes to the sustainability of Earth’s systems. Approximately 65% ...

Understanding the Variability in Soybean Nitrogen Fixation across Agroecosystems

USDA-ARS?s Scientific Manuscript database

Conventional farming practices have uncoupled carbon (C) and nitrogen (N) cycles through the application of inorganic N fertilizers applied in plant available forms at levels that saturate the system. As a result, extensive N losses via leaching and denitrification are having significant environment...

Symbiotic Nitrogen Fixation in the Fungus Gardens of Leaf-Cutter Ants

USDA-ARS?s Scientific Manuscript database

Bacteria-mediated acquisition of atmospheric dinitrogen by plants serves as a critical nitrogen source in terrestrial ecosystems, and through its key role in agriculture, this phenomenon has shaped the development of human civilizations. Here we show that, paralleling human agriculture, cultivation ...

Favoring the unfavored: Selective electrochemical nitrogen fixation using a reticular chemistry approach

PubMed Central

Lee, Hiang Kwee; Koh, Charlynn Sher Lin; Lee, Yih Hong; Liu, Chong; Phang, In Yee; Han, Xuemei; Tsung, Chia-Kuang; Ling, Xing Yi

2018-01-01

Electrochemical nitrogen-to-ammonia fixation is emerging as a sustainable strategy to tackle the hydrogen- and energy-intensive operations by Haber-Bosch process for ammonia production. However, current electrochemical nitrogen reduction reaction (NRR) progress is impeded by overwhelming competition from the hydrogen evolution reaction (HER) across all traditional NRR catalysts and the requirement for elevated temperature/pressure. We achieve both excellent NRR selectivity (~90%) and a significant boost to Faradic efficiency by 10 percentage points even at ambient operations by coating a superhydrophobic metal-organic framework (MOF) layer over the NRR electrocatalyst. Our reticular chemistry approach exploits MOF’s water-repelling and molecular-concentrating effects to overcome HER-imposed bottlenecks, uncovering the unprecedented electrochemical features of NRR critical for future theoretical studies. By favoring the originally unfavored NRR, we envisage our electrocatalytic design as a starting point for high-performance nitrogen-to-ammonia electroconversion directly from water vapor–abundant air to address increasing global demand of ammonia in (bio)chemical and energy industries. PMID:29536047

Inhibition of nitrogenase by oxygen in marine cyanobacteria controls the global nitrogen and oxygen cycles

NASA Astrophysics Data System (ADS)

Berman-Frank, I.; Chen, Y.-B.; Gerchman, Y.; Dismukes, G. C.; Falkowski, P. G.

2005-03-01

Cyanobacterial N2-fixation supplies the vast majority of biologically accessible inorganic nitrogen to nutrient-poor aquatic ecosystems. The process, catalyzed by the heterodimeric protein complex, nitrogenase, is thought to predate that of oxygenic photosynthesis. Remarkably, while the enzyme plays such a critical role in Earth's biogeochemical cycles, the activity of nitrogenase in cyanobacteria is markedly inhibited in vivo at a post-translational level by the concentration of O2 in the contemporary atmosphere leading to metabolic and biogeochemical inefficiency in N2 fixation. We illustrate this crippling effect with data from Trichodesmium spp. an important contributor of "new nitrogen" to the world's subtropical and tropical oceans. The enzymatic inefficiency of nitrogenase imposes a major elemental taxation on diazotrophic cyanobacteria both in the costs of protein synthesis and for scarce trace elements, such as iron. This restriction has, in turn, led to a global limitation of fixed nitrogen in the contemporary oceans and provides a strong biological control on the upper bound of oxygen concentration in Earth's atmosphere.

Effect of light intensity and nitrogen starvation on CO2 fixation and lipid/carbohydrate production of an indigenous microalga Scenedesmus obliquus CNW-N.

PubMed

Ho, Shih-Hsin; Chen, Chun-Yen; Chang, Jo-Shu

2012-06-01

Engineering strategies were applied to improve the CO(2) fixation rate and carbohydrate/lipid production of a Scenedesmus obliquus CNW-N isolate. The light intensity that promotes cell growth, carbohydrate/lipid productivity, and CO(2) fixation efficiency was identified. Nitrogen starvation was also employed to trigger the accumulation of lipid and carbohydrate. The highest productivity of biomass, lipid, and carbohydrate was 840.57 mg L(-1)d(-1), 140.35 mg L(-1)d(-1). The highest lipid and carbohydrate content was 22.4% (5-day N-starvation) and 46.65% (1-day N-starvation), respectively. The optimal CO(2) consumption rate was 1420.6 mg L(-1)d(-1). This performance is better than that reported in most other studies. Under nitrogen starvation, the microalgal lipid was mainly composed of C16/C18 fatty acid (around 90%), which is suitable for biodiesel synthesis. The carbohydrate present in the biomass was mainly glucose, accounting for 77-80% of total carbohydrates. This carbohydrate composition is also suitable for fermentative biofuels production (e.g., bioethanol and biobutanol). Copyright © 2011 Elsevier Ltd. All rights reserved.

Inoculation of Bacillus sphaericus UPMB-10 to Young Oil Palm and Measurement of Its Uptake of Fixed Nitrogen Using the 15N Isotope Dilution Technique

PubMed Central

Zakry, Fitri Abdul Aziz; Shamsuddin, Zulkifli H.; Rahim, Khairuddin Abdul; Zakaria, Zin Zawawi; Rahim, Anuar Abdul

2012-01-01

There are increasing applications of diazotrophic rhizobacteria in the sustainable agriculture system. A field experiment on young immature oil palm was conducted to quantify the uptake of N derived from N2 fixation by the diazotroph Bacillus sphaericus strain UPMB-10, using the 15N isotope dilution method. Eight months after 15N application, young immature oil palms that received 67% of standard N fertilizer application together with B. sphaericus inoculation had significantly lower 15N enrichment than uninoculated palms that received similar N fertilizers. The dilution of labeled N served as a marker for the occurrence of biological N2 fixation. The proportion of N uptake that was derived from the atmosphere was estimated as 63% on the whole plant basis. The inoculation process increased the N and dry matter yields of the palm leaflets and rachis significantly. Field planting of young, immature oil palm in soil inoculated with B. sphaericus UPMB-10 might mitigate inorganic fertilizer-N application through supplementation by biological nitrogen fixation. This could be a new and important source of nitrogen biofertilizer in the early phase of oil palm cultivation in the field. PMID:22446306

Molecular analysis of ureide accumulation under drought stress in Phaseolus vulgaris L.

PubMed

Alamillo, Josefa M; Díaz-Leal, Juan Luís; Sánchez-Moran, Ma Victoria; Pineda, Manuel

2010-11-01

Under water deficit, ureidic legumes accumulate ureides in plant tissues, and this accumulation has been correlated with the inhibition of nitrogen fixation. In this work we used a molecular approach to characterize ureide accumulation under drought stress in Phaseolus vulgaris. Accumulation of ureides, mainly allantoate, was found in roots, shoots and leaves, but only a limited transient increase was observed in nodules from drought-stressed plants. We show that ureide accumulation is regulated at the transcriptional level mainly through induction of allantoinase (ALN), whereas allantoate amidohydrolase (AAH), involved in allantoate degradation, was slightly reduced, indicating that inhibition of this enzyme, key in ureide breakdown in aerial tissues, is not the main cause of allantoate accumulation. Expression of the ureide metabolism genes analysed in this study was induced by abscisic acid (ABA), suggesting the involvement of this plant hormone in ureide accumulation. Moreover, we observed that increases of ureide levels in P. vulgaris drought-stressed tissues were similar in non-nodulated, nitrate-fed plants, and in plants cultured under nitrogen-fixation conditions. Our results indicate that ureide accumulation in response to water deficit is independent from de novo synthesis of ureides in nodules, and therefore uncoupled from nitrogen fixation. © 2010 Blackwell Publishing Ltd.

Cyanobacterial crusts linked to soil productivity under different grazing management practices in Northern Australia

NASA Astrophysics Data System (ADS)

Alchin, Bruce; Williams, Wendy

2015-04-01

In arid and semi-arid Australia, the central role of healthy soil ecosystems in broad-acre grazing lands may be attributed to the widespread presence of cyanobacterial crusts. In terms of soil nutrient cycling and stability their role is particularly crucial in a climate dominated by annual dry seasons and variable wet seasons. In this study, we aimed to measure the contribution of cyanobacteria to soil nutrient cycling under contrasting levels of disturbance associated with grazing management. Field sampling was carried out on six paired sites (twelve properties) located across an east-west 3,000 km transect that covered different rangeland types on grazing properties in northern Australia (Queensland, Northern Territory and Western Australia). At each location paired sites were established and two different management systems were assessed, cell-paddock rotations (25-400 ha) and continuous grazing (200-2,000 ha). Cyanobacterial soil crusts were recorded from all of the twelve sites and cyanobacteria with the capacity to fix nitrogen were found at ten of the twelve sites. The overall diversity of cyanobacteria varied from three to ten species under any type of grazing system. As field work was conducted in the dry season, it is likely that the diversity may be greater in the wet season than the initial data may indicate. The average cyanobacterial soil crust cover across soil surfaces, between grass tussocks, during the dry season was estimated to be 50.9% and, 42.6% in the early wet season. This reflected longer established crust cover (dry season) versus newly established crusts. There was a high level of variability in the biomass of cyanobacteria however; the grazing system did not have any marked effect on the biomass for any one rangeland type. The grazing system differences did not appear to significantly influence the diversity at any location except on a floodplain in the Pilbara (WA). Biological nitrogen fixation by cyanobacteria was recorded at all sites. Nitrogen fixation rates were significantly higher in the wet season samples compared to the dry season. Rates of nitrogen fixation, mineralisable nitrogen and cyanobacterial biomass were comparative to other studies both in Australia and globally. Eleven of the twelve sites had higher plant-available (mineralisable) nitrogen in the 0-1 cm depth compared to the 1-5 cm depth. Nitrogen isotopes showed that the nitrogen concentration found in the surface soils (0-1 cm) from five sites originated from cyanobacterial nitrogen fixation. At the remaining sites the isotopic signatures were slightly more positive, indicative of fractionation. The results have substantiated the link between cyanobacteria and their contribution to carbon and nitrogen cycling across the northern Australian rangelands. The data also highlights the variability between sites and management practices that influence biogeochemical processes that affect soil productivity.

High-nitrogen fixation rates in the particulate and dissolved pools in the Western Tropical Pacific (Solomon and Bismarck Seas)

NASA Astrophysics Data System (ADS)

Berthelot, H.; Benavides, M.; Moisander, P. H.; Grosso, O.; Bonnet, S.

2017-08-01

Dinitrogen (N2) fixation rates were investigated in the euphotic layer of the Bismarck and Solomon Seas using 15N2 incubation assays taking into account both the particulate and the dissolved pools. Average depth-integrated particulate N2 fixation rates were 203 (range 43-399) and 1396 (range 176-3132) μmol N m-2 d-1 in the Bismarck and Solomon Seas, respectively. In both seas, N2 fixation measured in the dissolved pool was similar to particulate N2 fixation, highlighting the potentially substantial underestimation of N2 fixation in oceanic budgets when only particulate N2 fixation is considered. Among the diazotroph phylotypes targeted using quantitative polymerase chain reaction amplification of nifH genes, Trichodesmium was the most abundant. Regression analyses suggest that it accounted for the major proportion of N2 fixation. However, unicellular cyanobacterial and non-cyanobacterial diazotrophs were also occasionally abundant. This study reports high pelagic N2 fixation rates and confirms that the Western Tropical South Pacific is a hot spot for marine N2 fixation.

Effects of irrigation and addition of nitrogen fertiliser on net ecosystem carbon balance for a grassland.

PubMed

Moinet, Gabriel Y K; Cieraad, Ellen; Turnbull, Matthew H; Whitehead, David

2017-02-01

The ability to quantify the impacts of changing management practices on the components of net ecosystem carbon balance (N B ) is required to forecast future changes in soil carbon stocks and potential feedbacks on atmospheric CO 2 concentrations. In this study we investigated seasonal changes on the components of net ecosystem carbon balance resulting from the application of irrigation and nitrogen fertiliser to a temperate grassland in New Zealand where we simulated grazing events. We made seasonal measurements of the components of N B using chamber measurements in field plots with and without irrigation and addition of nitrogen fertiliser. We developed models to determine the physiological responses of gross canopy photosynthesis (A), leaf respiration (R L ) and soil respiration (R S ) to soil and air temperature, soil water content and irradiance and we estimated annual N B for the first year after treatments were applied. Overall, irrigation and nitrogen addition had a synergistic effect to increase annual estimates of above-ground components of carbon balance (A, R L and carbon exported through simulated grazing, F export ), but there was no effect from adding nitrogen alone. Annual R S remained unchanged between treatments. The treatments resulted in increases in above-ground biomass production, but, with the high intensity of simulated grazing, these were not sufficient to offset ecosystem carbon losses, so all treatments remained a net source of carbon. There were no significant differences between treatments and annual N B ranged from -540gCm -2 y -1 for the treatment with no irrigation and no nitrogen addition and -284gCm -2 y -1 for the treatment with irrigation and nitrogen addition. Our findings from the first year of the treatments quantify the net benefits of addition of irrigation and nitrogen on increasing above-ground production for animal feed but show that this did not lead to a net increase carbon input to the soil. Copyright © 2016 Elsevier B.V. All rights reserved.

Molecular Ecological and Stable Isotopic Studies of Nitrogen Fixation in Modern Microbial Mats

NASA Technical Reports Server (NTRS)

Bebout, B. M.; Crumbliss, L. L.; DesMarais, D. J.; Hogan, M. E.; Omoregie, E.; Turk, K. A.; Zehr, J. P.

2003-01-01

Nitrogen is usually the element limiting biological productivity in the marine environment. Microbial mats, laminated microbial communities analogous to some of the oldest forms of life on Earth, are often the sites of high rates of N fixation (the energetically expensive conversion of atmospheric dinitrogen into a biologically useful form). The N fixing enzyme nitrogenase is generally considered to be of ancient origin, and is widely distributed throughout the Bacterial and Archaeal domains of life, indicating an important role for this process over evolutionary time. The stable isotopic signature of N fixation is purportedly recognizable in organic matter (ancient kerogens as well as present-day microbial mats) as a delta (15)N(sub organic) near zero. We studied two microbial mats exhibiting different rates of N fixation in order to better understand the impact of N fixation on the delta (15)N (sub organic) of the mats, as well as what organisms are important in this process. Mats dominated by the cyanobacterium Microcoleus chthonoplastes grow in permanently submerged hypersaline salterns, and exhibit low rates of N fixation, whereas mats dominated by the cyanobacterium Lyngbya spp grow in an intertidal area, and exhibit rates of N fixation an order of magnitude higher. To examine successional stages in mat growth, both developing and established mats at each location were sampled. PCR and RT-PCR based approaches were used to identify, respectively, the organisms containing nifH (one of the genes that encode nitrogenase) as well as those expressing nifH in these mats. Both mats exhibited a distinct diel cycle of N fixation, with highest rates occurring at night. The delta (15)N(sub organic) of the subtidal Microcoleus mats is near zero whereas the delta (15)N(sub organic) is slightly more positive (+ 2-3%), in the intertidal Lyngbya mats, an interesting difference in view of the fact that overall rates of activity in the intertidal mats are much higher that those in the submerged hypersaline mats. Developing mats in both the subtidal and intertidal locations had delta (15)N(sub organic) values very near those of the established mats. Further work is necessary in order to determine the importance of other transformations of nitrogen on the delta (15)N(sub organic) signature of the mats.

Overproduction of Hydrogen From an Anaerobic Bacterium

DTIC Science & Technology

2008-12-01

fixation of nitrogen ( Haber - Bosch process), mostly to produce fertilizer. Nitrogenase provides a catalytic alternative to the commercial fixation of...the culture and suggests a uniquely simple hydrogen reactor design based on renewable feedstocks. 1. INTRODUCTION Hydrogen is an ideal... renewable feedstocks. Clostridium phytofermentans is a recently- discovered anaerobic bacterium, reported to possess cellulase enzymes that degrade

Niche differentiation in nitrogen metabolism among methanotrophs within an operational taxonomic unit

PubMed Central

2014-01-01

Background The currently accepted thesis on nitrogenous fertilizer additions on methane oxidation activity assumes niche partitioning among methanotrophic species, with activity responses to changes in nitrogen content being dependent on the in situ methanotrophic community structure Unfortunately, widely applied tools for microbial community assessment only have a limited phylogenetic resolution mostly restricted to genus level diversity, and not to species level as often mistakenly assumed. As a consequence, intragenus or intraspecies metabolic versatility in nitrogen metabolism was never evaluated nor considered among methanotrophic bacteria as a source of differential responses of methane oxidation to nitrogen amendments. Results We demonstrated that fourteen genotypically different Methylomonas strains, thus distinct below the level at which most techniques assign operational taxonomic units (OTU), show a versatile physiology in their nitrogen metabolism. Differential responses, even among strains with identical 16S rRNA or pmoA gene sequences, were observed for production of nitrite and nitrous oxide from nitrate or ammonium, nitrogen fixation and tolerance to high levels of ammonium, nitrate, and hydroxylamine. Overall, reduction of nitrate to nitrite, nitrogen fixation, higher tolerance to ammonium than nitrate and tolerance and assimilation of nitrite were general features. Conclusions Differential responses among closely related methanotrophic strains to overcome inhibition and toxicity from high nitrogen loads and assimilation of various nitrogen sources yield competitive fitness advantages to individual methane-oxidizing bacteria. Our observations proved that community structure at the deepest phylogenetic resolution potentially influences in situ functioning. PMID:24708438

Niche differentiation in nitrogen metabolism among methanotrophs within an operational taxonomic unit.

PubMed

Hoefman, Sven; van der Ha, David; Boon, Nico; Vandamme, Peter; De Vos, Paul; Heylen, Kim

2014-04-04

The currently accepted thesis on nitrogenous fertilizer additions on methane oxidation activity assumes niche partitioning among methanotrophic species, with activity responses to changes in nitrogen content being dependent on the in situ methanotrophic community structure Unfortunately, widely applied tools for microbial community assessment only have a limited phylogenetic resolution mostly restricted to genus level diversity, and not to species level as often mistakenly assumed. As a consequence, intragenus or intraspecies metabolic versatility in nitrogen metabolism was never evaluated nor considered among methanotrophic bacteria as a source of differential responses of methane oxidation to nitrogen amendments. We demonstrated that fourteen genotypically different Methylomonas strains, thus distinct below the level at which most techniques assign operational taxonomic units (OTU), show a versatile physiology in their nitrogen metabolism. Differential responses, even among strains with identical 16S rRNA or pmoA gene sequences, were observed for production of nitrite and nitrous oxide from nitrate or ammonium, nitrogen fixation and tolerance to high levels of ammonium, nitrate, and hydroxylamine. Overall, reduction of nitrate to nitrite, nitrogen fixation, higher tolerance to ammonium than nitrate and tolerance and assimilation of nitrite were general features. Differential responses among closely related methanotrophic strains to overcome inhibition and toxicity from high nitrogen loads and assimilation of various nitrogen sources yield competitive fitness advantages to individual methane-oxidizing bacteria. Our observations proved that community structure at the deepest phylogenetic resolution potentially influences in situ functioning.

Global assessment of limitation to symbiotic nitrogen fixation by phosphorus availability in terrestrial ecosystems using a meta-analysis approach

NASA Astrophysics Data System (ADS)

Augusto, Laurent; Delerue, Florian; Gallet-Budynek, Anne; Achat, David L.

2013-09-01

nitrogen fixation (SNF) is the main natural source of nitrogen (N) in terrestrial ecosystems worldwide. Previous studies have shown that fixation of N by plants can be limited by the availability of phosphorus (P) in soils. We used global meta-analysis to investigate how P availability controls SNF. In experiments in which plants were grown in an artificial medium, severe P deficiencies in the nutritive solution ([PO4] < 5-42 μM) depressed SNF flux through both a direct decrease in the plant fixation rate (i.e., decreased N fixed per unit of plant biomass) and an indirect effect (i.e., through plant biomass). In most experiments with plants grown in soils, SNF was proportional to plant biomass and was consequently only indirectly limited by P. Some cases using unfertilized and weathered soils (ultisols or oxisols), where plants were particularly P stressed, were an exception with both direct and indirect P limitations. Our global analysis of the P-SNF relationship indicated that P bioavailability commonly limited SNF flux. We conclude that the main driver of in situ P limitation is indirect via limitation of plant growth, except in certain cases where both indirect and direct constraints may play a role. These cases of severe P deficiency may be mainly found in weathered tropical soils of Africa and South America, probably in unfertilized croplands which are depleted in P due to repeated biomass harvests.

Successful correction of tibial bone deformity through multiple surgical procedures, liquid nitrogen-pretreated bone tumor autograft, three-dimensional external fixation, and internal fixation in a patient with primary osteosarcoma: a case report.

PubMed

Takeuchi, Akihiko; Yamamoto, Norio; Shirai, Toshiharu; Nishida, Hideji; Hayashi, Katsuhiro; Watanabe, Koji; Miwa, Shinji; Tsuchiya, Hiroyuki

2015-12-07

In a previous report, we described a method of reconstruction using tumor-bearing autograft treated by liquid nitrogen for malignant bone tumor. Here we present the first case of bone deformity correction following a tumor-bearing frozen autograft via three-dimensional computerized reconstruction after multiple surgeries. A 16-year-old female student presented with pain in the left lower leg and was diagnosed with a low-grade central tibial osteosarcoma. Surgical bone reconstruction was performed using a tumor-bearing frozen autograft. Bone union was achieved at 7 months after the first surgical procedure. However, local tumor recurrence and lung metastases occurred 2 years later, at which time a second surgical procedure was performed. Five years later, the patient developed a 19° varus deformity and underwent a third surgical procedure, during which an osteotomy was performed using the Taylor Spatial Frame three-dimensional external fixation technique. A fourth corrective surgical procedure was performed in which internal fixation was achieved with a locking plate. Two years later, and 10 years after the initial diagnosis of tibial osteosarcoma, the bone deformity was completely corrected, and the patient's limb function was good. We present the first report in which a bone deformity due to a primary osteosarcoma was corrected using a tumor-bearing frozen autograft, followed by multiple corrective surgical procedures that included osteotomy, three-dimensional external fixation, and internal fixation.

Distribution and Magnitude of Dinitrogen Fixation in the Eastern Tropical North Pacific Oxygen Deficient Zone.

NASA Astrophysics Data System (ADS)

Selden, C.; Mulholland, M. R.; Widner, B.; Bernhardt, P. W.; Macías Tapia, A.; Jayakumar, A.

2016-12-01

The Eastern Tropical North Pacific Ocean (ETNP) hosts one of the world's three major open ocean oxygen deficient zones (ODZs). Hotspots for fixed nitrogen (N) loss processes, ODZs have classically been discounted as areas of significant dinitrogen (N2) fixation, the microbe-mediated reduction of N2 to ammonium (NH4+), which has historically been ascribed primarily to euphotic, nutrient-deplete tropical waters. Challenging this paradigm, active expression of nifH (the dinitrogen reductase structural gene) has recently been documented in the ETNP, Eastern Tropical South Pacific, and Arabian Sea ODZs, implying a closer coupling of fixed nitrogen input and loss processes than previously thought. Here, we report rates of N­2 fixation measured in the ETNP ODZ along vertical gradients of oxygen, light, and dissolved N concentrations. Detailed vertical profiles of N2 fixation rates and dissolved N concentrations made within the ODZ were compared with similar profiles from oxic waters outside the ODZ. In addition, different organic carbon sources were investigated as potential rate-limiting factors for N2 fixation in sub-euphotic waters. By establishing the magnitude and distribution of N­2 fixation in the ETNP ODZ, this study contributes to current understanding of N cycling in anoxic and aphotic waters, and serves to elucidate nuances in the global N budget, enabling more accurate biogeochemical modeling. Understanding these processes in present day ODZs is crucial for predicting how ongoing anthropogenic intensification of coastal ODZs will alter biogeochemical cycles in the future.

Nitrate and Ammonium Induced Photosynthetic Suppression in N-Limited Selenastrum minutum: II. Effects of NO(3) and NH(4) Addition to CO(2) Efflux in the Light.

PubMed

Birch, D G; Elrifi, I R; Turpin, D H

1986-11-01

The effects of nitrate and ammonium addition on net and gross photosynthesis, CO(2) efflux and the dissolved inorganic carbon compensation point of nitrogen-limited Selenastrum minutum Naeg. Collins (Chlorophyta) were studied. Cultures pulsed with nitrate or ammonium exhibited a marked decrease in both net and gross photosynthetic carbon fixation. During this period of suppression the specific activity of exogenous dissolved inorganic carbon decreased rapidly in comparison to control cells indicating an increase in the rate of CO(2) efflux in the light. The nitrate and ammmonium induced rates of CO(2) efflux were 31.0 and 33.8 micromoles CO(2) per milligram chlorophyll per hour, respectively, and represented 49 and 48% of the rate of gross photosynthesis. Nitrate addition to cells at dissolved inorganic carbon compensation point caused an increase in compensation point while ammonium had no effect. In the presence of the tricarboxylic acid cycle inhibitor fluoroacetate, the nitrate-induced change in compensation point was greatly reduced suggesting the source of this CO(2) was the tricarboxylic acid cycle. These results are consistent with the mechanism of N-induced photosynthetic suppression outlined by Elrifi and Turpin (1986 Plant Physiol 81: 273-279).

Global nitrogen budgets in cereals: A 50-year assessment for maize, rice, and wheat production systems.

PubMed

Ladha, J K; Tirol-Padre, A; Reddy, C K; Cassman, K G; Verma, Sudhir; Powlson, D S; van Kessel, C; de B Richter, Daniel; Chakraborty, Debashis; Pathak, Himanshu

2016-01-18

Industrially produced N-fertilizer is essential to the production of cereals that supports current and projected human populations. We constructed a top-down global N budget for maize, rice, and wheat for a 50-year period (1961 to 2010). Cereals harvested a total of 1551 Tg of N, of which 48% was supplied through fertilizer-N and 4% came from net soil depletion. An estimated 48% (737 Tg) of crop N, equal to 29, 38, and 25 kg ha(-1) yr(-1) for maize, rice, and wheat, respectively, is contributed by sources other than fertilizer- or soil-N. Non-symbiotic N2 fixation appears to be the major source of this N, which is 370 Tg or 24% of total N in the crop, corresponding to 13, 22, and 13 kg ha(-1) yr(-1) for maize, rice, and wheat, respectively. Manure (217 Tg or 14%) and atmospheric deposition (96 Tg or 6%) are the other sources of N. Crop residues and seed contribute marginally. Our scaling-down approach to estimate the contribution of non-symbiotic N2 fixation is robust because it focuses on global quantities of N in sources and sinks that are easier to estimate, in contrast to estimating N losses per se, because losses are highly soil-, climate-, and crop-specific.

Ultradian metabolic rhythm in the diazotrophic cyanobacterium Cyanothece sp. ATCC 51142.

PubMed

Červený, Jan; Sinetova, Maria A; Valledor, Luis; Sherman, Louis A; Nedbal, Ladislav

2013-08-06

The unicellular cyanobacterium Cyanothece sp. American Type Culture Collection (ATCC) 51142 is capable of performing oxygenic photosynthesis during the day and microoxic nitrogen fixation at night. These mutually exclusive processes are possible only by temporal separation by circadian clock or another cellular program. We report identification of a temperature-dependent ultradian metabolic rhythm that controls the alternating oxygenic and microoxic processes of Cyanothece sp. ATCC 51142 under continuous high irradiance and in high CO2 concentration. During the oxygenic photosynthesis phase, nitrate deficiency limited protein synthesis and CO2 assimilation was directed toward glycogen synthesis. The carbohydrate accumulation reduced overexcitation of the photosynthetic reactions until a respiration burst initiated a transition to microoxic N2 fixation. In contrast to the circadian clock, this ultradian period is strongly temperature-dependent: 17 h at 27 °C, which continuously decreased to 10 h at 39 °C. The cycle was expressed by an oscillatory modulation of net O2 evolution, CO2 uptake, pH, fluorescence emission, glycogen content, cell division, and culture optical density. The corresponding ultradian modulation was also observed in the transcription of nitrogenase-related nifB and nifH genes and in nitrogenase activities. We propose that the control by the newly identified metabolic cycle adds another rhythmic component to the circadian clock that reflects the true metabolic state depending on the actual temperature, irradiance, and CO2 availability.

Diversity and activity of nitrogen fixing archaea and bacteria associated with micro-environments of wetland rice

NASA Astrophysics Data System (ADS)

Schmidt, Hannes; Woebken, Dagmar

2017-04-01

Wetland rice is one of the world's most important crop plants. The cultivation on waterlogged paddy soils is strongly limited by nitrogen (N), which is typically supplied by industrial fertilizers that are not only costly but also exhibit hazardous effects on the environment. It has been reported that "Biological Nitrogen Fixation" through N2-fixing bacteria and archaea (diazotrophs) can alleviate the N-shortage in rice cultivation, thus carrying out an important ecosystem function. However, our understanding of the diversity and in situ N2 fixation activity of diazotrophs in flooded rice fields is still rudimentary. Moreover, knowledge on the impact of biochemical gradients established by root activity (i.e. exudation, radial oxygen loss) on the functioning of N-fixing microorganisms in paddy soil ecosystems is limited. We aimed at studying underlying processes on biologically relevant scales. Greenhouse studies were performed to identify key factors that control rice-diazotroph association and related N2 fixation activities. Paddy soils of different geographical origin were cultivated with two commercially used genotypes of wetland rice. Samples were separated into bulk soil, rhizosphere soil, rhizoplane, and roots at flowering stage of rice plant development. These samples were subjected to functional assays and various molecular biological techniques in order to analyze the associated diazotroph communities. Based on Illumina amplicon sequencing of nifH genes and transcripts, we show that the diversity and potential activity of diazotroph communities varies according to micro-environments. We will comparatively discuss the influence of (a) the soil microbial "seed bank" and (b) plant genotype in shaping the respective microbiomes and selecting for potentially active diazotrophs. Actual N2 fixation activities of soil-genotype combinations and micro-environments will be shown on the basis of incubation assays using 15N2-containing atmospheres. Areas of potential N-transfer between diazotrophs and rice roots will be presented via the detection and visualization of spatial colonization patterns of selected diazotrophic groups on rice rhizoplanes. Our approach will help to increase the understanding of the contribution of Biological Nitrogen Fixation to rice cultivation in paddy soil ecosystems.

N2 production and fixation in deep-tier burrows of Squilla empusa in muddy sediments of Great Peconic Bay

NASA Astrophysics Data System (ADS)

Waugh, Stuart; Aller, Robert C.

2017-11-01

Global marine N budgets often show deficits due to dominance of benthic N2 production relative to pelagic N2 fixation. Recent studies have argued that benthic N2 fixation in shallow water environments has been underestimated. In particular, N2 fixation associated with animal burrows may be significant as indicated by high rates of N2 fixation reported in muddy sands populated by the ghost shrimp, Neotrypaea californiensis (Bertics et al., 2010). We investigated whether N2 fixation occurs at higher rates in the burrow-walls of the deep-burrowing ( 0.5-4 m) mantis shrimp, Squilla empusa, compared to ambient, estuarine muds and measured seasonal in-situ N2 concentrations in burrow-water relative to bottom-water. Acetylene reduction assays showed lower N2 fixation in burrow-walls than in un-populated sediments, likely due to inhibitory effects of O2 on ethylene production. Dissolved N2 was higher in burrow-water than proximate bottom-water at all seasons, demonstrating a consistent balance of net N2 production relative to fixation in deep-tier biogenic structures.

What Is the True Nitrogenase Reaction? A Guided Approach

ERIC Educational Resources Information Center

Ipata, Piero L.; Pesi, Rossana

2015-01-01

Only diazotrophic bacteria, called "Rizhobia," living as symbionts in the root nodules of leguminous plants and certain free-living prokaryotic cells can fix atmospheric N[subscript 2]. In these microorganisms, nitrogen fixation is carried out by the nitrogenase protein complex. However, the reduction of nitrogen to ammonia has an…

Redesigning alfalfa for use in mixtures with forage grasses

USDA-ARS?s Scientific Manuscript database

A lush field consisting of a mixture of grass and legumes is the goal of many producers. Such a production system has many benefits. The most important in times of high fertilizer prices is the reduced need for nitrogen (N) because of the legume's capacity for biological nitrogen fixation. However, ...

N sub 2 -fixation by freshly isolated Nostoc from coralloid roots of the cycad Macrozamia riedlei (Fisch. ex Gaud. ) Gardn

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lindblad, P.; Atkins, C.A.; Pate, J.S.

1991-03-01

Nitrogenase (EC 1.7.99.2) activity (acetylene reduction) and nitrogen fixation ({sup 15}N{sub 2} fixation) were measured in cyanobacteria freshly isolated from the coralloid roots of Macrozamia riedlei (Fisch. ex Gaud.) Gardn. The data indicate that cyanobacteria within cycad coralloid roots are differentiated specifically for symbiotic functioning in a microaerobic environment. Specializations include a high heterocyst frequency, enhanced permeability to O{sub 2}, and a direct dependence on the cycad for substrates to support nitrogenase activity.

Disease-mediated declines in N-fixation inputs by Alnus tenuifolia to early-successional floodplains in interior and south-central Alaska

Treesearch

R.W. Ruess; J.M. McFarland; L.M. Trummer; J.K. Rohrs-Richey

2009-01-01

Atmospheric nitrogen (N) fixation by Alnus tenuifolia can account for up to 70 percent of the N accumulated during vegetation development along river flood plains in interior Alaska. We assessed disease incidence and related mortality of a recent outbreak or fungal stem cankers on A. tenuifolia across three regions in Alaska...

Bacterial N2-fixation in mangrove ecosystems: insights from a diazotroph-mangrove interaction.

PubMed

Alfaro-Espinoza, Gabriela; Ullrich, Matthias S

2015-01-01

Mangrove forests are highly productive ecosystems but represent low nutrient environments. Nitrogen availability is one of the main factors limiting mangrove growth. Diazotrophs have been identified as key organisms that provide nitrogen to these environments. N2-fixation by such organisms was found to be higher in the mangrove roots than in surrounding rhizosphere. Moreover, previous studies showed that mangroves grew better in the presence of N2-fixers indicating a potentially mutualistic relationship. However, the molecular signals and mechanisms that govern these interactions are still poorly understood. Here we present novel insights in the interaction of a diazotroph with a mangrove species to improve our understanding of the molecular and ecophysiological relationship between these two organisms under controlled conditions. Our results showed that Marinobacterium mangrovicola is a versatile organism capable of competing with other organisms to survive for long periods in mangrove soils. N2-fixation by this bacterium was up-regulated in the presence of mangrove roots, indicating a possible beneficial interaction. The increase in N2-fixation was limited to cells of the exponential growth phase suggesting that N2-fixation differs over the bacterial growth cycle. Bacterial transformants harboring a transcriptional nifH::gusA fusion showed that M. mangrovicola successfully colonized mangrove roots and simultaneously conducted N2-fixation. The colonization process was stimulated by the lack of an external carbon source suggesting a possible mutualistic relationship. M. mangrovicola represents an interesting genetically accessible diazotroph, which colonize mangrove roots and exhibit higher N2-fixation in the presence of mangrove roots. Consequently, we propose this microorganism as a tool to study molecular interactions between N2-fixers and mangrove plants and to better understand how changes in the environment could impact these important and relatively unknown interactions.

Moving on from rigid plant stoichiometry: Optimal canopy nitrogen allocation within a novel land surface model

NASA Astrophysics Data System (ADS)

Caldararu, S.; Kern, M.; Engel, J.; Zaehle, S.

2016-12-01

Despite recent advances in global vegetation models, we still lack the capacity to predict observed vegetation responses to experimental environmental changes such as elevated CO2, increased temperature or nutrient additions. In particular for elevated CO2 (FACE) experiments, studies have shown that this is related in part to the models' inability to represent plastic changes in nutrient use and biomass allocation. We present a newly developed vegetation model which aims to overcome these problems by including optimality processes to describe nitrogen (N) and carbon allocation within the plant. We represent nitrogen allocation to the canopy and within the canopy between photosynthetic components as an optimal processes which aims to maximize net primary production (NPP) of the plant. We also represent biomass investment into aboveground and belowground components (root nitrogen uptake , biological N fixation) as an optimal process that maximizes plant growth by considering plant carbon and nutrient demands as well as acquisition costs. The model can now represent plastic changes in canopy N content and chlorophyll and Rubisco concentrations as well as in belowground allocation both on seasonal and inter-annual time scales. Specifically, we show that under elevated CO2 conditions, the model predicts a lower optimal leaf N concentration, which, combined with a redistribution of leaf N between the Rubisco and chlorophyll components, leads to a continued NPP response under high CO2, where models with a fixed canopy stoichiometry would predicts a quick onset of N limitation. In general, our model aims to include physiologically-based plant processes and avoid arbitrarily imposed parameters and thresholds in order to improve our predictive capability of vegetation responses under changing environmental conditions.

Comparison of production-phase environmental impact metrics derived at the farm- and national-scale for United States agricultural commodities

NASA Astrophysics Data System (ADS)

Costello, Christine; Xue, Xiaobo; Howarth, Robert W.

2015-11-01

Agricultural production is critical for human survival and simultaneously contributes to ecosystem degradation. There is a need for transparent, rapid methods for evaluating the environmental impacts of agricultural production at the system-level in order to develop sustainable food supplies. We have developed a method for estimating the greenhouse gas (GHG), land use and reactive nitrogen inputs associated with the agricultural production phase of major crop and livestock commodities produced in the United States (US). Materials flow analysis (MFA) and life cycle assessment (LCA) techniques were applied to national inventory datasets. The net anthropogenic nitrogen inputs (NANI) toolbox served as the primary accounting tool for LCA and MFA. NANI was updated to create links between nitrogen fertilizer and nitrogen fixation associated with feed crops and animal food commodities. Results for the functional units kilogram (kg) of product and kg of protein for 2002 data fall within ranges of published LCA results from farm-scale studies across most metrics. Exceptions include eutrophication potential for milk and GHGs for chicken and eggs, these exceptions arise due to differing methods and boundary assumptions; suggestions for increasing agreement are identified. Land use for livestock commodities are generally higher than reported by other LCA studies due to the inclusion of all land identified as pasture or grazing land in the US in this study and given that most of the estimates from other LCAs were completed in Europe where land is less abundant. The method provides a view of the entire US agricultural system and could be applied to any year using publically available data. Additionally, utilizing a top-down approach reduces data collection and processing time making it possible to develop environmental inventory metrics rapidly for system-level decision-making.

Utilization of the terrestrial cyanobacteria

NASA Astrophysics Data System (ADS)

Katoh, Hiroshi; Tomita-Yokotani, Kaori; Furukawa, Jun; Kimura, Shunta; Yokoshima, Mika; Yamaguchi, Yuji; Takenaka, Hiroyuki

The terrestrial, N _{2}-fixing cyanobacterium, Nostoc commune has expected to utilize for agriculture, food and terraforming cause of its extracellular polysaccharide, desiccation tolerance and nitrogen fixation. Previously, the first author indicated that desiccation related genes were analyzed and the suggested that the genes were related to nitrogen fixation and metabolisms. In this report, we suggest possibility of agriculture, using the cyanobacterium. Further, we also found radioactive compounds accumulated N. commune (cyanobacterium) in Fukushima, Japan after nuclear accident. Thus, it is investigated to decontaminate radioactive compounds from the surface soil by the cyanobacterium and showed to accumulate radioactive compounds using the cyanobacterium. We will discuss utilization of terrestrial cyanobacteria under closed environment. Keyword: Desiccation, terrestrial cyanobacteria, bioremediation, agriculture

Nitrogen fixation is not the only trait that determines the success of tropical legumes during secondary succession

NASA Astrophysics Data System (ADS)

Gei, Maria G.; Powers, Jennifer S.

2017-04-01

Legumes trees are well represented throughout the entire precipitation gradient of tropical forests. Many of these species are able to fix atmospheric dinitrogen through symbiosis and offer a mechanism to overcome nitrogen limitation typical of initial stages of secondary forest succession. While it is often assumed the success of legumes is linked to their fixation ability, the variation of other functional traits within this large group has received considerably less attention. Here we assessed legume abundance in secondary forest plots in 42 Neotropical chronosequences (the 2ndFOR network) that span a broad gradient of precipitation regimes and identified those traits that are favored in distinct successional environments. Our main finding is that in young secondary dry forests (5-20 years), legumes that have the potential to fix nitrogen and have small leaflet size become exceptionally abundant (up to 17-99% relative basal area). We suggest that in those species, reduced leaf area could help regulate leaf temperature and minimize water loss, and the cost of reduced total leaf area may be compensated by high photosynthetic rates maximized with nitrogen obtained through fixation. Overall, our study underscores great functional heterogeneity within tropical legumes, which likely translates into diverse biogeochemical cycles. In addition, these results provide a useful framework for active restoration of degraded areas, as it identifies a group of species that accumulate carbon at fast rates under warm and dry environments, conditions that are expected to become more common in the tropics.

Influence of elevated CO2 concentrations on cell division and nitrogen fixation rates in the bloom-forming cyanobacterium Nodularia spumigena

NASA Astrophysics Data System (ADS)

Czerny, J.; Ramos, J. Barcelos E.; Riebesell, U.

2009-04-01

The surface ocean currently absorbs about one-fourth of the CO2 emitted to the atmosphere from human activities. As this CO2 dissolves in seawater, it reacts with seawater to form carbonic acid, increasing ocean acidity and shifting the partitioning of inorganic carbon species towards increased CO2 at the expense of CO32- concentrations. While the decrease in [CO32-] and/or increase in [H+] has been found to adversely affect many calcifying organisms, some photosynthetic organisms appear to benefit from increasing [CO2]. Among these is the cyanobacterium Trichodesmium, a predominant diazotroph (nitrogen-fixing) in large parts of the oligotrophic oceans, which responded with increased carbon and nitrogen fixation at elevated pCO2. With the mechanism underlying this CO2 stimulation still unknown, the question arises whether this is a common response of diazotrophic cyanobacteria. In this study we therefore investigate the physiological response of Nodularia spumigena, a heterocystous bloom-forming diazotroph of the Baltic Sea, to CO2-induced changes in seawater carbonate chemistry. N. spumigena reacted to seawater acidification/carbonation with reduced cell division rates and nitrogen fixation rates, accompanied by significant changes in carbon and phosphorus quota and elemental composition of the formed biomass. Possible explanations for the contrasting physiological responses of Nodularia compared to Trichodesmium may be found in the different ecological strategies of non-heterocystous (Trichodesmium) and heterocystous (Nodularia) cyanobacteria.

Nitrogen starvation strategies and photobioreactor design for enhancing lipid content and lipid production of a newly isolated microalga Chlorella vulgaris ESP-31: implications for biofuels.

PubMed

Yeh, Kuei-Ling; Chang, Jo-Shu

2011-11-01

Microalgae are recognized for serving as a sustainable source for biodiesel production. This study investigated the effect of nitrogen starvation strategies and photobioreactor design on the performance of lipid production and of CO(2) fixation of an indigenous microalga Chlorella vulgaris ESP-31. Comparison of single-stage and two-stage nitrogen starvation strategies shows that single-stage cultivation on basal medium with low initial nitrogen source concentration (i.e., 0.313 g/L KNO(3)) was the most effective approach to enhance microalgal lipid production, attaining a lipid productivity of 78 mg/L/d and a lipid content of 55.9%. The lipid productivity of C. vulgaris ESP-31 was further upgraded to 132.4 mg/L/d when it was grown in a vertical tubular photobioreactor with a high surface to volume ratio of 109.3 m(2)/m(3) . The high lipid productivity was also accompanied by fixation of 6.36 g CO(2) during the 10-day photoautotrophic growth with a CO(2) fixation rate of 430 mg/L/d. Analysis of fatty acid composition of the microalgal lipid indicates that over 65% of fatty acids in the microalgal lipid are saturated [i.e., palmitic acid (C16:0) and stearic acid (C18:0)] and monounsaturated [i.e., oleic acid (C18:1)]. This lipid quality is suitable for biodiesel production. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Net primary production and canopy nitrogen in a temperate forest landscape: an analysis using imaging spectroscopy, modeling and field data

Treesearch

Scott V. Ollinger; Marie-Louise Smith

2005-01-01

Understanding spatial patterns of net primary production (NPP) is central to the study of terrestrial ecosystems, but efforts are frequently hampered by a lack of spatial information regarding factors such as nitrogen availability and site history. Here, we examined the degree to which canopy nitrogen can serve as an indicator of patterns of NPP at the Bartlett...

Is the distribution of nitrogen-fixing cyanobacteria in the oceans related to temperature?

PubMed

Stal, Lucas J

2009-07-01

Approximately 50% of the global natural fixation of nitrogen occurs in the oceans supporting a considerable part of the new primary production. Virtually all nitrogen fixation in the ocean occurs in the tropics and subtropics where the surface water temperature is 25°C or higher. It is attributed almost exclusively to cyanobacteria. This is remarkable firstly because diazotrophic cyanobacteria are found in other environments irrespective of temperature and secondly because primary production in temperate and cold oceans is generally limited by nitrogen. Cyanobacteria are oxygenic phototrophic organisms that evolved a variety of strategies protecting nitrogenase from oxygen inactivation. Free-living diazotrophic cyanobacteria in the ocean are of the non-heterocystous type, namely the filamentous Trichodesmium and the unicellular groups A-C. I will argue that warm water is a prerequisite for these diazotrophic organisms because of the low-oxygen solubility and high rates of respiration allowing the organism to maintain anoxic conditions in the nitrogen-fixing cell. Heterocystous cyanobacteria are abundant in freshwater and brackish environments in all climatic zones. The heterocyst cell envelope is a tuneable gas diffusion barrier that optimizes the influx of both oxygen and nitrogen, while maintaining anoxic conditions inside the cell. It is not known why heterocystous cyanobacteria are absent from the temperate and cold oceans and seas.

Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense

PubMed Central

Sotomaior, P.; Araújo, L.M.; Nishikawa, C.Y.; Huergo, L.F.; Monteiro, R.A.; Pedrosa, F.O.; Chubatsu, L.S.; Souza, E.M.

2012-01-01

Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate. PMID:22983183

Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense.

PubMed

Sotomaior, P; Araújo, L M; Nishikawa, C Y; Huergo, L F; Monteiro, R A; Pedrosa, F O; Chubatsu, L S; Souza, E M

2012-12-01

Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate.

Photosynthetic strategies of two Mojave Desert shrubs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kleinkopf, G.E.; Hartsock, T.L.; Wallace, A.

1980-01-01

Photosynthetic production of two Mojave Desert shrubs was measured under natural growing conditions. Measurements of photosynthesis, transpiration, resistances to water vapor flux, soil moisture potential, and tissue water potential were made. Atriplex canescens (Pursh) Nutt., a member of the C/sub 4/ biochemical carbon dioxide fixation group was highly competitive in growth rate and production during conditions of adequate soil moisture. As soil moisture conditions declined to minus 40 bars, the net photosynthetic rate of Atriplex decreased to zero. However, the C/sub 3/ shrub species Larrea tridentata (Sesse and Moc. ex DC.) Cov. was able to maintain positive net photosynthetic productionmore » during conditions of high temperature and extreme low soil moisture through the major part of the season. The comparative advantages of the C/sub 4/ versus the C/sub 3/ pathway of carbon fixation was lost between these two species as the soil moisture potential declined to minus 40 bars. Desert plants have diffferent strategies for survival, one of the strategies being the C/sub 4/ biochemical carbon fixation pathway. However, many of the plants are members of the C/sub 3/ group. In this instance, the C/sub 4/ fixation pathway does not confer an added advantage to the productivity of the species in the Mojave Desert. Species distribution based on comparative photosynthetic production is discussed« less

Time-series resolution of gradual nitrogen starvation and its impact on photosynthesis in the cyanobacterium Synechocystis PCC 6803.

PubMed

Krasikov, Vladimir; Aguirre von Wobeser, Eneas; Dekker, Henk L; Huisman, Jef; Matthijs, Hans C P

2012-07-01

Sequential adaptation to nitrogen deprivation and ultimately to full starvation requires coordinated adjustment of cellular functions. We investigated changes in gene expression and cell physiology of the cyanobacterium Synechocystis PCC 6803 during 96 h of nitrogen starvation. During the first 6 h, the transcriptome showed activation of nitrogen uptake and assimilation systems and of the core nitrogen and carbon assimilation regulators. However, the nitrogen-deprived cells still grew at the same rate as the control and even showed transiently increased expression of phycobilisome genes. After 12 h, cell growth decreased and chlorosis started with degradation of the nitrogen-rich phycobilisomes. During this phase, the transcriptome showed suppression of genes for phycobilisomes, for carbon fixation and for de novo protein synthesis. Interestingly, photosynthetic activity of both photosystem I (PSI) and photosystem II was retained quite well. Excess electrons were quenched by the induction of terminal oxidase and hydrogenase genes, compensating for the diminished carbon fixation and nitrate reduction activity. After 48 h, the cells ceased most activities. A marked exception was the retained PSI gene transcription, possibly this supports the viability of Synechocystis cells and enables rapid recovery after relieving from nitrogen starvation. During early recovery, many genes changed expression, supporting the resumed cellular activity. In total, our results distinguished three phases during gradual nitrogen depletion: (1) an immediate response, (2) short-term acclimation and (3) long-term survival. This shows that cyanobacteria respond to nitrogen starvation by a cascade of physiological adaptations reflected by numerous changes in the transcriptome unfolding at different timescales. Copyright © Physiologia Plantarum 2012.

Constitutive expression of the nifA gene activates associative nitrogen fixation of Enterobacter gergoviae 57-7, an opportunistic endophytic diazotroph.

PubMed

An, Q; Dong, Y; Wang, W; Li, Y; Li, J

2007-09-01

This study was undertaken to investigate whether a nitrogen-fixing bacterium Enterobacter gergoviae 57-7, which was isolated from surface-sterilized maize (Zea mays L.) roots, can colonize in roots and whether constitutive expression of the nifA gene encoding the transcriptional activator of nitrogenase genes can activate nif gene expression in planta. Maize seedlings grown in an agar medium were inoculated with Ent. gergoviae strains containing the green fluorescent protein reporter gene. Root colonization and expression of the dinitrogenase reductase gene (nifH) by Ent. gergoviae were observed by confocal laser scanning microscopy. gfp-tagged Ent. gergoviae was observed to colonize predominantly in cortical aerenchyma of primary and lateral roots and in stellar parenchyma cells and xylem vessels of primary roots. In planta nifH :: gfp expression was not detected but after a constitutively expressed nifA gene was introduced into Ent. gergoviae. Enterobacter gergoviae 57-7 is an opportunistic endophyte because it can live in soil and colonize in maize roots in the gnotobiotic agar culture. In agreement with previous (15)N-dilution evidence that Ent. gergoviae 57-7 did not fix N(2) in association with maize in pots whereas a derivative E7 containing a constitutively expressed nifA gene promoted plant growth partly through associative nitrogen fixation, constitutive expression of the nifA gene can activate bacterial nif gene expression in planta. This study and our previous studies suggest that manipulation of the promoter of the nifA gene in a nitrogen-fixing bacterium having a high colonization competence is a practical and promising approach to achieve a stable associative nitrogen fixation for cereals.

Metabolic Adaptation, a Specialized Leaf Organ Structure and Vascular Responses to Diurnal N2 Fixation by Nostoc azollae Sustain the Astonishing Productivity of Azolla Ferns without Nitrogen Fertilizer.

PubMed

Brouwer, Paul; Bräutigam, Andrea; Buijs, Valerie A; Tazelaar, Anne O E; van der Werf, Adrie; Schlüter, Urte; Reichart, Gert-Jan; Bolger, Anthony; Usadel, Björn; Weber, Andreas P M; Schluepmann, Henriette

2017-01-01

Sustainable agriculture demands reduced input of man-made nitrogen (N) fertilizer, yet N 2 fixation limits the productivity of crops with heterotrophic diazotrophic bacterial symbionts. We investigated floating ferns from the genus Azolla that host phototrophic diazotrophic Nostoc azollae in leaf pockets and belong to the fastest growing plants. Experimental production reported here demonstrated N-fertilizer independent production of nitrogen-rich biomass with an annual yield potential per ha of 1200 kg -1 N fixed and 35 t dry biomass. 15 N 2 fixation peaked at noon, reaching 0.4 mg N g -1 dry weight h -1 . Azolla ferns therefore merit consideration as protein crops in spite of the fact that little is known about the fern's physiology to enable domestication. To gain an understanding of their nitrogen physiology, analyses of fern diel transcript profiles under differing nitrogen fertilizer regimes were combined with microscopic observations. Results established that the ferns adapted to the phototrophic N 2 -fixing symbionts N. azollae by (1) adjusting metabolically to nightly absence of N supply using responses ancestral to ferns and seed plants; (2) developing a specialized xylem-rich vasculature surrounding the leaf-pocket organ; (3) responding to N-supply by controlling transcripts of genes mediating nutrient transport, allocation and vasculature development. Unlike other non-seed plants, the Azolla fern clock is shown to contain both the morning and evening loops; the evening loop is known to control rhythmic gene expression in the vasculature of seed plants and therefore may have evolved along with the vasculature in the ancestor of ferns and seed plants.

Metabolic Adaptation, a Specialized Leaf Organ Structure and Vascular Responses to Diurnal N2 Fixation by Nostoc azollae Sustain the Astonishing Productivity of Azolla Ferns without Nitrogen Fertilizer

PubMed Central

Brouwer, Paul; Bräutigam, Andrea; Buijs, Valerie A.; Tazelaar, Anne O. E.; van der Werf, Adrie; Schlüter, Urte; Reichart, Gert-Jan; Bolger, Anthony; Usadel, Björn; Weber, Andreas P. M.; Schluepmann, Henriette

2017-01-01

Sustainable agriculture demands reduced input of man-made nitrogen (N) fertilizer, yet N2 fixation limits the productivity of crops with heterotrophic diazotrophic bacterial symbionts. We investigated floating ferns from the genus Azolla that host phototrophic diazotrophic Nostoc azollae in leaf pockets and belong to the fastest growing plants. Experimental production reported here demonstrated N-fertilizer independent production of nitrogen-rich biomass with an annual yield potential per ha of 1200 kg−1 N fixed and 35 t dry biomass. 15N2 fixation peaked at noon, reaching 0.4 mg N g−1 dry weight h−1. Azolla ferns therefore merit consideration as protein crops in spite of the fact that little is known about the fern’s physiology to enable domestication. To gain an understanding of their nitrogen physiology, analyses of fern diel transcript profiles under differing nitrogen fertilizer regimes were combined with microscopic observations. Results established that the ferns adapted to the phototrophic N2-fixing symbionts N. azollae by (1) adjusting metabolically to nightly absence of N supply using responses ancestral to ferns and seed plants; (2) developing a specialized xylem-rich vasculature surrounding the leaf-pocket organ; (3) responding to N-supply by controlling transcripts of genes mediating nutrient transport, allocation and vasculature development. Unlike other non-seed plants, the Azolla fern clock is shown to contain both the morning and evening loops; the evening loop is known to control rhythmic gene expression in the vasculature of seed plants and therefore may have evolved along with the vasculature in the ancestor of ferns and seed plants. PMID:28408911

Regulation of respiration and the oxygen diffusion barrier in soybean protect symbiotic nitrogen fixation from chilling-induced inhibition and shoots from premature senescence.

PubMed

van Heerden, Philippus D R; Kiddle, Guy; Pellny, Till K; Mokwala, Phatlane W; Jordaan, Anine; Strauss, Abram J; de Beer, Misha; Schlüter, Urte; Kunert, Karl J; Foyer, Christine H

2008-09-01

Symbiotic nitrogen fixation is sensitive to dark chilling (7 degrees C-15 degrees C)-induced inhibition in soybean (Glycine max). To characterize the mechanisms that cause the stress-induced loss of nodule function, we examined nodule structure, carbon-nitrogen interactions, and respiration in two soybean genotypes that differ in chilling sensitivity: PAN809 (PAN), which is chilling sensitive, and Highveld Top (HT), which is more chilling resistant. Nodule numbers were unaffected by dark chilling, as was the abundance of the nitrogenase and leghemoglobin proteins. However, dark chilling decreased nodule respiration rates, nitrogenase activities, and NifH and NifK mRNAs and increased nodule starch, sucrose, and glucose in both genotypes. Ureide and fructose contents decreased only in PAN nodules. While the chilling-induced decreases in nodule respiration persisted in PAN even after return to optimal temperatures, respiration started to recover in HT by the end of the chilling period. The area of the intercellular spaces in the nodule cortex and infected zone was greatly decreased in HT after three nights of chilling, an acclimatory response that was absent from PAN. These data show that HT nodules are able to regulate both respiration and the area of the intercellular spaces during chilling and in this way control the oxygen diffusion barrier, which is a key component of the nodule stress response. We conclude that chilling-induced loss of symbiotic nitrogen fixation in PAN is caused by the inhibition of respiration coupled to the failure to regulate the oxygen diffusion barrier effectively. The resultant limitations on nitrogen availability contribute to the greater chilling-induced inhibition of photosynthesis in PAN than in HT.

Regulation of Respiration and the Oxygen Diffusion Barrier in Soybean Protect Symbiotic Nitrogen Fixation from Chilling-Induced Inhibition and Shoots from Premature Senescence1[W][OA

PubMed Central

van Heerden, Philippus D.R.; Kiddle, Guy; Pellny, Till K.; Mokwala, Phatlane W.; Jordaan, Anine; Strauss, Abram J.; de Beer, Misha; Schlüter, Urte; Kunert, Karl J.; Foyer, Christine H.

2008-01-01

Symbiotic nitrogen fixation is sensitive to dark chilling (7°C–15°C)-induced inhibition in soybean (Glycine max). To characterize the mechanisms that cause the stress-induced loss of nodule function, we examined nodule structure, carbon-nitrogen interactions, and respiration in two soybean genotypes that differ in chilling sensitivity: PAN809 (PAN), which is chilling sensitive, and Highveld Top (HT), which is more chilling resistant. Nodule numbers were unaffected by dark chilling, as was the abundance of the nitrogenase and leghemoglobin proteins. However, dark chilling decreased nodule respiration rates, nitrogenase activities, and NifH and NifK mRNAs and increased nodule starch, sucrose, and glucose in both genotypes. Ureide and fructose contents decreased only in PAN nodules. While the chilling-induced decreases in nodule respiration persisted in PAN even after return to optimal temperatures, respiration started to recover in HT by the end of the chilling period. The area of the intercellular spaces in the nodule cortex and infected zone was greatly decreased in HT after three nights of chilling, an acclimatory response that was absent from PAN. These data show that HT nodules are able to regulate both respiration and the area of the intercellular spaces during chilling and in this way control the oxygen diffusion barrier, which is a key component of the nodule stress response. We conclude that chilling-induced loss of symbiotic nitrogen fixation in PAN is caused by the inhibition of respiration coupled to the failure to regulate the oxygen diffusion barrier effectively. The resultant limitations on nitrogen availability contribute to the greater chilling-induced inhibition of photosynthesis in PAN than in HT. PMID:18667725

Metabolic analysis of Chlorobium chlorochromatii CaD3 reveals clues of the symbiosis in ‘Chlorochromatium aggregatum'.

PubMed Central

Cerqueda-García, Daniel; Martínez-Castilla, León P; Falcón, Luisa I; Delaye, Luis

2014-01-01

A symbiotic association occurs in ‘Chlorochromatium aggregatum', a phototrophic consortium integrated by two species of phylogenetically distant bacteria composed by the green-sulfur Chlorobium chlorochromatii CaD3 epibiont that surrounds a central β-proteobacterium. The non-motile chlorobia can perform nitrogen and carbon fixation, using sulfide as electron donors for anoxygenic photosynthesis. The consortium can move due to the flagella present in the central β-protobacterium. Although Chl. chlorochromatii CaD3 is never found as free-living bacteria in nature, previous transcriptomic and proteomic studies have revealed that there are differential transcription patterns between the symbiotic and free-living status of Chl. chlorocromatii CaD3 when grown in laboratory conditions. The differences occur mainly in genes encoding the enzymatic reactions involved in nitrogen and amino acid metabolism. We performed a metabolic reconstruction of Chl. chlorochromatii CaD3 and an in silico analysis of its amino acid metabolism using an elementary flux modes approach (EFM). Our study suggests that in symbiosis, Chl. chlorochromatii CaD3 is under limited nitrogen conditions where the GS/GOGAT (glutamine synthetase/glutamate synthetase) pathway is actively assimilating ammonia obtained via N2 fixation. In contrast, when free-living, Chl. chlorochromatii CaD3 is in a condition of nitrogen excess and ammonia is assimilated by the alanine dehydrogenase (AlaDH) pathway. We postulate that ‘Chlorochromatium aggregatum' originated from a parasitic interaction where the N2 fixation capacity of the chlorobia would be enhanced by injection of 2-oxoglutarate from the β-proteobacterium via the periplasm. This consortium would have the advantage of motility, which is fundamental to a phototrophic bacterium, and the syntrophy of nitrogen and carbon sources. PMID:24285361

Efficient Nitrogen Fixation via a Redox-Flexible Single-Iron Site with Reverse-Dative Iron → Boron σ Bonding.

PubMed

Lu, Jun-Bo; Ma, Xue-Lu; Wang, Jia-Qi; Liu, Jin-Cheng; Xiao, Hai; Li, Jun

2018-05-10

Model systems of the FeMo cofactor of nitrogenase have been explored extensively in catalysis to gain insights into their ability for nitrogen fixation that is of vital importance to the human society. Here we investigate the trigonal pyramidal borane-ligand Fe complex by first-principles calculations, and find that the variation of oxidation state of Fe along the reaction path correlates with that of the reverse-dative Fe → B bonding. The redox-flexibility of the reverse-dative Fe → B bonding helps to provide an electron reservoir that buffers and stabilizes the evolution of Fe oxidation state, which is essential for forming the key intermediates of N 2 activation. Our work provides insights for understanding and optimizing homogeneous and surface single-atom catalysts with reverse-dative donating ligands for efficient dinitrogen fixation. The extension of this kind of molecular catalytic active center to heterogeneous catalysts with surface single-clusters is also discussed.

Transcriptional Activities of the Microbial Consortium Living with the Marine Nitrogen-Fixing Cyanobacterium Trichodesmium Reveal Potential Roles in Community-Level Nitrogen Cycling.

PubMed

Lee, Michael D; Webb, Eric A; Walworth, Nathan G; Fu, Fei-Xue; Held, Noelle A; Saito, Mak A; Hutchins, David A

2018-01-01

Trichodesmium is a globally distributed cyanobacterium whose nitrogen-fixing capability fuels primary production in warm oligotrophic oceans. Like many photoautotrophs, Trichodesmium serves as a host to various other microorganisms, yet little is known about how this associated community modulates fluxes of environmentally relevant chemical species into and out of the supraorganismal structure. Here, we utilized metatranscriptomics to examine gene expression activities of microbial communities associated with Trichodesmium erythraeum (strain IMS101) using laboratory-maintained enrichment cultures that have previously been shown to harbor microbial communities similar to those of natural populations. In enrichments maintained under two distinct CO 2 concentrations for ∼8 years, the community transcriptional profiles were found to be specific to the treatment, demonstrating a restructuring of overall gene expression had occurred. Some of this restructuring involved significant increases in community respiration-related transcripts under elevated CO 2 , potentially facilitating the corresponding measured increases in host nitrogen fixation rates. Particularly of note, in both treatments, community transcripts involved in the reduction of nitrate, nitrite, and nitrous oxide were detected, suggesting the associated organisms may play a role in colony-level nitrogen cycling. Lastly, a taxon-specific analysis revealed distinct ecological niches of consistently cooccurring major taxa that may enable, or even encourage, the stable cohabitation of a diverse community within Trichodesmium consortia. IMPORTANCE Trichodesmium is a genus of globally distributed, nitrogen-fixing marine cyanobacteria. As a source of new nitrogen in otherwise nitrogen-deficient systems, these organisms help fuel carbon fixation carried out by other more abundant photoautotrophs and thereby have significant roles in global nitrogen and carbon cycling. Members of the Trichodesmium genus tend to form large macroscopic colonies that appear to perpetually host an association of diverse interacting microbes distinct from the surrounding seawater, potentially making the entire assemblage a unique miniature ecosystem. Since its first successful cultivation in the early 1990s, there have been questions about the potential interdependencies between Trichodesmium and its associated microbial community and whether the host's seemingly enigmatic nitrogen fixation schema somehow involved or benefited from its epibionts. Here, we revisit these old questions with new technology and investigate gene expression activities of microbial communities living in association with Trichodesmium . Copyright © 2017 American Society for Microbiology.

Impact of diazotrophy on N stable isotope signatures of nitrate and particulate organic nitrogen: case studies in the north-eastern tropical Atlantic Ocean.

PubMed

Wannicke, Nicola; Liskow, Iris; Voss, Maren

2010-09-01

During two independent cruises in the north-eastern tropical Atlantic Ocean, we applied two different approaches to investigate the impact of diazotrophy on nitrogen stable isotope signatures in nitrate and particulate organic nitrogen (PON) of the food-web constituents. The first approach, used during the Poseidon cruise 348 in the Mauritanian upwelling, investigated the long-term influence of diazotrophy on the natural abundance of δ(15)N-NO(-)(3) and PON. The second approach, adopted during the Cape Verde field cruise, applied stable isotope tracer addition experiments. These served to determine the instantaneous transfer of diazotrophic N to the higher trophic level. Both approaches showed that N(2) fixation was compatible with the pattern and the magnitude of the isotopic depletion of dissolved NO(-)(3) during the Mauritanian upwelling cruise, as well as PON in zooplankton and phytoplankton during the Cape Verde cruises. An N-budget using (15)N incorporation rates and diazotrophic N(2) fixation rates showed that 6 % of the daily N(2) fixation was potentially taken up by the mesozooplankton community. Direct grazing accounted for 56 % of gross mesozooplanktonic N incorporation, while 46 % occurred due to channelling through the microbial loop.

[Effects of plastic film mulching and nitrogen application rate on net global warming potential in semiarid rain-fed maize cropland].

PubMed

Liu, Jian Can; Wang, Ze Lin; Yue, Shan Chao; Li, Shi Qing

2018-04-01

A one-year field experiment was conducted to evaluate the effects of plastic film mulching (FM) and nitrogen application rates applied to rain-fed maize fields on net global warming potential (Net GWP) and greenhouse gas intensity (GHGI) at the Changwu Agricultural and Ecological Experimental Station. Both GWP and GHGI were affected by the plastic film mulching and nitrogen application rate. Under the FM treatment, maize yield ranged from 1643 to 16699 kg·hm -2 , the net GWP (CO 2 -eq) ranged from 595 to 4376 kg·hm -2 ·a -1 , and the GHGI (CO 2 -eq) ranged from 213 to 358 kg·t -1 . The grain yield of maize, net GWP and GHGI for the UM (no mulching) treatment were 956 to 8821 kg·hm -2 , 342 to 4004 kg·hm -2 ·a -1 and 204 to 520 kg·t -1 , respectively. The results suggested that plastic film mulching could simultaneously improve grain yield and decrease GHGI in rain-fed cropland along with nitrogen fertilizer of 250 kg·hm -2 .

N2-fixing tropical legume evolution: a contributor to enhanced weathering through the Cenozoic?

PubMed Central

Hedin, Lars O.; Leake, Jonathan R.

2017-01-01

Fossil and phylogenetic evidence indicates legume-rich modern tropical forests replaced Late Cretaceous palm-dominated tropical forests across four continents during the early Cenozoic (58–42 Ma). Tropical legume trees can transform ecosystems via their ability to fix dinitrogen (N2) and higher leaf N compared with non-legumes (35–65%), but it is unclear how their evolutionary rise contributed to silicate weathering, the long-term sink for atmospheric carbon dioxide (CO2). Here we hypothesize that the increasing abundance of N2-fixing legumes in tropical forests amplified silicate weathering rates by increased input of fixed nitrogen (N) to terrestrial ecosystems via interrelated mechanisms including increasing microbial respiration and soil acidification, and stimulating forest net primary productivity. We suggest the high CO2 early Cenozoic atmosphere further amplified legume weathering. Evolution of legumes with high weathering rates was probably driven by their high demand for phosphorus and micronutrients required for N2-fixation and nodule formation. PMID:28814651

N2-fixing tropical legume evolution: a contributor to enhanced weathering through the Cenozoic?

PubMed

Epihov, Dimitar Z; Batterman, Sarah A; Hedin, Lars O; Leake, Jonathan R; Smith, Lisa M; Beerling, David J

2017-08-16

Fossil and phylogenetic evidence indicates legume-rich modern tropical forests replaced Late Cretaceous palm-dominated tropical forests across four continents during the early Cenozoic (58-42 Ma). Tropical legume trees can transform ecosystems via their ability to fix dinitrogen (N 2 ) and higher leaf N compared with non-legumes (35-65%), but it is unclear how their evolutionary rise contributed to silicate weathering, the long-term sink for atmospheric carbon dioxide (CO 2 ). Here we hypothesize that the increasing abundance of N 2 -fixing legumes in tropical forests amplified silicate weathering rates by increased input of fixed nitrogen (N) to terrestrial ecosystems via interrelated mechanisms including increasing microbial respiration and soil acidification, and stimulating forest net primary productivity. We suggest the high CO 2 early Cenozoic atmosphere further amplified legume weathering. Evolution of legumes with high weathering rates was probably driven by their high demand for phosphorus and micronutrients required for N 2 -fixation and nodule formation. © 2017 The Author(s).

Insect-mediated nitrogen dynamics in decomposing wood

Treesearch

Michael D. Ulyshen

2015-01-01

1.Wood decomposition is characterised by complex and poorly understood nitrogen (N) dynamics with unclear implications for forest nutrient cycling and productivity.Wood-dwelling microbes have developed unique strategies for coping with the N limitations imposed by their substrate, including the translocation of N into wood by cord-forming fungi and the fixation of...

A proteomic network for symbiotic nitrogen fixation efficiency in Bradyrhizobium elkanii

USDA-ARS?s Scientific Manuscript database

Rhizobia bacteroids colonize legumes and reduce N2 to NH3 in root nodules. The current model is that bacteroids avoid assimilating this NH3. Instead, the legume forms glutamine from it, the nitrogen of which is returned to the bacteroid as leucine, isoleucine, valine, dicarboxylates, and peptides. I...

Plant Guide: Searls' prairie clover (Dalea searlsiae [A. Gray] Barneby)

Treesearch

Loren St. John; Derek Tilley; Dan Ogle; Doug Johnson; Shaun Bushman

2011-01-01

Plants of Searls' prairie clover are grazed by many animals including rabbits, deer, sheep and cattle. This legume forms a symbiotic association with soil bacteria (Rhizobium spp.) that allows fixation of atmospheric nitrogen, which can enhance forage nitrogen content for wildlife and livestock. Flowers are visited by pollinators, primarily bees, during bloom from...

cDNA-AFLP analysis of differential gene expression related to cell chemotactic and encystment of Azospirillum brasilense.

PubMed

Li, Huamin; Cui, Yanhua; Wu, Lixian; Tu, Ran; Chen, Sanfeng

2011-12-20

Our previous study indicated org35 was involved in chemotaxis and interacted with nitrogen fixation transcriptional activator NifA via PAS domain. In order to reveal the role of org35 in nitrogen regulation, the downstream target genes of org35 were identified. We here report differentially expressed genes in org35 mutants comparing with wild type Sp7 by means of cDNA-AFLP. Four up-regulated transcript-derived fragments (TDFs) homologues of chemotaxis transduction proteins were found, including CheW, methyl-accepting chemotaxis protein and response regulator CheY-like receiver. Three distinct TDFs (AB46, AB58 and AB63) were similar to PHB de-polymerase C-terminus, cell shape-determining protein and flagellin domain protein. And 11 TDFs showed similarities with signal transduction proteins, including homologous protein of the nitrogen regulation protein NtrY and nitrate/nitrite response regulator protein NarL. These data suggested that the Azospirillum brasilense org35 was a multi-effecter and involved in chemotaxis, cyst development and regulation of nitrogen fixation. Copyright © 2010 Elsevier GmbH. All rights reserved.

Responses of the nitrogen-fixing aquatic fern Azolla to water contaminated with ciprofloxacin: Impacts on biofertilization.

PubMed

Gomes, Marcelo Pedrosa; de Brito, Júlio César Moreira; Carvalho Carneiro, Marília Mércia Lima; Ribeiro da Cunha, Mariem Rodrigues; Garcia, Queila Souza; Figueredo, Cleber Cunha

2018-01-01

We investigated the ability of the aquatic fern Azolla to take up ciprofloxacin (Cipro), as well as the effects of that antibiotic on the N-fixing process in plants grown in medium deprived (-N) or provided (+N) with nitrogen (N). Azolla was seen to accumulate Cipro at concentrations greater than 160 μg g -1 dry weight when cultivated in 3.05 mg Cipro l -1 , indicating it as a candidate for Cipro recovery from water. Although Cipro was not seen to interfere with the heterocyst/vegetative cell ratios, the antibiotic promoted changes with carbon and nitrogen metabolism in plants. Decreased photosynthesis and nitrogenase activity, and altered plant's amino acid profile, with decreases in cell N concentrations, were observed. The removal of N from the growth medium accentuated the deleterious effects of Cipro, resulting in lower photosynthesis, N-fixation, and assimilation rates, and increased hydrogen peroxide accumulation. Our results shown that Cipro may constrain the use of Azolla as a biofertilizer species due to its interference with nitrogen fixation processes. Copyright © 2017 Elsevier Ltd. All rights reserved.

Fixation Probability in a Haploid-Diploid Population

PubMed Central

Bessho, Kazuhiro; Otto, Sarah P.

2017-01-01

Classical population genetic theory generally assumes either a fully haploid or fully diploid life cycle. However, many organisms exhibit more complex life cycles, with both free-living haploid and diploid stages. Here we ask what the probability of fixation is for selected alleles in organisms with haploid-diploid life cycles. We develop a genetic model that considers the population dynamics using both the Moran model and Wright–Fisher model. Applying a branching process approximation, we obtain an accurate fixation probability assuming that the population is large and the net effect of the mutation is beneficial. We also find the diffusion approximation for the fixation probability, which is accurate even in small populations and for deleterious alleles, as long as selection is weak. These fixation probabilities from branching process and diffusion approximations are similar when selection is weak for beneficial mutations that are not fully recessive. In many cases, particularly when one phase predominates, the fixation probability differs substantially for haploid-diploid organisms compared to either fully haploid or diploid species. PMID:27866168

Competition for ammonia influences the structure of chemotrophic communities in geothermal springs.

PubMed

Hamilton, Trinity L; Koonce, Evangeline; Howells, Alta; Havig, Jeff R; Jewell, Talia; de la Torre, José R; Peters, John W; Boyd, Eric S

2014-01-01

Source waters sampled from Perpetual Spouter hot spring (pH 7.03, 86.4°C), Yellowstone National Park, WY, have low concentrations of total ammonia, nitrite, and nitrate, suggesting nitrogen (N) limitation and/or tight coupling of N cycling processes. Dominant small-subunit rRNA sequences in Perpetual Spouter source sediments are closely affiliated with the ammonia-oxidizing archaeon "Candidatus Nitrosocaldus yellowstonii" and the putatively nitrogen-fixing (diazotrophic) bacterium Thermocrinis albus, respectively, suggesting that these populations may interact at the level of the bioavailable N pool, specifically, ammonia. This hypothesis was evaluated by using a combination of geochemical, physiological, and transcriptomic analyses of sediment microcosms. Amendment of microcosms with allylthiourea, an inhibitor of ammonia oxidation, decreased rates of acetylene reduction (a proxy for N2 fixation) and nitrite production (a proxy for ammonia oxidation) and decreased transcript levels of structural genes involved in both nitrogen fixation (nifH) and ammonia oxidation (amoA). In contrast, amendment of microcosms with ammonia stimulated nitrite production and increased amoA transcript levels while it suppressed rates of acetylene reduction and decreased nifH transcript levels. Sequencing of amplified nifH and amoA transcripts from native sediments, as well as microcosms, at 2 and 4 h postamendment, indicates that the dominant and responsive populations involved in ammonia oxidation and N2 fixation are closely affiliated with Ca. Nitrosocaldus yellowstonii and T. albus, respectively. Collectively, these results suggest that ammonia-oxidizing archaea, such as Ca. Nitrosocaldus yellowstonii, have an apparent affinity for ammonia that is higher than that of the diazotrophs present in this ecosystem. Depletion of the bioavailable N pool through the activity of ammonia-oxidizing archaea likely represents a strong selective pressure for the inclusion of organisms capable of nitrogen fixation in geothermal communities. These observations help to explain the strong pattern in the codistribution of ammonia-oxidizing archaea and diazotrophs in circumneutral-to-alkaline geothermal springs.

Competition for Ammonia Influences the Structure of Chemotrophic Communities in Geothermal Springs

PubMed Central

Hamilton, Trinity L.; Koonce, Evangeline; Howells, Alta; Havig, Jeff R.; Jewell, Talia; de la Torre, José R.; Peters, John W.

2014-01-01

Source waters sampled from Perpetual Spouter hot spring (pH 7.03, 86.4°C), Yellowstone National Park, WY, have low concentrations of total ammonia, nitrite, and nitrate, suggesting nitrogen (N) limitation and/or tight coupling of N cycling processes. Dominant small-subunit rRNA sequences in Perpetual Spouter source sediments are closely affiliated with the ammonia-oxidizing archaeon “Candidatus Nitrosocaldus yellowstonii” and the putatively nitrogen-fixing (diazotrophic) bacterium Thermocrinis albus, respectively, suggesting that these populations may interact at the level of the bioavailable N pool, specifically, ammonia. This hypothesis was evaluated by using a combination of geochemical, physiological, and transcriptomic analyses of sediment microcosms. Amendment of microcosms with allylthiourea, an inhibitor of ammonia oxidation, decreased rates of acetylene reduction (a proxy for N2 fixation) and nitrite production (a proxy for ammonia oxidation) and decreased transcript levels of structural genes involved in both nitrogen fixation (nifH) and ammonia oxidation (amoA). In contrast, amendment of microcosms with ammonia stimulated nitrite production and increased amoA transcript levels while it suppressed rates of acetylene reduction and decreased nifH transcript levels. Sequencing of amplified nifH and amoA transcripts from native sediments, as well as microcosms, at 2 and 4 h postamendment, indicates that the dominant and responsive populations involved in ammonia oxidation and N2 fixation are closely affiliated with Ca. Nitrosocaldus yellowstonii and T. albus, respectively. Collectively, these results suggest that ammonia-oxidizing archaea, such as Ca. Nitrosocaldus yellowstonii, have an apparent affinity for ammonia that is higher than that of the diazotrophs present in this ecosystem. Depletion of the bioavailable N pool through the activity of ammonia-oxidizing archaea likely represents a strong selective pressure for the inclusion of organisms capable of nitrogen fixation in geothermal communities. These observations help to explain the strong pattern in the codistribution of ammonia-oxidizing archaea and diazotrophs in circumneutral-to-alkaline geothermal springs. PMID:24242238

Management matters: Testing a mitigation strategy of nitrous oxide emissions on managed grassland

NASA Astrophysics Data System (ADS)

Fuchs, Kathrin; Hörtnagl, Lukas; Eugster, Werner; Koller, Patrick; Käslin, Florian; Merbold, Lutz

2017-04-01

The magnitude of greenhouse gas (GHG) exchange between managed grasslands and the atmosphere depends besides climate predominantly on management practices. While natural or extensively managed grasslands are known to function as GHG sinks, intensively managed grasslands are characterized by substantial nitrous oxide (N2O) emissions diminishing their sink function. One potential N2O mitigation strategy is to reduce the required amount of nitrogen (N) fertilizer input by using biological nitrogen fixation (BNF) via legumes. However, the effect of legumes on nitrous oxide emissions is still not fully understood. In this study we quantify net GHG fluxes from two differently managed grassland parcels (mitigation, control) and relate our results to productivity (yields). In addition, we aim at revealing the influence of various driver variables on N2O exchange. Our experimental setup consisted of an eddy covariance tower that measured the net exchange of the three major anthropogenic GHGs, nitrous oxide (N2O), methane (CH4) and carbon dioxide (CO2). Both grassland parcels can be covered with this tower due to two prevailing wind directions. GHG flux measurements were accompanied by measurements of commonly known driver variables such as water filled pore space, soil temperature, soil oxygen concentrations and mineral N to disentangle the soil meteorological influence of N2O fluxes from human drivers. Following organic fertilizer application, we measured elevated N2O emissions (>1 nmol m-2 s-1) at the control parcel and unchanged N2O emissions at the treatment parcel. Net annual fluxes were 54% and 50% lower at the experimental parcel in 2015 and 2016, respectively. Annual yields did not significantly differ between parcels, but were slightly lower at the experimental parcel compared to the control parcel. Significantly lower nitrous oxide fluxes under experimental management indicate that nitrous oxide emissions can be effectively reduced at very low costs with a clover-based management. Long-term effects on the N budget, implications for the quality of animal feed as well as potential consequences at the farming system level (i.e. manure management) need further evaluation.

Changing nutrient stoichiometry affects phytoplankton production, DOP accumulation and dinitrogen fixation - a mesocosm experiment in the eastern tropical North Atlantic

NASA Astrophysics Data System (ADS)

Meyer, J.; Löscher, C. R.; Neulinger, S. C.; Reichel, A. F.; Loginova, A.; Borchard, C.; Schmitz, R. A.; Hauss, H.; Kiko, R.; Riebesell, U.

2016-02-01

Ocean deoxygenation due to climate change may alter redox-sensitive nutrient cycles in the marine environment. The productive eastern tropical North Atlantic (ETNA) upwelling region may be particularly affected when the relatively moderate oxygen minimum zone (OMZ) deoxygenates further and microbially driven nitrogen (N) loss processes are promoted. Consequently, water masses with a low nitrogen to phosphorus (N : P) ratio could reach the euphotic layer, possibly influencing primary production in those waters. Previous mesocosm studies in the oligotrophic Atlantic Ocean identified nitrate availability as a control of primary production, while a possible co-limitation of nitrate and phosphate could not be ruled out. To better understand the impact of changing N : P ratios on primary production and N2 fixation in the ETNA surface ocean, we conducted land-based mesocosm experiments with natural plankton communities and applied a broad range of N : P ratios (2.67-48). Silicic acid was supplied at 15 µmol L-1 in all mesocosms. We monitored nutrient drawdown, biomass accumulation and nitrogen fixation in response to variable nutrient stoichiometry. Our results confirmed nitrate to be the key factor determining primary production. We found that excess phosphate was channeled through particulate organic matter (POP) into the dissolved organic matter (DOP) pool. In mesocosms with low inorganic phosphate availability, DOP was utilized while N2 fixation increased, suggesting a link between those two processes. Interestingly this observation was most pronounced in mesocosms where nitrate was still available, indicating that bioavailable N does not necessarily suppress N2 fixation. We observed a shift from a mixed cyanobacteria-proteobacteria dominated active diazotrophic community towards a diatom-diazotrophic association of the Richelia-Rhizosolenia symbiosis. We hypothesize that a potential change in nutrient stoichiometry in the ETNA might lead to a general shift within the diazotrophic community, potentially influencing primary productivity and carbon export.

Modeling pathways of riverine nitrogen and phosphorus in the Baltic Sea

NASA Astrophysics Data System (ADS)

Radtke, H.; Neumann, T.; Voss, M.; Fennel, W.

2012-09-01

A better understanding of the fate of nutrients entering the Baltic Sea ecosystem is an important issue with implications for environmental management. There are two sources of nitrogen and phosphorus: riverine input and atmospheric deposition. In the case of nitrogen, the fixation of dinitrogen by diazotrophic bacteria represents a third source. From an analysis of stable nitrogen isotope ratios it was suggested that most of the riverine nitrogen is sequestered in the coastal rim, specifically along the southern Baltic Sea coast with its coarse sediments, whereas nitrogen from fixation dominates the central basins. However, pathways of nutrients and timescales between the input of the nutrients and their arrival in different basins are difficult to obtain from direct measurements. To elucidate this problem, we use a source attribution technique in a three-dimensional ecosystem model, ERGOM, to track nutrients originating from various rivers. An “age” variable is attributed to the marked elements to indicate their propagation speeds and residence times. In this paper, we specifically investigate the spreading of nitrogen and phosphorus from the riverine discharges of the Oder, Vistula, Neman and Daugava. We demonstrate which regions they are transported to and for how long they remain in the ecosystem. The model results show good agreement with source estimations from observed δ15N values in sediments. The model results suggest that 95% of nitrogen is lost by denitrification in sediments, after an average time of 1.4 years for riverine nitrogen. The residence time of riverine phosphorus is much longer and exceeds our simulated period of 35 years.

Exotic plant invasion alters nitrogen dynamics in an arid grassland

USGS Publications Warehouse

Evans, R.D.; Rimer, R.; Sperry, L.; Belnap, J.

2001-01-01

The introduction of nonnative plant species may decrease ecosystem stability by altering the availability of nitrogen (N) for plant growth. Invasive species can impact N availability by changing litter quantity and quality, rates of N2-fixation, or rates of N loss. We quantified the effects of invasion by the annual grass Bromus tectorum on N cycling in an arid grassland on the Colorado Plateau (USA). The invasion occurred in 1994 in two community types in an undisturbed grassland. This natural experiment allowed us to measure the immediate responses following invasion without the confounding effects of previous disturbance. Litter biomass and the C:N and lignin:N ratios were measured to determine the effects on litter dynamics. Long-term soil incubations (415 d) were used to measure potential microbial respiration and net N mineralization. Plant-available N was quantified for two years in situ with ion-exchange resin bags, and potential changes in rates of gaseous N loss were estimated by measuring denitrification enzyme activity. Bromus invasion significantly increased litter biomass, and Bromus litter had significantly greater C:N and lignin:N ratios than did native species. The change in litter quantity and chemistry decreased potential rates of net N mineralization in sites with Bromus by decreasing nitrogen available for microbial activity. Inorganic N was 50% lower on Hilaria sites with Bromus during the spring of 1997, but no differences were observed during 1998. The contrasting differences between years are likely due to moisture availability; spring precipitation was 15% greater than average during 1997, but 52% below average during spring of 1998. Bromus may cause a short-term decrease in N loss by decreasing substrate availability and denitrification enzyme activity, but N loss is likely to be greater in invaded sites in the long term because of increased fire frequency and greater N volatilization during fire. We hypothesize that the introduction of Bromus in conjunction with land-use change has established a series of positive feedbacks that will decrease N availability and alter species composition.

Nitrogen fixation in the activated sludge treatment of thermomechanical pulping wastewater: effect of dissolved oxygen.

PubMed

Slade, A H; Anderson, S M; Evans, B G

2003-01-01

N-ViroTech, a novel technology which selects for nitrogen-fixing bacteria as the bacteria primarily responsible for carbon removal, has been developed to treat nutrient limited wastewaters to a high quality without the addition of nitrogen, and only minimal addition of phosphorus. Selection of the operating dissolved oxygen level to maximise nitrogen fixation forms a key component of the technology. Pilot scale activated sludge treatment of a thermomechanical pulping wastewater was carried out in nitrogen-fixing mode over a 15 month period. The effect of dissolved oxygen was studied at three levels: 14% (Phase 1), 5% (Phase 2) and 30% (Phase 3). The plant was operated at an organic loading of 0.7-1.1 kg BOD5/m3/d, a solids retention time of approximately 10 d, a hydraulic retention time of 1.4 d and a F:M ratio of 0.17-0.23 mg BOD5/mg VSS/d. Treatment performance was very stable over the three dissolved oxygen operating levels. The plant achieved 94-96% BOD removal, 82-87% total COD removal, 79-87% soluble COD removal, and >99% total extractives removal. The lowest organic carbon removals were observed during operation at 30% DO but were more likely to be due to phosphorus limitation than operation at high dissolved oxygen, as there was a significant decrease in phosphorus entering the plant during Phase 3. Discharge of dissolved nitrogen, ammonium and oxidised nitrogen were consistently low (1.1-1.6 mg/L DKN, 0.1-0.2 mg/L NH4+-N and 0.0 mg/L oxidised nitrogen). Discharge of dissolved phosphorus was 2.8 mg/L, 0.1 mg/L and 0.6 mg/L DRP in Phases 1, 2 and 3 respectively. It was postulated that a population of polyphosphate accumulating bacteria developed during Phase 1. Operation at low dissolved oxygen during Phase 2 appeared to promote biological phosphorus uptake which may have been affected by raising the dissolved oxygen to 30% in Phase 3. Total nitrogen and phosphorus discharge was dependent on efficient secondary clarification, and improved over the course of the study as suspended solids discharge improved. Nitrogen fixation was demonstrated throughout the study using an acetylene reduction assay. Based on nitrogen balances around the plant, there was a 55, 354 and 98% increase in nitrogen during Phases 1, 2 and 3 respectively. There was a significant decrease in phosphorus between Phases 1 and 2, and Phase 3 of the study, as well as a significant increase in nitrogen between Phases 2 and 3 which masked the effect of changing the dissolved oxygen. Operation at low dissolved oxygen appeared to confer a competitive advantage to the nitrogen-fixing bacteria.

Metagenomic insights into strategies of carbon conservation and unusual sulfur biogeochemistry in a hypersaline Antarctic lake

PubMed Central

Yau, Sheree; Lauro, Federico M; Williams, Timothy J; DeMaere, Matthew Z; Brown, Mark V; Rich, John; Gibson, John AE; Cavicchioli, Ricardo

2013-01-01

Organic Lake is a shallow, marine-derived hypersaline lake in the Vestfold Hills, Antarctica that has the highest reported concentration of dimethylsulfide (DMS) in a natural body of water. To determine the composition and functional potential of the microbial community and learn about the unusual sulfur chemistry in Organic Lake, shotgun metagenomics was performed on size-fractionated samples collected along a depth profile. Eucaryal phytoflagellates were the main photosynthetic organisms. Bacteria were dominated by the globally distributed heterotrophic taxa Marinobacter, Roseovarius and Psychroflexus. The dominance of heterotrophic degradation, coupled with low fixation potential, indicates possible net carbon loss. However, abundant marker genes for aerobic anoxygenic phototrophy, sulfur oxidation, rhodopsins and CO oxidation were also linked to the dominant heterotrophic bacteria, and indicate the use of photo- and lithoheterotrophy as mechanisms for conserving organic carbon. Similarly, a high genetic potential for the recycling of nitrogen compounds likely functions to retain fixed nitrogen in the lake. Dimethylsulfoniopropionate (DMSP) lyase genes were abundant, indicating that DMSP is a significant carbon and energy source. Unlike marine environments, DMSP demethylases were less abundant, indicating that DMSP cleavage is the likely source of high DMS concentration. DMSP cleavage, carbon mixotrophy (photoheterotrophy and lithoheterotrophy) and nitrogen remineralization by dominant Organic Lake bacteria are potentially important adaptations to nutrient constraints. In particular, carbon mixotrophy relieves the extent of carbon oxidation for energy production, allowing more carbon to be used for biosynthetic processes. The study sheds light on how the microbial community has adapted to this unique Antarctic lake environment. PMID:23619305

Occurrence, structure, and nitrogen-fixation of root nodules of actinorhizal Arizona alder

Treesearch

J. O. Dawson; Gerald J. Gottfried; D. Hahn

2005-01-01

Actinorhizal plants are nodulated by the symbiotic, nitrogen-fixing actinomycete Frankia. The genus Alnus in the family Betulaceae is one of the 24 genera in 8 families of angiospermous plants that are actinorhizal. Arizona alder (Alnus oblongifolia Torr.) occurs in isolated populations associated with the watersheds of Madrean Sky Islands in the...

A Proteomic Network for Symbiotic Nitrogen Fixation Efficiency in Bradyrhizobium elkanii.

PubMed

Cooper, Bret; Campbell, Kimberly B; Beard, Hunter S; Garrett, Wesley M; Mowery, Joseph; Bauchan, Gary R; Elia, Patrick

2018-03-01

Rhizobia colonize legumes and reduce N 2 to NH 3 in root nodules. The current model is that symbiotic rhizobia bacteroids avoid assimilating this NH 3 . Instead, host legume cells form glutamine from NH 3 , and the nitrogen is returned to the bacteroid as dicarboxylates, peptides, and amino acids. In soybean cells surrounding bacteroids, glutamine also is converted to ureides. One problem for soybean cultivation is inefficiency in symbiotic N 2 fixation, the biochemical basis of which is unknown. Here, the proteomes of bacteroids of Bradyrhizobium elkanii USDA76 isolated from N 2 fixation-efficient Peking and -inefficient Williams 82 soybean nodules were analyzed by mass spectrometry. Nearly half of the encoded bacterial proteins were quantified. Efficient bacteroids produced greater amounts of enzymes to form Nod factors and had increased amounts of signaling proteins, transporters, and enzymes needed to generate ATP to power nitrogenase and to acquire resources. Parallel investigation of nodule proteins revealed that Peking had no significantly greater accumulation of enzymes needed to assimilate NH 3 than Williams 82. Instead, efficient bacteroids had increased amounts of enzymes to produce amino acids, including glutamine, and to form ureide precursors. These results support a model for efficient symbiotic N 2 fixation in soybean where the bacteroid assimilates NH 3 for itself.

Lon Protease of Azorhizobium caulinodans ORS571 Is Required for Suppression of reb Gene Expression

PubMed Central

Nakajima, Azusa; Tsukada, Shuhei; Siarot, Lowela; Ogawa, Tetsuhiro; Oyaizu, Hiroshi

2012-01-01

Bacterial Lon proteases play important roles in a variety of biological processes in addition to housekeeping functions. In this study, we focused on the Lon protease of Azorhizobium caulinodans, which can fix nitrogen both during free-living growth and in stem nodules of the legume Sesbania rostrata. The nitrogen fixation activity of an A. caulinodans lon mutant in the free-living state was not significantly different from that of the wild-type strain. However, the stem nodules formed by the lon mutant showed little or no nitrogen fixation activity. By microscopic analyses, two kinds of host cells were observed in the stem nodules formed by the lon mutant. One type has shrunken host cells containing a high density of bacteria, and the other type has oval or elongated host cells containing a low density or no bacteria. This phenotype is similar to a praR mutant highly expressing the reb genes. Quantitative reverse transcription-PCR analyses revealed that reb genes were also highly expressed in the lon mutant. Furthermore, a lon reb double mutant formed stem nodules showing higher nitrogen fixation activity than the lon mutant, and shrunken host cells were not observed in these stem nodules. These results suggest that Lon protease is required to suppress the expression of the reb genes and that high expression of reb genes in part causes aberrance in the A. caulinodans-S. rostrata symbiosis. In addition to the suppression of reb genes, it was found that Lon protease was involved in the regulation of exopolysaccharide production and autoagglutination of bacterial cells. PMID:22752172

Effect of Microgravity on Early Events of Biological Nitrogen Fixation in Medicago Truncatula: Initial Results from the SyNRGE Experiment

NASA Technical Reports Server (NTRS)

Stutte, Gary W.; Roberts, Michael S.

2011-01-01

SyNRGE (Symbiotic Nodulation in a Reduced Gravity Environment) was a sortie mission on STS-135 in the Biological Research in Canisters (BRIC) hardware to study the effect of microgravity on a plant-microbe symbiosis resulting in biological nitrogen fixation. Medicago truncatula, a model species of the legume family, was inoculated with its bacterial symbiont, Sinorhizobium meliloti, to observe early events associated with infection and nodulation in Petri Dish Fixation Units (PDFUs). Two sets of experiments were conducted in orbit and in 24-hour delayed ground controls. Experiment one was designed to determine if S. meliloti infect M. truncatula and initiate physiological changes associated with nodule formation. Roots of five-day-old M. truncatula cultivar Jemalong A17 (Enodll::gus) were inoculated 24 hr before launch with either S. meliloti strain 1021 or strain ABS7 and integrated into BRIC-PDFU hardware placed in a 4 C Cold Bag for launch on Atlantis. Inoculated plants and uninoculated controls were maintained in the dark at ambient temperature in the middeck of STS-135 for 11 days before fixation in RNAlater(tM) by crew activation of the PDFU. Experiment two was designed to determine if microgravity altered the process of bacterial infection and host plant nodule formation. Seeds of two M. truncatula cultivar Jemalong A17 lines, the Enodll::gus used in experiment 1, and SUNN, a super-nodulating mutant of A17, were germinated on orbit for 11 days in the middeck cabin and returned to Earth alive inside of BRIC-PDFU's at 4 C. S. meliloti strains 1021 and ABS7 were cultivated separately in broth culture on orbit and also returned to Earth alive. After landing, flight- and groundgrown plants and bacteria were transferred from BRIC-PDFU's into Nunc(tm) 4-well plates for reciprocity crosses. Rates of plant growth and nodule development on Buffered Nodulation Medium (lacking nitrogen) were measured for 14 days. Preliminary analysis' of Experiment 1 confirms that legumes and bacteria cultivated in space 'initiate the symbiotic interaction leading to nitrogen fixation and that bacteria retain the ability to form nodules on M. truncatula roots. Initial assessment of experiment 2 shows 100% seed germination and excellent bacterial growth in microgravity.

Tri-party underground symbiosis between a weevil, bacteria and a desert plant.

PubMed

Shelef, Oren; Helman, Yael; Friedman, Ariel-Leib-Leonid; Behar, Adi; Rachmilevitch, Shimon

2013-01-01

Inhabitants of arid ecosystems face severe nitrogen and water limitations. Inventive adaptations by organisms occupying such habitats are essential for survival. This study describes a tri-party symbiotic interaction between a plant (Salsola inermis), a beetle (Conorhynchus pistor), and a bacterium (Klebsiella pneumonia). The weevil survives by living within a mud structure affixed to the plant roots, thus benefiting from increased carbon and water, and refuge from predators and parasites. Active nitrogen-fixing bacteria harbored within the weevil's gut mediate this interaction, by supplying nitrogen to the system, which eventually promotes seed development. We studied the correlation between the weevil's existence and (i) root carbon and nitrogen content, (ii) soil water content and (iii) seed weight. Roots hosting weevils contained more nitrogen, heavier seeds and less carbon. In addition, water content was higher around the roots than in open spaces a short distance from the plant stem. Bacterial studies and nitrogen-fixation analyses, including molecular and chemical assays, indicated atmospheric nitrogen fixation in the larval stage and identified the bacterium. The coexistence of weevil and bacterial behavior coinciding with the plant's life cycle was revealed here by a long period of field observations. Out of over 60,000 known weevils, this is the only report of a weevil living most of its life underground without harming plants. The unique tri-party interaction described herein shows the important ecological role of desert plant roots and provides an example of a sustainable consortium of living organisms coping with the challenging desert environment.

Short-term N2 fixation kinetics in a moss-associated cyanobacteria.

PubMed

Jean, Marie-Eve; Cassar, Nicolas; Setzer, Cameron; Bellenger, Jean-Philippe

2012-08-21

N(2) fixation by moss-associated cyanobacteria plays an important role in the nitrogen cycling of terrestrial ecosystems. Recent studies have mainly focused on boreal ecosystems; little is known about such association in other ecosystems. Moss-associated cyanobacteria are subject to rapid changes (hourly or less) in environmental conditions that may affect N(2) fixation kinetics. Using a recently developed method (Acetylene Reduction Assays by Cavity ring-down laser Absorption Spectroscopy, ARACAS) with higher sensitivity and sampling frequency than the conventional method, we characterize short-term kinetics of N(2) fixation by cyanobacteria on moss carpets from warm and cold temperate forests. We report the identification of a heretofore unknown multispecies true-moss-cyanobacteria diazotrophic association. We demonstrate that short-term change in abiotic variables greatly influences N(2) fixation. We also show that difference in relative proportion of two epiphytic diazotrophs is consistent with divergent influences of temperature on their N(2) fixation kinetics. Further research is needed to determine whether this difference is consistent with a latitudinal trend.

Basin-wide N2 fixation in the deep waters of the Mediterranean Sea

NASA Astrophysics Data System (ADS)

Benavides, Mar; Bonnet, Sophie; Hernández, Nauzet; Martínez-Pérez, Alba María.; Nieto-Cid, Mar; Álvarez-Salgado, Xosé Antón; Baños, Isabel; Montero, María. F.; Mazuecos, Ignacio P.; Gasol, Josep M.; Osterholz, Helena; Dittmar, Thorsten; Berman-Frank, Ilana; Arístegui, Javier

2016-06-01

Recent findings indicate that N2 fixation is significant in aphotic waters, presumably due to heterotrophic diazotrophs depending on organic matter for their nutrition. However, the relationship between organic matter and heterotrophic N2 fixation remains unknown. Here we explore N2 fixation in the deep chlorophyll maximum and underneath deep waters across the whole Mediterranean Sea and relate it to organic matter composition, characterized by optical and molecular methods. Our N2 fixation rates were in the range of those previously reported for the euphotic zone of the Mediterranean Sea (up to 0.43 nmol N L-1 d-1) and were significantly correlated to the presence of relatively labile organic matter with fluorescence and molecular formula properties representative for peptides and unsaturated aliphatics and associated with the presence of more oxygenated ventilated water masses. Finally, and despite that the aphotic N2 fixation contributes largely to total water column diazotrophic activity (>50%), its contribution to overall nitrogen inputs to the basin is negligible (<0.5%).

NanoSIMS Analyses of Mo Indicate Nitrogenase Activity and Help Solve a N and C Fixation Puzzle in a Marine Cyanobacterium

NASA Astrophysics Data System (ADS)

Pett-Ridge, J.; Weber, P. K.; Finzi, J.; Hutcheon, I. D.; Capone, D. G.

2006-12-01

Diazotrophic cyanobacteria are capable of both CO2 and N2 fixation, yet must separate these two functions because the nitrogenase enzymes used in N2 fixation are strongly inhibited by O2 produced during photosynthesis. Some lineages, such as Anabaena, use specialized cells (heterocysts) to maintain functional segregation. However the mechanism of this segregation is poorly understood in Trichodesmium, a critical component of marine primary production in the tropical and subtropical North Atlantic. While some Trichodesmium studies suggest a temporal segregation of the nitrogen and carbon fixing processes, others indicate nitrogen fixation is spatially isolated in differentiated cells called diazocytes. In order to isolate the intracellular location of N fixation in both species, we used a combination of TEM, SEM and NanoSIMS analysis to map the distribution of C, N and Mo (a critical nitrogenase co-factor) isotopes in intact cells. NanoSIMS is a powerful surface analysis tool which combines nanometer-scale imaging resolution with the high sensitivity of mass spectrometry. Using cells grown in a 13CO^2 and 15N2 enriched atmosphere, our analyses indicate that in Anabaena, heterocysts are consistently enriched in Mo, and Mo accumulation suggests active N fixation (as opposed to N storage). In the non- heterocystous Trichodesmium, Mo is concentrated in sub-regions of individual cells, and is not associated with regions of N storage (cyanophycin granules). We suggest that NanoSIMS mapping of metal enzyme co- factors is a unique method of identifying physiological and morphological characteristics within individual bacterial cells. This combination of NanoSIMS analysis and high resolution microscopy allows isotopic analysis to be linked to morphological features and holds great promise for fine-scale studies of bacteria metabolism.

Effects of model structural uncertainty on carbon cycle projections: biological nitrogen fixation as a case study

NASA Astrophysics Data System (ADS)

Wieder, William R.; Cleveland, Cory C.; Lawrence, David M.; Bonan, Gordon B.

2015-04-01

Uncertainties in terrestrial carbon (C) cycle projections increase uncertainty of potential climate feedbacks. Efforts to improve model performance often include increased representation of biogeochemical processes, such as coupled carbon-nitrogen (N) cycles. In doing so, models are becoming more complex, generating structural uncertainties in model form that reflect incomplete knowledge of how to represent underlying processes. Here, we explore structural uncertainties associated with biological nitrogen fixation (BNF) and quantify their effects on C cycle projections. We find that alternative plausible structures to represent BNF result in nearly equivalent terrestrial C fluxes and pools through the twentieth century, but the strength of the terrestrial C sink varies by nearly a third (50 Pg C) by the end of the twenty-first century under a business-as-usual climate change scenario representative concentration pathway 8.5. These results indicate that actual uncertainty in future C cycle projections may be larger than previously estimated, and this uncertainty will limit C cycle projections until model structures can be evaluated and refined.

In silico characterization and transcriptomic analysis of nif family genes from Anabaena sp. PCC7120.

PubMed

Singh, Shilpi; Shrivastava, Alok Kumar

2017-10-01

In silico approaches in conjunction with morphology, nitrogenase activity, and qRT-PCR explore the impact of selected abiotic stressor such as arsenic, salt, cadmium, copper, and butachlor on nitrogen fixing (nif family) genes of diazotrophic cyanobacterium Anabaena sp. PCC7120. A total of 19 nif genes are present within the Anabaena genome that is involved in the process of nitrogen fixation. Docking studies revealed the interaction between these nif gene-encoded proteins and the selected abiotic stressors which were further validated through decreased heterocyst frequency, fragmentation of filaments, and downregulation of nitrogenase activity under these stresses indicating towards their toxic impact on nitrogen fixation potential of filamentous cyanobacterium Anabaena sp. PCC7120. Another appealing finding of this study is even though having similar binding energy and similar interacting residues between arsenic/salt and copper/cadmium to nif-encoded proteins, arsenic and cadmium are more toxic than salt and copper for nitrogenase activity of Anabaena which is crucial for growth and yield of rice paddy and soil reclamation.

Soluble carbohydrate allocation to roots, photosynthetic rate of leaves, and nitrate assimilation as affected by nitrogen stress and irradiance

NASA Technical Reports Server (NTRS)

Henry, L. T.; Raper, C. D. Jr

1991-01-01

Upon resupply of exogenous nitrogen to nitrogen-stressed plants, uptake rate of nitrogen is enhanced relative to nonstressed plants. Absorption of nitrogen presumably is dependent on availability of carbohydrates in the roots. A buildup in soluble carbohydrates thus should occur in roots of nitrogen-stressed plants, and upon resupply of exogenous nitrogen the increased uptake rate should be accompanied by a rapid decline in carbohydrates to prestress levels. To evaluate this relationship, three sets of tobacco plants growing in a complete hydroponic solution containing 1.0 mM NO3- were either continued in the complete solution for 21 d, transferred to a minus-nitrogen solution for 21 d, or transferred to a minus-nitrogen solution for 8-9 d and then returned to the 1.0 mM NO3- solution. These nitrogen treatments were imposed upon plants growing at photosynthetic photon flux densities of 700 and 350 micromoles m-2 s-1. Soluble carbohydrate levels in roots increased during onset of nitrogen stress to levels that were fourfold greater than in roots of non-stressed plants. Following resupply of external nitrogen, a rapid resumption of nitrogen uptake was accompanied by a decline in soluble carbohydrates in roots to levels characteristic of nonstressed plants. This pattern of soluble carbohydrate levels in roots during onset of and recovery from nitrogen stress occurred at both irradiance levels. The response of net photosynthetic rate to nitrogen stress could be expressed as a nonlinear function of concentration of reduced nitrogen in leaves. The net photosynthetic rate at a given concentration of reduced nitrogen, however, averaged 10% less at the lower than at the higher irradiance. The decline in net photosynthetic rate per unit of reduced nitrogen in leaves at the lower irradiance was accompanied by an increase in the nitrate fraction of total nitrogen in leaves from 20% at the higher irradiance to 38% at the lower irradiance.

RNA-Seq analysis of nodule development at five different developmental stages of soybean (Glycine max) inoculated with Bradyrhizobium japonicum strain 113-2

PubMed Central

Yuan, Song L.; Li, Rong; Chen, Hai F.; Zhang, Chan J.; Chen, Li M.; Hao, Qing N.; Chen, Shui L.; Shan, Zhi H.; Yang, Zhong L.; Zhang, Xiao J.; Qiu, De Z.; Zhou, Xin A.

2017-01-01

Nodule development directly affects nitrogen fixation efficiency during soybean growth. Although abundant genome-based information related to nodule development has been released and some studies have reported the molecular mechanisms that regulate nodule development, information on the way nodule genes operate in nodule development at different developmental stages of soybean is limited. In this report, notably different nodulation phenotypes in soybean roots inoculated with Bradyrhizobium japonicum strain 113-2 at five developmental stages (branching stage, flowering stage, fruiting stage, pod stage and harvest stage) were shown, and the expression of nodule genes at these five stages was assessed quantitatively using RNA-Seq. Ten comparisons were made between these developmental periods, and their differentially expressed genes were analysed. Some important genes were identified, primarily encoding symbiotic nitrogen fixation-related proteins, cysteine proteases, cystatins and cysteine-rich proteins, as well as proteins involving plant-pathogen interactions. There were no significant shifts in the distribution of most GO functional annotation terms and KEGG pathway enrichment terms between these five development stages. A cystatin Glyma18g12240 was firstly identified from our RNA-seq, and was likely to promote nodulation and delay nodule senescence. This study provides molecular material for further investigations into the mechanisms of nitrogen fixation at different soybean developmental stages. PMID:28169364

Using Synthetic Biology to Distinguish and Overcome Regulatory and Functional Barriers Related to Nitrogen Fixation

PubMed Central

Wang, Xia; Yang, Jian-Guo; Chen, Li; Wang, Ji-Long; Cheng, Qi; Dixon, Ray; Wang, Yi-Ping

2013-01-01

Biological nitrogen fixation is a complex process requiring multiple genes working in concert. To date, the Klebsiella pneumoniae nif gene cluster, divided into seven operons, is one of the most studied systems. Its nitrogen fixation capacity is subject to complex cascade regulation and physiological limitations. In this report, the entire K. pneumoniae nif gene cluster was reassembled as operon-based BioBrick parts in Escherichia coli. It provided ∼100% activity of native K. pneumoniae system. Based on the expression levels of these BioBrick parts, a T7 RNA polymerase–LacI expression system was used to replace the σ54-dependent promoters located upstream of nif operons. Expression patterns of nif operons were critical for the maximum activity of the recombinant system. By mimicking these expression levels with variable-strength T7-dependent promoters, ∼42% of the nitrogenase activity of the σ54-dependent nif system was achieved in E. coli. When the newly constructed T7-dependent nif system was challenged with different genetic and physiological conditions, it bypassed the original complex regulatory circuits, with minor physiological limitations. Therefore, we have successfully replaced the nif regulatory elements with a simple expression system that may provide the first step for further research of introducing nif genes into eukaryotic organelles, which has considerable potentials in agro-biotechnology. PMID:23935879

Functional Single-Cell Approach to Probing Nitrogen-Fixing Bacteria in Soil Communities by Resonance Raman Spectroscopy with 15N2 Labeling.

PubMed

Cui, Li; Yang, Kai; Li, Hong-Zhe; Zhang, Han; Su, Jian-Qiang; Paraskevaidi, Maria; Martin, Francis L; Ren, Bin; Zhu, Yong-Guan

2018-04-17

Nitrogen (N) fixation is the conversion of inert nitrogen gas (N 2 ) to bioavailable N essential for all forms of life. N 2 -fixing microorganisms (diazotrophs), which play a key role in global N cycling, remain largely obscure because a large majority are uncultured. Direct probing of active diazotrophs in the environment is still a major challenge. Herein, a novel culture-independent single-cell approach combining resonance Raman (RR) spectroscopy with 15 N 2 stable isotope probing (SIP) was developed to discern N 2 -fixing bacteria in a complex soil community. Strong RR signals of cytochrome c (Cyt c, frequently present in diverse N 2 -fixing bacteria), along with a marked 15 N 2 -induced Cyt c band shift, generated a highly distinguishable biomarker for N 2 fixation. 15 N 2 -induced shift was consistent well with 15 N abundance in cell determined by isotope ratio mass spectroscopy. By applying this biomarker and Raman imaging, N 2 -fixing bacteria in both artificial and complex soil communities were discerned and imaged at the single-cell level. The linear band shift of Cyt c versus 15 N 2 percentage allowed quantification of N 2 fixation extent of diverse soil bacteria. This single-cell approach will advance the exploration of hitherto uncultured diazotrophs in diverse ecosystems.

Significance of Herbaspirillum seropedicae inoculation and/or straw amendment on growth and dinitrogen fixation of wheat using 15N-dilution method.

PubMed

el-Komy, H M; Saad, O A; Hetta, A M

2003-01-01

The effect of Herbaspirillum seropedicae inoculation and/or maize straw (0, 5 and 10 Mg/hm2) amendment on the growth and N2 fixation of wheat was determined in pot experiments using 15N-dilution method. Inoculation resulted in accumulation of fixed nitrogen, and % N from atmosphere being 24.6 and 26.5% in wheat shoot and grain, respectively. Straw amendment reduced % Natm to 16.1 and 20.2% at high straw level (10 Mg/hm2). Rational nitrogen fertilization (180 kg N/hm2) completely inhibited N2 fixation by H. seropedicae inoculation. Bacterial inoculation increased dry shoot and grain yield up to 23 and 31%, respectively. The highest levels of shoot and grain dry mass (46.5 and 42.4%) were obtained by N-fertilization in both inoculated and uninoculated plants. Total shoot and grain N-yield increased irrespective of organic matter amendment by inoculation up to 9 and 25%, respectively. N-fertilized plants recorded a maximum increase in N-yield (57 and 51%). H. seropedicae was reisolated from inoculated wheat histosphere after harvesting (90 d from sowing). Neither organic matter nor mineral nitrogen applications had any marked effect on bacterial total counts colonizing wheat histosphere. Moreover, no symptoms of mottled stripe disease were observed on leaves and stems of inoculated plants.

The Medicago Genome Provides Insight into the Evolution of Rhizobial Symbioses

PubMed Central

Young, Nevin D.; Debellé, Frédéric; Oldroyd, Giles E. D.; Geurts, Rene; Cannon, Steven B.; Udvardi, Michael K.; Benedito, Vagner A.; Mayer, Klaus F. X.; Gouzy, Jérôme; Schoof, Heiko; Van de Peer, Yves; Proost, Sebastian; Cook, Douglas R.; Meyers, Blake C.; Spannagl, Manuel; Cheung, Foo; De Mita, Stéphane; Krishnakumar, Vivek; Gundlach, Heidrun; Zhou, Shiguo; Mudge, Joann; Bharti, Arvind K.; Murray, Jeremy D.; Naoumkina, Marina A.; Rosen, Benjamin; Silverstein, Kevin A. T.; Tang, Haibao; Rombauts, Stephane; Zhao, Patrick X.; Zhou, Peng; Barbe, Valérie; Bardou, Philippe; Bechner, Michael; Bellec, Arnaud; Berger, Anne; Bergès, Hélène; Bidwell, Shelby; Bisseling, Ton; Choisne, Nathalie; Couloux, Arnaud; Denny, Roxanne; Deshpande, Shweta; Dai, Xinbin; Doyle, Jeff; Dudez, Anne-Marie; Farmer, Andrew D.; Fouteau, Stéphanie; Franken, Carolien; Gibelin, Chrystel; Gish, John; Goldstein, Steven; González, Alvaro J.; Green, Pamela J.; Hallab, Asis; Hartog, Marijke; Hua, Axin; Humphray, Sean; Jeong, Dong-Hoon; Jing, Yi; Jöcker, Anika; Kenton, Steve M.; Kim, Dong-Jin; Klee, Kathrin; Lai, Hongshing; Lang, Chunting; Lin, Shaoping; Macmil, Simone L; Magdelenat, Ghislaine; Matthews, Lucy; McCorrison, Jamison; Monaghan, Erin L.; Mun, Jeong-Hwan; Najar, Fares Z.; Nicholson, Christine; Noirot, Céline; O’Bleness, Majesta; Paule, Charles R.; Poulain, Julie; Prion, Florent; Qin, Baifang; Qu, Chunmei; Retzel, Ernest F.; Riddle, Claire; Sallet, Erika; Samain, Sylvie; Samson, Nicolas; Sanders, Iryna; Saurat, Olivier; Scarpelli, Claude; Schiex, Thomas; Segurens, Béatrice; Severin, Andrew J.; Sherrier, D. Janine; Shi, Ruihua; Sims, Sarah; Singer, Susan R.; Sinharoy, Senjuti; Sterck, Lieven; Viollet, Agnès; Wang, Bing-Bing; Wang, Keqin; Wang, Mingyi; Wang, Xiaohong; Warfsmann, Jens; Weissenbach, Jean; White, Doug D.; White, Jim D.; Wiley, Graham B.; Wincker, Patrick; Xing, Yanbo; Yang, Limei; Yao, Ziyun; Ying, Fu; Zhai, Jixian; Zhou, Liping; Zuber, Antoine; Dénarié, Jean; Dixon, Richard A.; May, Gregory D.; Schwartz, David C.; Rogers, Jane; Quétier, Francis; Town, Christopher D.; Roe, Bruce A.

2011-01-01

Legumes (Fabaceae or Leguminosae) are unique among cultivated plants for their ability to carry out endosymbiotic nitrogen fixation with rhizobial bacteria, a process that takes place in a specialized structure known as the nodule. Legumes belong to one of the two main groups of eurosids, the Fabidae, which includes most species capable of endosymbiotic nitrogen fixation 1. Legumes comprise several evolutionary lineages derived from a common ancestor 60 million years ago (Mya). Papilionoids are the largest clade, dating nearly to the origin of legumes and containing most cultivated species 2. Medicago truncatula (Mt) is a long-established model for the study of legume biology. Here we describe the draft sequence of the Mt euchromatin based on a recently completed BAC-assembly supplemented with Illumina-shotgun sequence, together capturing ~94% of all Mt genes. A whole-genome duplication (WGD) approximately 58 Mya played a major role in shaping the Mt genome and thereby contributed to the evolution of endosymbiotic nitrogen fixation. Subsequent to the WGD, the Mt genome experienced higher levels of rearrangement than two other sequenced legumes, Glycine max (Gm) and Lotus japonicus (Lj). Mt is a close relative of alfalfa (M. sativa), a widely cultivated crop with limited genomics tools and complex autotetraploid genetics. As such, the Mt genome sequence provides significant opportunities to expand alfalfa’s genomic toolbox. PMID:22089132

Indicators of nitrate export from forested watersheds of the Chesapeake Bay region

Treesearch

Karl W. J. Williard

1997-01-01

Soil net nitrogen mineralization and nitrification rates were studied on nine relatively undisturbed, forested watersheds in an effort to explain the large variations in nitrate export in streamflow within the Chesapeake Bay region. The primary hypothesis tested was that nitrate export from the watersheds was positively associated with rates of net soil nitrogen...

Comparing Methods for Assessing Forest Soil Net Nitrogen Mineralization and Net Nitrification

Treesearch

S. S. Jefts; I. J. Fernandez; L.E. Rustad; D. B. Dail

2004-01-01

A variety of analytical techniques are used to evaluate rates of nitrogen (N) mineralization and nitrification in soils. The diversity of methods takes on added significance in forest ecosystem research where high soil heterogeneity and multiple soil horizons can make comparisons over time and space even more complex than in agricultural Ap horizons. This study...

Biological nitrogen fixation: rates, patterns and ecological controls in terrestrial ecosystems

USGS Publications Warehouse

Vitousek, Peter M.; Menge, Duncan N.L.; Reed, Sasha C.; Cleveland, Cory C.

2013-01-01

New techniques have identified a wide range of organisms with the capacity to carry out biological nitrogen fixation (BNF)—greatly expanding our appreciation of the diversity and ubiquity of N fixers—but our understanding of the rates and controls of BNF at ecosystem and global scales has not advanced at the same pace. Nevertheless, determining rates and controls of BNF is crucial to placing anthropogenic changes to the N cycle in context, and to understanding, predicting and managing many aspects of global environmental change. Here, we estimate terrestrial BNF for a pre-industrial world by combining information on N fluxes with 15N relative abundance data for terrestrial ecosystems. Our estimate is that pre-industrial N fixation was 58 (range of 40–100) Tg N fixed yr−1; adding conservative assumptions for geological N reduces our best estimate to 44 Tg N yr−1. This approach yields substantially lower estimates than most recent calculations; it suggests that the magnitude of human alternation of the N cycle is substantially larger than has been assumed.

Fixation Probability in a Haploid-Diploid Population.

PubMed

Bessho, Kazuhiro; Otto, Sarah P

2017-01-01

Classical population genetic theory generally assumes either a fully haploid or fully diploid life cycle. However, many organisms exhibit more complex life cycles, with both free-living haploid and diploid stages. Here we ask what the probability of fixation is for selected alleles in organisms with haploid-diploid life cycles. We develop a genetic model that considers the population dynamics using both the Moran model and Wright-Fisher model. Applying a branching process approximation, we obtain an accurate fixation probability assuming that the population is large and the net effect of the mutation is beneficial. We also find the diffusion approximation for the fixation probability, which is accurate even in small populations and for deleterious alleles, as long as selection is weak. These fixation probabilities from branching process and diffusion approximations are similar when selection is weak for beneficial mutations that are not fully recessive. In many cases, particularly when one phase predominates, the fixation probability differs substantially for haploid-diploid organisms compared to either fully haploid or diploid species. Copyright © 2017 by the Genetics Society of America.

Dynamics of N2 fixation and fate of diazotroph-derived nitrogen in a low nutrient low chlorophyll ecosystem: results from the VAHINE mesocosm experiment (New Caledonia)

NASA Astrophysics Data System (ADS)

Bonnet, S.; Berthelot, H.; Turk-Kubo, K.; Fawcett, S.; Rahav, E.; l'Helguen, S.; Berman-Frank, I.

2015-12-01

N2 fixation rates were measured daily in large (~ 50 m3) mesocosms deployed in the tropical South West Pacific coastal ocean (New Caledonia) to investigate the spatial and temporal dynamics of diazotrophy and the fate of diazotroph-derived nitrogen (DDN) in a low nutrient, low chlorophyll ecosystem. The mesocosms were intentionally fertilized with ~ 0.8 μM dissolved inorganic phosphorus (DIP) to stimulate diazotrophy. Bulk N2 fixation rates were replicable between the three mesocosms, averaged 18.5 ± 1.1 nmol N L-1 d-1 over the 23 days, and increased by a factor of two during the second half of the experiment (days 15 to 23) to reach 27.3 ± 1.0 nmol N L-1 d-1. These rates are higher than the upper range reported for the global ocean, indicating that the waters surrounding New Caledonia are particularly favourable for N2 fixation. During the 23 days of the experiment, N2 fixation rates were positively correlated with seawater temperature, primary production, bacterial production, standing stocks of particulate organic carbon, nitrogen and phosphorus, and alkaline phosphatase activity, and negatively correlated with DIP concentrations, DIP turnover time, nitrate, and dissolved organic nitrogen and phosphorus concentrations. The fate of DDN was investigated during the bloom of the unicellular diazotroph, UCYN-C, that occurred during the second half of the experiment. Quantification of diazotrophs in the sediment traps indicates that ~ 10 % of UCYN-C from the water column were exported daily to the traps, representing as much as 22.4 ± 5.5 % of the total POC exported at the height of the UCYN-C bloom. This export was mainly due to the aggregation of small (5.7 ± 0.8 μm) UCYN-C cells into large (100-500 μm) aggregates. During the same time period, a DDN transfer experiment based on high-resolution nanometer scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labelling revealed that 16 ± 6 % of the DDN was released to the dissolved pool and 21 ± 4 % was transferred to non-diazotrophic plankton, mainly picoplankton (18 ± 4 %) followed by diatoms (3 ± 2 %) within 24 h of incubation. This is consistent with the observed dramatic increase in picoplankton and diatom abundances, primary production, bacterial production and standing stocks of particulate organic carbon, nitrogen and phosphorus during the second half of the experiment in the mesocosms. These results offer insights into the fate of DDN during a bloom of UCYN-C in low nutrient, low chlorophyll ecosystems.

Nitrogen Cycling throughout Secondary Succession following Agricultural Disturbance in North-Central Virginia

NASA Astrophysics Data System (ADS)

Parisien, A.; Epstein, H. E.

2017-12-01

While much is known about the carbon cycle during succession that follows agricultural disturbance, less understood are the dynamics of the nitrogen cycle throughout secondary succession, and how plant-available nitrogen may or may not limit vegetation transitions and net primary productivity over time. Two chronosequences at the Blandy Experimental Farm in Boyce, north-central Virginia were examined to elucidate the complexities of the nitrogen cycle over a temporal successional gradient. Each chronosequence consists of one early, one mid, and one late secondary successional field ( 15 years, 30 years, and 100 years post agricultural abandonment, respectively). Five 10x10 m plots were established in each of the 6 fields for a total of 30 plots. Total soil nitrogen (and carbon) data were collected from soils to 30 cm depth at 10-cm intervals, and net nitrogen mineralization and nitrification were estimated using an in situ soil core with anion-cation exchange resin bag technique. Previous studies of carbon cycling at this location have indicated relatively constant soil CO2 efflux of approximately 1100 g C/m2, as well as increasing net primary production and therefore net ecosystem production, with time since abandonment. In addition, soil C and N, and the soil C:N ratio have been shown to increase from the early to late successional plots. Our current study marks the first comprehensive examination of soil nitrogen dynamics including mineralization and nitrification over a successional gradient at Blandy Farm. A thorough understanding of nitrogen dynamics during secondary succession is especially important in the southeastern United States, where a large portion of previously cultivated land has been abandoned over the past century, due to advances in farming efficiency and the move westward to more fertile soils. Much of the southeastern U.S. is now undergoing secondary succession, and quality data on the dynamics of nitrogen cycling during this procession can help guide future land management decisions and carbon cycling predictions.

Physiological and isotopic characteristics of nitrogen fixation by hyperthermophilic methanogens: Key insights into nitrogen anabolism of the microbial communities in Archean hydrothermal systems

NASA Astrophysics Data System (ADS)

Nishizawa, Manabu; Miyazaki, Junichi; Makabe, Akiko; Koba, Keisuke; Takai, Ken

2014-08-01

Hyperthermophilic hydrogenotrophic methanogens are considered to be one of the most predominant primary producers in hydrogen (H2)-abundant hydrothermal environments in the present-day ocean and throughout the history of the Earth. However, the nitrogen sources supporting the development of microbial communities in hydrothermal environments remain poorly understood. We have investigated, for the first time, methanogenic archaea commonly found in deep-sea hydrothermal environments to understand their physiological properties (growth kinetics, energetics, and metal requirements) and isotopic characteristics during the fixation of dinitrogen (N2), which is an abundant but less-bioavailable compound in hydrothermal fluids. Culture experiments showed that Methanocaldococcus strain (Mc 1-85N) (Topt = 85 °C) and Methanothermococcus strain (Mt 5-55N) (Topt = 55 °C) assimilated N2 and ammonium, but not nitrate. Previous phylogenetic studies have predicted that the Methanocaldococcus and Methanothermococcus lineages have nitrogenases, key enzymes for N2 fixation, with biochemically uncharacterised active site metal cofactors. We showed that Mt 5-55N required molybdenum for the nitrogenase to function, implying a molybdenum-bearing cofactor in the strain. Molybdenum also stimulated diazotrophic (i.e., N2-fixing) growth of Mc 1-85N, though further experiments are required to test whether the strain contains a molybdenum-dependent nitrogenase. Importantly, Mc 1-85N exhibited an apparently lower requirement of and higher tolerance to molybdenum and iron than Mt 5-55N. Furthermore, both strains produced more 15N-depleted biomass (-4‰ relative to N2) than that previously reported for diazotrophic photosynthetic prokaryotes. These results demonstrate that diazotrophic hyperthermophilic methanogens can be broadly distributed in seafloor and subseafloor hydrothermal environments, where the availability of transition metals is variable and where organic carbon, organic nitrogen, and ammonium are generally scarce. The emergence and function of diazotrophy, coupled with methanogenesis, in the early Earth is also consistent with the nitrogen isotopic records of 3.5 billion-year-old hydrothermal deposits.

Response of the nitrogen-fixing lichen Lobaria pulmonaria to phosphorus, molybdenum, and vanadium

USGS Publications Warehouse

Marks, Jade A; Pett-Ridge, Julie; Perakis, Steven S.; Allen, Jessica L; McCune, Bruce

2015-01-01

Nitrogen-fixing lichens (cyanolichens) are an important source of nitrogen (N) in Pacific Northwest forests, but limitation of lichen growth by elements essential for N fixation is poorly understood. To investigate how nutrient limitation may affect cyanolichen growth rates, we fertilized a tripartite cyanobacterial lichen (Lobaria pulmonaria) and a green algal non-nitrogen fixing lichen (Usnea longissima) with the micronutrients molybdenum (Mo) and vanadium (V), both known cofactors for enzymes involved in N fixation, and the macronutrient phosphorus (P). We then grew treated lichens in the field for one year in western Oregon, USA. Lichen growth was very rapid for both species and did not differ across treatments, despite a previous demonstration of P-limitation in L. pulmonaria at a nearby location. To reconcile these disparate findings, we analyzed P, Mo, and V concentrations, natural abundance δ15N isotopes, %N and change in thallus N in Lobaria pulmonaria from both growth experiments. Nitrogen levels in deposition and in lichens could not explain the large difference in growth or P limitation observed between the two studies. Instead, we provide evidence that local differences in P availability may have caused site-specific responses of Lobaria to P fertilization. In the previous experiment, Lobaria had low background levels of P, and treatment with P more than doubled growth. In contrast, Lobaria from the current experiment had much higher background P concentrations, similar to P-treated lichens in the previous experiment, consistent with the idea that ambient variation in P availability influences the degree of P limitation in cyanolichens. We conclude that insufficient P, Mo, and V did not limit the growth of either cyanolichens or chlorolichens at the site of the current experiment. Our findings point to the need to understand landscape-scale variation in P availability to cyanolichens, and its effect on spatial patterns of cyanolichen nutrient limitation and N fixation.

Glutamine Synthetase Is a Molecular Target of Nitric Oxide in Root Nodules of Medicago truncatula and Is Regulated by Tyrosine Nitration1[W][OA

PubMed Central

Melo, Paula M.; Silva, Liliana S.; Ribeiro, Isa; Seabra, Ana R.; Carvalho, Helena G.

2011-01-01

Nitric oxide (NO) is emerging as an important regulatory player in the Rhizobium-legume symbiosis, but its biological role in nodule functioning is still far from being understood. To unravel the signal transduction cascade and ultimately NO function, it is necessary to identify its molecular targets. This study provides evidence that glutamine synthetase (GS), a key enzyme for root nodule metabolism, is a molecular target of NO in root nodules of Medicago truncatula, being regulated by tyrosine (Tyr) nitration in relation to active nitrogen fixation. In vitro studies, using purified recombinant enzymes produced in Escherichia coli, demonstrated that the M. truncatula nodule GS isoenzyme (MtGS1a) is subjected to NO-mediated inactivation through Tyr nitration and identified Tyr-167 as the regulatory nitration site crucial for enzyme inactivation. Using a sandwich enzyme-linked immunosorbent assay, it is shown that GS is nitrated in planta and that its nitration status changes in relation to active nitrogen fixation. In ineffective nodules and in nodules fed with nitrate, two conditions in which nitrogen fixation is impaired and GS activity is reduced, a significant increase in nodule GS nitration levels was observed. Furthermore, treatment of root nodules with the NO donor sodium nitroprusside resulted in increased in vivo GS nitration accompanied by a reduction in GS activity. Our results support a role of NO in the regulation of nitrogen metabolism in root nodules and places GS as an important player in the process. We propose that the NO-mediated GS posttranslational inactivation is related to metabolite channeling to boost the nodule antioxidant defenses in response to NO. PMID:21914816

Pinus flexilis and Picea engelmannii share a simple and consistent needle endophyte microbiota with a potential role in nitrogen fixation

PubMed Central

Carrell, Alyssa A.; Frank, Anna C.

2014-01-01

Conifers predominantly occur on soils or in climates that are suboptimal for plant growth. This is generally attributed to symbioses with mycorrhizal fungi and to conifer adaptations, but recent experiments suggest that aboveground endophytic bacteria in conifers fix nitrogen (N) and affect host shoot tissue growth. Because most bacteria cannot be grown in the laboratory very little is known about conifer–endophyte associations in the wild. Pinus flexilis (limber pine) and Picea engelmannii (Engelmann spruce) growing in a subalpine, nutrient-limited environment are potential candidates for hosting endophytes with roles in N2 fixation and abiotic stress tolerance. We used 16S rRNA pyrosequencing to ask whether these conifers host a core of bacterial species that are consistently associated with conifer individuals and therefore potential mutualists. We found that while overall the endophyte communities clustered according to host species, both conifers were consistently dominated by the same phylotype, which made up 19–53% and 14–39% of the sequences in P. flexilis and P. engelmannii, respectively. This phylotype is related to Gluconacetobacter diazotrophicus and other N2 fixing acetic acid bacterial endophytes. The pattern observed for the P. flexilis and P. engelmannii needle microbiota—a small number of major species that are consistently associated with the host across individuals and species—is unprecedented for an endophyte community, and suggests a specialized beneficial endophyte function. One possibility is endophytic N fixation, which could help explain how conifers can grow in severely nitrogen-limited soil, and why some forest ecosystems accumulate more N than can be accounted for by known nitrogen input pathways. PMID:25071746

Tri-Party Underground Symbiosis between a Weevil, Bacteria and a Desert Plant

PubMed Central

Shelef, Oren; Helman, Yael; Friedman, Ariel-Leib-Leonid; Behar, Adi; Rachmilevitch, Shimon

2013-01-01

Inhabitants of arid ecosystems face severe nitrogen and water limitations. Inventive adaptations by organisms occupying such habitats are essential for survival. This study describes a tri-party symbiotic interaction between a plant (Salsola inermis), a beetle (Conorhynchus pistor), and a bacterium (Klebsiella pneumonia). The weevil survives by living within a mud structure affixed to the plant roots, thus benefiting from increased carbon and water, and refuge from predators and parasites. Active nitrogen-fixing bacteria harbored within the weevil's gut mediate this interaction, by supplying nitrogen to the system, which eventually promotes seed development. We studied the correlation between the weevil's existence and (i) root carbon and nitrogen content, (ii) soil water content and (iii) seed weight. Roots hosting weevils contained more nitrogen, heavier seeds and less carbon. In addition, water content was higher around the roots than in open spaces a short distance from the plant stem. Bacterial studies and nitrogen-fixation analyses, including molecular and chemical assays, indicated atmospheric nitrogen fixation in the larval stage and identified the bacterium. The coexistence of weevil and bacterial behavior coinciding with the plant's life cycle was revealed here by a long period of field observations. Out of over 60,000 known weevils, this is the only report of a weevil living most of its life underground without harming plants. The unique tri-party interaction described herein shows the important ecological role of desert plant roots and provides an example of a sustainable consortium of living organisms coping with the challenging desert environment. PMID:24244267

Loss of the nodule-specific cysteine rich peptide, NCR169, abolishes symbiotic nitrogen fixation in the Medicago truncatula dnf7 mutant.

PubMed

Horváth, Beatrix; Domonkos, Ágota; Kereszt, Attila; Szűcs, Attila; Ábrahám, Edit; Ayaydin, Ferhan; Bóka, Károly; Chen, Yuhui; Chen, Rujin; Murray, Jeremy D; Udvardi, Michael K; Kondorosi, Éva; Kaló, Péter

2015-12-08

Host compatible rhizobia induce the formation of legume root nodules, symbiotic organs within which intracellular bacteria are present in plant-derived membrane compartments termed symbiosomes. In Medicago truncatula nodules, the Sinorhizobium microsymbionts undergo an irreversible differentiation process leading to the development of elongated polyploid noncultivable nitrogen fixing bacteroids that convert atmospheric dinitrogen into ammonia. This terminal differentiation is directed by the host plant and involves hundreds of nodule specific cysteine-rich peptides (NCRs). Except for certain in vitro activities of cationic peptides, the functional roles of individual NCR peptides in planta are not known. In this study, we demonstrate that the inability of M. truncatula dnf7 mutants to fix nitrogen is due to inactivation of a single NCR peptide, NCR169. In the absence of NCR169, bacterial differentiation was impaired and was associated with early senescence of the symbiotic cells. Introduction of the NCR169 gene into the dnf7-2/NCR169 deletion mutant restored symbiotic nitrogen fixation. Replacement of any of the cysteine residues in the NCR169 peptide with serine rendered it incapable of complementation, demonstrating an absolute requirement for all cysteines in planta. NCR169 was induced in the cell layers in which bacteroid elongation was most pronounced, and high expression persisted throughout the nitrogen-fixing nodule zone. Our results provide evidence for an essential role of NCR169 in the differentiation and persistence of nitrogen fixing bacteroids in M. truncatula.

Loss of the nodule-specific cysteine rich peptide, NCR169, abolishes symbiotic nitrogen fixation in the Medicago truncatula dnf7 mutant

PubMed Central

Horváth, Beatrix; Domonkos, Ágota; Szűcs, Attila; Ábrahám, Edit; Ayaydin, Ferhan; Bóka, Károly; Chen, Yuhui; Chen, Rujin; Murray, Jeremy D.; Udvardi, Michael K.; Kondorosi, Éva; Kaló, Péter

2015-01-01

Host compatible rhizobia induce the formation of legume root nodules, symbiotic organs within which intracellular bacteria are present in plant-derived membrane compartments termed symbiosomes. In Medicago truncatula nodules, the Sinorhizobium microsymbionts undergo an irreversible differentiation process leading to the development of elongated polyploid noncultivable nitrogen fixing bacteroids that convert atmospheric dinitrogen into ammonia. This terminal differentiation is directed by the host plant and involves hundreds of nodule specific cysteine-rich peptides (NCRs). Except for certain in vitro activities of cationic peptides, the functional roles of individual NCR peptides in planta are not known. In this study, we demonstrate that the inability of M. truncatula dnf7 mutants to fix nitrogen is due to inactivation of a single NCR peptide, NCR169. In the absence of NCR169, bacterial differentiation was impaired and was associated with early senescence of the symbiotic cells. Introduction of the NCR169 gene into the dnf7-2/NCR169 deletion mutant restored symbiotic nitrogen fixation. Replacement of any of the cysteine residues in the NCR169 peptide with serine rendered it incapable of complementation, demonstrating an absolute requirement for all cysteines in planta. NCR169 was induced in the cell layers in which bacteroid elongation was most pronounced, and high expression persisted throughout the nitrogen-fixing nodule zone. Our results provide evidence for an essential role of NCR169 in the differentiation and persistence of nitrogen fixing bacteroids in M. truncatula. PMID:26401023

Microbial Ecology of Thermophilic Anaerobic Digestion. Final Report

DOE R&D Accomplishments Database

Zinder, Stephen H.

2000-04-15

This grant supported research on methanogenic archaea. The two major areas that were supported were conversion of acetic acid to methane and nitrogen fixation by Methanosarcina. Among the achievements of this research were the isolation of novel methanogenic cultures, elucidation of the pathways from acetate to methane, description of a specific DNA-binding complex in nitrogen fixing methanogens, and demonstration of an alternative nitrogenase in Methanosarcina.

Sustained effects of atmospheric [CO2] and nitrogen availability on forest soil CO2 efflux

Treesearch

A. Christopher Oishi; Sari Palmroth; Kurt H. Johnsen; Heather R. McCarthy; Ram Oren

2014-01-01

Soil CO2 efflux (Fsoil) is the largest source of carbon from forests and reflects primary productivity as well as how carbon is allocated within forest ecosystems. Through early stages of stand development, both elevated [CO2] and availability of soil nitrogen (N; sum of mineralization, deposition, and fixation) have been shown to increase gross primary productivity,...

Genome Sequence of Bradyrhizobium japonicum E109, One of the Most Agronomically Used Nitrogen-Fixing Rhizobacteria in Argentina

PubMed Central

Torres, Daniela; Revale, Santiago; Obando, Melissa; Maroniche, Guillermo; Paris, Gastón; Perticari, Alejandro; Vazquez, Martín; Wisniewski-Dyé, Florence; Martínez-Abarca, Francisco

2015-01-01

We present here the complete genome sequence of Bradyrhizobium japonicum strain E109, one of the most used rhizobacteria for soybean inoculation in Argentina since the 1970s. The genome consists of a 9.22-Mbp single chromosome and contains several genes related to nitrogen fixation, phytohormone biosynthesis, and a rhizospheric lifestyle. PMID:25700406

Effect of phosphate additive on the nitrogen transformation during pig manure composting.

PubMed

Wu, Juan; He, Shengzhou; Liang, Ying; Li, Guoxue; Li, Song; Chen, Shili; Nadeem, Faisal; Hu, Jingwei

2017-07-01

Previous studies revealed that phosphate, as an additive to composting, could significantly reduce NH 3 emission and nitrogen loss through change of pH and nitrogen fixation to form ammonium phosphate. However, few studies have explored the influence of pH change and phosphate additive on NO x - -N, NH 4 + -N, NH 3 , and N 2 O, which are dominate forms of nitrogen in composting. In this study, the equimolar H 3 PO 4 , H 2 SO 4 , and K 2 HPO 4 were added into pig manure composting to evaluate the effect of H + and PO 4 3- on nitrogen transformation. As a result, we reached the conclusion that pH displays significant influence on adsorption from PO 4 3- to NH 4 + . The NH 4 + -N concentration in H 3 PO 4 treatment kept over 3 g kg -1 DM (dry matter) which is obviously higher than that in H 2 SO 4 treatment, and NH 4 + -N concentration in K 2 HPO 4 treatment (pH>8.5) is lower than 0.5 g kg -1 DM because adsorption capacity of PO 4 3- is greatly weakened and NH 4 + -N rapidly transformed to NH 3 -N influenced by high pH value. The N 2 O emission of composting is significantly correlated with incomplete denitrification of NO x - -N, and PO 4 3- addition could raise NO x - -N contents to restrict denitrification and further to promote N 2 O emission. The study reveals the influence mechanism of phosphate additive to nitrogen transformation during composting, presents theoretical basis for additive selection in nitrogen fixation, and lays foundation for study about nitrogen circulation mechanism during composting.

[Characteristics of soil microorganisms and soil nutrients in different sand-fixation shrub plantations in Kubuqi Desert, China].

PubMed

Zhang, Li-Xin; Duan, Yu Xi; Wang, Bo; Wang, Wei Feng; Li, Xiao Jing; Liu, Jin Jie

2017-12-01

Three types of sand-fixation shrub plantations, including Artemisia ordosica + Hedysarum fruticosum, Caragana korshinskii and Salix psammophila, were selected in the eastern area of Kubuqi Desert to study the changes in soil microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN), quantities of soil microorganisms, contents of soil nutrients and the relations among these variables under the different plantation types and shifting sandy land. The restoration effects of each plantation type on soil quality were assessed by synthetic index method. The results showed that the contents of soil organic matter, total nitrogen and phosphorus, and available nitrogen and phosphorus under different plantations were all significantly greater than those under shifting sandy land, and the order of increase was A. ordosica + H. fruticosum ＞ C. korshinskii ＞ S. psammophila. The soil nutrient contents decreased with the increase of soil depth under all plantation types. The quantities of soil microorganisms and the contents of soil MBC and MBN under the plantations were higher at different degrees than those under shifting sandy land. MBC, MBN and the relative numbers of bacteria under A. ordosica+H. fruticosum plantation were higher than those under C. korshinskii plantation and S. psammophila plantation. The relative numbers of fungi and actinobacteria decreased in the order of C. korshinskii ＞ S. psammophila ＞ A. ordosica + H. fruticosum. The relative number of bacteria, MBC and MBN under the plantations were mainly affected by the contents of soil organic matter, total nitrogen, total phosphorus, available nitrogen, available phosphorus, as well as C/N, and the relative numbers of actinobacteria and fungi were primarily affected by the contents of soil total phosphorus, available nitrogen and available phosphorus. Soil quality was ranked in the order of A. ordosica + H. fruticosum ＞ C. korshinskii ＞ S. psammophila ＞ shifting sandy land. These results demonstrated that different sand-fixation shrub plantations could improve the quality of the desert soil and the A. ordosica + H. fruticosum plantation was the best for soil restoration and quality improvement in the desert.

Effects of Iron and Nitrogen Limitation on Sulfur Isotope Fractionation during Microbial Sulfate Reduction

PubMed Central

Ono, Shuhei; Bosak, Tanja

2012-01-01

Sulfate-reducing microbes utilize sulfate as an electron acceptor and produce sulfide that is depleted in heavy isotopes of sulfur relative to sulfate. Thus, the distribution of sulfur isotopes in sediments can trace microbial sulfate reduction (MSR), and it also has the potential to reflect the physiology of sulfate-reducing microbes. This study investigates the relationship between the availability of iron and reduced nitrogen and the magnitude of S-isotope fractionation during MSR by a marine sulfate-reducing bacterium, DMSS-1, a Desulfovibrio species, isolated from salt marsh in Cape Cod, MA. Submicromolar levels of iron increase sulfur isotope fractionation by about 50% relative to iron-replete cultures of DMSS-1. Iron-limited cultures also exhibit decreased cytochrome c-to-total protein ratios and cell-specific sulfate reduction rates (csSRR), implying changes in the electron transport chain that couples carbon and sulfur metabolisms. When DMSS-1 fixes nitrogen in ammonium-deficient medium, it also produces larger fractionation, but it occurs at faster csSRRs than in the ammonium-replete control cultures. The energy and reducing power required for nitrogen fixation may be responsible for the reverse trend between S-isotope fractionation and csSRR in this case. Iron deficiency and nitrogen fixation by sulfate-reducing microbes may lead to the large observed S-isotope effects in some euxinic basins and various anoxic sediments. PMID:23001667

New, national bottom-up estimate for tree-based biological ...

EPA Pesticide Factsheets

Nitrogen is a limiting nutrient in many ecosystems, but is also a chief pollutant from human activity. Quantifying human impacts on the nitrogen cycle and investigating natural ecosystem nitrogen cycling both require an understanding of the magnitude of nitrogen inputs from biological nitrogen fixation (BNF). A bottom-up approach to estimating BNF—scaling rates up from measurements to broader scales—is attractive because it is rooted in actual BNF measurements. However, bottom-up approaches have been hindered by scaling difficulties, and a recent top-down approach suggested that the previous bottom-up estimate was much too large. Here, we used a bottom-up approach for tree-based BNF, overcoming scaling difficulties with the systematic, immense (>70,000 N-fixing trees) Forest Inventory and Analysis (FIA) database. We employed two approaches to estimate species-specific BNF rates: published ecosystem-scale rates (kg N ha-1 yr-1) and published estimates of the percent of N derived from the atmosphere (%Ndfa) combined with FIA-derived growth rates. Species-specific rates can vary for a variety of reasons, so for each approach we examined how different assumptions influenced our results. Specifically, we allowed BNF rates to vary with stand age, N-fixer density, and canopy position (since N-fixation is known to require substantial light).Our estimates from this bottom-up technique are several orders of magnitude lower than previous estimates indicating

Dinitrogen Fixation Within and Adjacent to Oxygen Deficient Waters of the Eastern Tropical South Pacific Ocean

NASA Astrophysics Data System (ADS)

Widner, B.; Mulholland, M. R.; Bernhardt, P. W.; Chang, B. X.; Jayakumar, A.

2016-02-01

Recent work suggests that planktonic diazotrophs are geographically more widely distributed than previously thought including relatively warm (14-23oC) aphotic oxygenated pelagic waters and in aphotic waters within oxygen deficient zones. Because the volume of aphotic water in the ocean is large and may increase in the future, if dinitrogen (N2) fixation is widely occurring at sub-euphotic depths, this could result in a dramatic upward revision of global nitrogen (N) inputs via this process. N2 fixation rates were measured during a cruise in the Eastern Tropical South Pacific using stable isotope tracer techniques that account for slow gas dissolution. Results are compared with light, nutrient, and oxygen gradients (and necessarily temperature gradients). In addition, rates of N2 fixation made in vertical profiles within and above oxygen deficient waters are compared with those measured in vertical profiles adjacent to oxygen deficient waters. Results suggest that while rates of N2 fixation were measurable in deeper anoxic waters, volumetric N2 fixation rates were higher in surface waters.

A model for diurnal patterns of carbon fixation in a Precambrian microbial mat based on a modern analog

NASA Technical Reports Server (NTRS)

Rothschild, L. J.

1991-01-01

Microbial mat communities are one of the first and most prevalent biological communities known from the Precambrian fossil record. These fossil mat communities are found as laminated sedimentary rock structures called stromatolites. Using a modern microbial mat as an analog for Precambrian stromatolites, a study of carbon fixation during a diurnal cycle under ambient conditions was undertaken. The rate of carbon fixation depends primarily on the availability of light (consistent with photosynthetic carbon fixation) and inorganic carbon, and not nitrogen or phosphorus. Atmospheric PCO2 is thought to have decreased from 10 bars at 4 Ga (10(9) years before present) to approximately 10(-4) bars today, implying a change in the availability of inorganic carbon for carbon fixation. Experimental manipulation of levels of inorganic carbon to levels that may have been available to Precambrian mat communities resulted in increased levels of carbon fixation during daylight hours. Combining these data with models of daylength during the Precambrian, models are derived for diurnal patterns of photosynthetic carbon fixation in a Precambrian microbial mat community. The models suggest that, even in the face of shorter daylengths during the Precambrian, total daily carbon fixation has been declining over geological time, with most of the decrease having occurred during the Precambrian.

Heterotrophic N2-fixation contributes to nitrogen economy of a common wetland sedge, Schoenoplectus californicus.

PubMed

Rejmánková, Eliška; Sirová, Dagmara; Castle, Stephanie T; Bárta, Jiří; Carpenter, Heather

2018-01-01

A survey of the ecological variability within 52 populations of Schoenoplectus californicus (C.A. Mey.) Soják across its distributional range revealed that it is commonly found in nitrogen (N) limited areas, but rarely in phosphorus limited soils. We explored the hypothesis that S. californicus supplements its nitrogen demand by bacterial N2-fixation processes associated with its roots and rhizomes. We estimated N2-fixation of diazotrophs associated with plant rhizomes and roots from several locations throughout the species' range and conducted an experiment growing plants in zero, low, and high N additions. Nitrogenase activity in rhizomes and roots was measured using the acetylene reduction assay. The presence of diazotrophs was verified by the detection of the nifH gene. Nitrogenase activity was restricted to rhizomes and roots and it was two orders of magnitude higher in the latter plant organs (81 and 2032 nmol C2H4 g DW-1 d-1, respectively). Correspondingly, 40x more nifH gene copies were found on roots compared to rhizomes. The proportion of the nifH gene copies in total bacterial DNA was positively correlated with the nitrogenase activity. In the experiment, the contribution of fixed N to the plant N content ranged from 13.8% to 32.5% among clones from different locations. These are relatively high values for a non-cultivated plant and justify future research on the link between N-fixing bacteria and S. californicus production.

Iron deficiency increases growth and nitrogen-fixation rates of phosphorus-deficient marine cyanobacteria.

PubMed

Garcia, Nathan S; Fu, Feixue; Sedwick, Peter N; Hutchins, David A

2015-01-01

Marine dinitrogen (N2)-fixing cyanobacteria have large impacts on global biogeochemistry as they fix carbon dioxide (CO2) and fertilize oligotrophic ocean waters with new nitrogen. Iron (Fe) and phosphorus (P) are the two most important limiting nutrients for marine biological N2 fixation, and their availabilities vary between major ocean basins and regions. A long-standing question concerns the ability of two globally dominant N2-fixing cyanobacteria, unicellular Crocosphaera and filamentous Trichodesmium, to maintain relatively high N2-fixation rates in these regimes where both Fe and P are typically scarce. We show that under P-deficient conditions, cultures of these two cyanobacteria are able to grow and fix N2 faster when Fe deficient than when Fe replete. In addition, growth affinities relative to P increase while minimum concentrations of P that support growth decrease at low Fe concentrations. In Crocosphaera, this effect is accompanied by a reduction in cell sizes and elemental quotas. Relatively high growth rates of these two biogeochemically critical cyanobacteria in low-P, low-Fe environments such as those that characterize much of the oligotrophic ocean challenge the common assumption that low Fe levels can have only negative effects on marine primary producers. The closely interdependent influence of Fe and P on N2-fixing cyanobacteria suggests that even subtle shifts in their supply ratio in the past, present and future oceans could have large consequences for global carbon and nitrogen cycles.

Phylogeny of nodulation genes and symbiotic diversity of Acacia senegal (L.) Willd. and A. seyal (Del.) Mesorhizobium strains from different regions of Senegal.

PubMed

Bakhoum, Niokhor; Galiana, Antoine; Le Roux, Christine; Kane, Aboubacry; Duponnois, Robin; Ndoye, Fatou; Fall, Dioumacor; Noba, Kandioura; Sylla, Samba Ndao; Diouf, Diégane

2015-04-01

Acacia senegal and Acacia seyal are small, deciduous legume trees, most highly valued for nitrogen fixation and for the production of gum arabic, a commodity of international trade since ancient times. Symbiotic nitrogen fixation by legumes represents the main natural input of atmospheric N2 into ecosystems which may ultimately benefit all organisms. We analyzed the nod and nif symbiotic genes and symbiotic properties of root-nodulating bacteria isolated from A. senegal and A. seyal in Senegal. The symbiotic genes of rhizobial strains from the two Acacia species were closed to those of Mesorhizobium plurifarium and grouped separately in the phylogenetic trees. Phylogeny of rhizobial nitrogen fixation gene nifH was similar to those of nodulation genes (nodA and nodC). All A. senegal rhizobial strains showed identical nodA, nodC, and nifH gene sequences. By contrast, A. seyal rhizobial strains exhibited different symbiotic gene sequences. Efficiency tests demonstrated that inoculation of both Acacia species significantly affected nodulation, total dry weight, acetylene reduction activity (ARA), and specific acetylene reduction activity (SARA) of plants. However, these cross-inoculation tests did not show any specificity of Mesorhizobium strains toward a given Acacia host species in terms of infectivity and efficiency as stated by principal component analysis (PCA). This study demonstrates that large-scale inoculation of A. senegal and A. seyal in the framework of reafforestation programs requires a preliminary step of rhizobial strain selection for both Acacia species.

Investigating patterns of symbiotic nitrogen fixation during vegetation change from grassland to woodland using fine scale δ(15) N measurements.

PubMed

Soper, Fiona M; Boutton, Thomas W; Sparks, Jed P

2015-01-01

Biological nitrogen fixation (BNF) in woody plants is often investigated using foliar measurements of δ(15) N and is of particular interest in ecosystems experiencing increases in BNF due to woody plant encroachment. We sampled δ(15) N along the entire N uptake pathway including soil solution, xylem sap and foliage to (1) test assumptions inherent to the use of foliar δ(15) N as a proxy for BNF; (2) determine whether seasonal divergences occur between δ(15) Nxylem sap and δ(15) Nsoil inorganic N that could be used to infer variation in BNF; and (3) assess patterns of δ(15) N with tree age as indicators of shifting BNF or N cycling. Measurements of woody N-fixing Prosopis glandulosa and paired reference non-fixing Zanthoxylum fagara at three seasonal time points showed that δ(15) Nsoil inorganic N varied temporally and spatially between species. Fractionation between xylem and foliar δ(15) N was consistently opposite in direction between species and varied on average by 2.4‰. Accounting for these sources of variation caused percent nitrogen derived from fixation values for Prosopis to vary by up to ∼70%. Soil-xylem δ(15) N separation varied temporally and increased with Prosopis age, suggesting seasonal variation in N cycling and BNF and potential long-term increases in BNF not apparent through foliar sampling alone. © 2014 John Wiley & Sons Ltd.

Significance of Phosphoenolpyruvate Carboxylase during Ammonium Assimilation

PubMed Central

Guy, Robert D.; Vanlerberghe, Greg C.; Turpin, David H.

1989-01-01

The effect of N-assimilation on the partitioning of carbon fixation between phosphoenolpyruvate carboxylase (PEPcase) and ribulose bisphosphate carboxylase/oxygenase (Rubisco) was determined by measuring stable carbon isotope discrimination during photosynthesis by an N-limited green alga, Selenastrum minutum (Naeg.) Collins. This was facilitated by a two process model accounting for simultaneous CO2 fixation and respiratory CO2 release. Discrimination by control cells was consistent with the majority of carbon being fixed by Rubisco. During nitrogen assimilation however, discrimination was greatly reduced indicating an enhanced flux through PEPcase which accounted for upward of 70% of total carbon fixation. This shift toward anaplerotic metabolism supports a large increase in tricarboxylic acid cycle activity primarily between oxaloacetate and α-ketoglutarate thereby facilitating the provision of carbon skeletons for amino acid synthesis. This provides an example of a unique set of conditions under which anaplerotic carbon fixation by PEPcase exceeds photosynthetic carbon fixation by Rubisco in a C3 organism. Images Figure 6 PMID:16666678

Significance of Phosphoenolpyruvate Carboxylase during Ammonium Assimilation: Carbon Isotope Discrimination in Photosynthesis and Respiration by the N-Limited Green Alga Selenastrum minutum.

PubMed

Guy, R D; Vanlerberghe, G C; Turpin, D H

1989-04-01

The effect of N-assimilation on the partitioning of carbon fixation between phosphoenolpyruvate carboxylase (PEPcase) and ribulose bisphosphate carboxylase/oxygenase (Rubisco) was determined by measuring stable carbon isotope discrimination during photosynthesis by an N-limited green alga, Selenastrum minutum (Naeg.) Collins. This was facilitated by a two process model accounting for simultaneous CO(2) fixation and respiratory CO(2) release. Discrimination by control cells was consistent with the majority of carbon being fixed by Rubisco. During nitrogen assimilation however, discrimination was greatly reduced indicating an enhanced flux through PEPcase which accounted for upward of 70% of total carbon fixation. This shift toward anaplerotic metabolism supports a large increase in tricarboxylic acid cycle activity primarily between oxaloacetate and alpha-ketoglutarate thereby facilitating the provision of carbon skeletons for amino acid synthesis. This provides an example of a unique set of conditions under which anaplerotic carbon fixation by PEPcase exceeds photosynthetic carbon fixation by Rubisco in a C(3) organism.

Soybean nodulation and symbiotic nitrogen fixation in response to soil compaction and mulching

NASA Astrophysics Data System (ADS)

Siczek, A.; Lipiec, J.

2009-04-01

Symbiotic nitrogen fixation by legume crops such as soybean plays a key role in supplying nitrogen for agricultural systems. In symbiotic associations with Bradyrhizobium japonicum soybean can fix up to 200 kg N ha-1 yr-1. This reduces the need for expensive and often environmentally harmful because of leaching nitrogen fertilization. However both soybean nodulation and nitrogen fixation are sensitive to soil conditions. One of the critical soil constraints is soil compaction. Increasing use of heavy equipment and intensive cropping in modern agriculture leads to excessive soil compaction. Compaction often is found as a result of field operations that have to be performed in a very short period of time and when soils are wet and more susceptible to compaction. This results in unfavourable water content, temperature, aeration, pore size distribution, strength for plant growth and microbial activity. The surface mulching can alleviate the adverse effect of the environmental factors on soil by decreasing fluctuation of soil temperature, increasing moisture by controlling evaporation from the soil surface, decreasing bulk density, preventing soil crusting. The effect of mulch on soil conditions largely depends on soil compaction and weather conditions during growing season. The positive effect of the straw mulch on soil moisture has been seen under seasons with insufficient rainfalls. However thicker layers of mulch can act as diffusion barrier, especially when the mulch is wet. Additionally, low soil temperature prevalent during early spring under mulch can impede development of nodule, nodule size and delay onset of nodulation. The aim of this study was to determine the effect of the straw mulch on nodulation and nitrogen fixation of soybean in variously compacted soil. The experimental field was 192 m2and was divided into three parts composed of 6 micro-plots with area 7 m2. Three degrees of soil compaction obtained in each field part through tractor passes were compared: low, medium and heavy (0, 3 or 5 passes, respectively). This resulted in a wide range of soil bulk density (1.2 to 1.65 Mg m-3) that can occur in the arable fields. To obtain uniform conditions for seed germination and initial seedling growth the entire plot area was tilled with a cultivator-harrow to a depth of 5 cm after soil compaction. Soybean "Aldana" seeds inoculated with B. japonicum were sown in rows with spacing of 0.3 m. After sowing half of each micro-plot was mulched with wheat straw (0.5 kg m-2) and another one - not. Nodulation was evaluated by using the parameters of nodule number and nodule weight and acetylene reduction assay was used for the measurement of nitrogenase activity. Number of nodules on root system under mulched and not mulched soil was the highest in not compacted and medium compacted soil, respectively and the lowest - in most compacted soil with mulch. Nitrogenase activity ( mol C2H4 h-1 plant-1) decreased as soil compaction increased but the more pronounced tendency and higher values were obtained in mulched plots. The results indicate that mulching in some range of soil compaction can improve soybean nodulation and nitrogen fixation. This work was funded in part by the Polish Ministry of Science and Higher Education (Grant No N N310 149635).

Arctic shelves as platforms for biogeochemical activity: Nitrogen and carbon transformations in the Chukchi Sea, Alaska

NASA Astrophysics Data System (ADS)

Hardison, Amber K.; McTigue, Nathan D.; Gardner, Wayne S.; Dunton, Kenneth H.

2017-10-01

Continental shelves comprise <5% of global ocean area but may account for a disproportionate 30% of primary production, 80% of organic matter burial, and >50% of marine denitrification. The Hanna Shoal region, part of the continental shelf system in the northeast Chukchi Sea, Alaska, is recognized for its high biodiversity and productivity. We investigated the role of sediments in organic matter decomposition and nutrient cycling at five stations on the shallow Hanna Shoal. In particular, we asked (1) how much sediment organic matter is remineralized in the Chukchi Sea, and what factors drive this degradation, (2) do sediments function as a net source for fixed nitrogen (thus fueling primary production in the overlying water), or as a net sink for fixed nitrogen (thereby removing it from the system), and (3) what is the balance between sediment NH4+ uptake and regeneration, and what factors drive NH4+ cycling? We conducted dark sediment core incubations to measure sediment O2 consumption, net N2 and nutrient (NH4+, NO3-, NO2-, PO43-) fluxes, and rates of sediment NH4+ cycling, including uptake and regeneration. Rates of sediment O2 consumption and NH4+ and PO43- efflux suggest that high organic matter remineralization rates occurred in these cold (-2 °C) sediments. We estimated that total organic carbon remineralization accounted for 20-57% of summer export production measured on the Chukchi Shelf. Net N2 release was the dominant nitrogen flux, indicating that sediments acted as a net sink for bioavailable nitrogen via denitrification. Organic carbon remineralization via denitrification accounted for 6-12% of summer export production, which made up 25% of the total organic carbon oxidized in Hanna Shoal sediments. These shallow, productive Arctic shelves are ;hotspots; for organic matter remineralization.

Global climate change and terrestrial net primary production

NASA Technical Reports Server (NTRS)

Melillo, Jerry M.; Mcguire, A. D.; Kicklighter, David W.; Moore, Berrien, III; Vorosmarty, Charles J.; Schloss, Annette L.

1993-01-01

A process-based model was used to estimate global patterns of net primary production and soil nitrogen cycling for contemporary climate conditions and current atmospheric CO2 concentration. Over half of the global annual net primary production was estimated to occur in the tropics, with most of the production attributable to tropical evergreen forest. The effects of CO2 doubling and associated climate changes were also explored. The responses in tropical and dry temperate ecosystems were dominated by CO2, but those in northern and moist temperate ecosystems reflected the effects of temperature on nitrogen availability.

Azolla filiculoides Nitrogenase Activity Decrease Induced by Inoculation with Chlamydomonas sp. †

PubMed Central

Habte, Mitiku

1986-01-01

Experiments were conducted to determine the influence of Chlamydomonas sp. on nitrogen fixation (C2H2 → C2H4) in Azolla filiculoides and on the nitrogen fixation and growth of free-living Anabaena azollae 2B organisms. Inoculation of azolla medium with Chlamydomonas sp. was associated with decreased nitrogenase activity in A. filiculoides and with increases in the density of a fungal population identified as Acremonium sp. Subsequent inoculation of azolla medium with this fungus was also accompanied by a significant decrease in nitrogenase activity of A. filiculoides. However, the extent of depression of nitrogenase activity was significantly higher when azolla medium was inoculated with Chlamydomonas sp. than when it was inoculated with Acremonium sp. Inoculation of nitrogen-free Stanier medium with either Acremonium sp. or Chlamydomonas sp. did not adversely affect the growth or nitrogenase activity of free-living A. azollae. Decreased nitrogenase activity in A. filiculoides is apparently related to the adverse influence of the green alga and the fungus on the macrosymbiont. The mechanisms that might be involved are discussed. PMID:16347211

[Rice endogenous nitrogen fixing and growth promoting bacterium Herbaspirillum seropedicae DX35].

PubMed

Wang, Xiucheng; Cao, Yanhua; Tang, Xue; Ma, Xiaotong; Gao, Jusheng; Zhang, Xiaoxia

2014-03-04

To screen efficient nitrogen fixation endophytes from rice and to analyze their growth-promoting properties. We isolated strains from the roots of rice in the field where it has a rice-rice-green manure rotation system for 30 years. Efficient strains were screened by acetylene reduction assay. Phylogenetic analysis is based on 16S rRNA gene, nifH gene and the composition of fatty acid. In addition, we also detected the ability of indole acetic acid secretion through the Salkowski colorimetric method, measured the production of siderophore through the blue plate assay and detected phosphate solubilization, to analyze the growth-promoting properties. A total of 48 strains were isolated, in which DX35 has the highest nitrogenase activity. It belongs to Herbaspirillum seropedicae after identification. Its nitrogenase activity (181.21 nmol C2H4/(mg protein x h)) was 10 times as much as the reference strain Azotobacter chroococcum ACCC10006. In addition, it also can secrete siderophore and solubilize phosphorus. Strain DX35, belonging to Herbaspirillum seropedicae, is an efficient nitrogen fixation endophytes.

Exploring the effects of black mangrove (Avicennia germinans) expansions on nutrient cycling in smooth cordgrass (Spartina alterniflora) marsh sediments of southern Louisiana, USA

NASA Astrophysics Data System (ADS)

Henry, K. M.; Twilley, R. R.

2011-12-01

Located at the northernmost extent of mangroves in the Gulf of Mexico, coastal Louisiana (LA) provides an excellent opportunity to study the effects of a climate-induced vegetation shift on nutrient cycling within an ecosystem. Climate throughout the Gulf Coast region is experiencing a general warming trend and scientists predict both hotter summers (+1.5 to 4 °C) and warmer winters (+1.5 to 5.5 °C) by 2100. Over the last two decades, mild winter temperatures have facilitated the expansion of black mangrove trees (Avicennia germinans) into the smooth cordgrass (Spartina alterniflora) along parts of the LA coast. Due to differences in morphology and physiology between these two species, the expansion of Avicennia has the potential to greatly alter sediment biogeochemistry, especially nutrient cycling. With such an extensive history of coastal nutrient enrichment and eutrophication in the Mississippi River delta, it is important to understand how nutrient cycling, retention, and removal in this region will be affected by this climate-induced vegetation expansion. We examined the effect of this species shift on porewater salinity, sulfide, and dissolved inorganic nutrient concentrations (nitrite, nitrate, ammonium, and phosphate) as well as sediment oxidation-reduction potential, bulk density, and nutrient content (carbon, nitrogen, phosphorus). We also measured net dinitrogen (N2:Ar), oxygen, and dissolved inorganic nutrient fluxes on intact, non-vegetated sediment cores collected from both Spartina and Avicennia habitats. Spartina sediments were more reducing, with higher concentrations of sulfides and ammonium. We found no significant difference between Spartina and Avicennia sediment dinitrogen, oxygen, or dissolved inorganic nutrient fluxes. Net dinitrogen fluxes for both habitat types were predominately positive, indicating higher rates of denitrification than nitrogen fixation at these sites. Sediments were primarily a nitrate sink, but functioned as both a source and sink of nitrite, ammonium, and phosphate depending on the season and light conditions. Further sediment analysis showed no significant difference in bulk density, carbon, nitrogen, or phosphorus content between Spartina and Avicennia sediments. Marine sediments high in bulk density and phosphorus content and low carbon and nitrogen content dominated the top several centimeters in both Spartina and Avicennia habitats. These surprising but reassuring results suggest that in a region where allochthonous sediment input dominates organic accretion from the primary producers, the climate-induced shift from Spartina to Avicennia will have little to no affect on littoral nutrient cycling.

Lightning - Estimates of the rates of energy dissipation and nitrogen fixation

NASA Technical Reports Server (NTRS)

Borucki, W. J.; Chameides, W. L.

1984-01-01

The nitrogen needed by plants can normally not be directly obtained from the nitrogen present in molecular form in the atmosphere. The reason for this situation is related to the great energy required to break the N-N bond. Only a few organisms, such as algae and certain bacteria, can 'fix' nitrogen. An abiological process for breaking the N-N bond is provided by lightning. The present investigation is concerned with this possibility. It is found that lightning produces approximately 2.6 x 10 to the 9th kg N per year. There are, however, uncertainties, which are mainly related to the energy of a lightning flash.

[CAM in Tillandsia usneoides: Studies on the pathway of carbon and the dependency of CO2-exchange on light intensity, temperature and water content of the plant].

PubMed

Kluge, M; Lange, O L; Eichmann, M V; Schmid, R

1973-12-01

Tillandsia usneoides, in the common sense a non-succulent plant, exhibits CO2 exchange characterized by net CO2 dark fixation during the night and depression of CO2 exchange during the day. Malate has been demonstrated to accumulate during CO2 dark fixation and to be converted to carbohydrates in light. Thus, T. usneoides exhibits CAM like typical succulents.Net CO2 uptake during the day is increased with net CO2 output being suppressed in duration of time and extent when light intensity increases. Furthermore, a slight increase in CO2 fixation during the following night can be observed if the plants were treated with high light intensity during the previous day.Curves of CO2 exchange typical for CAM are obtained if T. usneoides is kept at 15°C and 20°C. Lower temperature tend to increase CO2 uptake during the day and to inhibit CO2 dark fixation. Temperatures higher than 20°C favour loss of CO2 by respiration, which becomes apparent during the whole day and night at 30°C and higher temperatures. Thus, T. usneoides gains carbon only at temperatures well below 25°C.Net CO2 uptake during the day occurs only in moist plant material and is inhibited in plants cept under water stress conditions. However, CO2 uptake during the night is clearly favoured if the plants dry out. Therefore dry plants gain more carbon than moist ones.Curves of CO2 exchange typical for CAM were also obtained with 13 other species of the genus Tillandsia.The exhibition of CAM by the non-succulent T. usneoides calls for a new definition of the term "succulence" if it is to remain useful in characterizing this metabolic pathway. Because CO2-fixing cells of T. usneoides possess relatively large vacuoles and are relatively poor in chloroplasts, they resembles the assimilatory cells of typical CAM-exhibiting succulents. Therefore, if "succulence" only means the capacity of big vacuoles to store malate, the assimilatory cells in T. usneoides are succulent. It seems to be useful to investigate parameters which would allow a definition of the term "succulence" on the level of the cell rather than on the level of the whole plant or plant organs.

Coupled nutrient cycling determines tropical forest trajectory under elevated CO2.

NASA Astrophysics Data System (ADS)

Bouskill, N.; Zhu, Q.; Riley, W. J.

2017-12-01

Tropical forests have a disproportionate capacity to affect Earth's climate relative to their areal extent. Despite covering just 12 % of land surface, tropical forests account for 35 % of global net primary productivity and are among the most significant of terrestrial carbon stores. As atmospheric CO2 concentrations increase over the next century, the capacity of tropical forests to assimilate and sequester anthropogenic CO2 depends on limitation by multiple factors, including the availability of soil nutrients. Phosphorus availability has been considered to be the primary factor limiting metabolic processes within tropical forests. However, recent evidence points towards strong spatial and temporal co-limitation of tropical forests by both nitrogen and phosphorus. Here, we use the Accelerated Climate Modeling for Energy (ACME) Land Model (ALMv1-ECA-CNP) to examine how nutrient cycles interact and affect the trajectory of the tropical forest carbon sink under, (i) external nutrient input, (ii) climate (iii) elevated CO2, and (iv) a combination of 1-3. ALMv1 includes recent theoretical advances in representing belowground competition between roots, microbes and minerals for N and P uptake, explicit interactions between the nitrogen and phosphorus cycles (e.g., phosphatase production and nitrogen fixation), the dynamic internal allocation of plant N and P resources, and the integration of global datasets of plant physiological traits. We report nutrient fertilization (N, P, N+P) predictions for four sites in the tropics (El Verde, Puerto Rico, Barro Colorado Island, Panama, Manaus, Brazil and the Osa Peninsula, Coast Rica) to short-term nutrient fertilization (N, P, N+P), and benchmarking of the model against a meta-analysis of forest fertilization experiments. Subsequent simulations focus on the interaction of the carbon, nitrogen, and phosphorus cycles across the tropics with a focus on the implications of coupled nutrient cycling and the fate of the tropical forest carbon sink. Our results highlight the importance of transient CNP allocation, leaf-level stoichiometric controls on photosynthesis, and trade-offs between above and belowground plant investments.

Genome Sequence of Bradyrhizobium japonicum E109, One of the Most Agronomically Used Nitrogen-Fixing Rhizobacteria in Argentina.

PubMed

Torres, Daniela; Revale, Santiago; Obando, Melissa; Maroniche, Guillermo; Paris, Gastón; Perticari, Alejandro; Vazquez, Martín; Wisniewski-Dyé, Florence; Martínez-Abarca, Francisco; Cassán, Fabricio

2015-02-19

We present here the complete genome sequence of Bradyrhizobium japonicum strain E109, one of the most used rhizobacteria for soybean inoculation in Argentina since the 1970s. The genome consists of a 9.22-Mbp single chromosome and contains several genes related to nitrogen fixation, phytohormone biosynthesis, and a rhizospheric lifestyle. Copyright © 2015 Torres et al.

Modelling the combined impacts of climate change and direct anthropogenic drivers on the ecosystem of the northwest European continental shelf

NASA Astrophysics Data System (ADS)

Wakelin, Sarah L.; Artioli, Yuri; Butenschön, Momme; Allen, J. Icarus; Holt, Jason T.

2015-12-01

The potential response of the marine ecosystem of the northwest European continental shelf to climate change under a medium emissions scenario (SRES A1B) is investigated using the coupled hydrodynamics-ecosystem model POLCOMS-ERSEM. Changes in the near future (2030-2040) and the far future (2082-2099) are compared to the recent past (1983-2000). The sensitivity of the ecosystem to potential changes in multiple anthropogenic drivers (river nutrient loads and benthic trawling) in the near future is compared to the impact of changes in climate. With the exception of the biomass of benthic organisms, the influence of the anthropogenic drivers only exceeds the impact of climate change in coastal regions. Increasing river nitrogen loads has a limited impact on the ecosystem whilst reducing river nitrogen and phosphate concentrations affects net primary production (netPP) and phytoplankton and zooplankton biomass. Direct anthropogenic forcing is seen to mitigate/amplify the effects of climate change. Increasing river nitrogen has the potential to amplify the effects of climate change at the coast by increasing netPP. Reducing river nitrogen and phosphate mitigates the effects of climate change for netPP and the biomass of small phytoplankton and large zooplankton species but amplifies changes in the biomass of large phytoplankton and small zooplankton.

Transcriptome analysis of two recombinant inbred lines of common bean contrasting for symbiotic nitrogen fixation

PubMed Central

Kamfwa, Kelvin; Zhao, Dongyan; Kelly, James D.

2017-01-01

Common bean (Phaseolus vulgaris L.) fixes atmospheric nitrogen (N2) through symbiotic nitrogen fixation (SNF) at levels lower than other grain legume crops. An understanding of the genes and molecular mechanisms underlying SNF will enable more effective strategies for the genetic improvement of SNF traits in common bean. In this study, transcriptome profiling was used to identify genes and molecular mechanisms underlying SNF differences between two common bean recombinant inbred lines that differed in their N-fixing abilities. Differential gene expression and functional enrichment analyses were performed on leaves, nodules and roots of the two lines when grown under N-fixing and non-fixing conditions. Receptor kinases, transmembrane transporters, and transcription factors were among the differentially expressed genes identified under N-fixing conditions, but not under non-fixing conditions. Genes up-regulated in the stronger nitrogen fixer, SA36, included those involved in molecular functions such as purine nucleoside binding, oxidoreductase and transmembrane receptor activities in nodules, and transport activity in roots. Transcription factors identified in this study are candidates for future work aimed at understanding the functional role of these genes in SNF. Information generated in this study will support the development of gene-based markers to accelerate genetic improvement of SNF in common bean. PMID:28192540

Microgravity Effects on the Early Events of Biological Nitrogen Fixation in Medicago Truncatula: Results from the SyNRGE Experiment

NASA Astrophysics Data System (ADS)

Stutte, Gary W.; Roberts, Michael S.

2013-02-01

SyNRGE (Symbiotic Nodulation in a Reduced Gravity Environment) was a sortie mission on STS-135 in the Biological Research in Canisters (BRIC) hardware to study the effect of μg on a plant-microbe symbiosis resulting in biological nitrogen fixation. Medicago truncatula, a model species for the legume family, was inoculated with its bacterial symbiont, Sinorhizobium meliloti, to observe early biomolecular events associated with infection and nodulation in Petri Dish Fixation Units (PDFU’s). Two sets of experiments were conducted in orbit and in 24-hour delayed ground controls. Experiments were designed to determine if S. meliloti would infect M. truncatula and initiate biomolecular changes associated with nodule formation and if the μg environment altered the host plant and/or bacteria to induce nodule formation upon return to 1g. Initial analysis results demonstrate that the legumes and bacteria cultivated in μg have potential to develop a symbiotic interaction, but suggest that μg alters their ability to form nodules upon return to 1g. (Research supported by NASA ESMD/ Advance Capabilities Division grant NNX10AR09A)

Engineering strategies for simultaneous enhancement of C-phycocyanin production and CO2 fixation with Spirulina platensis.

PubMed

Chen, Chun-Yen; Kao, Pei-Chun; Tsai, Chia-Jung; Lee, Duu-Jong; Chang, Jo-Shu

2013-10-01

Spirulina platensis produces nutraceutical product C-phycocyanin (C-PC) and simultaneously mitigates CO2 emissions during its growth. Using a designed flat-type photobioreactor, the S. platensis biomass production was markedly enhanced, leading to a CO2 removal rate and a biomass concentration of 0.23 g/L/d and 2.25 g/L, respectively. The cell growth, CO2 fixation rate and C-PC production of S. platensis were investigated when it was cultivated under different irradiation conditions. As the light intensity increased from 100 to 700 μmol/m(2)/s, the overall biomass productivity, CO2 consumption rate and maximal C-PC productivity increased significantly to 0.74, 1.53 and 0.11 g/L/d, respectively. After determining the suitable light intensity, the nitrogen concentration was also adjusted to further enhance the performance of CO2 fixation and C-PC production. The results show that with an optimal nitrogen concentration of 0.045 M, the CO2 consumption rate and maximal C-PC productivity were further increased to 1.58 and 0.13 g/L/d, respectively. Copyright © 2013 Elsevier Ltd. All rights reserved.

The Influence of the Host Plant Is the Major Ecological Determinant of the Presence of Nitrogen-Fixing Root Nodule Symbiont Cluster II Frankia Species in Soil

PubMed Central

Battenberg, Kai; Wren, Jannah A.; Hillman, Janell; Edwards, Joseph; Huang, Liujing

2016-01-01

ABSTRACT The actinobacterial genus Frankia establishes nitrogen-fixing root nodule symbioses with specific hosts within the nitrogen-fixing plant clade. Of four genetically distinct subgroups of Frankia, cluster I, II, and III strains are capable of forming effective nitrogen-fixing symbiotic associations, while cluster IV strains generally do not. Cluster II Frankia strains have rarely been detected in soil devoid of host plants, unlike cluster I or III strains, suggesting a stronger association with their host. To investigate the degree of host influence, we characterized the cluster II Frankia strain distribution in rhizosphere soil in three locations in northern California. The presence/absence of cluster II Frankia strains at a given site correlated significantly with the presence/absence of host plants on the site, as determined by glutamine synthetase (glnA) gene sequence analysis, and by microbiome analysis (16S rRNA gene) of a subset of host/nonhost rhizosphere soils. However, the distribution of cluster II Frankia strains was not significantly affected by other potential determinants such as host-plant species, geographical location, climate, soil pH, or soil type. Rhizosphere soil microbiome analysis showed that cluster II Frankia strains occupied only a minute fraction of the microbiome even in the host-plant-present site and further revealed no statistically significant difference in the α-diversity or in the microbiome composition between the host-plant-present or -absent sites. Taken together, these data suggest that host plants provide a factor that is specific for cluster II Frankia strains, not a general growth-promoting factor. Further, the factor accumulates or is transported at the site level, i.e., beyond the host rhizosphere. IMPORTANCE Biological nitrogen fixation is a bacterial process that accounts for a major fraction of net new nitrogen input in terrestrial ecosystems. Transfer of fixed nitrogen to plant biomass is especially efficient via root nodule symbioses, which represent evolutionarily and ecologically specialized mutualistic associations. Frankia spp. (Actinobacteria), especially cluster II Frankia spp., have an extremely broad host range, yet comparatively little is known about the soil ecology of these organisms in relation to the host plants and their rhizosphere microbiomes. This study reveals a strong influence of the host plant on soil distribution of cluster II Frankia spp. PMID:27795313

Facets of diazotrophy in the oxygen minimum zone waters off Peru

PubMed Central

Loescher, Carolin R; Großkopf, Tobias; Desai, Falguni D; Gill, Diana; Schunck, Harald; Croot, Peter L; Schlosser, Christian; Neulinger, Sven C; Pinnow, Nicole; Lavik, Gaute; Kuypers, Marcel M M; LaRoche, Julie; Schmitz, Ruth A

2014-01-01

Nitrogen fixation, the biological reduction of dinitrogen gas (N2) to ammonium (NH4+), is quantitatively the most important external source of new nitrogen (N) to the open ocean. Classically, the ecological niche of oceanic N2 fixers (diazotrophs) is ascribed to tropical oligotrophic surface waters, often depleted in fixed N, with a diazotrophic community dominated by cyanobacteria. Although this applies for large areas of the ocean, biogeochemical models and phylogenetic studies suggest that the oceanic diazotrophic niche may be much broader than previously considered, resulting in major implications for the global N-budget. Here, we report on the composition, distribution and abundance of nifH, the functional gene marker for N2 fixation. Our results show the presence of eight clades of diazotrophs in the oxygen minimum zone (OMZ) off Peru. Although proteobacterial clades dominated overall, two clusters affiliated to spirochaeta and archaea were identified. N2 fixation was detected within OMZ waters and was stimulated by the addition of organic carbon sources supporting the view that non-phototrophic diazotrophs were actively fixing dinitrogen. The observed co-occurrence of key functional genes for N2 fixation, nitrification, anammox and denitrification suggests that a close spatial coupling of N-input and N-loss processes exists in the OMZ off Peru. The wide distribution of diazotrophs throughout the water column adds to the emerging view that the habitat of marine diazotrophs can be extended to low oxygen/high nitrate areas. Furthermore, our statistical analysis suggests that NO2− and PO43− are the major factors affecting diazotrophic distribution throughout the OMZ. In view of the predicted increase in ocean deoxygenation resulting from global warming, our findings indicate that the importance of OMZs as niches for N2 fixation may increase in the future. PMID:24813564

Facets of diazotrophy in the oxygen minimum zone waters off Peru.

PubMed

Loescher, Carolin R; Großkopf, Tobias; Desai, Falguni D; Gill, Diana; Schunck, Harald; Croot, Peter L; Schlosser, Christian; Neulinger, Sven C; Pinnow, Nicole; Lavik, Gaute; Kuypers, Marcel M M; LaRoche, Julie; Schmitz, Ruth A

2014-11-01

Nitrogen fixation, the biological reduction of dinitrogen gas (N2) to ammonium (NH4(+)), is quantitatively the most important external source of new nitrogen (N) to the open ocean. Classically, the ecological niche of oceanic N2 fixers (diazotrophs) is ascribed to tropical oligotrophic surface waters, often depleted in fixed N, with a diazotrophic community dominated by cyanobacteria. Although this applies for large areas of the ocean, biogeochemical models and phylogenetic studies suggest that the oceanic diazotrophic niche may be much broader than previously considered, resulting in major implications for the global N-budget. Here, we report on the composition, distribution and abundance of nifH, the functional gene marker for N2 fixation. Our results show the presence of eight clades of diazotrophs in the oxygen minimum zone (OMZ) off Peru. Although proteobacterial clades dominated overall, two clusters affiliated to spirochaeta and archaea were identified. N2 fixation was detected within OMZ waters and was stimulated by the addition of organic carbon sources supporting the view that non-phototrophic diazotrophs were actively fixing dinitrogen. The observed co-occurrence of key functional genes for N2 fixation, nitrification, anammox and denitrification suggests that a close spatial coupling of N-input and N-loss processes exists in the OMZ off Peru. The wide distribution of diazotrophs throughout the water column adds to the emerging view that the habitat of marine diazotrophs can be extended to low oxygen/high nitrate areas. Furthermore, our statistical analysis suggests that NO2(-) and PO4(3-) are the major factors affecting diazotrophic distribution throughout the OMZ. In view of the predicted increase in ocean deoxygenation resulting from global warming, our findings indicate that the importance of OMZs as niches for N2 fixation may increase in the future.

Temporal and spatial variability of biological nitrogen fixation off the upwelling system of central Chile (35-38.5°S)

NASA Astrophysics Data System (ADS)

Fernandez, Camila; González, Maria Lorena; Muñoz, Claudia; Molina, Veronica; Farias, Laura

2015-05-01

Although N2 fixation could represent a supplementary source of bioavailable nitrogen in coastal upwelling areas and underlying oxygen minimum zones (OMZs), the limited data available prevent assessing its variability and biogeochemical significance. Here we report the most extensive N2 fixation data set gathered to date in the upwelling area off central Chile (36°S). It covers interannual to high frequency time scales in an area of about 82,500 km2 in the eastern South Pacific (ESP). Because heterotrophic N2 fixation may be regulated by DOM availability in the ESP, we conducted experiments at different oxygen conditions and included DOM amendments in order to test diazotrophic activity. Rates in the euphotic zone showed strong temporal variability which resulted in values reaching 0.5 nmol L-1 d-1 in 2006 (average 0.32 ± 0.17 nmol L-1 d-1) and up to 126.8 nmol L-1 d-1 (average 24.75 ± 37.9 nmol L-1 d-1) in 2011. N2 fixation in subsurface suboxic conditions (1.5 ± 1.16 nmol L-1 d-1) also occurred mainly during late summer and autumn while virtually absent in winter. The diversity of diazotrophs was dominated by heterotrophs, with higher richness in surface compared to OMZ waters. Rates in oxygen depleted conditions could exceed values obtained in the euphotic layer, but rates were not dependent on the availability of dissolved organic matter. N2 fixation also showed a positive correlation with total chlorophyll and the C:N ratio of phytoplankton, but not to the P excess compared to N. We conclude that the diazotrophic community responds to the composition of phytoplankton rather than the extent of N deficiency and the availability of bulk DOM in this system.

Nitrogen fixation sustained productivity in the wake of the Palaeoproterozoic Great Oxygenation Event.

PubMed

Luo, Genming; Junium, Christopher K; Izon, Gareth; Ono, Shuhei; Beukes, Nicolas J; Algeo, Thomas J; Cui, Ying; Xie, Shucheng; Summons, Roger E

2018-03-07

The marine nitrogen cycle is dominated by redox-controlled biogeochemical processes and, therefore, is likely to have been revolutionised in response to Earth-surface oxygenation. The details, timing, and trajectory of nitrogen cycle evolution, however, remain elusive. Here we couple nitrogen and carbon isotope records from multiple drillcores through the Rooihoogte-Timeball Hill Formations from across the Carletonville area of the Kaapvaal Craton where the Great Oxygenation Event (GOE) and its aftermath are recorded. Our data reveal that aerobic nitrogen cycling, featuring metabolisms involving nitrogen oxyanions, was well established prior to the GOE and that ammonium may have dominated the dissolved nitrogen inventory. Pronounced signals of diazotrophy imply a stepwise evolution, with a temporary intermediate stage where both ammonium and nitrate may have been scarce. We suggest that the emergence of the modern nitrogen cycle, with metabolic processes that approximate their contemporary balance, was retarded by low environmental oxygen availability.

Utilization of ammonium as a nitrogen source: effects of ambient acidity on growth and nitrogen accumulation by soybean

NASA Technical Reports Server (NTRS)

Tolley-Henry, L.; Raper, C. D. Jr; Raper CD, J. r. (Principal Investigator)

1986-01-01

Dry matter accumulation of plants utilizing NH4+ as the sole nitrogen source generally is less than that of plants receiving NO3- unless acidity of the root-zone is controlled at a pH of about 6.0. To test the hypothesis that the reduction in growth is a consequence of nitrogen stress within the plant in response to effects of increased acidity during uptake of NH4+ by roots, nonnodulated soybean plants (Glycine max [L.] Merr. cv Ransom) were grown for 24 days in flowing nutrient culture containing 1.0 millimolar NH4+ as the nitrogen source. Acidities of the culture solutions were controlled at pH 6.1, 5.1, and 4.1 +/- 0.1 by automatic additions of 0.01 N H2SO4 or Ca(OH)2. Plants were sampled at intervals of 3 to 4 days for determination of dry matter and nitrogen accumulation. Rates of NH4+ uptake per gram root dry weight were calculated from these data. Net CO2 exchange rates per unit leaf area were measured on attached leaves by infrared gas analysis. When acidity of the culture solution was increased from pH 6.1 to 5.1, dry matter and nitrogen accumulation were reduced by about 40% within 14 days. Net CO2 exchange rates per unit leaf area, however, were not affected, and the decreased growth was associated with a reduction in rates of appearance and expansion of new leaves. The uptake rates of NH4+ per gram root were about 25% lower throughout the 24 days at pH 5.1 than at 6.1. A further increase in solution acidity from pH 5.1 to 4.1 resulted in cessation of net dry matter production and appearance of new leaves within 10 days. Net CO2 exchange rates per unit leaf area declined rapidly until all viable leaves had abscised by 18 days. Uptake rates of NH4+, which were initially about 50% lower at pH 4.1 than at 6.1 continued to decline with time of exposure until net uptake ceased at 10 days. Since these responses also are characteristic of the sequence of responses that occur during onset and progression of a nitrogen stress, they corroborate our hypothesis.

Diversity of free-Living nitrogen fixing Streptomyces in soils of the badlands of South Dakota.

PubMed

Dahal, Bibha; NandaKafle, Gitanjali; Perkins, Lora; Brözel, Volker S

2017-01-01

Biological Nitrogen Fixation is critical for ecosystem productivity. Select members of Bacteria and Archaea express a nitrogenase enzyme complex that reduces atmospheric nitrogen to ammonia. Several nitrogen fixing bacteria form symbiotic associations with plants, but free-living diazotrophs also contribute a substantial amount of nitrogen to ecosystems. The aim of this study was to isolate and characterize free-living diazotrophs in arid lands of South Dakota Badlands. Samples were obtained from sod tables and the surrounding base in spring and fall. Diazotrophs were isolated on solid nitrogen free medium (NFM) under hypoxic conditions, and their16S rRNA and nifH genes sequenced. nifH was also amplified directly from soil DNA extracts. The 16S rRNA gene data indicated a diversity of putative free-living diazotrophs across 4 phyla (Actinomycetes, Proteobacteria, Bacteroidetes, and Firmicutes), but ∼50% of these clustered with Streptomyces. These Streptomyces isolates grew in liquid NFM in an ammonia-depleted environment. Only 5 of these yielded a nifH gene product using the PolF/PolR primer set. Four of these aligned with nifH of the cyanobacteria Scytonema and Nostoc, and the other one aligned with nifH of Bradyrhizobium. Six selected Streptomyces isolates, three of which were nifH positive by PCR, all indicated 15 N 2 incorporation, providing strong support of nitrogen fixation. All nifH amplicons from soil DNA extract resembled Cyanobacteria. This is the first known report of diazotrophic Streptomyces, other than the thermophilic, autotrophic S. thermoautotrophicus. nifH genes of these Streptomyces were related to those from Cyanobacteria. It is possible that the cyanobacteria-like nifH amplicons obtained from soil DNA were associated with Streptomyces. Copyright © 2016 Elsevier GmbH. All rights reserved.

Sources of organic nitrogen at the serpentinite-hosted Lost City hydrothermal field.

PubMed

Lang, S Q; Früh-Green, G L; Bernasconi, S M; Butterfield, D A

2013-03-01

The reaction of ultramafic rocks with water during serpentinization at moderate temperatures results in alkaline fluids with high concentrations of reduced chemical compounds such as hydrogen and methane. Such environments provide unique habitats for microbial communities capable of utilizing these reduced compounds in present-day and, possibly, early Earth environments. However, these systems present challenges to microbial communities as well, particularly due to high fluid pH and possibly the availability of essential nutrients such as nitrogen. Here we investigate the source and cycling of organic nitrogen at an oceanic serpentinizing environment, the Lost City hydrothermal field (30°N, Mid-Atlantic Ridge). Total hydrolizable amino acid (THAA) concentrations in the fluids range from 736 to 2300 nm and constitute a large fraction of the dissolved organic carbon (2.5-15.1%). The amino acid distributions, and the relative concentrations of these compounds across the hydrothermal field, indicate they most likely derived from chemolithoautotrophic production. Previous studies have identified the presence of numerous nitrogen fixation genes in the fluids and the chimneys. Organic nitrogen in actively venting chimneys has δ(15) N values as low as 0.1‰ which is compatible with biological nitrogen fixation. Total hydrolizable amino acids in the chimneys are enriched in (13) C by 2-7‰ compared to bulk organic matter. The distribution and absolute δ(13) C(THAA) values are compatible with a chemolithoautotrophic source, an attribution also supported by molar organic C/N ratios in most active chimneys (4.1-5.5) which are similar to those expected for microbial communities. In total, these data indicate nitrogen is readily available to microbial communities at Lost City. © 2013 Blackwell Publishing Ltd.

Rubisco activity and regulation as targets for crop improvement

USDA-ARS?s Scientific Manuscript database

Rubisco (ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase) enables net carbon fixation through the carboxylation of RuBP. However, some characteristics of Rubisco make it surprisingly inefficient and compromise photosynthetic productivity. For example, Rubisco catalyses a wasteful reaction wit...

Effect of organic matter supplementation on nitrogen transformations in soils. I. Chemical and bacteriological changes.

PubMed

Abd-el-Malek, Y; Monib, M; Hosny, I; Girgis, S A

1979-01-01

The effect of supplementation with different organic materials on nitrogen transformations and on certain bacterial groups in soil was studied. Addition of wide C/N ratio organic matter, sawdust and maize stalks prevented NO3-N from being lost through leaching out or dentrification and favoured the development of Azotobacter and N2-fixing clostridia that in turn resulted in marked gains in nitrogen through N2-fixation. Nitrifying bacteria were adversely affected. Application of such materials together with high amounts of NH4NO3 lessened nitrogen losses in drainage water but increased losses through denitrification. Nitrogen-rich organic matter resulted in higher losses in nitrates from soils in comparison to those of wide C/N ratio organic materials.

The USA Nr Inventory: Dominant Sources and Primary Transport Pathways

NASA Astrophysics Data System (ADS)

Sabo, R. D.; Clark, C.; Sobota, D. J.; Compton, J.; Cooter, E. J.; Schwede, D. B.; Bash, J. O.; Rea, A.; Dobrowolski, J. P.

2016-12-01

Efforts to mitigate the deleterious effects of excess reactive nitrogen (Nr) on human health and ecosystem goods and service while ensuring food, biofuel, and fiber availability, is one of the most pressing environmental management challenges of this century. Effective management of Nr requires up to date inventories that quantitatively characterize the sources, transport, and transformation of Nr through the environment. The inherent complexity of the nitrogen cycle, however, through multiple exchange points across air, water, and terrestrial media, renders such inventories difficult to compile and manage. Previous Nr Inventories are for 2002 and 2007, and used data sources that have since been improved. Thus, this recent inventory will substantially advance the methodology across many sectors of the inventory (e.g. deposition and biological fixation in crops and natural systems) and create a recent snapshot that is sorely needed for policy planning and trends analysis. Here we use a simple mass balance approach to estimate the input-output budgets for all United States Geologic Survey Hydrologic Unit Code-8 watersheds. We focus on a recent year (i.e. 2012) to update the Nr Inventory, but apply the analytical approach for multiple years where possible to assess trends through time. We also compare various sector estimates using multiple methodologies. Assembling datasets that account for new Nr inputs into watersheds (e.g., atmospheric NOy deposition, food imports, biologic N fixation) and internal fluxes of recycled Nr (e.g., manure, Nr emmissions/volatilization) provide an unprecedented, data driven computation of N flux. Input-output budgets will offer insight into 1) the dominant sources of Nr in a watershed (e.g., food imports, atmospheric N deposition, or fertilizer), 2) the primary loss pathways for Nr (e.g., crop N harvest, volatilization/emissions), and 3) what watersheds are net sources versus sinks of Nr. These insights will provide needed clarity for managers looking to minimize the loss of Nr to atmospheric and aquatic compartments, while also providing a foundational database for researchers assessing the dominant controls of N retention and loss in natural and anthropogenically dominated ecosystems. Disclaimer: Views expressed are the authors' and not views or polices of the U.S.EPA.

Transcriptomic Analysis and Microscopic Observations in the Cyanobacterium UCYN-A during Diel Cycles

NASA Astrophysics Data System (ADS)

Muñoz-Marin, M. D. C.; Farnelid, H.; Zehr, J. P.

2016-02-01

Candidatus Atelocyanobacterium thalassa (UCYN-A) is a nitrogen-fixing marine cyanobacterium recently recognized for its widespread distribution and significant contributions to oceanic nitrogen (N2)-fixation. UCYN-A is a group of related cyanobacteria that are symbiotic with a single-celled eukaryotic phytoplankter, the haptophyte Braarudosphaera bigelowii. UCYN-A fixes N2 and expresses nitrogenase during the day. Since the nitrogenase is inactivated by oxygen evolved through photosynthesis, most cyanobacteria use temporal or spatial separation of photosynthesis and N2 fixation. Genomic studies revealed that UCYN-A lacks the entire PSII apparatus (photosystem II). The lack of oxygenic photosynthesis at least partially explains why they can fix nitrogen during the day, although the host is a photoautotroph. However, UCYN-A has retained photosystem I (PSI), and PSI activity may be important in the energetics of N2 fixation in the symbiosis. Because UCYN-A lacks photosystem II, which normally supplies electrons to photosystem I from water, UCYN-A needs alternative electron donors if it uses photosystem I to make the reductant NADPH. In order to determine if UCYN-A expresses photosynthetic genes and which other proteins may be involved with energy metabolism, we developed a whole genome array to examine gene transcription over the diel cycle in two strains. Our results show that there is a temporal separation of the expression of photosynthesis genes from the expression of nitrogenase genes. Moreover, the transcription profile of NADH dehydrogenases and hydrogenases suggest they may be involved as alternative electron donors for the N2 fixation. In addition, we used a double-CARD-FISH (Catalyzed Reporter Deposition-Fluorescence in situ Hybridization) assay to study cell division of the host and symbiont during diel cycles in relation to UCYN-A gene expression carried out during the transcriptomic analysis. These results help us move toward a deeper understanding of the metabolism of this unusual cyanobacterium and the differences in environmental adaptations between these two strains.

Evidence for high-temperature in situ nifH transcription in an alkaline hot spring of Lower Geyser Basin, Yellowstone National Park.

PubMed

Loiacono, Sara T; Meyer-Dombard, D'Arcy R; Havig, Jeff R; Poret-Peterson, Amisha T; Hartnett, Hilairy E; Shock, Everett L

2012-05-01

Genes encoding nitrogenase (nifH) were amplified from sediment and photosynthetic mat samples collected in the outflow channel of Mound Spring, an alkaline thermal feature in Yellowstone National Park. Results indicate the genetic capacity for nitrogen fixation over the entire range of temperatures sampled (57.2°C to 80.2°C). Amplification of environmental nifH transcripts revealed in situ expression of nifH genes at temperatures up to 72.7°C. However, we were unable to amplify transcripts of nifH at the higher-temperature locations (> 72.7°C). These results indicate that microbes at the highest temperature sites contain the genetic capacity to fix nitrogen, yet either do not express nifH or do so only transiently. Field measurements of nitrate and ammonium show fixed nitrogen limitation as temperature decreases along the outflow channel, suggesting nifH expression in response to the downstream decrease in bioavailable nitrogen. Nitrogen stable isotope values of Mound Spring sediment communities further support geochemical and genetic data. DNA and cDNA nifH amplicons form several unique phylogenetic clades, some of which appear to represent novel nifH sequences in both photosynthetic and chemosynthetic microbial communities. This is the first report of in situ nifH expression in strictly chemosynthetic zones of terrestrial (non-marine) hydrothermal systems, and sets a new upper temperature limit (72.7°C) for nitrogen fixation in alkaline, terrestrial hydrothermal environments. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

Deepwater Nitrogen Fixation: Who's Doing it, Where, and Why?

NASA Astrophysics Data System (ADS)

Montoya, J. P.; Weber, S.; Vogts, A.; Voss, M.; Saxton, M.; Joye, S. B.

2016-02-01

Nitrogen availability frequently limits marine primary production and N2-fixation plays an important role in supporting biological production in surface waters of many oligotrophic regions. Although subsurface waters typically contain high concentrations of nitrate and other nutrients, measurements from a variety of oceanic settings show measurable, and at times high rates of N2-fixation in deep, dark waters below the mixed layer. We have explored the distribution of N2-fixation throughout the water column of the Gulf of Mexico (GoM) during a series of cruises beginning shortly after the Deepwater Horizon (DWH) spill in 2010 and continuing at roughly annual intervals. These cruises allowed us to sample oligotrophic waters across a range of depths, and to explore the connections between the C and N cycles mediated by release of oil and gas (petrocarbon) from natural seeps as well as anthropogenic sources (e.g., the DWH). We used stable isotope abundances (15N and 13C) in particles and zooplankton in combination with experimental measurements of N2-fixation and CH4 assimilation to assess the contribution of oil- and gas-derived C to the pelagic food web, and the impact of CH4 releases on the pelagic C and N cycles. Our isotopic measurements document the movement of petrocarbon into the pelagic food web, and our experiments revealed that high rates of N2-fixation were widespread in deep water immediately after the DWH incident, and restricted to the vicinity of natural seeps in subsequent years. Unfortunately, these approaches provided no insight into the organisms actually responsible for N2-fixation and CH4-assimilation. We used nano-scale Secondary Ion Mass Spectrometry (nanoSIMS) to image the organisms responsible for these processes, and molecular approaches to explore the diversity of methanotrophs and diazotrophs present in the system. The ability to resolve isotopic distributions on the scale of individual cells is a critical part of bridging the gap between molecular approaches that identify organisms, and biogeochemical techniques that allow us to measure the activity of communities.

In situ Expression of Functional Genes Reveals Nitrogen Cycling at High Temperatures in Terrestrial Hydrothermal Systems

NASA Astrophysics Data System (ADS)

Loiacono, S. T.; Meyer-Dombard, D. R.

2011-12-01

An essential element for life, nitrogen occurs in all living organisms and is critical for the synthesis of amino acids, proteins, nucleic acids, and other forms of biomass. Thus, nitrogen cycling likely plays a vital role in microbial metabolic processes as well as nutrient availability. For microorganisms in "extreme" environments, this means developing adaptations that allow them to survive in harsh conditions and still perform the metabolisms essential to sustain life. Recent studies have screened biofilms and thermal sediments of Yellowstone National Park (YNP) thermal features for the presence of nifH genes, which code for a key enzyme in the nitrogen fixation process [1-4]. Furthermore, analysis of nitrogen isotopes in biofilms across a temperature and chemical gradient revealed that nitrogen fixation likely varies across the chemosynthetic/photosynthetic ecotone [5]. Although research has evaluated and confirmed the presence of nifH genes in various thermophilic microbial communities, the existence of a gene in the DNA of an organism does not verify its use. Instead, other methods, such as culturing, isotope tracer assays, and gene expression studies are required to provide direct evidence of biological nitrogen fixation. Culturing and isotope tracer approaches have successfully revealed high-temperature biological nitrogen fixation in both marine hydrothermal vent microbial communities [6] and in acidic, terrestrial hydrothermal sediment [3]. Transcriptomics-based techniques (using mRNA extracted from samples to confirm in situ expression of targeted genes) have been much more limited in number, and only a few studies have, to date, investigated in situ expression of the nifH gene in thermophilic microbial communities [2, 7]. This study explores the presence and expression of nifH genes in several features of the Lower Geyser Basin (LGB) of YNP. Nucleic acids from chemosynthetic and photosynthetic microbial communities were extracted and then amplified using (reverse-transcription) polymerase chain reaction to identify the presence and expression of nifH genes, and resultant (RT-)PCR product was cloned and sequenced. Results reveal high-temperature in situ expression of nifH in select LGB features [7] which is, to the authors' knowledge, the first direct evidence of nifH transcription in the chemosynthetic zones of terrestrial hydrothermal systems. Results also indicate the presence of novel nifH sequences and allow phylogenetic comparison of nifH genes along geochemical gradients within individual hot spring features and between various thermal features in the LGB. Collectively, these results provide evidence for microbial adaptations that have led to the ability to support basic metabolic processes under "extreme" conditions. [1] Hall et al., 2008. AEM 74: 4910-4922. [2] Steunou et al., 2008. The ISME Journal 2: 364-378. [3] Hamilton et al., 2011. Microb Ecol DOI 10.1007/s00248-011-9824-9. [4] Raymond et al., 2008. EOS Trans AGU. Abstract B14A-03. [5] Havig et al., 2010. J Geophys Res-Biogeo 116: G01005. [6] Mehta & Baross, 2006. Science 314: 1783-1786. [7] Loiacono et al., 2011. Submitted FEMS Microbiol Ecol.

The Paradox of Excess Nitrogen in Boreal Peatlands: Biogeochemical Gaps in Nitrogen Cycling Revealed

NASA Astrophysics Data System (ADS)

Vile, M. A.; Prsa, T.; Wieder, R.; Lamers, L. P.

2011-12-01

Globally, peatlands cover 3-4 % of the Earth's land surface (over 4 million km 2, yet they store 25-30 % of the world's soil carbon (C) and 9-16% of the world's soil nitrogen (N, 8-15 Pg) in peat. As in other terrestrial ecosystems, the cycling of C and N is closely linked, especially for ombrotrophic bogs. Bogs receive nutrient and water exclusively from the atmosphere, which ensures an N-limited, nutrient-poor habitat. In Alberta, NW Canada, peatlands have received exceptionally low atmospheric inputs of N (< 1 7 kg/ha/yr) from their first introduction on the landscape ~ 7000 yrs bp, up to the present time. Paradoxically, despite these low inputs of atmospheric N deposition, bases on 210-fixation Pb dating of peat cores, we have shown that over the past 50 years these bogs have accumulated approximately 11-21 times more N in peat than can be explained by inputs of atmospheric N. A likely missing input is N2-fixation from cyanobacteria associated with Sphagnum mosses, however this process has been largely overlooked in boreal peatlands. Here we demonstrate the importance of N2-fixation in explaining the high accumulation rates of N found in unpolluted, boreal bogs of western Canada. Calibrated (using theoretical ratio of 1.5-3:1) rates of N2-fixation for 4 bogs in northern Alberta ranged from 1.6 to 8.0 ± 0.7 kg/ha/yr, indicating that 42-58 % of the N accumulated over in peat, can be attributed to biological N2-fixation. Although most of northern Alberta's peatlands continue to receive exceptionally low atmospheric N deposition rates, over the last 3 decades, rapid development and industrial expansion of Alberta's Oil Sands Mining (OSM) potentially threaten the pristine nature of peatlands through regionally elevated deposition of N-compounds (NOx). Prior to OSM, N inputs to bogs were limited exclusively to (1) biological N fixation, and (2) bulk background deposition. We examined the response of peatlands located in the OSM area to enhanced N deposition. Despite the large accumulation rates of N in peat, mean N:P ratios in Sphagnum moss capitula (11.0 ± 3.4; mean ± stdev) suggest that peat of boreal western Canada is still severely N limited and not limited by phosphorus. Collectively, these data underscore the severity of N-limitation in pristine bogs and their potential sensitivity to increased N inputs from oils sands mining. Additionally, because the majority of the data generated for N stress in peatlands is from eastern Canada and western Europe, we stress the need to encompass the response of bogs to N deposition within the bounds of the low N deposition gradient. We postulate the loss of symbiosis between Sphagnum and N-fixing microorganisms (cyanobacteria, bacteria) in nitrogen-polluted areas, and indicate its consequences at the species level (trade-off) and ecosystem level (including C sequestration).

Molecular Determinants of a Symbiotic Chronic Infection

PubMed Central

Gibson, Katherine E.; Kobayashi, Hajime

2009-01-01

Rhizobial bacteria colonize legume roots for the purpose of biological nitrogen fixation. A complex series of events, coordinated by host and bacterial signal molecules, underlie the development of this symbiotic interaction. Rhizobia elicit de novo formation of a novel root organ within which they establish a chronic intracellular infection. Legumes permit rhizobia to invade these root tissues while exerting control over the infection process. Once rhizobia gain intracellular access to their host, legumes also strongly influence the process of bacterial differentiation that is required for nitrogen fixation. Even so, symbiotic rhizobia play an active role in promoting their goal of host invasion and chronic persistence by producing a variety of signal molecules that elicit changes in host gene expression. In particular, rhizobia appear to advocate for their access to the host by producing a variety of signal molecules capable of suppressing a general pathogen defense response. PMID:18983260

Temperature and Nutrients Interact to Control Nitrogen Fixation in a Subalpine Stream: An Experimental Examination

NASA Astrophysics Data System (ADS)

Marcarelli, A. M.

2005-05-01

To test the importance of factors controlling N-fixation in subalpine streams, I conducted a stream-side mesocosm experiment with epilithic communities and nutrient diffusing substrates (NDS) to test how temperature and nutrients interact to influence algal communities. Within two days, warm temperature (18°C) stimulated N-fixation by Calothrix in the epilithic community 2X above cold temperature (13°C), indicating a strong physiological response. Community responses measured on NDS indicated that cold-water diatoms dominated by day 45 in the cold treatment, while diatoms containing N-fixing endosymbionts dominated only in warm treatments with added phosphorus. There was a significant interaction between nutrient supply and temperature on N-fixation rates in the experiment. On nutrient controls, warm temperature boosted fixation 2X above cold temperature, but when P was added, temperature increased fixation 20X. This study indicates that N-fixation is stimulated both by temperature and nutrients in this stream, but the magnitude of response to phosphorus was much greater than to temperature. Furthermore, our results support the hypothesis that biological characteristics in streams, including community structure and biogeochemical processes, can be altered in complex ways by disturbances like grazing and logging that alter multiple controlling factors simultaneously.

Dinitrogen and Cyanide Fixation by Methane Seep Microorganisms Revealed by FISH- SIMS And Implications for AOM Productivity and Nitrogenase Evolution

NASA Astrophysics Data System (ADS)

Dekas, A.; Orphan, V.

2008-12-01

The anaerobic oxidation of methane (AOM), mediated by methane oxidizing archaea (ANME) and sulfate reducing bacterial symbionts (SRB), minimizes the flux of methane from marine sediment to the overlying water column. Understanding the factors determining AOM productivity, and particularly the rates of methane catabolism and anabolism, is of interest to both modern and ancient investigations of climate and bulk carbon isotopic change. It has been hypothesized that nitrogen availability in methane seeps is temporally variable, and that the seep biomass may be at least partially nitrogen limited. The recent finding of nif genes, those necessary for the production of nitrogenase, in enrichments of ANME and SRB consortia suggested that the organisms mediating AOM have the potential to fix dinitrogen. In the present study we incubated methane seep sediment with nitrogen-deplete artificial marine media and a headspace of methane (CH4) and either 15N-labeled dinitrogen (15N2), cyanide (C15N-), or ammonia (15NH3) in order to (1) test the ability of these currently unculturable microorganisms to fix nitrogen and other triple bonded substrates, (2) investigate which AOM partner was responsible for the fixation, (3) compare growth rates on different nitrogen sources, and (4) characterize the phylogeny of these methane seep-associated nitrogenases. Fluorescence in situ hybridization coupled to nano-scale Secondary Ion Mass Spectroscopy imaging (FISH-SIMS) revealed incorporation of 15N into ANME and SRB biomass of up to 0.06 15N fractional abundance in the 15N2 incubation, and up to 0.02 in the C15N- incubation, after 6 and 4 months, respectively. This represents a nearly ten-fold enrichment of 15N compared to the measured natural 15N fractional abundance (0.0036). The NanoSIMS ion images of ANME/SRB aggregates from 15N2 incubations show evidence for 15N enrichment in both partners with the highest incorporation of 15N within the methanotrophic ANME cells. Cyanide incubations revealed a more heterogeneous pattern of 15N distribution, with localized zones of enrichment within both the SRB and the ANME biomass. From these findings, two alternative explanations are considered: (1) both partners are capable of nitrogenase production, but express the nif genes under different conditions, and (2) the distribution of fixed nitrogen within the ANME and SRB is driven by intimate metabolic coupling and resource sharing, with only one partner serving as the primary diazotroph. In incubations with 15NH3, the AOM biomass 15N fractional abundance was nearly 1.0 after 6 months, demonstrating a much faster growth rate when NH3 rather than N2 or CN- is the nitrogen source, consistent with what is observed in other diazotrophic organisms. Nitrogenase genes recovered from these incubations primarily were associated with the nifH group III clade, but the majority were diverged from known nifH sequences. This suggests that novel nitrogenases are responsible for the N2 fixation observed, and the poor substrate specificity and the potential use as a CN- detoxification mechanism could imply that they are similar to the first nitrogenases. The finding that nitrogen fixation occurs within these potentially ancient organisms therefore may provide a window for examining the history and functional diversity of nitrogenase, and the variable growth rates depending on nitrogen substrate could have implications for AOM productivity through time.

Synthesis of hydrogen cyanide under simulated hydrothermal conditions

NASA Astrophysics Data System (ADS)

Pinedo-González, Paulina

Nitrogen is a fundamental element for life, where is present in structural (e.g., proteins), catalytic (e.g., enzymes and ribozymes), energy transfer (e.g., ATP) and information storage (RNA and DNA) biomolecules. Atmospheric and planetary models suggest that nitrogen was abundant in the early atmospheres of Earth as dinitrogen (N2 ), an inert gas under normal atmospheric conditions. To be available for prebiotic synthesis it must be converted into hydrogen cyanide (HCN), ammonia (NH3 ) and/or nitric oxide (NO), in a process referred to as nitrogen fixation. Due to the strength of the triple bond in N2 , nitrogen fixation, while thermodynamically favored is kinetically restricted. In a reducing atmosphere dominated by CH4 -N2 , thunderstorm lightning efficiently produces HCN and NH3 (Stribling and Miller, 1987). Nevertheless, photochemical and geochemical constraints strongly suggest that the early atmosphere was weakly reducing, dominated by CO2 and N2 with traces of CH4 , CO, and H2 (Kasting, 1993). Under these conditions, HCN is no longer synthesized in the lightning channel and instead NO is formed (Navarro-Gonźlez, et al., 2001). In volcanic plumes, where magmatic gases a were more reducing than in the atmosphere, NO can also be formed by the lava heat (Mather et al., 2004) or volcanic lightning (Navarro-Gonźlez et al., 1998). Surprisingly, dinitrogen can be a reduced to NH3 in hydrothermal systems (Brandes et al., 1998), but the formation of HCN and its derivates were not investigated. The present work explores the possibility of the formation of HCN as well as other nitrile derivatives catalyzed by mineral surfaces in hydrothermal vents. To simulate a hydrothermal atmosphere, the experiments were carried out in a stainless steel Parr R minireactor with a 0.1 M NH4 HCO3 solution (200 ml) with or without a mineral surface exposed at 1 bar at temperatures ranging from 100 to 375° C. Different mineral matrices are been investigated. Our preliminary results have been conducted with pyrite and quantified by headspace-gas chromatography-mass spectrometry. These results indicate that catalysis induced by the mineral surface under hydrothermal conditions does not stop with the production of HCN, but the reaction continues leading to more complex nitriles. The experiments also reveal a clear trend between time and the production of more complex molecules, which are measurable by the chromatographic method. Brandes, J.A., Boctor, N.Z., Cody, G.D., Cooper, B. A., Hazen, R. M. and Yoder Jr, H.S. (1998). Abiotic nitrogen reduction on the early Earth. Nature 395, 365-367. Kasting J.F. (1993) Earth's early atmosphere. Science 259, 920-926. Mather, T.A., Pyle, D.M., and Allen, A.G. (2004) Volcanic source of fixed nitrogen in the early Earth's atmosphere. Geology 32, 905-908. Navarro-Gonźlez, R., Molina, M.J. and. Molina, L.T. (1998) Nitrogen fixation by volcanic a lightning in the early Earth. Geophys. Res. Lett. 25, 3123-3126. Navarro-Gonźlez, R., McKay, C.P. and Nna Mvondo, D. ( 2001) A possible nitrogen crisis for a Archean life due to reduced nitrogen fixation by lightning. Nature 412, 61-64. Stribling, R., and Miller, S.L. (1987) Energy yields for the hydrogen cyanide and formaldehyde synthesis: the HCN and amino acid concentrations in the primitive ocean. Origins Life 17, 261-273.

The novel regulatory ncRNA, NfiS, optimizes nitrogen fixation via base pairing with the nitrogenase gene nifK mRNA in Pseudomonas stutzeri A1501

PubMed Central

Zhan, Yuhua; Yan, Yongliang; Deng, Zhiping; Chen, Ming; Lu, Wei; Lu, Chao; Shang, Liguo; Yang, Zhimin; Zhang, Wei; Wang, Wei; Li, Yun; Ke, Qi; Lu, Jiasi; Xu, Yuquan; Zhang, Liwen; Xie, Zhihong; Cheng, Qi; Elmerich, Claudine; Lin, Min

2016-01-01

Unlike most Pseudomonas, the root-associated bacterium Pseudomonas stutzeri A1501 fixes nitrogen after the horizontal acquisition of a nitrogen-fixing (nif) island. A genome-wide search for small noncoding RNAs (ncRNAs) in P. stutzeri A1501 identified the novel P. stutzeri-specific ncRNA NfiS in the core genome, whose synthesis was significantly induced under nitrogen fixation or sorbitol stress conditions. The expression of NfiS was RNA chaperone Hfq-dependent and activated by the sigma factor RpoN/global nitrogen activator NtrC/nif-specific activator NifA regulatory cascade. The nfiS-deficient mutant displayed reduced nitrogenase activity, as well as increased sensitivity to multiple stresses, such as osmotic and oxidative stresses. Secondary structure prediction and complementation studies confirmed that a stem-loop structure was essential for NfiS to regulate the nitrogenase gene nifK mRNA synthesis and thus nitrogenase activity. Microscale thermophoresis and physiological analysis showed that NfiS directly pairs with nifK mRNA and ultimately enhances nitrogenase activity by increasing the translation efficiency and the half-life of nifK mRNA. Our data also suggest structural and functional divergence of NfiS evolution in diazotrophic and nondiazotrophic backgrounds. It is proposed that NfiS was recruited by nifK mRNA as a novel regulator to integrate the horizontally acquired nif island into host global networks. PMID:27407147

MtMOT1.2 is responsible for molybdate supply to Medicago truncatula nodules.

PubMed

Gil-Díez, Patricia; Tejada-Jiménez, Manuel; León-Mediavilla, Javier; Wen, Jiangqi; Mysore, Kirankumar S; Imperial, Juan; González-Guerrero, Manuel

2018-06-25

Symbiotic nitrogen fixation in legume root nodules requires a steady supply of molybdenum for synthesis of the iron-molybdenum cofactor of nitrogenase. This nutrient has to be provided by the host plant from the soil, crossing several symplastically disconnected compartments through molybdate transporters, including members of the MOT1 family. MtMOT1.2 is a Medicago truncatula MOT1 family member located in the endodermal cells in roots and nodules. Immunolocalization of a tagged MtMOT1.2 indicates that it is associated to the plasma membrane and to intracellular membrane systems, where it would be transporting molybdate towards the cytosol, as indicated in yeast transport assays. Loss-of-function mot1.2-1 mutant showed reduced growth compared to wild-type plants when nitrogen fixation was required, but not when nitrogen was provided as nitrate. While no effect on molybdenum-dependent nitrate reductase activity was observed, nitrogenase activity was severely affected, explaining the observed difference of growth depending on nitrogen source. This phenotype was the result of molybdate not reaching the nitrogen-fixing nodules, since genetic complementation with a wild-type MtMOT1.2 gene or molybdate-fortification of the nutrient solution, both restored wild-type levels of growth and nitrogenase activity. These results support a model in which MtMOT1.2 would mediate molybdate delivery by the vasculature into the nodules. This article is protected by copyright. All rights reserved.

The Contamination of Commercial 15N2 Gas Stocks with 15N–Labeled Nitrate and Ammonium and Consequences for Nitrogen Fixation Measurements

PubMed Central

Dabundo, Richard; Lehmann, Moritz F.; Treibergs, Lija; Tobias, Craig R.; Altabet, Mark A.; Moisander, Pia H.; Granger, Julie

2014-01-01

We report on the contamination of commercial 15-nitrogen (15N) N2 gas stocks with 15N-enriched ammonium, nitrate and/or nitrite, and nitrous oxide. 15N2 gas is used to estimate N2 fixation rates from incubations of environmental samples by monitoring the incorporation of isotopically labeled 15N2 into organic matter. However, the microbial assimilation of bioavailable 15N-labeled N2 gas contaminants, nitrate, nitrite, and ammonium, is liable to lead to the inflation or false detection of N2 fixation rates. 15N2 gas procured from three major suppliers was analyzed for the presence of these 15N-contaminants. Substantial concentrations of 15N-contaminants were detected in four Sigma-Aldrich 15N2 lecture bottles from two discrete batch syntheses. Per mole of 15N2 gas, 34 to 1900 µmoles of 15N-ammonium, 1.8 to 420 µmoles of 15N-nitrate/nitrite, and ≥21 µmoles of 15N-nitrous oxide were detected. One 15N2 lecture bottle from Campro Scientific contained ≥11 µmoles of 15N-nitrous oxide per mole of 15N2 gas, and no detected 15N-nitrate/nitrite at the given experimental 15N2 tracer dilutions. Two Cambridge Isotopes lecture bottles from discrete batch syntheses contained ≥0.81 µmoles 15N-nitrous oxide per mole 15N2, and trace concentrations of 15N-ammonium and 15N-nitrate/nitrite. 15N2 gas equilibrated cultures of the green algae Dunaliella tertiolecta confirmed that the 15N-contaminants are assimilable. A finite-differencing model parameterized using oceanic field conditions typical of N2 fixation assays suggests that the degree of detected 15N-ammonium contamination could yield inferred N2 fixation rates ranging from undetectable, <0.01 nmoles N L−1 d−1, to 530 nmoles N L−1 d−1, contingent on experimental conditions. These rates are comparable to, or greater than, N2 fixation rates commonly detected in field assays. These results indicate that past reports of N2 fixation should be interpreted with caution, and demonstrate that the purity of commercial 15N2 gas must be ensured prior to use in future N2 fixation rate determinations. PMID:25329300

The contamination of commercial 15N2 gas stocks with 15N-labeled nitrate and ammonium and consequences for nitrogen fixation measurements.

PubMed

Dabundo, Richard; Lehmann, Moritz F; Treibergs, Lija; Tobias, Craig R; Altabet, Mark A; Moisander, Pia H; Granger, Julie

2014-01-01

We report on the contamination of commercial 15-nitrogen (15N) N2 gas stocks with 15N-enriched ammonium, nitrate and/or nitrite, and nitrous oxide. 15N2 gas is used to estimate N2 fixation rates from incubations of environmental samples by monitoring the incorporation of isotopically labeled 15N2 into organic matter. However, the microbial assimilation of bioavailable 15N-labeled N2 gas contaminants, nitrate, nitrite, and ammonium, is liable to lead to the inflation or false detection of N2 fixation rates. 15N2 gas procured from three major suppliers was analyzed for the presence of these 15N-contaminants. Substantial concentrations of 15N-contaminants were detected in four Sigma-Aldrich 15N2 lecture bottles from two discrete batch syntheses. Per mole of 15N2 gas, 34 to 1900 µmoles of 15N-ammonium, 1.8 to 420 µmoles of 15N-nitrate/nitrite, and ≥21 µmoles of 15N-nitrous oxide were detected. One 15N2 lecture bottle from Campro Scientific contained ≥11 µmoles of 15N-nitrous oxide per mole of 15N2 gas, and no detected 15N-nitrate/nitrite at the given experimental 15N2 tracer dilutions. Two Cambridge Isotopes lecture bottles from discrete batch syntheses contained ≥0.81 µmoles 15N-nitrous oxide per mole 15N2, and trace concentrations of 15N-ammonium and 15N-nitrate/nitrite. 15N2 gas equilibrated cultures of the green algae Dunaliella tertiolecta confirmed that the 15N-contaminants are assimilable. A finite-differencing model parameterized using oceanic field conditions typical of N2 fixation assays suggests that the degree of detected 15N-ammonium contamination could yield inferred N2 fixation rates ranging from undetectable, <0.01 nmoles N L(-1) d(-1), to 530 nmoles N L(-1) d(-1), contingent on experimental conditions. These rates are comparable to, or greater than, N2 fixation rates commonly detected in field assays. These results indicate that past reports of N2 fixation should be interpreted with caution, and demonstrate that the purity of commercial 15N2 gas must be ensured prior to use in future N2 fixation rate determinations.

Utilization of the terrestrial cyanobacterial sheet

NASA Astrophysics Data System (ADS)

Katoh, Hiroshi; Tomita-Yokotani, Kaori; Furukawa, Jun; Kimura, Shunta; Yamaguchi, Yuji; Takenaka, Hiroyuki; Kohno, Nobuyuki

2016-07-01

The terrestrial nitrogen-fixing cyanobacterium, Nostoc commune, is living ranging from polar to desert. N. commune makes visible colonies composed extracellular polymeric substances. N. commune has expected to utilize for agriculture, food and terraforming cause of its extracellular polysaccharide, desiccation tolerance and nitrogen fixation. To exhibit the potential abilities, the N. commune sheet is made to use convenient and evaluated by plant growth and radioactive accumulation. We will discuss utilization of terrestrial cyanobacteria under closed environment.

Relation ofSpodoptera eridania choice to tannins and protein oflotus corniculatus.

PubMed

Briggs, M A

1990-05-01

Plant secondary compounds such as tannins may influence herbivore choice. To determine if herbivory was influenced by tannin concentration,Spodoptera eridania larvae were given a choice ofLotus corniculatus plants whose chemical profiles were altered by fertilization. Herbivores chose plants that had been grown with symbiotic nitrogen fixation as their only nitrogen source more often than fertilized plants. Choice was related to protein concentration, but not to tannin concentration.

Iron depletion affects nitrogenase activity and expression of nifH and nifA genes in Herbaspirillum seropedicae.

PubMed

Rosconi, Federico; Souza, Emanuel M; Pedrosa, Fabio O; Platero, Raúl A; González, Cecilia; González, Marcela; Batista, Silvia; Gill, Paul R; Fabiano, Elena R

2006-05-01

Herbaspirillum seropedicae Z67 is a nitrogen-fixing bacterium able to colonize the rhizosphere and the interior of several plants. As iron is a key element for nitrogen fixation, we examined the response of this microorganism to iron deficiency under nitrogen fixing conditions. We identified a H. seropedicae exbD gene that was induced in response to iron limitation and is involved in iron homeostasis. We found that an exbD mutant grown in iron-chelated medium is unable to fix nitrogen. Moreover, we provide evidence that expression of the nifH and nifA genes is iron dependent in a H. seropedicae genetic background.

EnviroAtlas - Cultivated biological nitrogen fixation in agricultural lands by 12-digit HUC in the Conterminous United States, 2006

EPA Pesticide Factsheets

This EnviroAtlas dataset contains data on the mean cultivated biological nitrogen fixation (C-BNF) in cultivated crop and hay/pasture lands per 12-digit Hydrologic Unit (HUC) in 2006. Nitrogen (N) inputs from the cultivation of legumes, which possess a symbiotic relationship with N-fixing bacteria, were calculated with a recently developed model relating county-level yields of various leguminous crops with BNF rates. We accessed county-level data on annual crop yields for soybeans (Glycine max L.), alfalfa (Medicago sativa L.), peanuts (Arachis hypogaea L.), various dry beans (Phaseolus, Cicer, and Lens spp.), and dry peas (Pisum spp.) for 2006 from the USDA Census of Agriculture (http://www.agcensus.usda.gov/index.php). We estimated the yield of the non-alfalfa leguminous component of hay as 32% of the yield of total non-alfalfa hay (http://www.agcensus.usda.gov/index.php). Annual rates of C-BNF by crop type were calculated using a model that relates yield to C-BNF. We assume yield data reflect differences in soil properties, water availability, temperature, and other local and regional factors that can influence root nodulation and rate of N fixation. We distributed county-specific, C-BNF rates to cultivated crop and hay/pasture lands delineated in the 2006 National Land Cover Database (30 x 30 m pixels) within the corresponding county. C-BNF data described here represent an average input to a typical agricultural land type within a county, i.e., they are not

Regional constraints to biological nitrogen fixation in post-fire forest communities

USGS Publications Warehouse

Yelenik, Stephanie; Perakis, Steven S.; Hibbs, David

2013-01-01

Biological nitrogen fixation (BNF) is a key ecological process that can restore nitrogen (N) lost in wildfire and shape the pace and pattern of post-fire forest recovery. To date, there is limited information on how climate and soil fertility interact to influence different pathways of BNF in early forest succession. We studied asymbiotic (forest floor and soil) and symbiotic (the shrub Ceanothus integerrimus) BNF rates across six sites in the Klamath National Forest, California, USA. We used combined gradient and experimental phosphorus (P) fertilization studies to explore cross-site variation in BNF rates and then related these rates to abiotic and biotic variables. We estimate that our measured BNF rates 22 years after wildfire (6.1–12.1 kg N·ha-1·yr-1) are unlikely to fully replace wildfire N losses. We found that asymbiotic BNF is P limited, although this is not the case for symbiotic BNF in Ceanothus. In contrast, Ceanothus BNF is largely driven by competition from other vegetation: in high-productivity sites with high potential evapotranspiration (Et), shrub biomass is suppressed as tree biomass increases. Because shrub biomass governed cross-site variation in Ceanothus BNF, this competitive interaction led to lower BNF in sites with high productivity and Et. Overall, these results suggest that the effects of nutrients play a larger role in driving asymbiotic than symbiotic fixation across our post-fire sites. However, because symbiotic BNF is 8–90x greater than asymbiotic BNF, it is interspecific plant competition that governs overall BNF inputs in these forests.

Biology Notes.

ERIC Educational Resources Information Center

School Science Review, 1982

1982-01-01

Presents procedures, exercises, demonstrations, and information on a variety of biology topics including labeling systems, biological indicators of stream pollution, growth of lichens, reproductive capacity of bulbous buttercups, a straw balance to measure transpiration, interaction of fungi, osmosis, and nitrogen fixation and crop production. (DC)

The NifA-RpoN Regulon of Mesorhizobium loti Strain R7A and Its Symbiotic Activation by a Novel LacI/GalR-Family Regulator

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sullivan, John T.; Brown, Steven D.; Ronson, Clive W.

Mesorhizobium loti is the microsymbiont of Lotus species, including the model legume L. japonicus. M. loti differs from other rhizobia in that it contains two copies of the key nitrogen fixation regulatory gene nifA, nifA1 and nifA2, both of which are located on the symbiosis island ICEMlSymR7A. M. loti R7A also contains two rpoN genes, rpoN1 located on the chromosome outside of ICEMlSymR7A and rpoN2 that is located on ICEMlSymR7A. The aims of the current work were to establish how nifA expression was activated in M. loti and to characterise the NifA-RpoN regulon. The nifA2 and rpoN2 genes were essentialmore » for nitrogen fixation whereas nifA1 and rpoN1 were dispensable. Expression of nifA2 was activated, possibly in response to an inositol derivative, by a novel regulator of the LacI/GalR family encoded by the fixV gene located upstream of nifA2. Other than the well-characterized nif/fix genes, most NifA2-regulated genes were not required for nitrogen fixation although they were strongly expressed in nodules. The NifA-regulated nifZ and fixU genes, along with nifQ which was not NifA-regulated, were required in M. loti for a fully effective symbiosis although they are not present in some other rhizobia. The NifA-regulated gene msi158 that encodes a porin was also required for a fully effective symbiosis. Several metabolic genes that lacked NifA-regulated promoters were strongly expressed in nodules in a NifA2-dependent manner but again mutants did not have an overt symbiotic phenotype. In summary, many genes encoded on ICEMlSymR7A were strongly expressed in nodules but not free-living rhizobia, but were not essential for symbiotic nitrogen fixation. It seems likely that some of these genes have functional homologues elsewhere in the genome and that bacteroid metabolism may be sufficiently plastic to adapt to loss of certain enzymatic functions.« less

The NifA-RpoN Regulon of Mesorhizobium loti Strain R7A and Its Symbiotic Activation by a Novel LacI/GalR-Family Regulator

PubMed Central

Sullivan, John T.; Brown, Steven D.; Ronson, Clive W.

2013-01-01

Mesorhizobium loti is the microsymbiont of Lotus species, including the model legume L. japonicus. M. loti differs from other rhizobia in that it contains two copies of the key nitrogen fixation regulatory gene nifA, nifA1 and nifA2, both of which are located on the symbiosis island ICEMlSymR7A. M. loti R7A also contains two rpoN genes, rpoN1 located on the chromosome outside of ICEMlSymR7A and rpoN2 that is located on ICEMlSymR7A. The aims of the current work were to establish how nifA expression was activated in M. loti and to characterise the NifA-RpoN regulon. The nifA2 and rpoN2 genes were essential for nitrogen fixation whereas nifA1 and rpoN1 were dispensable. Expression of nifA2 was activated, possibly in response to an inositol derivative, by a novel regulator of the LacI/GalR family encoded by the fixV gene located upstream of nifA2. Other than the well-characterized nif/fix genes, most NifA2-regulated genes were not required for nitrogen fixation although they were strongly expressed in nodules. The NifA-regulated nifZ and fixU genes, along with nifQ which was not NifA-regulated, were required in M. loti for a fully effective symbiosis although they are not present in some other rhizobia. The NifA-regulated gene msi158 that encodes a porin was also required for a fully effective symbiosis. Several metabolic genes that lacked NifA-regulated promoters were strongly expressed in nodules in a NifA2-dependent manner but again mutants did not have an overt symbiotic phenotype. In summary, many genes encoded on ICEMlSymR7A were strongly expressed in nodules but not free-living rhizobia, but were not essential for symbiotic nitrogen fixation. It seems likely that some of these genes have functional homologues elsewhere in the genome and that bacteroid metabolism may be sufficiently plastic to adapt to loss of certain enzymatic functions. PMID:23308282

Annual Removal of Aboveground Plant Biomass Alters Soil Microbial Responses to Warming

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xue, Kai; Yuan, Mengting M.; Xie, Jianping

Clipping (i.e., harvesting aboveground plant biomass) is common in agriculture and for bioenergy production. However, microbial responses to clipping in the context of climate warming are poorly understood. We investigated the interactive effects of grassland warming and clipping on soil properties and plant and microbial communities, in particular, on microbial functional genes. Clipping alone did not change the plant biomass production, but warming and clipping combined increased the C4 peak biomass by 47% and belowground net primary production by 110%. Clipping alone and in combination with warming decreased the soil carbon input from litter by 81% and 75%, respectively. Withmore » less carbon input, the abundances of genes involved in degrading relatively recalcitrant carbon increased by 38% to 137% in response to either clipping or the combined treatment, which could weaken long-term soil carbon stability and trigger positive feedback with respect to warming. Clipping alone also increased the abundance of genes for nitrogen fixation, mineralization, and denitrification by 32% to 39%. Such potentially stimulated nitrogen fixation could help compensate for the 20% decline in soil ammonium levels caused by clipping alone and could contribute to unchanged plant biomass levels. Moreover, clipping tended to interact antagonistically with warming, especially with respect to effects on nitrogen cycling genes, demonstrating that single-factor studies cannot predict multifactorial changes. These results revealed that clipping alone or in combination with warming altered soil and plant properties as well as the abundance and structure of soil microbial functional genes. Aboveground biomass removal for biofuel production needs to be reconsidered, as the long-term soil carbon stability may be weakened. IMPORTANCE Global change involves simultaneous alterations, including those caused by climate warming and land management practices (e.g., clipping). Data on the interactive effects of warming and clipping on ecosystems remain elusive, particularly in microbial ecology. This study found that clipping alters microbial responses to warming and demonstrated the effects of antagonistic interactions between clipping and warming on microbial functional genes. Clipping alone or combined with warming enriched genes degrading relatively recalcitrant carbon, likely reflecting the decreased quantity of soil carbon input from litter, which could weaken long-term soil C stability and trigger positive warming feedback. These results have important implications in assessing and predicting the consequences of global climate change and indicate that the removal of aboveground biomass for biofuel production may need to be reconsidered.« less

Annual Removal of Aboveground Plant Biomass Alters Soil Microbial Responses to Warming

DOE PAGES

Xue, Kai; Yuan, Mengting M.; Xie, Jianping; ...

2016-09-27

Clipping (i.e., harvesting aboveground plant biomass) is common in agriculture and for bioenergy production. However, microbial responses to clipping in the context of climate warming are poorly understood. We investigated the interactive effects of grassland warming and clipping on soil properties and plant and microbial communities, in particular, on microbial functional genes. Clipping alone did not change the plant biomass production, but warming and clipping combined increased the C4 peak biomass by 47% and belowground net primary production by 110%. Clipping alone and in combination with warming decreased the soil carbon input from litter by 81% and 75%, respectively. Withmore » less carbon input, the abundances of genes involved in degrading relatively recalcitrant carbon increased by 38% to 137% in response to either clipping or the combined treatment, which could weaken long-term soil carbon stability and trigger positive feedback with respect to warming. Clipping alone also increased the abundance of genes for nitrogen fixation, mineralization, and denitrification by 32% to 39%. Such potentially stimulated nitrogen fixation could help compensate for the 20% decline in soil ammonium levels caused by clipping alone and could contribute to unchanged plant biomass levels. Moreover, clipping tended to interact antagonistically with warming, especially with respect to effects on nitrogen cycling genes, demonstrating that single-factor studies cannot predict multifactorial changes. These results revealed that clipping alone or in combination with warming altered soil and plant properties as well as the abundance and structure of soil microbial functional genes. Aboveground biomass removal for biofuel production needs to be reconsidered, as the long-term soil carbon stability may be weakened. IMPORTANCE Global change involves simultaneous alterations, including those caused by climate warming and land management practices (e.g., clipping). Data on the interactive effects of warming and clipping on ecosystems remain elusive, particularly in microbial ecology. This study found that clipping alters microbial responses to warming and demonstrated the effects of antagonistic interactions between clipping and warming on microbial functional genes. Clipping alone or combined with warming enriched genes degrading relatively recalcitrant carbon, likely reflecting the decreased quantity of soil carbon input from litter, which could weaken long-term soil C stability and trigger positive warming feedback. These results have important implications in assessing and predicting the consequences of global climate change and indicate that the removal of aboveground biomass for biofuel production may need to be reconsidered.« less

Thousand Year Archives of the Bulk and Compound-Specific δ15N of Export Production From the North Pacific Subtropical Gyre Indicate Increasing Nitrogen Fixation Over the Past 150 Years

NASA Astrophysics Data System (ADS)

Sherwood, O.; Batista, F. C.; Brown, J. T.; Guilderson, T. P.; McCarthy, M.

2012-12-01

Stable nitrogen isotopic analysis of amino acids (δ15N-AA) preserved in proteins has emerged as a powerful new tool to explore trophic levels and nutrient cycling in nature. To date, little has been done to explore δ15N-AA in paleo-studies of the marine nitrogen cycle. We analysed the bulk and AA-specific δ15N in the long-lived, deep-sea, proteinaceous coral Gerardia. By feeding on sinking particulate organic matter, proteinaceous corals integrate the biogeochemical signature of recently exported production within discrete skeletal growth layers. Sub-decadal resolution time-series records spanning the time period 1000 AD to present were generated from specimens of Gerardia collected from the main Hawaiian Islands, Cross Seamount, and French Frigate Shoals in the North Pacific Subtropical Gyre (NPSG). Records of bulk δ15N from the three different locations, geographically separated by up to 1000 km, showed remarkably similar long term trends. Bulk δ15N remained relatively stable from ~1000-1850 years AD, and then decreased by a total of 2 ‰ from ~1850 AD to the present. The δ15N-AA of the "trophic" group of amino acids indicated no significant changes in trophic level or microbial re-synthesis of export production over this time period. The δ15N of "source" amino acids was significantly correlated with corresponding values of bulk δ15N, with the δ15N of phenylalanine decreasing from 4.2 to 2.1‰. The latter value is similar to recent measurements of subsurface nitrate δ15N near Hawaii, suggesting that the δ15N of phenylalanine may be used to quantitatively track changes in the isotopic signature of nitrate at the base of the food web. Using a simple isotopic mass balance between upwelled nitrate and nitrogen fixation we calculate a 30% increase in nitrogen fixation in the NPSG since ~1850. These results provide invaluable long-term context for recent observations, and highlight profound changes in the marine biogeochemical cycling of nitrogen over the past 150 years in this climatically-sensitive area of the world ocean.

The Co-Distribution of Nitrifying Archaea and Diazotrophic Bacteria in Geothermal Springs

NASA Astrophysics Data System (ADS)

Hamilton, T. L.; Jewell, T. N. M.; de la Torre, J. R.; Boyd, E. S.

2014-12-01

Microbial processes that regulate availability of nutrients play key roles in shaping community composition. All life requires fixed nitrogen (N), and its bioavailability is what often limits ecosystem productivity. Biological nitrogen fixation, or the reduction of dinitrogen (N2) to ammonia (NH3), is a keystone process in N limited ecosystems, providing nitrogen for members of the community. N2 fixing organisms likely represent a 'bottom up control' on the structure of communities that develop in N limited environments. N2 fixation is catalyzed by a limited number of metabolically diverse bacteria and some methanogenic archaea and occurs in a variety of physically and geochemically diverse environments. Nitrification, or the sequential oxidation of NH4+ to nitrite (NO2-) and ultimately nitrate (NO3-), is catalyzed by several lineages of Proteobacteria at temperatures of < 62°C and by members of the Thaumarcheota at temperatures up to 90°C. Nitrification can thus be considered a 'top down control' on the structure of communities that develop in N limited environments. Our research in Yellowstone National Park (YNP) reveals a strong correspondence between the distribution of ammonia oxidizing archaea (AOA) and nitrogen fixing aquificae (NFA) in nitrogen-limited geothermal hot springs over large environmental gradients. Based on the physiology of AOA and NFA, we propose that the strong co-distributional pattern results from interspecies interactions, namely competition for bioavailable ammonia. Our recent work has shown that in springs where the niche dimension of AOA and NFA overlap (e.g., Perpetual Spouter; pH 7.1, 86.4°C), the dissimilar affinities for NH4 result in AOA metabolism maintaining a low NH4(T) pool and selecting for inclusion of NFA during the assembly of these communities. Here, we examine in situ physiological interactions of AOA and NFA, tracking changes in transcript levels of key genes involved in nitrogen metabolism and carbon fixation of these organisms in springs where the niche dimension of AOA and NFA overlap (e.g., Perpetual Spouter). These data suggest affinity for substrate and electron donor use play key roles in structuring the biodiversity of this hydrothermal community, and likely influences the structure of other N limited hydrothermal and non-hydrothermal ecosystems.

Substantial nitrogen pollution embedded in international trade

NASA Astrophysics Data System (ADS)

Oita, Azusa; Malik, Arunima; Kanemoto, Keiichiro; Geschke, Arne; Nishijima, Shota; Lenzen, Manfred

2016-02-01

Anthropogenic emissions of reactive nitrogen to the atmosphere and water bodies can damage human health and ecosystems. As a measure of a nation’s contribution to this potential damage, a country’s nitrogen footprint has been defined as the quantity of reactive nitrogen emitted during the production, consumption and transportation of commodities consumed within that country, whether those commodities are produced domestically or internationally. Here we use global emissions databases, a global nitrogen cycle model, and a global input-output database of domestic and international trade to calculate the nitrogen footprints for 188 countries as the sum of emissions of ammonia, nitrogen oxides and nitrous oxide to the atmosphere, and of nitrogen potentially exportable to water bodies. Per-capita footprints range from under 7 kg N yr-1 in some developing countries to over 100 kg N yr-1 in some wealthy nations. Consumption in China, India, the United States and Brazil is responsible for 46% of global emissions. Roughly a quarter of the global nitrogen footprint is from commodities that were traded across country borders. The main net exporters have significant agricultural, food and textile exports, and are often developing countries, whereas important net importers are almost exclusively developed economies. We conclude that substantial local nitrogen pollution is driven by demand from consumers in other countries.

A comparative study on phyllosphere nitrogen fixation by newly isolated Corynebacterium sp. & Flavobacterium sp. and their potentialities as biofertilizer.

PubMed

Giri, S; Pati, B R

2004-01-01

A number of nitrogen fixing bacteria has been isolated from forest phyllosphere on the basis of nitrogenase activity. Among them two best isolates are selected and identified as Corynebacterium sp. AN1 & Flavobacterium sp. TK2 able to reduce 88 and 132 n mol of acetylene (10(8)cells(-1)h(-1)) respectively. They were grown in large amount and sprayed on the phyllosphere of maize plants as a substitute for nitrogenous fertilizer. Marked improvements in growth and total nitrogen content of the plant have been observed by the application of these nitrogen-fixing bacteria. An average 30-37% increase in yield was obtained, which is nearer to chemical fertilizer treatment. Comparatively better effect was obtained by application of Flavobacterium sp.

Diversity and Contributions to Nitrogen Cycling and Carbon Fixation of Soil Salinity Shaped Microbial Communities in Tarim Basin

PubMed Central

Ren, Min; Zhang, Zhufeng; Wang, Xuelian; Zhou, Zhiwei; Chen, Dong; Zeng, Hui; Zhao, Shumiao; Chen, Lingling; Hu, Yuanliang; Zhang, Changyi; Liang, Yunxiang; She, Qunxin; Zhang, Yi; Peng, Nan

2018-01-01

Arid and semi-arid regions comprise nearly one-fifth of the earth's terrestrial surface. However, the diversities and functions of their soil microbial communities are not well understood, despite microbial ecological importance in driving biogeochemical cycling. Here, we analyzed the geochemistry and microbial communities of the desert soils from Tarim Basin, northwestern China. Our geochemical data indicated half of these soils are saline. Metagenomic analysis showed that bacterial phylotypes (89.72% on average) dominated the community, with relatively small proportions of Archaea (7.36%) and Eukaryota (2.21%). Proteobacteria, Firmicutes, Actinobacteria, and Euryarchaeota were most abundant based on metagenomic data, whereas genes attributed to Proteobacteria, Actinobacteria, Euryarchaeota, and Thaumarchaeota most actively transcribed. The most abundant phylotypes (Halobacterium, Halomonas, Burkholderia, Lactococcus, Clavibacter, Cellulomonas, Actinomycetospora, Beutenbergia, Pseudomonas, and Marinobacter) in each soil sample, based on metagenomic data, contributed marginally to the population of all microbial communities, whereas the putative halophiles, which contributed the most abundant transcripts, were in the majority of the active microbial population and is consistent with the soil salinity. Sample correlation analyses according to the detected and active genotypes showed significant differences, indicating high diversity of microbial communities among the Tarim soil samples. Regarding ecological functions based on the metatranscriptomic data, transcription of genes involved in various steps of nitrogen cycling, as well as carbon fixation, were observed in the tested soil samples. Metatranscriptomic data also indicated that Thaumarchaeota are crucial for ammonia oxidation and Proteobacteria play the most important role in other steps of nitrogen cycle. The reductive TCA pathway and dicarboxylate-hydroxybutyrate cycle attributed to Proteobacteria and Crenarchaeota, respectively, were highly represented in carbon fixation. Our study reveals that the microbial communities could provide carbon and nitrogen nutrients for higher plants in the sandy saline soils of Tarim Basin. PMID:29593680

The Micro-RNA172c-APETALA2-1 Node as a Key Regulator of the Common Bean-Rhizobium etli Nitrogen Fixation Symbiosis1[OPEN

PubMed Central

Nova-Franco, Bárbara; Íñiguez, Luis P.; Valdés-López, Oswaldo; Leija, Alfonso; Fuentes, Sara I.; Ramírez, Mario; Paul, Sujay

2015-01-01

Micro-RNAs are recognized as important posttranscriptional regulators in plants. The relevance of micro-RNAs as regulators of the legume-rhizobia nitrogen-fixing symbiosis is emerging. The objective of this work was to functionally characterize the role of micro-RNA172 (miR172) and its conserved target APETALA2 (AP2) transcription factor in the common bean (Phaseolus vulgaris)-Rhizobium etli symbiosis. Our expression analysis revealed that mature miR172c increased upon rhizobial infection and continued increasing during nodule development, reaching its maximum in mature nodules and decaying in senescent nodules. The expression of AP2-1 target showed a negative correlation with miR172c expression. A drastic decrease in miR172c and high AP2-1 mRNA levels were observed in ineffective nodules. Phenotypic analysis of composite bean plants with transgenic roots overexpressing miR172c or a mutated AP2-1 insensitive to miR172c cleavage demonstrated the pivotal regulatory role of the miR172 node in the common bean-rhizobia symbiosis. Increased miR172 resulted in improved root growth, increased rhizobial infection, increased expression of early nodulation and autoregulation of nodulation genes, and improved nodulation and nitrogen fixation. In addition, these plants showed decreased sensitivity to nitrate inhibition of nodulation. Through transcriptome analysis, we identified 114 common bean genes that coexpressed with AP2-1 and proposed these as being targets for transcriptional activation by AP2-1. Several of these genes are related to nodule senescence, and we propose that they have to be silenced, through miR172c-induced AP2-1 cleavage, in active mature nodules. Our work sets the basis for exploring the miR172-mediated improvement of symbiotic nitrogen fixation in common bean, the most important grain legume for human consumption. PMID:25739700

The micro-RNA72c-APETALA2-1 node as a key regulator of the common bean-Rhizobium etli nitrogen fixation symbiosis.

PubMed

Nova-Franco, Bárbara; Íñiguez, Luis P; Valdés-López, Oswaldo; Alvarado-Affantranger, Xochitl; Leija, Alfonso; Fuentes, Sara I; Ramírez, Mario; Paul, Sujay; Reyes, José L; Girard, Lourdes; Hernández, Georgina

2015-05-01

Micro-RNAs are recognized as important posttranscriptional regulators in plants. The relevance of micro-RNAs as regulators of the legume-rhizobia nitrogen-fixing symbiosis is emerging. The objective of this work was to functionally characterize the role of micro-RNA172 (miR172) and its conserved target APETALA2 (AP2) transcription factor in the common bean (Phaseolus vulgaris)-Rhizobium etli symbiosis. Our expression analysis revealed that mature miR172c increased upon rhizobial infection and continued increasing during nodule development, reaching its maximum in mature nodules and decaying in senescent nodules. The expression of AP2-1 target showed a negative correlation with miR172c expression. A drastic decrease in miR172c and high AP2-1 mRNA levels were observed in ineffective nodules. Phenotypic analysis of composite bean plants with transgenic roots overexpressing miR172c or a mutated AP2-1 insensitive to miR172c cleavage demonstrated the pivotal regulatory role of the miR172 node in the common bean-rhizobia symbiosis. Increased miR172 resulted in improved root growth, increased rhizobial infection, increased expression of early nodulation and autoregulation of nodulation genes, and improved nodulation and nitrogen fixation. In addition, these plants showed decreased sensitivity to nitrate inhibition of nodulation. Through transcriptome analysis, we identified 114 common bean genes that coexpressed with AP2-1 and proposed these as being targets for transcriptional activation by AP2-1. Several of these genes are related to nodule senescence, and we propose that they have to be silenced, through miR172c-induced AP2-1 cleavage, in active mature nodules. Our work sets the basis for exploring the miR172-mediated improvement of symbiotic nitrogen fixation in common bean, the most important grain legume for human consumption. © 2015 American Society of Plant Biologists. All Rights Reserved.

Nitrogen-Fixing Nodules Are an Important Source of Reduced Sulfur, Which Triggers Global Changes in Sulfur Metabolism in Lotus japonicus

PubMed Central

Kalloniati, Chrysanthi; Krompas, Panagiotis; Udvardi, Michael K.; Flemetakis, Emmanouil

2015-01-01

We combined transcriptomic and biochemical approaches to study rhizobial and plant sulfur (S) metabolism in nitrogen (N) fixing nodules (Fix+) of Lotus japonicus, as well as the link of S-metabolism to symbiotic nitrogen fixation and the effect of nodules on whole-plant S-partitioning and metabolism. Our data reveal that N-fixing nodules are thiol-rich organs. Their high adenosine 5′-phosphosulfate reductase activity and strong 35S-flux into cysteine and its metabolites, in combination with the transcriptional upregulation of several rhizobial and plant genes involved in S-assimilation, highlight the function of nodules as an important site of S-assimilation. The higher thiol content observed in nonsymbiotic organs of N-fixing plants in comparison to uninoculated plants could not be attributed to local biosynthesis, indicating that nodules are an important source of reduced S for the plant, which triggers whole-plant reprogramming of S-metabolism. Enhanced thiol biosynthesis in nodules and their impact on the whole-plant S-economy are dampened in plants nodulated by Fix− mutant rhizobia, which in most respects metabolically resemble uninoculated plants, indicating a strong interdependency between N-fixation and S-assimilation. PMID:26296963

Expression of the nifH Gene of a Herbaspirillum Endophyte in Wild Rice Species: Daily Rhythm during the Light-Dark Cycle

PubMed Central

You, Mu; Nishiguchi, Tomohiro; Saito, Asami; Isawa, Tsuyoshi; Mitsui, Hisayuki; Minamisawa, Kiwamu

2005-01-01

The expression of nitrogenase genes of Herbaspirillum sp. B501 associated in shoot (leaf and stem) of wild rice, Oryza officinalis, was studied by means of reverse transcription-PCR (RT-PCR) targeted at the nifH gene. RT-PCR analyses indicate that nifH transcript was detected exclusively from nitrogen-fixing cells of gfp-tagged strain B501gfp1 in both free-living and endophytic states by using a constitutive gfp gene transcript as a positive control. Transcription of nifH and nitrogen fixation in free-living cells were induced maximally at a 2% O2 concentration and repressed in free air (21% O2). nifH transcription was monitored in the endophytic cells by using total RNA extracted from B501gfp1-inoculated wild rice plants during daily light-dark cycles. The level of nifH transcription in planta varied dramatically, with a maximum during the light period. Moreover, the light radiation enhanced nifH expression even in free-living cells grown in culture. These results suggest that in planta nitrogen fixation by the endophyte shows a daily rhythm determined by the plant's light environment. PMID:16332801

Lignin Modification Leads to Increased Nodule Numbers in Alfalfa1[C][W][OPEN

PubMed Central

Gallego-Giraldo, Lina; Bhattarai, Kishor; Pislariu, Catalina I.; Nakashima, Jin; Jikumaru, Yusuke; Kamiya, Yuji; Udvardi, Michael K.; Monteros, Maria J.; Dixon, Richard A.

2014-01-01

Reduction of lignin levels in the forage legume alfalfa (Medicago sativa) by down-regulation of the monolignol biosynthetic enzyme hydroxycinnamoyl coenzyme A:shikimate hydroxycinnamoyl transferase (HCT) results in strongly increased digestibility and processing ability of lignocellulose. However, these modifications are often also associated with dwarfing and other changes in plant growth. Given the importance of nitrogen fixation for legume growth, we evaluated the impact of constitutively targeted lignin modification on the belowground organs (roots and nodules) of alfalfa plants. HCT down-regulated alfalfa plants exhibit a striking reduction in root growth accompanied by an unexpected increase in nodule numbers when grown in the greenhouse or in the field. This phenotype is associated with increased levels of gibberellins and certain flavonoid compounds in roots. Although HCT down-regulation reduced biomass yields in both the greenhouse and field experiments, the impact on the allocation of nitrogen to shoots or roots was minimal. It is unlikely, therefore, that the altered growth phenotype of reduced-lignin alfalfa is a direct result of changes in nodulation or nitrogen fixation efficiency. Furthermore, HCT down-regulation has no measurable effect on carbon allocation to roots in either greenhouse or 3-year field trials. PMID:24406794

Prone to fix: Resilience of the active nitrogen-fixing rice root microbiome

NASA Astrophysics Data System (ADS)

Hurek, Thomas; Sabale, Mugdha; Sarkar, Abhijit; Pees, Tobias; Reinhold-Hurek, Barbara

2016-04-01

Due to water consumption, many lowland rice areas in Asia are undergoing a transition that involves adoption of new management strategies, with crop rotations encompassing a non-flooded crop, including maize. Shifting from flooded to non-flooded cropping is likely to affect microbial nitrogen cycling. For analysis of the root-associated microbiome of rice and maize in response to flooding or nitrogen fertilizer, we combine methods of microbial ecology (Next-Generation sequencing of amplicons), and a reductionist approach with pure cultures of the endophytic diazotroph Azoarus sp.. Field plots of the ICON project (Introducing non-flooded crops in rice-dominated landscapes: Impact on Carbon, nitrogen and water budgets) at the International Rice Research Institute in the Philippines were analyzed. Root-associated activity of nitrogenase gene expression was assessed by quantitative RT-PCR of nifH. For rice, expression levels were surprisingly stable, in response to non-flooded versus flooded conditions, or in response to conventional nitrogen fertilizer applications versus lack of N-fertilizer. In contrast, the active diazotrophic population of maize roots was not resistant to N-fertilization, nifH expression strongly decreased. Concordant changes in the diazotrophic resident or active communities were detected by nifH amplicon sequence analysis, based on bacterial DNA or mRNA, respectively. For high-resolution analyses of the endobiome in gnotobiotic culture, we developed a dual fluorescence reporter system for Azoarcus sp. BH72 which allows to quantify and visualize epi- and endophytic gene expression by concfocal microscopy (CLSM). This allowed us to demonstrate sites of active nitrogen fixation (gene expression) in association with rice roots. We confirmed that at low nitrogen fertilizer levels, endophytic nifH gene expression persisted in rice roots, while it was repressed in maize roots. This supports our observation of remarkable stability of nitrogen fixation in association with rice roots.

Dynamics of N2 fixation and fate of diazotroph-derived nitrogen in a low-nutrient, low-chlorophyll ecosystem: results from the VAHINE mesocosm experiment (New Caledonia)

NASA Astrophysics Data System (ADS)

Bonnet, Sophie; Berthelot, Hugo; Turk-Kubo, Kendra; Fawcett, Sarah; Rahav, Eyal; L'Helguen, Stéphane; Berman-Frank, Ilana

2016-05-01

N2 fixation rates were measured daily in large (˜ 50 m3) mesocosms deployed in the tropical southwest Pacific coastal ocean (New Caledonia) to investigate the temporal variability in N2 fixation rates in relation with environmental parameters and study the fate of diazotroph-derived nitrogen (DDN) in a low-nutrient, low-chlorophyll ecosystem. The mesocosms were fertilized with ˜ 0.8 µM dissolved inorganic phosphorus (DIP) to stimulate diazotrophy. Bulk N2 fixation rates were replicable between the three mesocosms, averaged 18.5 ± 1.1 nmol N L-1 d-1 over the 23 days, and increased by a factor of 2 during the second half of the experiment (days 15 to 23) to reach 27.3 ± 1.0 nmol N L-1 d-1. These later rates measured after the DIP fertilization are higher than the upper range reported for the global ocean. During the 23 days of the experiment, N2 fixation rates were positively correlated with seawater temperature, primary production, bacterial production, standing stocks of particulate organic carbon (POC), nitrogen (PON) and phosphorus (POP), and alkaline phosphatase activity, and negatively correlated with DIP concentrations, DIP turnover time, nitrate, and dissolved organic nitrogen and phosphorus concentrations. The fate of DDN was investigated during a bloom of the unicellular diazotroph UCYN-C that occurred during the second half of the experiment. Quantification of diazotrophs in the sediment traps indicates that ˜ 10 % of UCYN-C from the water column was exported daily to the traps, representing as much as 22.4 ± 5.5 % of the total POC exported at the height of the UCYN-C bloom. This export was mainly due to the aggregation of small (5.7 ± 0.8 µm) UCYN-C cells into large (100-500 µm) aggregates. During the same time period, a DDN transfer experiment based on high-resolution nanometer-scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labeling revealed that 16 ± 6 % of the DDN was released to the dissolved pool and 21 ± 4 % was transferred to non-diazotrophic plankton, mainly picoplankton (18 ± 4 %) followed by diatoms (3 ± 2 %). This is consistent with the observed dramatic increase in picoplankton and diatom abundances, primary production, bacterial production, and standing stocks of POC, PON, and POP in the mesocosms during the second half of the experiment. These results offer insights into the fate of DDN during a bloom of UCYN-C in low-nutrient, low-chlorophyll ecosystems.

Soil nitrogen dynamics as an indicator for longleaf pine restoration

Treesearch

George L. McCaskill; Shibu Jose; Ashvini Chauhan; Andrew V. Ogram

2017-01-01

Assessing the status of soil nutrients with their corresponding microbial communities provides important information about degraded soils during the restoration of coastal wet pine forests. Net nitrogen mineralization, nitrogen-oxidizing bacteria (NOB), and soil microbial biomass were compared with patch-derived volume along a 110-year longleaf pine (Pinus...

NITROGEN DEPOSITION AND ORGANIC MATTER MANIPULATIONS AFFECT GROSS AND NET NITROGEN TRANSFORMATIONS IN TWO TEMPERATE FORESTS SOILS

EPA Science Inventory

Soil nitrogen transformations are intricately linked to carbon transformations. We utilized two existing organic matter manipulation sites in western Oregon, USA and Hungary to investigate these linkages. Our questions were: 1) Does the quantity and quality of organic matter af...

Photoperiod length paces the temporal orchestration of cell cycle and carbon-nitrogen metabolism in Crocosphaera watsonii.

PubMed

Dron, Anthony; Rabouille, Sophie; Claquin, Pascal; Talec, Amélie; Raimbault, Virginie; Sciandra, Antoine

2013-12-01

We analysed the effect of photoperiod length (PPL) (16:8 and 8:16 h of light-dark regime, named long and short PPL, respectively) on the temporal orchestration of the two antagonistic, carbon and nitrogen acquisitions in the unicellular, diazotrophic cyanobacterium Crocosphaera watsonii strain WH8501 growing diazotrophically. Carbon and nitrogen metabolism were monitored at high frequency, and their patterns were compared with the cell cycle progression. The oxygen-sensitive N2 fixation process occurred mainly during the dark period, where photosynthesis cannot take place, inducing a light-dark cycle of cellular C : N ratio. Examination of circadian patterns in the cell cycle revealed that cell division occurred during the midlight period, (8 h and 4 h into the light in the long and short PPL conditions, respectively), thus timely separated from the energy-intensive diazotrophic process. Results consistently show a nearly 5 h time lag between the end of cell division and the onset of N2 fixation. Shorter PPLs affected DNA compaction of C. watsonii cells and also led to a decrease in the cell division rate. Therefore, PPL paces the growth of C. watsonii: a long PPL enhances cell division while a short PPL favours somatic growth (biomass production) with higher carbon and nitrogen cell contents. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

Electron transfer protein complexes in the thylakoid membranes of heterocysts from the cyanobacterium Nostoc punctiforme.

PubMed

Cardona, Tanai; Battchikova, Natalia; Zhang, Pengpeng; Stensjö, Karin; Aro, Eva-Mari; Lindblad, Peter; Magnuson, Ann

2009-04-01

Filamentous, heterocystous cyanobacteria are capable of nitrogen fixation and photoautotrophic growth. Nitrogen fixation takes place in heterocysts that differentiate as a result of nitrogen starvation. Heterocysts uphold a microoxic environment to avoid inactivation of nitrogenase, e.g. by downregulation of oxygenic photosynthesis. The ATP and reductant requirement for the nitrogenase reaction is considered to depend on Photosystem I, but little is known about the organization of energy converting membrane proteins in heterocysts. We have investigated the membrane proteome of heterocysts from nitrogen fixing filaments of Nostoc punctiforme sp. PCC 73102, by 2D gel electrophoresis and mass spectrometry. The membrane proteome was found to be dominated by the Photosystem I and ATP-synthase complexes. We could identify a significant amount of assembled Photosystem II complexes containing the D1, D2, CP43, CP47 and PsbO proteins from these complexes. We could also measure light-driven in vitro electron transfer from Photosystem II in heterocyst thylakoid membranes. We did not find any partially disassembled Photosystem II complexes lacking the CP43 protein. Several subunits of the NDH-1 complex were also identified. The relative amount of NDH-1M complexes was found to be higher than NDH-1L complexes, which might suggest a role for this complex in cyclic electron transfer in the heterocysts of Nostoc punctiforme.

A Dicarboxylate Transporter, LjALMT4, Mainly Expressed in Nodules of Lotus japonicus.

PubMed

Takanashi, Kojiro; Sasaki, Takayuki; Kan, Tomohiro; Saida, Yuka; Sugiyama, Akifumi; Yamamoto, Yoko; Yazaki, Kazufumi

2016-07-01

Legume plants can establish symbiosis with soil bacteria called rhizobia to obtain nitrogen as a nutrient directly from atmospheric N2 via symbiotic nitrogen fixation. Legumes and rhizobia form nodules, symbiotic organs in which fixed-nitrogen and photosynthetic products are exchanged between rhizobia and plant cells. The photosynthetic products supplied to rhizobia are thought to be dicarboxylates but little is known about the movement of dicarboxylates in the nodules. In terms of dicarboxylate transporters, an aluminum-activated malate transporter (ALMT) family is a strong candidate responsible for the membrane transport of carboxylates in nodules. Among the seven ALMT genes in the Lotus japonicus genome, only one, LjALMT4, shows a high expression in the nodules. LjALMT4 showed transport activity in a Xenopus oocyte system, with LjALMT4 mediating the efflux of dicarboxylates including malate, succinate, and fumarate, but not tricarboxylates such as citrate. LjALMT4 also mediated the influx of several inorganic anions. Organ-specific gene expression analysis showed LjALMT4 mRNA mainly in the parenchyma cells of nodule vascular bundles. These results suggest that LjALMT4 may not be involved in the direct supply of dicarboxylates to rhizobia in infected cells but is responsible for supplying malate as well as several anions necessary for symbiotic nitrogen fixation, via nodule vasculatures.

The nitrate-reduction gene cluster components exert lineage-dependent contributions to optimization of Sinorhizobium symbiosis with soybeans.

PubMed

Liu, Li Xue; Li, Qin Qin; Zhang, Yun Zeng; Hu, Yue; Jiao, Jian; Guo, Hui Juan; Zhang, Xing Xing; Zhang, Biliang; Chen, Wen Xin; Tian, Chang Fu

2017-12-01

Receiving nodulation and nitrogen fixation genes does not guarantee rhizobia an effective symbiosis with legumes. Here, variations in gene content were determined for three Sinorhizobium species showing contrasting symbiotic efficiency on soybeans. A nitrate-reduction gene cluster absent in S. sojae was found to be essential for symbiotic adaptations of S. fredii and S. sp. III. In S. fredii, the deletion mutation of the nap (nitrate reductase), instead of nir (nitrite reductase) and nor (nitric oxide reductase), led to defects in nitrogen-fixation (Fix - ). By contrast, none of these core nitrate-reduction genes were required for the symbiosis of S. sp. III. However, within the same gene cluster, the deletion of hemN1 (encoding oxygen-independent coproporphyrinogen III oxidase) in both S. fredii and S. sp. III led to the formation of nitrogen-fixing (Fix + ) but ineffective (Eff - ) nodules. These Fix + /Eff - nodules were characterized by significantly lower enzyme activity of glutamine synthetase indicating rhizobial modulation of nitrogen-assimilation by plants. A distant homologue of HemN1 from S. sojae can complement this defect in S. fredii and S. sp. III, but exhibited a more pleotropic role in symbiosis establishment. These findings highlighted the lineage-dependent optimization of symbiotic functions in different rhizobial species associated with the same host. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Diazotroph Diversity in the Sea Ice, Melt Ponds, and Surface Waters of the Eurasian Basin of the Central Arctic Ocean.

PubMed

Fernández-Méndez, Mar; Turk-Kubo, Kendra A; Buttigieg, Pier L; Rapp, Josephine Z; Krumpen, Thomas; Zehr, Jonathan P; Boetius, Antje

2016-01-01

The Eurasian basin of the Central Arctic Ocean is nitrogen limited, but little is known about the presence and role of nitrogen-fixing bacteria. Recent studies have indicated the occurrence of diazotrophs in Arctic coastal waters potentially of riverine origin. Here, we investigated the presence of diazotrophs in ice and surface waters of the Central Arctic Ocean in the summer of 2012. We identified diverse communities of putative diazotrophs through targeted analysis of the nifH gene, which encodes the iron protein of the nitrogenase enzyme. We amplified 529 nifH sequences from 26 samples of Arctic melt ponds, sea ice and surface waters. These sequences resolved into 43 clusters at 92% amino acid sequence identity, most of which were non-cyanobacterial phylotypes from sea ice and water samples. One cyanobacterial phylotype related to Nodularia sp. was retrieved from sea ice, suggesting that this important functional group is rare in the Central Arctic Ocean. The diazotrophic community in sea-ice environments appear distinct from other cold-adapted diazotrophic communities, such as those present in the coastal Canadian Arctic, the Arctic tundra and glacial Antarctic lakes. Molecular fingerprinting of nifH and the intergenic spacer region of the rRNA operon revealed differences between the communities from river-influenced Laptev Sea waters and those from ice-related environments pointing toward a marine origin for sea-ice diazotrophs. Our results provide the first record of diazotrophs in the Central Arctic and suggest that microbial nitrogen fixation may occur north of 77°N. To assess the significance of nitrogen fixation for the nitrogen budget of the Arctic Ocean and to identify the active nitrogen fixers, further biogeochemical and molecular biological studies are needed.

Diazotroph Diversity in the Sea Ice, Melt Ponds, and Surface Waters of the Eurasian Basin of the Central Arctic Ocean

PubMed Central

Fernández-Méndez, Mar; Turk-Kubo, Kendra A.; Buttigieg, Pier L.; Rapp, Josephine Z.; Krumpen, Thomas; Zehr, Jonathan P.; Boetius, Antje

2016-01-01

The Eurasian basin of the Central Arctic Ocean is nitrogen limited, but little is known about the presence and role of nitrogen-fixing bacteria. Recent studies have indicated the occurrence of diazotrophs in Arctic coastal waters potentially of riverine origin. Here, we investigated the presence of diazotrophs in ice and surface waters of the Central Arctic Ocean in the summer of 2012. We identified diverse communities of putative diazotrophs through targeted analysis of the nifH gene, which encodes the iron protein of the nitrogenase enzyme. We amplified 529 nifH sequences from 26 samples of Arctic melt ponds, sea ice and surface waters. These sequences resolved into 43 clusters at 92% amino acid sequence identity, most of which were non-cyanobacterial phylotypes from sea ice and water samples. One cyanobacterial phylotype related to Nodularia sp. was retrieved from sea ice, suggesting that this important functional group is rare in the Central Arctic Ocean. The diazotrophic community in sea-ice environments appear distinct from other cold-adapted diazotrophic communities, such as those present in the coastal Canadian Arctic, the Arctic tundra and glacial Antarctic lakes. Molecular fingerprinting of nifH and the intergenic spacer region of the rRNA operon revealed differences between the communities from river-influenced Laptev Sea waters and those from ice-related environments pointing toward a marine origin for sea-ice diazotrophs. Our results provide the first record of diazotrophs in the Central Arctic and suggest that microbial nitrogen fixation may occur north of 77°N. To assess the significance of nitrogen fixation for the nitrogen budget of the Arctic Ocean and to identify the active nitrogen fixers, further biogeochemical and molecular biological studies are needed. PMID:27933047

Measuring N2 Pressure Using Cyanobacteria Discipline: Geomicrobiology

NASA Technical Reports Server (NTRS)

Silverman, Shaelyn N.; Kopf, Sebastian; Gordon, Richard; Bebout, Brad M.; Som, Sanjoy

2017-01-01

The evolution of Earth's atmosphere has been governed by biological evolution. Dinitrogen (N2) has been a major constituent of Earth's atmosphere throughout the planet's history, yet only a few constraints exist for the partial pressure of N2 (pN2). In this study we evaluate two new potential proxies for pN2: the physical spacing between heterocysts and the isotopic signature of nitrogen fixation in filamentous cyanobacteria. Heterocyst-forming filamentous cyanobacteria are some of the oldest photosynthetic microorganisms on Earth, and debated fossilized specimens have been found in sedimentary rocks as old as 2 Ga. These organisms overcome nitrogen limitation in their aqueous environment through cellular differentiation along their filaments. The specialized cells that develop, known as heterocysts, fix the nitrogen and laterally distribute it to neighboring cells along the filaments. Because the concentration of the dissolved N2 available to the filaments correlates directly with pN2, any preservable physiological response of the organism to the changed N2 availability constitutes a potential proxy for pN2. In the laboratory, we have examined how pN2 is reflected in the heterocyst spacing pattern and in the isotopic signature of nitrogen fixation by subjecting the representative species Anabaena cylindrica and Anabaena variabilis to different N2 partial pressures during growth at constant temperature and lighting (in media free of combined nitrogen). We show experimentally that the distance between heterocysts and the nitrogen isotope fractionation measured in bulk biomass reflect the pN2 experienced by Anabaena cylindrica. Current work is investigating these responses in Anabaena variabilis. When heterocystous cyanobacteria fossilize, these morphological and isotopic signatures should preserve information about pN2 at that time. Application of this relationship to the rock record may provide a paleoproxy to complement the two existing geobarometers.

Algal-mediated ecosystem exchanges in the Eel River drainage network: towards photogrammetric mapping of color to function

NASA Astrophysics Data System (ADS)

Power, M. E.; Welter, J.; Furey, P.; Lowe, R.; Finlay, J. C.; Hondzo, M.; Limm, M.; Bode, C.; Dietrich, W. E.

2009-12-01

Seasonal algal proliferations in river networks are typically short-lived (weeks-months) but spatially extensive. They mediate important ecological and biogeochemical exchanges within and between ecosystems. We are investigating correspondence of assemblage color with ecosystem function in the nitrogen-limited Eel River of northern California. During summer base flow following winter floods, Eel algal assemblages are dominated by the green macroalga Cladophora glomerata. New growths are green, but blooms turn yellow as Cladophora filaments are colonized by epiphytic diatoms (Cocconeis spp.). Later, proliferations turn rust colored as epiphytic assemblages became dominated by Epithemia spp., diatoms that contain nitrogen-fixing cyanobacterial endosymbionts. Epithemia-encrusted Cladophora occurs at and downstream of reaches draining > 100 km2 (where summer inundated average channel widths > 25 m), coinciding with a threshold increase in concentration of total dissolved nitrogen. Areal nitrogen fixation rates are 14x higher in rusty algal proliferations than in green, and 3-4x higher than in yellow Cladophora mats. Corresponding increases in insect emergence suggest that nitrogen fixed by cyanobacterial endosymbionts is highly edible. Rates of biomass emergence from rusty Cladophora mats are 12-17 times greater than from green mats, and 8-10 times greater from rusty than from yellow Cladophora mats, because larger taxa emerge from rusty mats (Chironominae versus Ceratopogonidae in yellow mats). Photogrammetric detection of spatial coverage and color changes in algal proliferations may help us track nitrogen fluxes they mediate (riverine loading from the atmosphere via fixation, river to the watershed return via insect emergence) that link riverine to aerial, watershed, and potentially nearshore marine ecosystems at reach to basin scales.

Cyanobacterial nitrogenases: phylogenetic diversity, regulation and functional predictions

PubMed Central

Esteves-Ferreira, Alberto A.; Cavalcanti, João Henrique Frota; Vaz, Marcelo Gomes Marçal Vieira; Alvarenga, Luna V.; Nunes-Nesi, Adriano; Araújo, Wagner L.

2017-01-01

Abstract Cyanobacteria is a remarkable group of prokaryotic photosynthetic microorganisms, with several genera capable of fixing atmospheric nitrogen (N2) and presenting a wide range of morphologies. Although the nitrogenase complex is not present in all cyanobacterial taxa, it is spread across several cyanobacterial strains. The nitrogenase complex has also a high theoretical potential for biofuel production, since H2 is a by-product produced during N2 fixation. In this review we discuss the significance of a relatively wide variety of cell morphologies and metabolic strategies that allow spatial and temporal separation of N2 fixation from photosynthesis in cyanobacteria. Phylogenetic reconstructions based on 16S rRNA and nifD gene sequences shed light on the evolutionary history of the two genes. Our results demonstrated that (i) sequences of genes involved in nitrogen fixation (nifD) from several morphologically distinct strains of cyanobacteria are grouped in similarity with their morphology classification and phylogeny, and (ii) nifD genes from heterocytous strains share a common ancestor. By using this data we also discuss the evolutionary importance of processes such as horizontal gene transfer and genetic duplication for nitrogenase evolution and diversification. Finally, we discuss the importance of H2 synthesis in cyanobacteria, as well as strategies and challenges to improve cyanobacterial H2 production. PMID:28323299

Nitrogen fixation and hydrogen metabolism in cyanobacteria.

PubMed

Bothe, Hermann; Schmitz, Oliver; Yates, M Geoffrey; Newton, William E

2010-12-01

This review summarizes recent aspects of (di)nitrogen fixation and (di)hydrogen metabolism, with emphasis on cyanobacteria. These organisms possess several types of the enzyme complexes catalyzing N(2) fixation and/or H(2) formation or oxidation, namely, two Mo nitrogenases, a V nitrogenase, and two hydrogenases. The two cyanobacterial Ni hydrogenases are differentiated as either uptake or bidirectional hydrogenases. The different forms of both the nitrogenases and hydrogenases are encoded by different sets of genes, and their organization on the chromosome can vary from one cyanobacterium to another. Factors regulating the expression of these genes are emerging from recent studies. New ideas on the potential physiological and ecological roles of nitrogenases and hydrogenases are presented. There is a renewed interest in exploiting cyanobacteria in solar energy conversion programs to generate H(2) as a source of combustible energy. To enhance the rates of H(2) production, the emphasis perhaps needs not to be on more efficient hydrogenases and nitrogenases or on the transfer of foreign enzymes into cyanobacteria. A likely better strategy is to exploit the use of radiant solar energy by the photosynthetic electron transport system to enhance the rates of H(2) formation and so improve the chances of utilizing cyanobacteria as a source for the generation of clean energy.