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

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.

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

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.

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.

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

"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.

Nitrogen Fixation in the Intertidal Sediments of the Yangtze Estuary: Occurrence and Environmental Implications

NASA Astrophysics Data System (ADS)

Hou, Lijun; Wang, Rong; Yin, Guoyu; Liu, Min; Zheng, Yanling

2018-03-01

Nitrogen fixation is a microbial-mediated process converting atmospheric dinitrogen gas to biologically available ammonia or other molecules, and it plays an important role in regulating nitrogen budgets in coastal marine ecosystems. In this study, nitrogen fixation in the intertidal sediments of the Yangtze Estuary was investigated using nitrogen isotope tracing technique. The abundance of nitrogen fixation functional gene (nifH) was also quantified. The measured rates of sediment nitrogen fixation ranged from 0.37 to 7.91 nmol N g-1 hr-1, while the abundance of nifH gene varied from 2.28 × 106 to 1.28 × 108 copies g-1 in the study area. The benthic nitrogen fixation was correlated closely to the abundance of nifH gene and was affected significantly by salinity, pH, and availability of sediment organic carbon and ammonium. It is estimated that sediment nitrogen fixation contributed approximately 9.3% of the total terrigenous inorganic nitrogen transported annually into the Yangtze estuarine and coastal environment. This result implies that the occurrence of benthic nitrogen fixation acts as an important internal source of reactive nitrogen and to some extent exacerbates nitrogen pollution in this aquatic ecosystem.

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).

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.).

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

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...

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.

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 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.

[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.

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 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.

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.

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.

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.

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

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

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, ...

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

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

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

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

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.

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

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.

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

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.

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.

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

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

An antimicrobial peptide essential for bacterial survival in the nitrogen-fixing symbiosis.

PubMed

Kim, Minsoo; Chen, Yuhui; Xi, Jiejun; Waters, Christopher; Chen, Rujin; Wang, Dong

2015-12-08

In the nitrogen-fixing symbiosis between legume hosts and rhizobia, the bacteria are engulfed by a plant cell membrane to become intracellular organelles. In the model legume Medicago truncatula, internalization and differentiation of Sinorhizobium (also known as Ensifer) meliloti is a prerequisite for nitrogen fixation. The host mechanisms that ensure the long-term survival of differentiating intracellular bacteria (bacteroids) in this unusual association are unclear. The M. truncatula defective nitrogen fixation4 (dnf4) mutant is unable to form a productive symbiosis, even though late symbiotic marker genes are expressed in mutant nodules. We discovered that in the dnf4 mutant, bacteroids can apparently differentiate, but they fail to persist within host cells in the process. We found that the DNF4 gene encodes NCR211, a member of the family of nodule-specific cysteine-rich (NCR) peptides. The phenotype of dnf4 suggests that NCR211 acts to promote the intracellular survival of differentiating bacteroids. The greatest expression of DNF4 was observed in the nodule interzone II-III, where bacteroids undergo differentiation. A translational fusion of DNF4 with GFP localizes to the peribacteroid space, and synthetic NCR211 prevents free-living S. meliloti from forming colonies, in contrast to mock controls, suggesting that DNF4 may interact with bacteroids directly or indirectly for its function. Our findings indicate that a successful symbiosis requires host effectors that not only induce bacterial differentiation, but also that maintain intracellular bacteroids during the host-symbiont interaction. The discovery of NCR211 peptides that maintain bacterial survival inside host cells has important implications for improving legume crops.

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.

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.

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

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

A mechanistic, globally-applicable model of plant nitrogen uptake, retranslocation and fixation

NASA Astrophysics Data System (ADS)

Fisher, J. B.; Tan, S.; Malhi, Y.; Fisher, R. A.; Sitch, S.; Huntingford, C.

2008-12-01

Nitrogen is one of the nutrients that can most limit plant growth, and nitrogen availability may be a controlling factor on biosphere responses to climate change. We developed a plant nitrogen assimilation model based on a) advective transport through the transpiration stream, b) retranslocation whereby carbon is expended to resorb nitrogen from leaves, c) active uptake whereby carbon is expended to acquire soil nitrogen, and d) biological nitrogen fixation whereby carbon is expended for symbiotic nitrogen fixers. The model relies on 9 inputs: 1) net primary productivity (NPP), 2) plant C:N ratio, 3) available soil nitrogen, 4) root biomass, 5) transpiration rate, 6) saturated soil depth,7) leaf nitrogen before senescence, 8) soil temperature, and 9) ability to fix nitrogen. A carbon cost of retranslocation is estimated based on leaf nitrogen and compared to an active uptake carbon cost based on root biomass and available soil nitrogen; for nitrogen fixers both costs are compared to a carbon cost of fixation dependent on soil temperature. The NPP is then allocated to optimize growth while maintaining the C:N ratio. The model outputs are total plant nitrogen uptake, remaining NPP available for growth, carbon respired to the soil and updated available soil nitrogen content. We test and validate the model (called FUN: Fixation and Uptake of Nitrogen) against data from the UK, Germany and Peru, and run the model under simplified scenarios of primary succession and climate change. FUN is suitable for incorporation into a land surface scheme of a General Circulation Model and will be coupled with a soil model and dynamic global vegetation model as part of a land surface model (JULES).

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.

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.

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

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.

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.

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...

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.

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

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.

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

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...

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.

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)

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.

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.

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.

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.

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.

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.

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 (...

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...

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

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

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

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.

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

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

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.

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.

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.

Understanding Nitrogen Fixation

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

Paul J. Chirik

. The synthesis of ammonia, NH{sub 3}, from its elements, H{sub 2} and N{sub 2}, via the venerable Haber-Bosch process is one of the most significant technological achievements of the past century. Our research program seeks to discover new transition metal reagents and catalysts to disrupt the strong N {triple_bond} N bond in N{sub 2} and create new, fundamental chemical linkages for the construction of molecules with application as fuels, fertilizers and fine chemicals. With DOE support, our group has discovered a mild method for ammonia synthesis in solution as well as new methods for the construction of nitrogen-carbon bonds directly from N{sub 2}. Ideally these achievements will evolve into more efficient nitrogen fixation schemes that circumvent the high energy demands of industrial ammonia synthesis. Industrially, atmospheric nitrogen enters the synthetic cycle by the well-established Haber-Bosch process whereby N{sub 2} is hydrogenated to ammonia at high temperature and pressure. The commercialization of this reaction represents one of the greatest technological achievements of the 20th century as Haber-Bosch ammonia is responsible for supporting approximately 50% of the world's population and serves as the source of half of the nitrogen in the human body. The extreme reaction conditions required for an economical process have significant energy consequences, consuming 1% of the world's energy supply mostly in the form of pollution-intensive coal. Moreover, industrial H{sub 2} synthesis via the water gas shift reaction and the steam reforming of methane is fossil fuel intensive and produces CO{sub 2} as a byproduct. New synthetic methods that promote this thermodynamically favored transformation ({Delta}G{sup o} = -4.1 kcal/mol) under milder conditions or completely obviate it are therefore desirable. Most nitrogen-containing organic molecules are derived from ammonia (and hence rely on the Haber-Bosch and H{sub 2} synthesis processes) and direct synthesis from

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...

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

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...

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

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

Contribution of dinitrogen fixation to bacterial and primary productivity in the Gulf of Aqaba (Red Sea)

NASA Astrophysics Data System (ADS)

Rahav, E.; Herut, B.; Mulholland, M. R.; Voß, B.; Stazic, D.; Steglich, C.; Hess, W. R.; Berman-Frank, I.

2013-06-01

We evaluated the seasonal contribution of heterotrophic and autotrophic diazotrophy to the total dinitrogen (N2) fixation in a representative pelagic station in the northern Gulf of Aqaba in early spring when the water column was mixed and during summer under full thermal stratification. N2 fixation rates were low during the mixed period (˜ 0.1 nmol N L-1 d-1) and were significantly coupled with both primary and bacterial productivity. During the stratified period N2 fixation rates were four-fold higher (˜ 0.4 nmol N L-1 d-1) and were significantly correlated solely with bacterial productivity. Furthermore, while experimental enrichment of seawater by phosphorus (P) enhanced bacterial productivity and N2 fixation rates during both seasons primary productivity was stimulated by P only in the early spring. Metatranscriptomic analyses from the stratified period identified the major diazotrophic contributors as related to heterotrophic prokaryotes from the Euryarchaeota and Desulfobacterales (Deltaproteobacteria) or Chlorobiales (Chlorobia). Moreover, during this season, experimental amendments to seawater applying a combination of the photosynthetic inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and a mixture of amino acids increased both bacterial productivity and N2 fixation rates. Our findings from the northern Gulf of Aqaba indicate a~shift in the diazotrophic community from phototrophic and heterotrophic populations, including small blooms of the cyanobacterium Trichodesmium, in winter/early spring, to predominantly heterotrophic diazotrophs in summer that may be both P and carbon limited as the additions of P and amino acids illustrated.

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...

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

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.

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

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

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 ...

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.

Carbon fixation in sediments of Sino-Pacific seas-differential contributions of bacterial and archaeal domains

NASA Astrophysics Data System (ADS)

Das, Anindita; Cao, Wenrui; Zhang, Hongjie; Saren, Gaowa; Jiang, Mingyu; Yu, Xinke

2017-11-01

Oceanic stretches experiencing perpetual darkness and extreme limitation of utilizable organic matter often rely on chemosynthetic carbon (C)-fixation. However, C-fixation is not limited to carbon-deplete environments alone but might also occur in varying degrees in carbon-replete locales depending on the nature and concentration of utilizable carbon, electron donors and acceptors. Quantification of microbial C-fixation and relative contribution of domains bacteria and archaea are therefore crucial. The present experiment estimates the differential rates of C-fixation by archaea and bacteria along with the effects of different electron donors. Four Sino-Pacific marine sediments from Bashi strait (Western Pacific Warm Pool), East China Sea, South China Sea and Okinawa Trough were examined. Total microbial C-uptake was estimated by doping of aqueous NaH14CO3. Total bacterial C-uptake was measured by blocking archaeal metabolism using inhibitor GC7. Archaeal contribution was estimated by subtracting total bacterial from total microbial C-uptake. Effect of electron donor addition was analyzed by spiking with ammonium, sulfide, and reduced metals. Results suggested that C-fixation in marine sediments was not the function of archaea alone, which was in contrast to results from several recent publications. C-fixing bacteria are also equally active. Often in spite of great effort of one domain to fix carbon, the system does not become net C-fixing due to equal and opposite C-releasing activity of the other domain. Thus a C-releasing bacterial or archaeal community can become C-fixing with the change of nature and concentration of electron donors.

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

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.

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.

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.

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

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.

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

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.

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.

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...

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

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)

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 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

Regulation of Development and Nitrogen Fixation in Anabaena

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

James W. Golden

2008-10-17

The regulation of development and cellular differentiation is important for all multicellular organisms. The nitrogen-fixing filamentous cyanobacterium Anabaena (also Nostoc) sp. PCC 7120 (hereafter Anabaena) provides a model of multicellular microbial development and pattern formation. Anabaena reduces N2 to ammonia in specialized terminally differentiated cells called heterocysts. A one-dimensional developmental pattern of single heterocysts regularly spaced along filaments of photosynthetic vegetative cells is established to form a multicellular organism composed of these two interdependent cell types. This multicellular growth pattern, the distinct phylogeny of cyanobacteria, and the suspected antiquity of heterocyst development make this an important model system. Our long-termmore » goal is to understand the regulatory network required for heterocyst development and nitrogen fixation. This project is focused on two key aspects of heterocyst regulation: one, the mechanism by which HetR controls the initiation of differentiation, and two, the cis and trans acting factors required for expression of the nitrogen-fixation (nif) genes. HetR is thought to be a central regulator of heterocyst development but the partners and mechanisms involved in this regulation are unknown. Our recent results indicate that PatS and other signals that regulate heterocyst pattern cannot interact, directly or indirectly, with a R223W mutant of HetR. We plan to use biochemical and genetic approaches to identify proteins that interact with the HetR protein, which will help reveal the mechanisms underlying its regulation of development. Our second goal is to determine how the nif genes are expressed. It is important to understand the mechanisms controlling nif genes since they represent the culmination of the differentiation process and the essence of heterocyst function. The Anabaena genome lacks the genes required for expression of nif genes present in other organisms such as rpo

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

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.

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

Endophytic Colonization and In Planta Nitrogen Fixation by a Herbaspirillum sp. Isolated from Wild Rice Species

PubMed Central

Elbeltagy, Adel; Nishioka, Kiyo; Sato, Tadashi; Suzuki, Hisa; Ye, Bin; Hamada, Toru; Isawa, Tsuyoshi; Mitsui, Hisayuki; Minamisawa, Kiwamu

2001-01-01

Nitrogen-fixing bacteria were isolated from the stems of wild and cultivated rice on a modified Rennie medium. Based on 16S ribosomal DNA (rDNA) sequences, the diazotrophic isolates were phylogenetically close to four genera: Herbaspirillum, Ideonella, Enterobacter, and Azospirillum. Phenotypic properties and signature sequences of 16S rDNA indicated that three isolates (B65, B501, and B512) belong to the Herbaspirillum genus. To examine whether Herbaspirillum sp. strain B501 isolated from wild rice, Oryza officinalis, endophytically colonizes rice plants, the gfp gene encoding green fluorescent protein (GFP) was introduced into the bacteria. Observations by fluorescence stereomicroscopy showed that the GFP-tagged bacteria colonized shoots and seeds of aseptically grown seedlings of the original wild rice after inoculation of the seeds. Conversely, for cultivated rice Oryza sativa, no GFP fluorescence was observed for shoots and only weak signals were observed for seeds. Observations by fluorescence and electron microscopy revealed that Herbaspirillum sp. strain B501 colonized mainly intercellular spaces in the leaves of wild rice. Colony counts of surface-sterilized rice seedlings inoculated with the GFP-tagged bacteria indicated significantly more bacterial populations inside the original wild rice than in cultivated rice varieties. Moreover, after bacterial inoculation, in planta nitrogen fixation in young seedlings of wild rice, O. officinalis, was detected by the acetylene reduction and 15N2 gas incorporation assays. Therefore, we conclude that Herbaspirillum sp. strain B501 is a diazotrophic endophyte compatible with wild rice, particularly O. officinalis. PMID:11679357

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

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

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.

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

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

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.

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.

Metatranscriptomic and metagenomic description of the bacterial nitrogen metabolism in waste water wet oxidation effluents.

PubMed

Crovadore, Julien; Soljan, Vice; Calmin, Gautier; Chablais, Romain; Cochard, Bastien; Lefort, François

2017-10-01

Anaerobic digestion is a common method for reducing the amount of sludge solids in used waters and enabling biogas production. The wet oxidation process (WOX) improves anaerobic digestion by converting carbon into methane through oxidation of organic compounds. WOX produces effluents rich in ammonia, which must be removed to maintain the activity of methanogens. Ammonia removal from WOX could be biologically operated by aerobic granules. To this end, granulation experiments were conducted in 2 bioreactors containing an activated sludge (AS). For the first time, the dynamics of the microbial community structure and the expression levels of 7 enzymes of the nitrogen metabolism in such active microbial communities were followed in regard to time by metagenomics and metatranscriptomics. It was shown that bacterial communities adapt to the wet oxidation effluent by increasing the expression level of the nitrogen metabolism, suggesting that these biological activities could be a less costly alternative for the elimination of ammonia, resulting in a reduction of the use of chemicals and energy consumption in sewage plants. This study reached a strong sequencing depth (from 4.4 to 7.6 Gb) and enlightened a yet unknown diversity of the microorganisms involved in the nitrogen pathway. Moreover, this approach revealed the abundance and expression levels of specialised enzymes involved in nitrification, denitrification, ammonification, dissimilatory nitrate reduction to ammonium (DNRA) and nitrogen fixation processes in AS.

Nitrogen starvation affects bacterial adhesion to soil

PubMed Central

Borges, Maria Tereza; Nascimento, Antônio Galvão; Rocha, Ulisses Nunes; Tótola, Marcos Rogério

2008-01-01

One of the main factors limiting the bioremediation of subsoil environments based on bioaugmentation is the transport of selected microorganisms to the contaminated zones. The characterization of the physiological responses of the inoculated microorganisms to starvation, especially the evaluation of characteristics that affect the adhesion of the cells to soil particles, is fundamental to anticipate the success or failure of bioaugmentation. The objective of this study was to investigate the effect of nitrogen starvation on cell surface hydrophobicity and cell adhesion to soil particles by bacterial strains previously characterized as able to use benzene, toluene or xilenes as carbon and energy sources. The strains LBBMA 18-T (non-identified), Arthrobacter aurescens LBBMA 98, Arthrobacter oxydans LBBMA 201, and Klebsiella sp. LBBMA 204–1 were used in the experiments. Cultivation of the cells in nitrogen-deficient medium caused a significant reduction of the adhesion to soil particles by all the four strains. Nitrogen starvation also reduced significantly the strength of cell adhesion to the soil particles, except for Klebsiella sp. LBBMA 204–1. Two of the four strains showed significant reduction in cell surface hydrophobicity. It is inferred that the efficiency of bacterial transport through soils might be potentially increased by nitrogen starvation. PMID:24031246

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

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

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.

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.

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.

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

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.

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.

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

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

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

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.

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.

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

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

Uncoupling between dinitrogen fixation and primary productivity in the eastern Mediterranean Sea

NASA Astrophysics Data System (ADS)

Rahav, Eyal; Herut, Barak; Stambler, Noga; Bar-Zeev, Edo; Mulholland, Margaret R.; Berman-Frank, Ilana

2013-03-01

In the nitrogen (N)-impoverished photic zones of many oceanic regions, prokaryotic organisms fixing atmospheric dinitrogen (N2; diazotrophs) supply an essential source of new nitrogen and fuel primary production. We measured dinitrogen fixation and primary productivity (PP) during the thermally stratified summer period in different water regimes of the oligotrophic eastern Mediterranean Sea, including the Cyprus Eddy and the Rhodes Gyre. Low N2 fixation rates were measured (0.8-3.2 µmol N m-2 d-1) excluding 10-fold higher rates in the Rhodes Gyre and Cyprus Eddy ( 20 µmol N m-2 d-1). The corresponding PP increased from east to west (200-2500 µmol C m-2 d-1), with relatively higher productivity recorded in the Rhodes Gyre and Cyprus Eddy (2150 and 2300 µmol C m-2 d-1, respectively). These measurements demonstrate that N2 fixation in the photic zone of the eastern Mediterranean Sea contributes only negligibly by direct inputs to PP (i.e., cyanobacterial diazotrophs) and is in fact uncoupled from PP. By contrast, N2 fixation is significantly coupled to bacterial productivity and to net heterotrophic areas, suggesting that heterotrophic N2 fixation may in fact be significant in this ultraoligotrophic system. This is further substantiated by the high N2 fixation rates we measured from aphotic depths and by the results of phylogenetic analysis in other studies showing an abundance of heterotrophic diazotrophs.

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.

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

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

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

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.

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

Potential for Nitrogen Fixation and Nitrification in the Granite-Hosted Subsurface at Henderson Mine, CO

PubMed Central

Swanner, Elizabeth D.; Templeton, Alexis S.

2011-01-01

The existence of life in the deep terrestrial subsurface is established, yet few studies have investigated the origin of nitrogen that supports deep life. Previously, 16S rRNA gene surveys cataloged a diverse microbial community in subsurface fluids draining from boreholes 3000 feet deep at Henderson Mine, CO, USA (Sahl et al., 2008). The prior characterization of the fluid chemistry and microbial community forms the basis for the further investigation here of the source of NH4+. The reported fluid chemistry included N2, NH4+ (5–112 μM), NO2− (27–48 μM), and NO3− (17–72 μM). In this study, the correlation between low NH4+ concentrations in dominantly meteoric fluids and higher NH4+ in rock-reacted fluids is used to hypothesize that NH4+ is sourced from NH4+-bearing biotite. However, biotite samples from the host rocks and ore-body minerals were analyzed by Fourier transform infrared (FTIR) microscopy and none-contained NH4+. However, the nitrogenase-encoding gene nifH was successfully amplified from DNA of the fluid sample with high NH4+, suggesting that subsurface microbes have the capability to fix N2. If so, unregulated nitrogen fixation may account for the relatively high NH4+ concentrations in the fluids. Additionally, the amoA and nxrB genes for archaeal ammonium monooxygenase and nitrite oxidoreductase, respectively, were amplified from the high NH4+ fluid DNA, while bacterial amoA genes were not. Putative nitrifying organisms are closely related to ammonium-oxidizing Crenarchaeota and nitrite-oxidizing Nitrospira detected in other subsurface sites based upon 16S rRNA sequence analysis. Thermodynamic calculations underscore the importance of NH4+ as an energy source in a subsurface nitrification pathway. These results suggest that the subsurface microbial community at Henderson is adapted to the low nutrient and energy environment by their capability of fixing nitrogen, and that fixed nitrogen may support subsurface biomass via

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.

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

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

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.

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.

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

Restoration using Azolla imbricata increases nitrogen functional bacterial groups and genes in soil.

PubMed

Lu, Xiao-Ming; Lu, Peng-Zhen; Yang, Ke

2017-05-01

Microbial groups are major factors that influence soil function. Currently, there is a lack of studies on microbial functional groups. Although soil microorganisms play an important role in the nitrogen cycle, systematic studies of the effects of environmental factors on microbial populations in relation to key metabolic processes in the nitrogen cycle are seldom reported. In this study, we conducted a systematic analysis of the changes in nitrogen functional groups in mandarin orange garden soil treated with Azolla imbricata. The structures of the major functional bacterial groups and the functional gene abundances involved in key processes of the soil nitrogen cycle were analyzed using high-throughput sequencing (HTS) and quantitative real-time PCR, respectively. The results indicated that returning A. imbricata had an important influence on the composition of soil nitrogen functional bacterial communities. Treatment with A. imbricata increased the diversity of the nitrogen functional bacteria. The abundances of nitrogen functional genes were significantly higher in the treated soil compared with the control soil. Both the diversity of the major nitrogen functional bacteria (nifH bacteria, nirK bacteria, and narG bacteria) and the abundances of nitrogen functional genes in the soil showed significant positive correlations with the soil pH, the organic carbon content, available nitrogen, available phosphorus, and NH 4 + -N and NO 3 - -N contents. Treatment with 12.5 kg fresh A. imbricata per mandarin orange tree was effective to improve the quality of the mandarin orange garden soil. This study analyzed the mechanism of the changes in functional bacterial groups and genes involved in key metabolic processes of the nitrogen cycle in soil treated by A. imbricata.

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....

RELATIONS BETWEEN BACTERIAL NITROGEN METABOLISM AND GROWTH EFFICIENCY IN AN ESTUARINE AND AN OPEN-WATER ECOSYSTEM

EPA Science Inventory

Bacterial uptake or release of dissolved nitrogen compounds (amino nitrogen, urea, ammonium and nitrate) were examined in 0.8 |m filtered water from an estuary (Santa Rosa Sound [SRS], northwestern Florida) and an open-water location in the Gulf of Mexico [GM]. The bacterial nutr...

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

Woody encroachment impacts on ecosystem nitrogen cycling: fixation, storage and gas loss

NASA Astrophysics Data System (ADS)

Soper, F.; Sparks, J. P.

2016-12-01

Woody encroachment is a pervasive land cover change throughout the tropics and subtropics. Encroachment is frequently catalyzed by nitrogen (N)-fixing trees and the resulting N inputs have the potential to alter whole-ecosystem N cycling, accumulation and loss. In the southern US, widespread encroachment by legume Prosopis glandulosa is associated with increased soil total N storage, inorganic N concentrations, and net mineralization and nitrification rates. To better understand the effects of this process on ecosystem N cycling, we investigated patterns of symbiotic N fixation, N accrual and soil N trace gas and N2 emissions during Prosopis encroachment into the southern Rio Grande Plains. Analyses of d15N in foliage, xylem sap and plant-available soil N suggested that N fixation rates vary seasonally, inter-annually and as a function of plant age and abiotic conditions. Applying a small-scale mass balance model to soil N accrual around individual trees (accounting for atmospheric inputs, and gas and hydrologic losses) generated current fixation estimates of 11 kg N ha-1 yr-1, making symbiotic fixation the largest input of N to the ecosystem. However, soil N accrual and increased cycling rates did not translate into increased N gas losses. Two years of field measurements of a complete suite of N trace gases (ammonia, nitrous oxide, nitric oxide and other oxidized N compounds) found no difference in flux between upland Prosopis groves and adjacent unencroached grasslands. Total emissions average 0.56-0.65 kg N ha-1 yr-1, comparable to other southern US grasslands. Lab incubations suggested that N2 losses are likely to be low, with field oxygen conditions not usually conducive to denitrification. Taken together, results suggest that this ecosystem is currently experiencing a period of significant net N accrual, driven by fixation under ongoing encroachment. Given the large scale of woody legume encroachment in the USA, this process is likely to contribute

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.

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.

Bacterial quorum sensing and nitrogen cycling in rhizosphere soil

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

DeAngelis, K.M.; Lindow, S.E.; Firestone, M.K.

2008-10-01

Plant photosynthate fuels carbon-limited microbial growth and activity, resulting in increased rhizosphere nitrogen (N)-mineralization. Most soil organic N is macromolecular (chitin, protein, nucleotides); enzymatic depolymerization is likely rate-limiting for plant N accumulation. Analyzing Avena (wild oat) planted in microcosms containing sieved field soil, we observed increased rhizosphere chitinase and protease specific activities, bacterial cell densities, and dissolved organic nitrogen (DON) compared to bulk soil. Low-molecular weight DON (<3000 Da) was undetectable in bulk soil but comprised 15% of rhizosphere DON. Extracellular enzyme production in many bacteria requires quorum sensing (QS), cell-density dependent group behavior. Because proteobacteria are considered major rhizospheremore » colonizers, we assayed the proteobacterial QS signals acyl-homoserine lactones (AHLs), which were significantly increased in the rhizosphere. To investigate the linkage between soil signaling and N cycling, we characterized 533 bacterial isolates from Avena rhizosphere: 24% had chitinase or protease activity and AHL production; disruption of QS in 7 of 8 eight isolates disrupted enzyme activity. Many {alpha}-Proteobacteria were newly found with QS-controlled extracellular enzyme activity. Enhanced specific activities of N-cycling enzymes accompanied by bacterial density-dependent behaviors in rhizosphere soil gives rise to the hypothesis that QS could be a control point in the complex process of rhizosphere N-mineralization.« less

Deep-Sea Archaea Fix and Share Nitrogen in Methane-Consuming Microbial Consortia

NASA Astrophysics Data System (ADS)

Dekas, Anne E.; Poretsky, Rachel S.; Orphan, Victoria J.

2009-10-01

Nitrogen-fixing (diazotrophic) microorganisms regulate productivity in diverse ecosystems; however, the identities of diazotrophs are unknown in many oceanic environments. Using single-cell-resolution nanometer secondary ion mass spectrometry images of 15N incorporation, we showed that deep-sea anaerobic methane-oxidizing archaea fix N2, as well as structurally similar CN-, and share the products with sulfate-reducing bacterial symbionts. These archaeal/bacterial consortia are already recognized as the major sink of methane in benthic ecosystems, and we now identify them as a source of bioavailable nitrogen as well. The archaea maintain their methane oxidation rates while fixing N2 but reduce their growth, probably in compensation for the energetic burden of diazotrophy. This finding extends the demonstrated lower limits of respiratory energy capable of fueling N2 fixation and reveals a link between the global carbon, nitrogen, and sulfur cycles.

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.

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.

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

Responsiveness of soil nitrogen fractions and bacterial communities to afforestation in the Loess Hilly Region (LHR) of China

NASA Astrophysics Data System (ADS)

Ren, Chengjie; Sun, Pingsheng; Kang, Di; Zhao, Fazhu; Feng, Yongzhong; Ren, Guangxin; Han, Xinhui; Yang, Gaihe

2016-06-01

In the present paper, we investigated the effects of afforestation on nitrogen fractions and microbial communities. A total of 24 soil samples were collected from farmland (FL) and three afforested lands, namely Robinia pseudoacacia L (RP), Caragana korshinskii Kom (CK), and abandoned land (AL), which have been arable for the past 40 years. Quantitative PCR and Illumina sequencing of 16S rRNA genes were used to analyze soil bacterial abundance, diversity, and composition. Additionally, soil nitrogen (N) stocks and fractions were estimated. The results showed that soil N stock, N fractions, and bacterial abundance and diversity increased following afforestation. Proteobacteria, Acidobacteria, and Actinobacteria were the dominant phyla of soil bacterial compositions. Overall, soil bacterial compositions generally changed from Actinobacteria (Acidobacteria)-dominant to Proteobacteria-dominant following afforestation. Soil N fractions, especially for dissolved organic nitrogen (DON), were significantly correlated with most bacterial groups and bacterial diversity, while potential competitive interactions between Proteobacteria (order Rhizobiales) and Cyanobacteria were suggested. In contrast, nitrate nitrogen (NO3--N) influenced soil bacterial compositions less than other N fractions. Therefore, the present study demonstrated that bacterial diversity and specific species respond to farmland-to-forest conversion and hence have the potential to affect N dynamic processes in the Loess Plateau.

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

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

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

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

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.

Coliform Bacteria and Nitrogen Fixation in Pulp and Paper Mill Effluent Treatment Systems

PubMed Central

Gauthier, Francis; Neufeld, Josh D.; Driscoll, Brian T.; Archibald, Frederick S.

2000-01-01

The majority of pulp and paper mills now biotreat their combined effluents using activated sludge. On the assumption that their wood-based effluents have negligible fixed N, and that activated-sludge microorganisms will not fix significant N, these mills routinely spend large amounts adding ammonia or urea to their aeration tanks (bioreactors) to permit normal biomass growth. N2 fixation in seven Eastern Canadian pulp and paper mill effluent treatment systems was analyzed using acetylene reduction assays, quantitative nitrogenase (nifH) gene probing, and bacterial isolations. In situ N2 fixation was undetectable in all seven bioreactors but was present in six associated primary clarifiers. One primary clarifier was studied in greater detail. Approximately 50% of all culturable cells in the clarifier contained nifH, of which >90% were Klebsiella strains. All primary-clarifier coliform bacteria growing on MacConkey agar were identified as klebsiellas, and all those probed contained nifH. In contrast, analysis of 48 random coliform isolates from other mill water system locations showed that only 24 (50%) possessed the nifH gene, and only 13 (27%) showed inducible N2-fixing activity. Thus, all the pulp and paper mill primary clarifiers tested appeared to be sites of active N2 fixation (0.87 to 4.90 mg of N liter−1 day−1) and a microbial community strongly biased toward this activity. This may also explain why coliform bacteria, especially klebsiellas, are indigenous in pulp and paper mill water systems. PMID:11097883

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.

Transcriptional Analysis of an Ammonium-Excreting Strain of Azotobacter vinelandii Deregulated for Nitrogen Fixation

PubMed Central

Plunkett, Mary H.; Natarajan, Velmurugan; Mus, Florence; Knutson, Carolann M.; Peters, John W.

2017-01-01

ABSTRACT Biological nitrogen fixation is accomplished by a diverse group of organisms known as diazotrophs and requires the function of the complex metalloenzyme nitrogenase. Nitrogenase and many of the accessory proteins required for proper cofactor biosynthesis and incorporation into the enzyme have been characterized, but a complete picture of the reaction mechanism and key cellular changes that accompany biological nitrogen fixation remain to be fully elucidated. Studies have revealed that specific disruptions of the antiactivator-encoding gene nifL result in the deregulation of the nif transcriptional activator NifA in the nitrogen-fixing bacterium Azotobacter vinelandii, triggering the production of extracellular ammonium levels approaching 30 mM during the stationary phase of growth. In this work, we have characterized the global patterns of gene expression of this high-ammonium-releasing phenotype. The findings reported here indicated that cultures of this high-ammonium-accumulating strain may experience metal limitation when grown using standard Burk's medium, which could be amended by increasing the molybdenum levels to further increase the ammonium yield. In addition, elevated levels of nitrogenase gene transcription are not accompanied by a corresponding dramatic increase in hydrogenase gene transcription levels or hydrogen uptake rates. Of the three potential electron donor systems for nitrogenase, only the rnf1 gene cluster showed a transcriptional correlation to the increased yield of ammonium. Our results also highlight several additional genes that may play a role in supporting elevated ammonium production in this aerobic nitrogen-fixing model bacterium. IMPORTANCE The transcriptional differences found during stationary-phase ammonium accumulation show a strong contrast between the deregulated (nifL-disrupted) and wild-type strains and what was previously reported for the wild-type strain under exponential-phase growth conditions. These results

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

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.

Long-Term Nitrogen Amendment Alters the Diversity and Assemblage of Soil Bacterial Communities in Tallgrass Prairie

PubMed Central

Todd, Timothy C.; Blair, John M.; Herman, Michael A.

2013-01-01

Anthropogenic changes are altering the environmental conditions and the biota of ecosystems worldwide. In many temperate grasslands, such as North American tallgrass prairie, these changes include alteration in historically important disturbance regimes (e.g., frequency of fires) and enhanced availability of potentially limiting nutrients, particularly nitrogen. Such anthropogenically-driven changes in the environment are known to elicit substantial changes in plant and consumer communities aboveground, but much less is known about their effects on soil microbial communities. Due to the high diversity of soil microbes and methodological challenges associated with assessing microbial community composition, relatively few studies have addressed specific taxonomic changes underlying microbial community-level responses to different fire regimes or nutrient amendments in tallgrass prairie. We used deep sequencing of the V3 region of the 16S rRNA gene to explore the effects of contrasting fire regimes and nutrient enrichment on soil bacterial communities in a long-term (20 yrs) experiment in native tallgrass prairie in the eastern Central Plains. We focused on responses to nutrient amendments coupled with two extreme fire regimes (annual prescribed spring burning and complete fire exclusion). The dominant bacterial phyla identified were Proteobacteria, Verrucomicrobia, Bacteriodetes, Acidobacteria, Firmicutes, and Actinobacteria and made up 80% of all taxa quantified. Chronic nitrogen enrichment significantly impacted bacterial community diversity and community structure varied according to nitrogen treatment, but not phosphorus enrichment or fire regime. We also found significant responses of individual bacterial groups including Nitrospira and Gammaproteobacteria to long-term nitrogen enrichment. Our results show that soil nitrogen enrichment can significantly alter bacterial community diversity, structure, and individual taxa abundance, which have important

Organic carbon and nitrogen availability determine bacterial community composition in paddy fields of the Indo-Gangetic plain.

PubMed

Kumar, Arvind; Rai, Lal Chand

2017-07-01

Soil quality is an important factor and maintained by inhabited microorganisms. Soil physicochemical characteristics determine indigenous microbial population and rice provides food security to major population of the world. Therefore, this study aimed to assess the impact of physicochemical variables on bacterial community composition and diversity in conventional paddy fields which could reflect a real picture of the bacterial communities operating in the paddy agro-ecosystem. To fulfill the objective; soil physicochemical characterization, bacterial community composition and diversity analysis was carried out using culture-independent PCR-DGGE method from twenty soils distributed across eight districts. Bacterial communities were grouped into three clusters based on UPGMA cluster analysis of DGGE banding pattern. The linkage of measured physicochemical variables with bacterial community composition was analyzed by canonical correspondence analysis (CCA). CCA ordination biplot results were similar to UPGMA cluster analysis. High levels of species-environment correlations (0.989 and 0.959) were observed and the largest proportion of species data variability was explained by total organic carbon (TOC), available nitrogen, total nitrogen and pH. Thus, results suggest that TOC and nitrogen are key regulators of bacterial community composition in the conventional paddy fields. Further, high diversity indices and evenness values demonstrated heterogeneity and co-abundance of the bacterial communities.

Do foliar endophytic bacteria fix nitrogen?

NASA Astrophysics Data System (ADS)

Kueppers, L. M.; Moyes, A. B.; Frank, C.; Pett-Ridge, J.; Carper, D.; Vandehey, N.; O'Neil, J.; Dekas, A.

2015-12-01

Endophytic microorganisms - bacteria and fungi that live inside healthy plant tissue - are a relatively unexplored source of functional diversity in natural ecosystems. Prior to modern sequencing technology, detecting uncultured endophytic bacteria and assessing their putative functions was challenging. However, recent work has revealed a remarkable diversity of as yet non-culturable endophytic taxa and is beginning to identify functional roles within plant microbiomes. We recently examined bacterial communities in the foliage of a long-lived, high-elevation conifer species, limber pine (Pinus flexilis), and discovered a community strongly dominated by acetic acid bacteria (Acetobacteraceae), with several taxa closely related to known nitrogen fixers. Given limber pine's status as a pioneer species that is able to grow in low fertility soils, we hypothesized that this bacterial community has a potential functional role in fixing atmospheric nitrogen, providing a source of this limiting nutrient to the host tree. We used the radioisotope 13N2 to confirm that N2 rapidly diffuses into pine needles, where it could potentially be fixed. With an acetylene reduction assay we confirmed nitrogenase enzyme activity inside excised twigs 4 times over a growing season, and estimate potential rates of N2 fixation at 0.1 nmol N2 g needle-1 hr-1. Scaled to the stand level, this N input could be on the order of ~20 mg N m-2 d-1 over a growing season. While these rates are low, the long lifespan of individual trees (~1000 years) makes them biologically meaningful. Still, measured rates of acetylene reduction and bulk 15N2 incorporation are quite variable in space and time. Much work remains to better characterize the plant-microbial interactions in this system, including the rates of nitrogen fixation and their variability over the growing season, across edaphic conditions, among host species, and through plant development; and to determine which community members are responsible

Evolutionary Dynamics of Nitrogen Fixation in the Legume–Rhizobia Symbiosis

PubMed Central

Fujita, Hironori; Aoki, Seishiro; Kawaguchi, Masayoshi

2014-01-01

The stabilization of host–symbiont mutualism against the emergence of parasitic individuals is pivotal to the evolution of cooperation. One of the most famous symbioses occurs between legumes and their colonizing rhizobia, in which rhizobia extract nutrients (or benefits) from legume plants while supplying them with nitrogen resources produced by nitrogen fixation (or costs). Natural environments, however, are widely populated by ineffective rhizobia that extract benefits without paying costs and thus proliferate more efficiently than nitrogen-fixing cooperators. How and why this mutualism becomes stabilized and evolutionarily persists has been extensively discussed. To better understand the evolutionary dynamics of this symbiosis system, we construct a simple model based on the continuous snowdrift game with multiple interacting players. We investigate the model using adaptive dynamics and numerical simulations. We find that symbiotic evolution depends on the cost–benefit balance, and that cheaters widely emerge when the cost and benefit are similar in strength. In this scenario, the persistence of the symbiotic system is compatible with the presence of cheaters. This result suggests that the symbiotic relationship is robust to the emergence of cheaters, and may explain the prevalence of cheating rhizobia in nature. In addition, various stabilizing mechanisms, such as partner fidelity feedback, partner choice, and host sanction, can reinforce the symbiotic relationship by affecting the fitness of symbionts in various ways. This result suggests that the symbiotic relationship is cooperatively stabilized by various mechanisms. In addition, mixed nodule populations are thought to encourage cheater emergence, but our model predicts that, in certain situations, cheaters can disappear from such populations. These findings provide a theoretical basis of the evolutionary dynamics of legume–rhizobia symbioses, which is extendable to other single-host, multiple

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.

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.

Biological nitrogen fixation in sugar cane: A key to energetically viable biofuel production

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

Boddey, R.M.

1995-05-01

The advantages of producing biofuels to replace fossil energy sources are derived from the fact that the energy accumulated in the biomass in captured directly from photosynthesis and is thus renewable, and that the cycle of carbon dioxide fixation by the crop, followed by burning of the fuel makes no overall contribution to atmospheric CO{sub 2} or, consequently, to global warming. However, these advantages are negated if large quantities of fossil fuels need to be used to grow or process the biofuel crop. In this regard, the Brazilian bioethanol program, based on the fermentation/distillation of sugar cane juice, is particularlymore » favorable, not only because the crop is principally hand harvested, but also because of the low nitrogen fertilizer use on sugar cane in Brazil. Recent {sup 15}N and N balance studies have shown that in some Brazilian cane varieties, high yields are possible without N fertilization because the plants are able to obtain large contributions of nitrogen from plant-associated biological N{sub 2} fixation (BNF). The N{sub 2}-fixing acid-tolerant bacterium Acetobacter diazotrophicus was first found to occur within roots, stems, and leaves of sugar cane. Subsequently, two species of Herbaspirillum also have been found to occur within the interior of all sugar cane tissues. The discovery of these, and other N{sub 2}-fixing bacteria that survive poorly in soil but thrive within plant tissue (endophytic bacteria), may account for the high BNF contributions observed in sugar cane. Further study of this system should allow the gradual elimination of N fertilizer use on sugar cane, at least in Brazil, and opens up the possibility of the extension of this efficient N{sub 2}-fixing system to cereal and other crops with consequent immense potential benefits to tropical agriculture. 44 refs., 9 figs., 4 tabs.« less

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

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.

Effect of chronic oil pollution on salt-marsh nitrogen fixation (acetylene redution). [Spartina alterniflora

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

Thomson, A.D.; Webb, K.L.

1984-03-01

Annual acetylene reduction rates associated with intertidal communities in a chronically oil polluted Virgina salt marsh were compared to rates measured in an undisturbed marsh. Chronic oil treatment resulted in visible damage to the higher plants of the Spartina alterniflora zones; however, vegetation-associated acetylene reduction was not different from the untreated control. Sediment rates generally were affected little by oil application, except during the summer when rates in the median tidal elevation zones were considerably higher than those of the control. Acetylene reduction occurred in all transects, each of which extended from upper mudflat to the Spartina patens zone. Intertidalmore » sediment acetylene reduction was patchy, both spatially and seasonally. Estimated rates were greatest near the surface; free-living bacterial N/sub 2/ fixation activity averaged 2.23 mg N per m/sup 2/ per d (range = undetectable to 365 mg N per m/sup 2/ per d) in the untreated and 3.17 mg N per m/sup 2/ per d (range = undetectable to 564 mg N per m/sup 2/ per d) in the oil-treated marsh during the year. Vegetation-associated N/sub 2/ fixation activity yielded highest overall mean rates (156 mg N per M/sub 2/ per d). The seasonal pattern of sediment and vegetation-associated fixation may be controlled by temperature and availability of oxidizable substrates. 39 references, 2 figures, 5 tables.« less

Metagenomic assessment of the potential microbial nitrogen pathways in the rhizosphere of a mediterranean forest after a wildfire.

PubMed

Cobo-Díaz, José F; Fernández-González, Antonio J; Villadas, Pablo J; Robles, Ana B; Toro, Nicolás; Fernández-López, Manuel

2015-05-01

Wildfires are frequent in the forests of the Mediterranean Basin and have greatly influenced this ecosystem. Changes to the physical and chemical properties of the soil, due to fire and post-fire conditions, result in alterations of both the bacterial communities and the nitrogen cycle. We explored the effects of a holm oak forest wildfire on the rhizospheric bacterial communities involved in the nitrogen cycle. Metagenomic data of the genes involved in the nitrogen cycle showed that both the undisturbed and burned rhizospheres had a conservative nitrogen cycle with a larger number of sequences related to the nitrogen incorporation pathways and a lower number for nitrogen output. However, the burned rhizosphere showed a statistically significant increase in the number of sequences for nitrogen incorporation (allantoin utilization and nitrogen fixation) and a significantly lower number of sequences for denitrification and dissimilatory nitrite reductase subsystems, possibly in order to compensate for nitrogen loss from the soil after burning. The genetic potential for nitrogen incorporation into the ecosystem was assessed through the diversity of the nitrogenase reductase enzyme, which is encoded by the nifH gene. We found that nifH gene diversity and richness were lower in burned than in undisturbed rhizospheric soils. The structure of the bacterial communities involved in the nitrogen cycle showed a statistically significant increase of Actinobacteria and Firmicutes phyla after the wildfire. Both approaches showed the important role of gram-positive bacteria in the ecosystem after a wildfire.

Inactivation of Lactobacillus rhamnosus GG by fixation modifies its probiotic properties.

PubMed

Markowicz, C; Kubiak, P; Grajek, W; Schmidt, M T

2016-01-01

Probiotics are microorganisms that have beneficial effects on the host and are safe for oral intake in a suitable dose. However, there are situations in which the administration of living microorganisms poses a risk for immunocompromised host. The objective of this study was to evaluate the influence of several fixation methods on selected biological properties of Lactobacillus rhamnosus GG that are relevant to its probiotic action. Fixation of the bacterial cells with ethanol, 2-propanol, glutaraldehyde, paraformaldehyde, and heat treatment resulted in a significant decrease of alkaline phosphatase, peroxidase, and β-galactosidase activities. Most of the fixation procedures reduced bacterial cell hydrophobicity and increased adhesion capacity. The fixation procedures resulted in a different perception of the bacterial cells by enterocytes, which was shown as changes in gene expression in enterocytes. The results show that some procedures of inactivation allow a fraction of the enzymatic activity to be maintained. The adhesion properties of the bacterial cells were enhanced, but the response of enterocytes to fixed cells was different than to live bacteria. Inactivation allows maintenance and modification of some of the properties of the bacterial cells.

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.

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.

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

Functional Genomics Approaches to Studying Symbioses between Legumes and Nitrogen-Fixing Rhizobia.

PubMed

Lardi, Martina; Pessi, Gabriella

2018-05-18

Biological nitrogen fixation gives legumes a pronounced growth advantage in nitrogen-deprived soils and is of considerable ecological and economic interest. In exchange for reduced atmospheric nitrogen, typically given to the plant in the form of amides or ureides, the legume provides nitrogen-fixing rhizobia with nutrients and highly specialised root structures called nodules. To elucidate the molecular basis underlying physiological adaptations on a genome-wide scale, functional genomics approaches, such as transcriptomics, proteomics, and metabolomics, have been used. This review presents an overview of the different functional genomics approaches that have been performed on rhizobial symbiosis, with a focus on studies investigating the molecular mechanisms used by the bacterial partner to interact with the legume. While rhizobia belonging to the alpha-proteobacterial group (alpha-rhizobia) have been well studied, few studies to date have investigated this process in beta-proteobacteria (beta-rhizobia).

Stable Isotope Evidence of Variation in Nitrogen Fixation by Cyanobacteria in Coastal Ecosystems

NASA Astrophysics Data System (ADS)

Paul, V.; Clementz, M.

2006-12-01

Increased nutrient loading via both natural and anthropogenic factors has been reported as one possible mechanism for the recent increase in the occurrence and intensity of harmful algal blooms (HAB) in coastal ecosystems. Influx of iron, phosphorous, and organic carbon have proven to be significant stimulating factors for HAB, since the benthic cyanobacteria that often make up these blooms are capable of nitrogen-fixation and require these nutrients for this process as well as photosynthesis. These cyanobacteria can switch to direct uptake of dissolved inorganic nitrogen (DIN), however, when concentrations are high enough to energetically favor this source, suggesting that high nitrogen input may also stimulate HAB. Given the distinct isotope differences between atmospheric N2 (0‰) and anthropogenic sources of DIN (>6‰), measurement of the δ15N composition of cyanobacteria can provide a means of gauging the relative significance of anthropogenic versus atmospheric nitrogen to the growth of these blooms. Likewise, the δ13C composition of these primary producers is controlled by the δ13C composition of the DIC, and can be a second tracer of anthropogenic influx into marine ecosystems. A combined approach using both isotope tracers was employed to determine the significance of anthropogenic nitrogen on HAB in subtropical/tropical coastal marine ecosystems. Samples of cyanobacteria and associated macroalgae were collected from three coastal sites in Guam (Facpi Point, Tanguisson, and Ypao Beach), one locality in Hawaii, and three sites in southern Florida (Pepper Park, Fort Lauderdale, Florida Keys). Following removal of marine carbonates via an acid rinse, the δ13C and δ15N values were determined for each species. Cyanobacterial δ15N values ranged from -2.3‰ to 7.7‰ with the highest values reported from sites in Guam. Only cyanobacteria sampled from Hawaii showed no isotope evidence of an anthropogenic source for nitrogen. A strong negative correlation

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

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

Heterotrophic bacterial production and metabolic balance during the VAHINE mesocosm experiment in the New Caledonia lagoon

NASA Astrophysics Data System (ADS)

Van Wambeke, F.; Pfreundt, U.; Barani, A.; Berthelot, H.; Moutin, T.; Rodier, M.; Hess, W. R.; Bonnet, S.

2015-12-01

N2 fixation fuels ~ 50 % of new primary production in the oligotrophic South Pacific Ocean. The VAHINE mesocosm experiment designed to track the fate of diazotroph derived nitrogen (DDN) in the New Caledonia lagoon. Here, we examined the temporal dynamics of heterotrophic bacterial production during this experiment. Three replicate large-volume (~ 50 m3) mesocosms were deployed and were intentionally fertilized with dissolved inorganic phosphorus (DIP) to stimulate N2 fixation. We specifically examined relationships between N2 fixation rates and primary production, determined bacterial growth efficiency and established carbon budgets of the system from the DIP fertilization to the end of the experiment (days 5-23). Heterotrophic bacterioplankton production (BP) and alkaline phosphatase activity (APA) were statistically higher during the second phase of the experiment (P2: days 15-23), when chlorophyll biomass started to increase compared to the first phase (P1: days 5-14). Among autotrophs, Synechococcus abundances increased during P2, possibly related to its capacity to assimilate leucine and to produce alkaline phosphatase. Bacterial growth efficiency based on the carbon budget was notably higher than generally cited for oligotrophic environments (27-43 %), possibly due to a high representation of proteorhodopsin-containing organisms within the picoplanctonic community. The carbon budget showed that the main fate of gross primary production (particulate + dissolved) was respiration (67 %), and export through sedimentation (17 %). BP was highly correlated with particulate primary production and chlorophyll biomass during both phases of the experiment but slightly correlated, and only during P2 phase, with N2 fixation rates. Our results suggest that most of the DDN reached the heterotrophic bacterial community through indirect processes, like mortality, lysis and grazing.

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.

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

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

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

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.

Temporal changes in the diazotrophic bacterial communities associated with Caribbean sponges Ircinia stroblina and Mycale laxissima

PubMed Central

Zhang, Fan; Vicente, Jan; Hill, Russell T.

2014-01-01

Sponges that harbor microalgal or, cyanobacterial symbionts may benefit from photosynthetically derived carbohydrates, which are rich in carbon but devoid of nitrogen, and may therefore encounter nitrogen limitation. Diazotrophic communities associated with two Caribbean sponges, Ircinia strobilina and Mycale laxissima were studied in a time series during which three individuals of each sponge were collected in four time points (5:00 AM, 12:00 noon, 5:00 PM, 10:00 PM). nifH genes were successfully amplified from the corresponding gDNA and cDNA pools and sequenced by high throughput 454 amplicon sequencing. In both sponges, over half the nifH transcripts were classified as from cyanobacteria and the remainder from heterotrophic bacteria. We found various groups of bacteria actively expressing the nifH gene during the entire day-night cycle, an indication that the nitrogen fixation potential was fully exploited by different nitrogen fixing bacteria groups associated with their hosts. This study showed for the first time the dynamic changes in the activity of the diazotrophic bacterial communities in marine sponges. Our study expands understanding of the diazotrophic groups that contribute to the fixed nitrogen pool in the benthic community. Sponge bacterial community-associated diazotrophy may have an important impact on the nitrogen biogeochemical cycle in the coral reef ecosystem. PMID:25389420

Temporal changes in the diazotrophic bacterial communities associated with Caribbean sponges Ircinia stroblina and Mycale laxissima.

PubMed

Zhang, Fan; Vicente, Jan; Hill, Russell T

2014-01-01

Sponges that harbor microalgal or, cyanobacterial symbionts may benefit from photosynthetically derived carbohydrates, which are rich in carbon but devoid of nitrogen, and may therefore encounter nitrogen limitation. Diazotrophic communities associated with two Caribbean sponges, Ircinia strobilina and Mycale laxissima were studied in a time series during which three individuals of each sponge were collected in four time points (5:00 AM, 12:00 noon, 5:00 PM, 10:00 PM). nifH genes were successfully amplified from the corresponding gDNA and cDNA pools and sequenced by high throughput 454 amplicon sequencing. In both sponges, over half the nifH transcripts were classified as from cyanobacteria and the remainder from heterotrophic bacteria. We found various groups of bacteria actively expressing the nifH gene during the entire day-night cycle, an indication that the nitrogen fixation potential was fully exploited by different nitrogen fixing bacteria groups associated with their hosts. This study showed for the first time the dynamic changes in the activity of the diazotrophic bacterial communities in marine sponges. Our study expands understanding of the diazotrophic groups that contribute to the fixed nitrogen pool in the benthic community. Sponge bacterial community-associated diazotrophy may have an important impact on the nitrogen biogeochemical cycle in the coral reef ecosystem.

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.

The influence of woody encroachment on the nitrogen cycle: fixation, storage and gas loss

NASA Astrophysics Data System (ADS)

Soper, F.; Sparks, J. P.

2015-12-01

Woody encroachment is a pervasive land cover change throughout the tropics and subtropics. Encroachment is frequently catalyzed by nitrogen (N)-fixing trees and the resulting N inputs potentially alter whole-ecosystem N cycling, accumulation and loss. In the southern US, widespread encroachment by legume Prosopis glandulosa is associated with increased soil total N storage, inorganic N concentrations, and net mineralization and nitrification rates. To better understand the effects of this process on ecosystem N cycling, we investigated patterns of symbiotic N fixation, N accrual and soil N trace gas and N2 emissions during Prosopis encroachment into the southern Rio Grande Plains. Analyses of d15N in foliage, xylem sap and plant-available soil N suggested that N fixation rates increase with tree age and are influenced by abiotic conditions. A model of soil N accrual around individual trees, accounting for atmospheric inputs and gas losses, generates lifetimes N fixation estimates of up to 9 kg for a 100-year-old tree and current rates of 7 kg N ha-1 yr-1. However, these N inputs and increased soil cycling rates do not translate into increased N gas losses. Two years of field measurements of a complete suite of N trace gases (ammonia, nitrous oxide, nitric oxide and other oxidized N compounds) found no difference in flux between upland Prosopis groves and adjacent unencroached grasslands. Total emissions for both land cover types average 0.56-0.65 kg N ha-1 yr-1, comparable to other southern US grasslands. Additional lab experiments suggested that N2 losses are low and that field oxygen conditions are not usually conducive to denitrification. Taken together, results suggest that this ecosystem is currently experiencing a period of net N accrual under ongoing encroachment.

Key role of symbiotic dinitrogen fixation in tropical forest secondary succession

NASA Astrophysics Data System (ADS)

Batterman, Sarah A.; Hedin, Lars O.; van Breugel, Michiel; Ransijn, Johannes; Craven, Dylan J.; Hall, Jefferson S.

2013-10-01

Forests contribute a significant portion of the land carbon sink, but their ability to sequester CO2 may be constrained by nitrogen, a major plant-limiting nutrient. Many tropical forests possess tree species capable of fixing atmospheric dinitrogen (N2), but it is unclear whether this functional group can supply the nitrogen needed as forests recover from disturbance or previous land use, or expand in response to rising CO2 (refs 6, 8). Here we identify a powerful feedback mechanism in which N2 fixation can overcome ecosystem-scale deficiencies in nitrogen that emerge during periods of rapid biomass accumulation in tropical forests. Over a 300-year chronosequence in Panama, N2-fixing tree species accumulated carbon up to nine times faster per individual than their non-fixing neighbours (greatest difference in youngest forests), and showed species-specific differences in the amount and timing of fixation. As a result of fast growth and high fixation, fixers provided a large fraction of the nitrogen needed to support net forest growth (50,000kg carbon per hectare) in the first 12years. A key element of ecosystem functional diversity was ensured by the presence of different N2-fixing tree species across the entire forest age sequence. These findings show that symbiotic N2 fixation can have a central role in nitrogen cycling during tropical forest stand development, with potentially important implications for the ability of tropical forests to sequester CO2.

Key role of symbiotic dinitrogen fixation in tropical forest secondary succession.

PubMed

Batterman, Sarah A; Hedin, Lars O; van Breugel, Michiel; Ransijn, Johannes; Craven, Dylan J; Hall, Jefferson S

2013-10-10

Forests contribute a significant portion of the land carbon sink, but their ability to sequester CO2 may be constrained by nitrogen, a major plant-limiting nutrient. Many tropical forests possess tree species capable of fixing atmospheric dinitrogen (N2), but it is unclear whether this functional group can supply the nitrogen needed as forests recover from disturbance or previous land use, or expand in response to rising CO2 (refs 6, 8). Here we identify a powerful feedback mechanism in which N2 fixation can overcome ecosystem-scale deficiencies in nitrogen that emerge during periods of rapid biomass accumulation in tropical forests. Over a 300-year chronosequence in Panama, N2-fixing tree species accumulated carbon up to nine times faster per individual than their non-fixing neighbours (greatest difference in youngest forests), and showed species-specific differences in the amount and timing of fixation. As a result of fast growth and high fixation, fixers provided a large fraction of the nitrogen needed to support net forest growth (50,000 kg carbon per hectare) in the first 12 years. A key element of ecosystem functional diversity was ensured by the presence of different N2-fixing tree species across the entire forest age sequence. These findings show that symbiotic N2 fixation can have a central role in nitrogen cycling during tropical forest stand development, with potentially important implications for the ability of tropical forests to sequester CO2.

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.

A single evolutionary innovation drives the deep evolution of symbiotic N2-fixation in angiosperms

PubMed Central

Werner, Gijsbert D. A.; Cornwell, William K.; Sprent, Janet I.; Kattge, Jens; Kiers, E. Toby

2014-01-01

Symbiotic associations occur in every habitat on earth, but we know very little about their evolutionary histories. Current models of trait evolution cannot adequately reconstruct the deep history of symbiotic innovation, because they assume homogenous evolutionary processes across millions of years. Here we use a recently developed, heterogeneous and quantitative phylogenetic framework to study the origin of the symbiosis between angiosperms and nitrogen-fixing (N2) bacterial symbionts housed in nodules. We compile the largest database of global nodulating plant species and reconstruct the symbiosis’ evolution. We identify a single, cryptic evolutionary innovation driving symbiotic N2-fixation evolution, followed by multiple gains and losses of the symbiosis, and the subsequent emergence of ‘stable fixers’ (clades extremely unlikely to lose the symbiosis). Originating over 100 MYA, this innovation suggests deep homology in symbiotic N2-fixation. Identifying cryptic innovations on the tree of life is key to understanding the evolution of complex traits, including symbiotic partnerships. PMID:24912610

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.

Tricalcium phosphate solubilization and nitrogen fixation by newly isolated Aneurinibacillus aneurinilyticus CKMV1 from rhizosphere of Valeriana jatamansi and its growth promotional effect.

PubMed

Chauhan, Anjali; Guleria, Shiwani; Balgir, Praveen P; Walia, Abhishek; Mahajan, Rishi; Mehta, Preeti; Shirkot, Chand Karan

Aneurinibacillus aneurinilyticus strain CKMV1 was isolated from rhizosphere of Valeriana jatamansi and possessed multiple plant growth promoting traits like production of phosphate solubilization (260mg/L), nitrogen fixation (202.91nmolethylenemL -1 h -1 ), indole-3-acetic acid (IAA) (8.1μg/mL), siderophores (61.60%), HCN (hydrogen cyanide) production and antifungal activity. We investigated the ability of isolate CKMV1 to solubilize insoluble P via mechanism of organic acid production. High-performance liquid chromatography (HPLC) study showed that isolate CKMV1 produced mainly gluconic (1.34%) and oxalic acids. However, genetic evidences for nitrogen fixation and phosphate solubilization by organic acid production have been reported first time for A. aneurinilyticus strain CKMV1. A unique combination of glucose dehydrogenase (gdh) gene and pyrroloquinoline quinone synthase (pqq) gene, a cofactor of gdh involved in phosphate solubilization has been elucidated. Nitrogenase (nif H) gene for nitrogen fixation was reported from A. aneurinilyticus. It was notable that isolate CKMV1 exhibited highest antifungal against Sclerotium rolfsii (93.58%) followed by Fusarium oxysporum (64.3%), Dematophora necatrix (52.71%), Rhizoctonia solani (91.58%), Alternaria sp. (71.08%) and Phytophthora sp. (71.37%). Remarkable increase was observed in seed germination (27.07%), shoot length (42.33%), root length (52.6%), shoot dry weight (62.01%) and root dry weight (45.7%) along with NPK (0.74, 0.36, 1.82%) content of tomato under net house condition. Isolate CKMV1 possessed traits related to plant growth promotion, therefore, could be a potential candidate for the development of biofertiliser or biocontrol agent and this is the first study to include the Aneurinibacillus as PGPR. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

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.

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.

The fixation effect of a silk fibroin-bacterial cellulose composite plate in segmental defects of the zygomatic arch: an experimental study.

PubMed

Lee, Jung Min; Kim, Ji Heui; Lee, Ok Joo; Park, Chan Hum

2013-06-01

Bioresorbable fixation systems have been popular for the treatment of facial fractures. However, their mechanical properties are uncertain and complications have been reported. To overcome these problems, we developed a bioresorbable fixation plate using a composite of silk fibroin and bacterial cellulose (SF-BC) with biodegradability and increased biocompatibility. To investigate the regenerative effect of the bioresorbable SF-BC fixation plate on zygomatic arch defects in rats. In vivo animal study. The SF-BC composite plate had a tensile strength similar to that of a polylactic acid plate and a tight, pore-free microstructure. Bilateral segmental bone defects (2 mm in length) were created in the zygomatic arches of adult rats. One side was fixed with the SF-BC composite plate, and the other side was left without fixation. Academic research laboratory. Fifteen adult Sprague-Dawley rats. Fixation of the zygomatic arch defect with the SF-BC composite plate. Micro-computed tomography and histological evaluation of bone samples. Gross inspection revealed no specific complication. At 1, 2, 4, and 8 postoperative weeks, the zygomatic arches were explored by micro-computed tomography and histological examination. Control sides did not heal completely and showed bony degeneration and necrosis during the 8-week follow-up. However, we observed new bone formation in sides treated with the SF-BC composite plate, and bony defects were completely healed within 8 weeks. The SF-BC composite plate is a potential candidate for a new bioresorbable fixation system. Our composite material could considerably shorten bone regeneration time. Additional study of the control of biodegradability and mechanical properties of SF-BC composite plates and a comparative study with the resorbable plates currently in use should be undertaken.

Improving tribological and anti-bacterial properties of titanium external fixation pins through surface ceramic conversion.

PubMed

Dong, Huan; Mukinay, Tatiana; Li, Maojun; Hood, Richard; Soo, Sein Leung; Cockshott, Simon; Sammons, Rachel; Li, Xiaoying

2017-01-01

In this study, an advanced ceramic conversion surface engineering technology has been applied for the first time to self-drilling Ti6Al4V external fixation pins to improve their performance in terms of biomechanical, bio-tribological and antibacterial properties. Systematic characterisation of the ceramic conversion treated Ti pins was carried out using Scanning electron microscope, X-ray diffraction, Glow-discharge optical emission spectroscopy, nano- and micro-indentation and scratching; the biomechanical and bio-tribological properties of the surface engineered Ti pins were evaluated by insertion into high density bone simulation material; and the antibacterial behaviour was assessed with Staphylococcus aureus NCTC 6571. The experimental results have demonstrated that the surfaces of Ti6Al4V external fixation pins were successfully converted into a TiO 2 rutile layer (~2 μm in thickness) supported by an oxygen hardened case (~15 μm in thickness) with very good bonding due to the in-situ conversion nature. The maximum insertion force and temperature were reduced from 192N and 31.2 °C when using the untreated pins to 182N and 26.1 °C when the ceramic conversion treated pins were tested. This is mainly due to the significantly increased hardness (more than three times) and the effectively enhanced wear resistance of the cutting edge of the self-drilling Ti pins following the ceramic conversion treatment. The antibacterial tests also revealed that there was a significantly reduced number of bacteria isolated from the ceramic conversion treated pins compared to the untreated pins of around 50 % after 20 h incubation, P < 0.01 (0.0024). The results reported are encouraging and could pave the way towards high-performance anti-bacterial titanium external fixation pins with reduced pin-track infection and pin loosing.

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.

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.

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

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

A novel endo-hydrogenase activity recycles hydrogen produced by nitrogen fixation.

PubMed

Ng, Gordon; Tom, Curtis G S; Park, Angela S; Zenad, Lounis; Ludwig, Robert A

2009-01-01

Nitrogen (N(2)) fixation also yields hydrogen (H(2)) at 1:1 stoichiometric amounts. In aerobic diazotrophic (able to grow on N(2) as sole N-source) bacteria, orthodox respiratory hupSL-encoded hydrogenase activity, associated with the cell membrane but facing the periplasm (exo-hydrogenase), has nevertheless been presumed responsible for recycling such endogenous hydrogen. As shown here, for Azorhizobium caulinodans diazotrophic cultures open to the atmosphere, exo-hydrogenase activity is of no consequence to hydrogen recycling. In a bioinformatic analysis, a novel seven-gene A. caulinodans hyq cluster encoding an integral-membrane, group-4, Ni,Fe-hydrogenase with homology to respiratory complex I (NADH: quinone dehydrogenase) was identified. By analogy, Hyq hydrogenase is also integral to the cell membrane, but its active site faces the cytoplasm (endo-hydrogenase). An A. caulinodans in-frame hyq operon deletion mutant, constructed by "crossover PCR", showed markedly decreased growth rates in diazotrophic cultures; normal growth was restored with added ammonium--as expected of an H(2)-recycling mutant phenotype. Using A. caulinodans hyq merodiploid strains expressing beta-glucuronidase as promoter-reporter, the hyq operon proved strongly and specifically induced in diazotrophic culture; as well, hyq operon induction required the NIFA transcriptional activator. Therefore, the hyq operon is constituent of the nif regulon. Representative of aerobic N(2)-fixing and H(2)-recycling alpha-proteobacteria, A. caulinodans possesses two respiratory Ni,Fe-hydrogenases: HupSL exo-hydrogenase activity drives exogenous H(2) respiration, and Hyq endo-hydrogenase activity recycles endogenous H(2), specifically that produced by N(2) fixation. To benefit human civilization, H(2) has generated considerable interest as potential renewable energy source as its makings are ubiquitous and its combustion yields no greenhouse gases. As such, the reversible, group-4 Ni,Fe-hydrogenases, such

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.

Facultative anoxygenic photosynthesis in cyanobacteria driven by arsenite and sulfide with evidence for the support of nitrogen fixation

NASA Astrophysics Data System (ADS)

Wolfe-Simon, F.; Hoeft, S. E.; Baesman, S. M.; Oremland, R. S.

2010-12-01

The rise in atmospheric oxygen (O2) over geologic time is attributed to the evolution and widespread proliferation of oxygenic photosynthesis in cyanobacteria. However, cyanobacteria maintain a metabolic flexibility that may not always result in O2 release. In the environment, cyanobacteria may use a variety of alternative electron donors rather than water that are known to be used by other anoxygenic phototrophs (eg. purple sulfur bacteria) including reduced forms of sulfur, iron, nitrogen, and arsenic. Recent evidence suggests cyanobacteria actively take advantage of at least a few of these alternatives. We used a classical Winogradsky approach to enrich for cyanobacteria from the high salinity, elevated pH and arsenic-enriched waters of Mono Lake (CA). Experiments, optimized for cyanobacteria, revealed light-dependent, anaerobic arsenite-oxidation in sub-cultured sediment-free enrichments dominated by a filamentous cyanobacteria. We isolated and identified the dominant member of this enrichment to be a member of the Oscillatoriales by 16S rDNA. Addition of 1 mM arsenite induced facultative anoxygenic photosynthesis under continuous and circadian light. This isolate also oxidized sulfide under the same light-based conditions. Aerobic conditions elicited no arsenite oxidation in the light or dark and the isolate grew as a typical cyanobacterium using oxygenic photosynthesis. Under near-infrared light (700 nm) there was a direct correlation of enhanced growth with an increase in the rate arsenite or sulfide oxidation suggesting the use of photosystem I. Additionally, to test the wide-spread nature of this metabolism in the Oscillatoriales, we followed similar arsenite- and sulfide-driven facultative anoxygenic photosynthesis as well as nitrogen fixation (C2H2 reduction) in the axenic isolate Oscillatoria sp. CCMP 1731. Future characterization includes axenic isolation of the Mono Lake Oscillatoria sp. as well as the arsenite oxidase responsible for electron

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.

Soil Carbon-Fixation Rates and Associated Bacterial Diversity and Abundance in Three Natural Ecosystems.

PubMed

Lynn, Tin Mar; Ge, Tida; Yuan, Hongzhao; Wei, Xiaomeng; Wu, Xiaohong; Xiao, Keqing; Kumaresan, Deepak; Yu, San San; Wu, Jinshui; Whiteley, Andrew S

2017-04-01

CO 2 assimilation by autotrophic microbes is an important process in soil carbon cycling, and our understanding of the community composition of autotrophs in natural soils and their role in carbon sequestration of these soils is still limited. Here, we investigated the autotrophic C incorporation in soils from three natural ecosystems, i.e., wetland (WL), grassland (GR), and forest (FO) based on the incorporation of labeled C into the microbial biomass. Microbial assimilation of 14 C ( 14 C-MBC) differed among the soils from three ecosystems, accounting for 14.2-20.2% of 14 C-labeled soil organic carbon ( 14 C-SOC). We observed a positive correlation between the cbbL (ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) large-subunit gene) abundance, 14 C-SOC level, and 14 C-MBC concentration confirming the role of autotrophic bacteria in soil carbon sequestration. Distinct cbbL-bearing bacterial communities were present in each soil type; form IA and form IC RubisCO-bearing bacteria were most abundant in WL, followed by GR soils, with sequences from FO soils exclusively derived from the form IC clade. Phylogenetically, the diversity of CO 2 -fixing autotrophs and CO oxidizers differed significantly with soil type, whereas cbbL-bearing bacterial communities were similar when assessed using coxL. We demonstrate that local edaphic factors such as pH and salinity affect the C-fixation rate as well as cbbL and coxL gene abundance and diversity. Such insights into the effect of soil type on the autotrophic bacterial capacity and subsequent carbon cycling of natural ecosystems will provide information to enhance the sustainable management of these important natural ecosystems.

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 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 regulation of nitrogen fixation

NASA Astrophysics Data System (ADS)

Menge, D. N.

2012-12-01

I am not a proponent of including more detail in models simply because it makes them more realistic. More complexity increases the difficulty of model interpretation, so it only makes sense to include complexity if its benefit exceeds its costs. Biological nitrogen (N) fixation (BNF) is one process for which I feel the benefits of including greater complexity far outweigh the costs. I don't think that just because I work on BNF; I work on BNF because I think that. BNF, a microbial process carried out by free-living and symbiotic microbes, is the dominant N input to many ecosystems, the primary mechanism by which N deficiency can feed back to N inputs, and a main mechanism by which N surplus can develop. The dynamics of BNF, therefore, have huge implications for the rate of carbon uptake and the extent of CO2 fertilization, as well as N export to waterways and N2O emissions to the atmosphere. Unfortunately, there are serious deficiencies in our understanding of BNF. One main deficiency in our understanding is the extent to which various symbiotic N fixing organisms respond to imbalanced nutrition. Theory suggests that these responses, which I will call "strategies," have fundamental consequences for N fixer niches and ecosystem-level N and C cycling. Organisms that fix N regardless of whether they need it, a strategy that I will call "obligate," occupy post-disturbance niches and rapidly lead to N surplus. On the contrary, organisms that only fix as much N as they need, a "facultative" strategy, can occupy a wider range of successional niches, do not produce surplus N, and respond more rapidly to increased atmospheric CO2. In this talk I will show new results showing that consideration of these strategies could on its own explain the latitudinal distribution of symbiotic N fixing trees in North America. Specifically, the transition in N-fixing tree abundance from ~10% of basal area south of 35° latitude to ~1% of basal area north of 35° latitude that we observe

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.

Root nodule symbiosis in Lotus japonicus drives the establishment of distinctive rhizosphere, root, and nodule bacterial communities.

PubMed

Zgadzaj, Rafal; Garrido-Oter, Ruben; Jensen, Dorthe Bodker; Koprivova, Anna; Schulze-Lefert, Paul; Radutoiu, Simona

2016-12-06

Lotus japonicus has been used for decades as a model legume to study the establishment of binary symbiotic relationships with nitrogen-fixing rhizobia that trigger root nodule organogenesis for bacterial accommodation. Using community profiling of 16S rRNA gene amplicons, we reveal that in Lotus, distinctive nodule- and root-inhabiting communities are established by parallel, rather than consecutive, selection of bacteria from the rhizosphere and root compartments. Comparative analyses of wild-type (WT) and symbiotic mutants in Nod factor receptor5 (nfr5), Nodule inception (nin) and Lotus histidine kinase1 (lhk1) genes identified a previously unsuspected role of the nodulation pathway in the establishment of different bacterial assemblages in the root and rhizosphere. We found that the loss of nitrogen-fixing symbiosis dramatically alters community structure in the latter two compartments, affecting at least 14 bacterial orders. The differential plant growth phenotypes seen between WT and the symbiotic mutants in nonsupplemented soil were retained under nitrogen-supplemented conditions that blocked the formation of functional nodules in WT, whereas the symbiosis-impaired mutants maintain an altered community structure in the nitrogen-supplemented soil. This finding provides strong evidence that the root-associated community shift in the symbiotic mutants is a direct consequence of the disabled symbiosis pathway rather than an indirect effect resulting from abolished symbiotic nitrogen fixation. Our findings imply a role of the legume host in selecting a broad taxonomic range of root-associated bacteria that, in addition to rhizobia, likely contribute to plant growth and ecological performance.

Root nodule symbiosis in Lotus japonicus drives the establishment of distinctive rhizosphere, root, and nodule bacterial communities

PubMed Central

Zgadzaj, Rafal; Garrido-Oter, Ruben; Jensen, Dorthe Bodker; Koprivova, Anna; Schulze-Lefert, Paul; Radutoiu, Simona

2016-01-01

Lotus japonicus has been used for decades as a model legume to study the establishment of binary symbiotic relationships with nitrogen-fixing rhizobia that trigger root nodule organogenesis for bacterial accommodation. Using community profiling of 16S rRNA gene amplicons, we reveal that in Lotus, distinctive nodule- and root-inhabiting communities are established by parallel, rather than consecutive, selection of bacteria from the rhizosphere and root compartments. Comparative analyses of wild-type (WT) and symbiotic mutants in Nod factor receptor5 (nfr5), Nodule inception (nin) and Lotus histidine kinase1 (lhk1) genes identified a previously unsuspected role of the nodulation pathway in the establishment of different bacterial assemblages in the root and rhizosphere. We found that the loss of nitrogen-fixing symbiosis dramatically alters community structure in the latter two compartments, affecting at least 14 bacterial orders. The differential plant growth phenotypes seen between WT and the symbiotic mutants in nonsupplemented soil were retained under nitrogen-supplemented conditions that blocked the formation of functional nodules in WT, whereas the symbiosis-impaired mutants maintain an altered community structure in the nitrogen-supplemented soil. This finding provides strong evidence that the root-associated community shift in the symbiotic mutants is a direct consequence of the disabled symbiosis pathway rather than an indirect effect resulting from abolished symbiotic nitrogen fixation. Our findings imply a role of the legume host in selecting a broad taxonomic range of root-associated bacteria that, in addition to rhizobia, likely contribute to plant growth and ecological performance. PMID:27864511

Compartmentalized microbial composition, oxygen gradients and nitrogen fixation in the gut of Odontotaenius disjunctus.

PubMed

Ceja-Navarro, Javier A; Nguyen, Nhu H; Karaoz, Ulas; Gross, Stephanie R; Herman, Donald J; Andersen, Gary L; Bruns, Thomas D; Pett-Ridge, Jennifer; Blackwell, Meredith; Brodie, Eoin L

2014-01-01

Coarse woody debris is an important biomass pool in forest ecosystems that numerous groups of insects have evolved to take advantage of. These insects are ecologically important and represent useful natural analogs for biomass to biofuel conversion. Using a range of molecular approaches combined with microelectrode measurements of oxygen, we have characterized the gut microbiome and physiology of Odontotaenius disjunctus, a wood-feeding beetle native to the eastern United States. We hypothesized that morphological and physiological differences among gut regions would correspond to distinct microbial populations and activities. In fact, significantly different communities were found in the foregut (FG), midgut (MG)/posterior hindgut (PHG) and anterior hindgut (AHG), with Actinobacteria and Rhizobiales being more abundant toward the FG and PHG. Conversely, fermentative bacteria such as Bacteroidetes and Clostridia were more abundant in the AHG, and also the sole region where methanogenic Archaea were detected. Although each gut region possessed an anaerobic core, micron-scale profiling identified radial gradients in oxygen concentration in all regions. Nitrogen fixation was confirmed by (15)N2 incorporation, and nitrogenase gene (nifH) expression was greatest in the AHG. Phylogenetic analysis of nifH identified the most abundant transcript as related to Ni-Fe nitrogenase of a Bacteroidetes species, Paludibacter propionicigenes. Overall, we demonstrate not only a compartmentalized microbiome in this beetle digestive tract but also sharp oxygen gradients that may permit aerobic and anaerobic metabolism to occur within the same regions in close proximity. We provide evidence for the microbial fixation of N2 that is important for this beetle to subsist on woody biomass.

Comparison of the Rhizosphere Bacterial Communities of Zigongdongdou Soybean and a High-Methionine Transgenic Line of This Cultivar

PubMed Central

Ji, Jun; Wu, Haiying; Meng, Fang; Zhang, Mingrong; Zheng, Xiaobo; Wu, Cunxiang; Zhang, Zhengguang

2014-01-01

Previous studies have shown that methionine from root exudates affects the rhizosphere bacterial population involved in soil nitrogen fixation. A transgenic line of Zigongdongdou soybean cultivar (ZD91) that expresses Arabidopsis cystathionine γ-synthase resulting in an increased methionine production was examined for its influence to the rhizosphere bacterial population. Using 16S rRNA gene-based pyrosequencing analysis of the V4 region and DNA extracted from bacterial consortia collected from the rhizosphere of soybean plants grown in an agricultural field at the pod-setting stage, we characterized the populational structure of the bacterial community involved. In total, 87,267 sequences (approximately 10,908 per sample) were analyzed. We found that Acidobacteria, Proteobacteria, Bacteroidetes, Actinobacteria, Chloroflexi, Planctomycetes, Gemmatimonadetes, Firmicutes, and Verrucomicrobia constitute the dominant taxonomic groups in either the ZD91 transgenic line or parental cultivar ZD, and that there was no statistically significant difference in the rhizosphere bacterial community structure between the two cultivars. PMID:25079947

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...

[Automated analysis of bacterial preparations manufactured on automatic heat fixation and staining equipment].

PubMed

2012-01-01

Heat fixation of preparations was made in the fixation bath designed by EMKO (Russia). Programmable "Emkosteiner" (EMKO, Russia) was used for trial staining. Reagents set Micko-GRAM-NITsF was applied for Gram's method of staining. It was demostrated that automatic smear fixation equipment and programmable staining ensure high-quality imaging (1% chromaticity variation) good enough for standardization of Gram's staining of microbial preparations.

The impact of post-fire salvage logging on microbial nitrogen cyclers in Mediterranean forest soil.

PubMed

Pereg, Lily; Mataix-Solera, Jorge; McMillan, Mary; García-Orenes, Fuensanta

2018-04-01

Forest fires are a regular occurrence in the Mediterranean basin. High severity fires and post-fire management can affect biological, chemical and physical properties of soil, including the composition and abundance of soil microbial communities. Salvage logging is a post-fire management strategy, which involves the removal of burnt wood from land after a fire. The main objective of this work was to evaluate the impact of post-fire salvage logging and microaggregation on soil microbial communities, specifically on the abundance of nitrogen cyclers and, thus, the potential of the soil for microbial nitrogen cycling. The abundance of nitrogen cyclers was assessed by quantification of microbial nitrogen cycling genes in soil DNA, including nifH (involved in nitrogen fixation), nirS/K and nosZ (involved in denitrification), amoA-B and amoA-Arch (involved in bacterial and archaeal nitrification, respectively). It was demonstrated that salvage logging reduced bacterial load post-fire when compared to tree retention control and resulted in significant changes to the abundance of functional bacteria involved in nitrogen cycling. Microbial gene pools involved in various stages of the nitrogen cycle were larger in control soil than in soil subjected to post-fire salvage logging and were significantly correlated with organic matter, available phosphorous, nitrogen and aggregate stability. The microaggregate fraction of the soil, which has been associated with greater organic carbon, was shown to be a hotspot for nitrogen cyclers particularly under salvage logging. The impact of post-fire management strategies on soil microbial communities needs to be considered in relation to maintaining ecosystem productivity, resilience and potential impact on climate change. Copyright © 2017 Elsevier B.V. All rights reserved.

[Nitrogen fixation potential of biological soil crusts in Heidaigou open coal mine, Inner Mongolia, China].

PubMed

Zhang, Peng; Huang, Lei; Hu, Yi-gang; Zhao, Yang; Wu, Yong-chen

2016-02-01

Nitrogen limitation is common in terrestrial ecosystems, and it is particularly severe in damaged ecosystems in arid regions. Biological soil crusts (BSCs) , as a crucial component of recovered vegetation, play a vital role in nitrogen fixation during the ecological restoration processes of damaged ecosystems in arid and semi-arid regions. In this study, two dominant types of BSCs (i.e., cyanobacterial-algal crusts and moss crusts) that are widely distributed in the re-vegetated area of Heidaigou open pit coal mine were investigated. Samples were collected in the field and their nitrogenase activities (NA) were measured in the laboratory. The responses of NA to different hydro-thermal factors and the relationships between NA and herbs in addition to crust coverage were analyzed. The results indicated that BSCs under reconstructed vegetation at different succession stages, abandoned land and natural vegetation showed values of NA ranging from 9 to 150 µmol C2H4 . m-2 . h-1, and the NA value of algae crust (77 µmol C2H4 . m-2 . h-1) was markedly higher than that of moss crust (17 µmol C2H4 . m-2 . h-1). In the re-vegetated area, cyanobacterial-algal crust and moss crust under shrub-herb had higher NA values than those of crusts under arbor-shrnb and arbor-shrub-herb. The relationship between NA of the two BSCs and soil relative water content (10% - 100%) as well as culture temperature (5-45 °C) were of quadratic function. With elevated water content and cultural temperature, the NA values increased at the initial stage and then decreased, and reached the maximum value at 25 °C of cultural temperature and 60% or 80% of relative water content. The NA of cyanobacterial-algal crust had a significant quadratic function with herb coverage, as NA declined when herb coverage was higher than 20%. A significant negative correlation was observed between the NA of moss crusts and herb coverage. The NA values of the two types of BSCs had a significant positive correlation

Long-term nitrogen fertilization decreases bacterial diversity and favors the growth of Actinobacteria and Proteobacteria in agro-ecosystems across the globe

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

Dai, Zhongmin; Su, Weiqin; Chen, Huaihai

Long-term Elevated nitrogen (N) input from anthropogenic sources may cause soil acidification and decrease crop yield, yet the response of the belowground microbial community to long-term N input and the input of N combined with phosphorus (P) and potassium (K) is still poorly understood. Here, we explored the effect of long-term N and NPK fertilization on soil bacterial diversity and community composition using meta-analysis of a global dataset. Nitrogen fertilization decreased soil pH, and increased soil organic carbon (C) and available N contents. Bacterial taxonomic diversity was decreased by N fertilization alone, but was increased by NPK fertilization. The effectmore » of N fertilization on bacterial diversity depends on soil texture and water management, but independent of crop type or N application rate. Both soil pH and organic C content were positively related to changes in bacterial diversity under N fertilization, while soil organic C was the dominant factor determining changes in bacterial diversity under NPK fertilization. Microbial biomass C decreased with decreasing bacterial diversity under long-term N fertilization. Nitrogen fertilization increased the relative abundance of copiotrophic bacteria (i.e. Proteobacteria and Actinobacteria), but reduced the abundance of oligotrophic taxa (i.e. Acidobacteria), consistent with the general life history strategy theory for bacteria. The relative abundance of Proteobacteria was also increased by NPK fertilization. The positive correlation between N application rate and the relative abundance of Actinobacteria indicates that increased N availability favored the growth of Actinobacteria. This first global analysis of long-term N and NPK fertilization effect on bacterial diversity and community composition suggests that N input decreases bacterial diversity but favors the growth of copiotrophic bacteria, providing a reference for nutrient management strategies for maintaining belowground microbial diversity

Long-term nitrogen fertilization decreases bacterial diversity and favors the growth of Actinobacteria and Proteobacteria in agro-ecosystems across the globe

DOE PAGES

Dai, Zhongmin; Su, Weiqin; Chen, Huaihai; ...

2018-04-25

Long-term Elevated nitrogen (N) input from anthropogenic sources may cause soil acidification and decrease crop yield, yet the response of the belowground microbial community to long-term N input and the input of N combined with phosphorus (P) and potassium (K) is still poorly understood. Here, we explored the effect of long-term N and NPK fertilization on soil bacterial diversity and community composition using meta-analysis of a global dataset. Nitrogen fertilization decreased soil pH, and increased soil organic carbon (C) and available N contents. Bacterial taxonomic diversity was decreased by N fertilization alone, but was increased by NPK fertilization. The effectmore » of N fertilization on bacterial diversity depends on soil texture and water management, but independent of crop type or N application rate. Both soil pH and organic C content were positively related to changes in bacterial diversity under N fertilization, while soil organic C was the dominant factor determining changes in bacterial diversity under NPK fertilization. Microbial biomass C decreased with decreasing bacterial diversity under long-term N fertilization. Nitrogen fertilization increased the relative abundance of copiotrophic bacteria (i.e. Proteobacteria and Actinobacteria), but reduced the abundance of oligotrophic taxa (i.e. Acidobacteria), consistent with the general life history strategy theory for bacteria. The relative abundance of Proteobacteria was also increased by NPK fertilization. The positive correlation between N application rate and the relative abundance of Actinobacteria indicates that increased N availability favored the growth of Actinobacteria. This first global analysis of long-term N and NPK fertilization effect on bacterial diversity and community composition suggests that N input decreases bacterial diversity but favors the growth of copiotrophic bacteria, providing a reference for nutrient management strategies for maintaining belowground microbial diversity

Diversity and distribution of the endophytic bacterial community at different stages of Eucalyptus growth.

PubMed

Miguel, Paulo Sérgio Balbino; de Oliveira, Marcelo Nagem Valério; Delvaux, Júlio César; de Jesus, Guilherme Luiz; Borges, Arnaldo Chaer; Tótola, Marcos Rogério; Neves, Júlio César Lima; Costa, Maurício Dutra

2016-06-01

The relationships between plants and endophytic bacteria significantly contribute to plant health and yield. However, the microbial diversity in leaves of Eucalyptus spp. is still poorly characterized. Here, we investigated the endophytic diversity in leaves of hybrid Eucalyptus grandis x E. urophylla (Eucalyptus "urograndis") by using culture-independent and culture-dependent approaches, to better understand their ecology in leaves at different stages of Eucalyptus development, including bacteria with N2 fixation potential. Firmicutes, Proteobacteria (classes alpha-, beta- and gamma-) and Actinobacteria were identified in the Eucalyptus "urograndis" endophytic bacterial community. Within this community, the species Novosphingobium barchaimii, Rhizobium grahamii, Stenotrophomonas panacihumi, Paenibacillus terrigena, P. darwinianus and Terrabacter lapilli represent the first report these bacteria as endophytes. The diversity of the total endophytic bacteria was higher in the leaves from the 'field' (the Shannon-Wiener index, 2.99), followed by the indices obtained in the 'clonal garden' (2.78), the 'recently out from under shade (2.68), 'under shade' (2.63) and 'plants for dispatch' (2.51). In contrast, for diazotrophic bacteria, the highest means of these indices were obtained from the leaves of plants in the 'under shade' (2.56), 'recently out from under shade (2.52)' and 'field' stages (2.54). The distribution of the endophytic bacterial species in Eucalyptus was distinct and specific to the development stages under study, and many of the species had the potential for nitrogen fixation, raising the question of whether these bacteria could contribute to overall nitrogen metabolism of Eucalyptus.

Comparative assessment of nitrogen fixation methodologies, conducted in the oligotrophic North Pacific Ocean.

PubMed

Wilson, Samuel T; Böttjer, Daniela; Church, Matthew J; Karl, David M

2012-09-01

Resolution of the nitrogen (N) cycle in the marine environment requires an accurate assessment of dinitrogen (N(2)) fixation. We present here an update on progress in conducting field measurements of acetylene reduction (AR) and (15)N(2) tracer assimilation in the oligotrophic North Pacific Subtropical Gyre (NPSG). The AR assay was conducted on discrete seawater samples using a headspace analysis system, followed by quantification of ethylene (C(2)H(4)) with a reducing compound photodetector. The rates of C(2)H(4) production were measurable for nonconcentrated seawater samples after an incubation period of 3 to 4 h. The (15)N(2) tracer measurements compared the addition of (15)N(2) as a gas bubble and dissolved as (15)N(2) enriched seawater. On all sampling occasions and at all depths, a 2- to 6-fold increase in the rate of (15)N(2) assimilation was measured when (15)N(2)-enriched seawater was added to the seawater sample compared to the addition of (15)N(2) as a gas bubble. In addition, we show that the (15)N(2)-enriched seawater can be prepared prior to its use with no detectable loss (<1.7%) of dissolved (15)N(2) during 4 weeks of storage, facilitating its use in the field. The ratio of C(2)H(4) production to (15)N(2) assimilation varied from 7 to 27 when measured simultaneously in surface seawater samples. Collectively, the modifications to the AR assay and the (15)N(2) assimilation technique present opportunities for more accurate and high frequency measurements (e.g., diel scale) of N(2) fixation, providing further insight into the contribution of different groups of diazotrophs to the input of N in the global oceans.

Improving nitrogen utilization efficiency of aquaponics by introducing algal-bacterial consortia.

PubMed

Fang, Yingke; Hu, Zhen; Zou, Yina; Zhang, Jian; Zhu, Zhuoran; Zhang, Jianda; Nie, Lichao

2017-12-01

Aquaponics is a promising technology combining aquaculture with hydroponics. In this study, algal-bacterial consortia were introduced into aquaponics, i.e., algal-bacterial based aquaponics (AA), to improve the nitrogen utilization efficiency (NUE) of aquaponics. The results showed that the NUE of AA was 13.79% higher than that of media-based aquaponics (MA). In addition, higher NO 3 - removal by microalgae assimilation led to better water quality in AA, which made up for the deficiencies of poor aquaponic management of nitrate. As a result of lower NO 3 - concentrations and dramatically higher dissolved oxygen (DO) concentrations caused by microalgae photosynthesis in the photobioreactor, the N 2 O emission of AA was 89.89% lower than that of MA, although nosZ gene abundance in MA's hydroponic bed was approximately 30 times over that in AA. Considering the factors mentioned above, AA would improve the sustainability of aquaponics and have a good application foreground. Copyright © 2017 Elsevier Ltd. All rights reserved.

Long-term nitrogen fertilization decreases bacterial diversity and favors the growth of Actinobacteria and Proteobacteria in agro-ecosystems across the globe.

PubMed

Dai, Zhongmin; Su, Weiqin; Chen, Huaihai; Barberán, Albert; Zhao, Haochun; Yu, Mengjie; Yu, Lu; Brookes, Philip C; Schadt, Christopher W; Chang, Scott X; Xu, Jianming

2018-04-12

Long-term elevated nitrogen (N) input from anthropogenic sources may cause soil acidification and decrease crop yield, yet the response of the belowground microbial community to long-term N input alone or in combination with phosphorus (P) and potassium (K) is poorly understood. We explored the effect of long-term N and NPK fertilization on soil bacterial diversity and community composition using meta-analysis of a global dataset. Nitrogen fertilization decreased soil pH, and increased soil organic carbon (C) and available N contents. Bacterial taxonomic diversity was decreased by N fertilization alone, but was increased by NPK fertilization. The effect of N fertilization on bacterial diversity varied with soil texture and water management, but was independent of crop type or N application rate. Changes in bacterial diversity were positively related to both soil pH and organic C content under N fertilization alone, but only to soil organic C under NPK fertilization. Microbial biomass C decreased with decreasing bacterial diversity under long-term N fertilization. Nitrogen fertilization increased the relative abundance of Proteobacteria and Actinobacteria, but reduced the abundance of Acidobacteria, consistent with the general life history strategy theory for bacteria. The positive correlation between N application rate and the relative abundance of Actinobacteria indicates that increased N availability favored the growth of Actinobacteria. This first global analysis of long-term N and NPK fertilization that differentially affects bacterial diversity and community composition provides a reference for nutrient management strategies for maintaining belowground microbial diversity in agro-ecosystems worldwide. © 2018 John Wiley & Sons Ltd.

Interspecific Plant Interactions Reflected in Soil Bacterial Community Structure and Nitrogen Cycling in Primary Succession

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

Knelman, Joseph E.; Graham, Emily B.; Prevéy, Janet S.

Past research demonstrating the importance plant-microbe interactions as drivers of ecosystem succession has focused on how plants condition soil microbial communities, impacting subsequent plant performance and successional trajectories in plant community assembly. These studies, however, largely treat microbial communities as a black box. In this study we sought to examine how emblematic shifts from early-successional Alnus sinuata (alder) to late successional Picea sitchensis (spruce) in primary succession may be reflected in specific belowground changes in bacterial community structure and nitrogen cycling related to the interaction of these two plants. We examined early successional alder-conditioned soils in a glacial forefield tomore » delineate how alders alter the soil microbial community with increasing dominance. Further, we assessed the impact of late-successional spruce plants on these early-successional alder-conditioned microbiomes and related nitrogen cycling through a leachate addition microcosm experiment. We show how increasingly abundant alder select for particular bacterial taxa. Additionally, we found that spruce leachate drives shifts in the relative abundance of major taxa of bacteria in alder-influenced soils, including declines in those that are enriched by alder. We found these effects to be spruce-specific, beyond a general leachate effect. Our work also demonstrates a unique influence of spruce on ammonium availability. Our results show that spruce leachate addition more strongly structures bacterial communities than alders (less dispersion in bacterial community beta diversity). Such insights bolster theory relating the importance of plant-microbe interactions with late-successional plants and interspecific plant interactions more generally.« less

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.

Nitrogen-fixing bacteria with multiple plant growth-promoting activities enhance growth of tomato and red pepper.

PubMed

Islam, Md Rashedul; Sultana, Tahera; Joe, M Melvin; Yim, Woojong; Cho, Jang-Cheon; Sa, Tongmin

2013-12-01

As a suitable alternative to chemical fertilizers, the application of plant growth-promoting rhizobacteria has been increasing in recent years due to their potential to be used as biofertilizers. In the present work, 13 nitrogen-fixing bacterial strains belonging to 11 different genera were tested for their PGP attributes. All of the strains were positive for 1-aminocyclopropane-1-carboxylate deaminase (ACCD), indole-3-acetic acid (IAA), salicylic acid, and ammonia production while negative for cellulase, pectinase, and hydrocyanic acid production. The strains Pseudomonas sp. RFNB3 and Serratia sp. RFNB14 were the most effective in solubilizing both tri-calcium phosphate and zinc oxide. In addition, all strains except Pseudomonas sp. RFNB3 were able to oxidize sulfur, and six strains were positive for siderophore synthesis. Each strain tested in this study possesses at least four PGP properties in addition to nitrogen fixation. Nine strains were selected based on their multiple PGP potential, particularly ACCD and IAA production, and evaluated for their effects on early growth of tomato and red pepper under gnotobiotic conditions. Bacterial inoculation considerably influenced root and shoot length, seedling vigor, and dry biomass of the two crop plants. Three strains that demonstrated substantial effects on plant performance were further selected for greenhouse trials with red pepper, and among them Pseudomonas sp. RFNB3 resulted in significantly higher plant height (26%) and dry biomass (28%) compared to control. The highest rate of nitrogen fixation, as determined by acetylene reduction assay, occurred in Novosphingobium sp. RFNB21 inoculated red pepper root (49.6 nM of ethylene/h/g of dry root) and rhizosphere soil (41.3 nM of ethylene/h/g of dry soil). Inoculation with nitrogen-fixing bacteria significantly increased chlorophyll content, and the uptake of different macro- and micro-nutrient contents enhancing also in red pepper shoots, in comparison with

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 ...

Interspecific Plant Interactions Reflected in Soil Bacterial Community Structure and Nitrogen Cycling in Primary Succession.

PubMed

Knelman, Joseph E; Graham, Emily B; Prevéy, Janet S; Robeson, Michael S; Kelly, Patrick; Hood, Eran; Schmidt, Steve K

2018-01-01

Past research demonstrating the importance plant-microbe interactions as drivers of ecosystem succession has focused on how plants condition soil microbial communities, impacting subsequent plant performance and plant community assembly. These studies, however, largely treat microbial communities as a black box. In this study, we sought to examine how emblematic shifts from early successional Alnus viridus ssp. sinuata (Sitka alder) to late successional Picea sitchensis (Sitka spruce) in primary succession may be reflected in specific belowground changes in bacterial community structure and nitrogen cycling related to the interaction of these two plants. We examined early successional alder-conditioned soils in a glacial forefield to delineate how alders alter the soil microbial community with increasing dominance. Further, we assessed the impact of late-successional spruce plants on these early successional alder-conditioned microbiomes and related nitrogen cycling through a leachate addition microcosm experiment. We show how increasingly abundant alder select for particular bacterial taxa. Additionally, we found that spruce leachate significantly alters the composition of these microbial communities in large part by driving declines in taxa that are enriched by alder, including bacterial symbionts. We found these effects to be spruce specific, beyond a general leachate effect. Our work also demonstrates a unique influence of spruce on ammonium availability. Such insights bolster theory relating the importance of plant-microbe interactions with late-successional plants and interspecific plant interactions more generally.

Interspecific Plant Interactions Reflected in Soil Bacterial Community Structure and Nitrogen Cycling in Primary Succession

PubMed Central

Knelman, Joseph E.; Graham, Emily B.; Prevéy, Janet S.; Robeson, Michael S.; Kelly, Patrick; Hood, Eran; Schmidt, Steve K.

2018-01-01

Past research demonstrating the importance plant–microbe interactions as drivers of ecosystem succession has focused on how plants condition soil microbial communities, impacting subsequent plant performance and plant community assembly. These studies, however, largely treat microbial communities as a black box. In this study, we sought to examine how emblematic shifts from early successional Alnus viridus ssp. sinuata (Sitka alder) to late successional Picea sitchensis (Sitka spruce) in primary succession may be reflected in specific belowground changes in bacterial community structure and nitrogen cycling related to the interaction of these two plants. We examined early successional alder-conditioned soils in a glacial forefield to delineate how alders alter the soil microbial community with increasing dominance. Further, we assessed the impact of late-successional spruce plants on these early successional alder-conditioned microbiomes and related nitrogen cycling through a leachate addition microcosm experiment. We show how increasingly abundant alder select for particular bacterial taxa. Additionally, we found that spruce leachate significantly alters the composition of these microbial communities in large part by driving declines in taxa that are enriched by alder, including bacterial symbionts. We found these effects to be spruce specific, beyond a general leachate effect. Our work also demonstrates a unique influence of spruce on ammonium availability. Such insights bolster theory relating the importance of plant–microbe interactions with late-successional plants and interspecific plant interactions more generally. PMID:29467741

Bacterial interactions in the rhizosphere of seagrass communities in shallow coastal lagoons.

PubMed

Donnelly, A P; Herbert, R A

1998-12-01

Rooted phanerogam communities in the shallow intertidal and subtidal coastal zone represent productive and healthy ecosystems. Inorganic nutrients are assimilated into seagrass biomass. Much of the organic matter resulting from moribund seagrass is rapidly mineralized, principally by bacteria. The microbial community of the rhizosphere is also highly active due to the supply of organic matter released during photosynthesis. This active sediment community plays an important role through carbon, nitrogen and phosphorous cycling in maintaining the stability and productivity of seagrass meadows. Over the last two decades, however, seagrass meadows in European coastal areas have declined due to increasing pollution. As eutrophication advances a trasition occurs from rooted phanerogram dominated communities to planktonic algal blooms and/or cyanobacterial blooms. Such changes represent the decline of a stable, high biodiversity habitat to an unstable one dominated by a few species. These changes of community structure can occur rapidly once the internal nutrient and organic matter control cycles are exceeded. A field investigation was undertaken to establish the spatial distribution of bacterial populations of Zostera noltii colonized and uncolonized sediment in the Bassin d'Arcachon, France. Bacteria were enumerated using both plate count and MPN techniques for different functional groups as well as determining the total bacterial populations present. Nitrogen fixation, ammonification, sulphate reduction rates, as well as alkaline phosphatase activity were also determined. Colonization of the Z. noltii roots and rhizomes was studied by light and scanning electron microscopy. Results confirmed that higher bacterial populations were present in the rhizosphere of Z. noltii compared to uncolonized sediments. Furthermore, electron microscopy identified the rhizome as the main site of colonization for a diverse range of morphological groups of bacteria. Sulphate reducing

Nitrogen removal capacity and bacterial community dynamics of a Canon biofilter system at different organic matter concentrations.

PubMed

García-Ruiz, María J; Maza-Márquez, Paula; González-López, Jesús; Osorio, Francisco

2018-02-01

Three Canon bench-scale bioreactors with a volume of 2 L operating in parallel were configured as submerged biofilters. In the present study we investigated the effects of a high ammonium concentration (320 mgNH 4 + · L -1 ) and different concentrations of organic matter (0, 100 and 400 mgCOD·L -1 ) on the nitrogen removal capacity and the bacterial community structure. After 60 days, the Canon biofilters operated properly under concentrations of 0 and 100 mgCOD·L -1 of organic matter, with nitrogen removal efficiencies up to 85%. However, a higher concentration of organic matter (400 mgCOD·L -1 ) produced a partial inhibition of nitrogen removal (68.1% efficiency). The addition of higher concentrations of organic matter a modified the bacterial community structure in the Canon biofilter, increasing the proliferation of heterotrophic bacteria related to the genera of Thauera, Longilinea, Ornatilinea, Thermomarinilinea, unclassified Chlorobiales and Denitratisoma. However, heterotrophic bacteria co-exist with Nitrosomonas and Candidatus Scalindua. Thus, our study confirms the co-existence of different microbial activities (AOB, Anammox and denitrification) and the adaptation of a fixed-biofilm system to different concentrations of organic matter. Copyright © 2017 Elsevier Ltd. All rights reserved.

Distinct summer and winter bacterial communities in the active layer of Svalbard permafrost revealed by DNA- and RNA-based analyses

DOE PAGES

Schostag, Morten; Stibal, Marek; Jacobsen, Carsten S.; ...

2015-04-30

The active layer of soil overlaying permafrost in the Arctic is subjected to dramatic annual changes in temperature and soil chemistry, which likely affect bacterial activity and community structure. We studied seasonal variations in the bacterial community of active layer soil from Svalbard (78°N) by co-extracting DNA and RNA from 12 soil cores collected monthly over a year. PCR amplicons of 16S rRNA genes (DNA) and reverse transcribed transcripts (cDNA) were quantified and sequenced to test for the effect of low winter temperature and seasonal variation in concentration of easily degradable organic matter on the bacterial communities. The copy numbermore » of 16S rRNA genes and transcripts revealed no distinct seasonal changes indicating potential bacterial activity during winter despite soil temperatures well below -10°C. Multivariate statistical analysis of the bacterial diversity data (DNA and cDNA libraries) revealed a season-based clustering of the samples, and, e.g., the relative abundance of potentially active Cyanobacteria peaked in June and Alphaproteobacteria increased over the summer and then declined from October to November. The structure of the bulk (DNA-based) community was significantly correlated with pH and dissolved organic carbon, while the potentially active (RNA-based) community structure was not significantly correlated with any of the measured soil parameters. A large fraction of the 16S rRNA transcripts was assigned to nitrogen-fixing bacteria (up to 24% in June) and phototrophic organisms (up to 48% in June) illustrating the potential importance of nitrogen fixation in otherwise nitrogen poor Arctic ecosystems and of phototrophic bacterial activity on the soil surface.« less

Distinct summer and winter bacterial communities in the active layer of Svalbard permafrost revealed by DNA- and RNA-based analyses

PubMed Central

Schostag, Morten; Stibal, Marek; Jacobsen, Carsten S.; Bælum, Jacob; Taş, Neslihan; Elberling, Bo; Jansson, Janet K.; Semenchuk, Philipp; Priemé, Anders

2015-01-01

The active layer of soil overlaying permafrost in the Arctic is subjected to dramatic annual changes in temperature and soil chemistry, which likely affect bacterial activity and community structure. We studied seasonal variations in the bacterial community of active layer soil from Svalbard (78°N) by co-extracting DNA and RNA from 12 soil cores collected monthly over a year. PCR amplicons of 16S rRNA genes (DNA) and reverse transcribed transcripts (cDNA) were quantified and sequenced to test for the effect of low winter temperature and seasonal variation in concentration of easily degradable organic matter on the bacterial communities. The copy number of 16S rRNA genes and transcripts revealed no distinct seasonal changes indicating potential bacterial activity during winter despite soil temperatures well below −10°C. Multivariate statistical analysis of the bacterial diversity data (DNA and cDNA libraries) revealed a season-based clustering of the samples, and, e.g., the relative abundance of potentially active Cyanobacteria peaked in June and Alphaproteobacteria increased over the summer and then declined from October to November. The structure of the bulk (DNA-based) community was significantly correlated with pH and dissolved organic carbon, while the potentially active (RNA-based) community structure was not significantly correlated with any of the measured soil parameters. A large fraction of the 16S rRNA transcripts was assigned to nitrogen-fixing bacteria (up to 24% in June) and phototrophic organisms (up to 48% in June) illustrating the potential importance of nitrogen fixation in otherwise nitrogen poor Arctic ecosystems and of phototrophic bacterial activity on the soil surface. PMID:25983731

Culture-independent analysis of hydrocarbonoclastic bacterial communities in environmental samples during oil-bioremediation.

PubMed

Dashti, Narjes; Ali, Nedaa; Salamah, Samar; Khanafer, Majida; Al-Shamy, Ghada; Al-Awadhi, Husain; Radwan, Samir S

2018-04-15

To analyze microbial communities in environmental samples, this study combined Denaturing Gradient Gel Electrophoresis of amplified 16S rRNA-genes in total genomic DNA extracts from those samples with gene sequencing. The environmental samples studied were oily seawater and soil samples, that had been bioaugmented with natural materials rich in hydrocarbonoclastic bacteria. This molecular approach revealed much more diverse bacterial taxa than the culture-dependent method we had used in an earlier study for the analysis of the same samples. The study described the dynamics of bacterial communities during bioremediation. The main limitation associated with this molecular approach, namely of not distinguishing hydrocarbonoclastic taxa from others, was overcome by consulting the literature for the hydrocarbonoclastic potential of taxa related to those identified in this study. By doing so, it was concluded that the hydrocarbonoclastic bacterial taxa were much more diverse than those captured by the culture-dependent approach. The molecular analysis also revealed the frequent occurrence of nifH-genes in the total genomic DNA extracts of all the studied environmental samples, which reflects a nitrogen-fixation potential. Nitrogen fertilization is long known to enhance microbial oil-bioremediation. The study revealed that bioaugmentation using plant rhizospheres or soil with long history of oil-pollution was more effective in oil-removal in the desert soil than in seawater microcosms. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Difference of nitrogen-cycling microbes between shallow bay and deep-sea sediments in the South China Sea.

PubMed

Yu, Tiantian; Li, Meng; Niu, Mingyang; Fan, Xibei; Liang, Wenyue; Wang, Fengping

2018-01-01

In marine sediments, microorganisms are known to play important roles in nitrogen cycling; however, the composition and quantity of microbes taking part in each process of nitrogen cycling are currently unclear. In this study, two different types of marine sediment samples (shallow bay and deep-sea sediments) in the South China Sea (SCS) were selected to investigate the microbial community involved in nitrogen cycling. The abundance and composition of prokaryotes and seven key functional genes involved in five processes of the nitrogen cycle [nitrogen fixation, nitrification, denitrification, dissimilatory nitrate reduction to ammonium (DNRA), and anaerobic ammonia oxidation (anammox)] were presented. The results showed that a higher abundance of denitrifiers was detected in shallow bay sediments, while a higher abundance of microbes involved in ammonia oxidation, anammox, and DNRA was found in the deep-sea sediments. Moreover, phylogenetic differentiation of bacterial amoA, nirS, nosZ, and nrfA sequences between the two types of sediments was also presented, suggesting environmental selection of microbes with the same geochemical functions but varying physiological properties.

Carboxysomes: metabolic modules for CO 2 fixation

DOE PAGES

Turmo, Aiko; Gonzalez-Esquer, Cesar Raul; Kerfeld, Cheryl A.

2017-08-14

The carboxysome is a bacterial microcompartment encapsulating the enzymes carbonic anhydrase and ribulose-1,5-bisphosphate carboxylase/oxygenase. As the site of CO 2 fixation, it serves an essential role in the carbon dioxide concentrating mechanism of many chemoautotrophs and all cyanobacteria. Carboxysomes and other bacterial microcompartments self-assemble through specific protein–protein interactions that are typically mediated by conserved protein domains. In this review, we frame our current understanding of carboxysomes in the context of their component protein domains with their inherent function as the ‘building blocks’ of carboxysomes. These building blocks are organized in genetic modules (conserved chromosomal loci) that encode for carboxysomes andmore » ancillary proteins essential for the integration of the organelle with the rest of cellular metabolism. This conceptual framework provides the foundation for ‘plug-and-play’ engineering of carboxysomes as CO 2 fixation modules in a variety of biotechnological applications.« less

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

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

Mixed, short rotation culture of red alder and black cottonwood: growth, coppicing, nitrogen fixation, and allelopathy

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

Heilman, P.; Stettler, R.F.

1985-01-01

Alnus rubra seedlings were grown in a 1:1 mixture at a spacing of 1.2 x 1.2 m with 28 Populus clones (25 clones pf P. trichocarpa, 2 of P. deltoides x P. trichocarpa, and one P. deltoides x P. nigra) in a study established in W. Washington in March 1979. Trees were harvested at 4 yr old. At harvest, average heights were: pure Populus, 10.2 m; Populus in the mixed stand 11.0 m; and alder 8.4 m. Most Populus sprouted satisfactorily after harvest (6.6 shoots/plant when pure, 7.6 shoots/plant in the mixture), but alder sprouted poorly (3.6 shoots/plant). Above-ground biomassmore » at harvest was 15.9 t/ha p.a. for the mixture and 16.7 t/ha p.a. for pure Populus, although the mixture had been more productive at 2 yr. Nitrogenase activity (nitrogen fixation as measured by acetylene reduction) of alder declines in the 4th season; competition was the most important factor influencing this decline. Soil N content had no effect on fixation. A pot study showed that ground Populus leaf and litter material inhibited the growth of red alder seedlings, although soil collected from Populus plots had no effect. Results indicated that allelopathy is probably a minor factor under field conditions, at most, and that growing mixed stands may, on balance, be beneficial. 20 references.« less

Effect of light on N2 fixation and net nitrogen release of Trichodesmium in a field study

NASA Astrophysics Data System (ADS)

Lu, Yangyang; Wen, Zuozhu; Shi, Dalin; Chen, Mingming; Zhang, Yao; Bonnet, Sophie; Li, Yuhang; Tian, Jiwei; Kao, Shuh-Ji

2018-01-01

Dinitrogen fixation (NF) by marine cyanobacteria is an important pathway to replenish the oceanic bioavailable nitrogen inventory. Light is the key to modulating NF; however, field studies investigating the light response curve (NF-I curve) of NF rate and the effect of light on diazotroph-derived nitrogen (DDN) net release are relatively sparse in the literature, hampering prediction using models. A dissolution method was applied using uncontaminated 15N2 gas to examine how the light changes may influence the NF intensity and DDN net release in the oligotrophic ocean. Experiments were conducted at stations with diazotrophs dominated by filamentous cyanobacterium Trichodesmium spp. in the western Pacific and the South China Sea. The effect of light on carbon fixation (CF) was measured in parallel using the 13C tracer method specifically for a station characterized by Trichodesmium bloom. Both NF-I and CF-I curves showed a Ik (light saturation coefficient) range of 193 to 315 µE m-2 s-1, with light saturation at around 400 µE m-2 s-1. The proportion of DDN net release ranged from ˜ 6 to ˜ 50 %, suggesting an increasing trend as the light intensity decreased. At the Trichodesmium bloom station, we found that the CF / NF ratio was light-dependent and the ratio started to increase as light was lower than the carbon compensation point of 200 µE m-2 s-1. Under low-light stress, Trichodesmium physiologically preferred to allocate more energy for CF to alleviate the intensive carbon consumption by respiration; thus, there is a metabolism tradeoff between CF and NF pathways. Results showed that short-term ( < 24 h) light change modulates the physiological state, which subsequently determined the C / N metabolism and DDN net release by Trichodesmium. Reallocation of energy associated with the variation in light intensity would be helpful for prediction of the global biogeochemical cycle of N by models involving Trichodesmium blooms.

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

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

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...

Initial nitrogen enrichment conditions determines variations in nitrogen substrate utilization by heterotrophic bacterial isolates.

PubMed

Ghosh, Suchismita; Ayayee, Paul A; Valverde-Barrantes, Oscar J; Blackwood, Christopher B; Royer, Todd V; Leff, Laura G

2017-04-04

The nitrogen (N) cycle consists of complex microbe-mediated transformations driven by a variety of factors, including diversity and concentrations of N compounds. In this study, we examined taxonomic diversity and N substrate utilization by heterotrophic bacteria isolated from streams under complex and simple N-enrichment conditions. Diversity estimates differed among isolates from the enrichments, but no significant composition were detected. Substrate utilization and substrate range of bacterial assemblages differed within and among enrichments types, and not simply between simple and complex N-enrichments. N substrate use patterns differed between isolates from some complex and simple N-enrichments while others were unexpectedly similar. Taxonomic composition of isolates did not differ among enrichments and was unrelated to N use suggesting strong functional redundancy. Ultimately, our results imply that the available N pool influences physiology and selects for bacteria with various abilities that are unrelated to their taxonomic affiliation.

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.

On the Relationship Between Hydrogen Saturation in the Tropical Atlantic Ocean and Nitrogen Fixation by the Symbiotic Diazotroph UCYN-A

NASA Astrophysics Data System (ADS)

Moore, R. M.; Grefe, I.; Zorz, J.; Shan, S.; Thompson, K.; Ratten, J.; LaRoche, J.

2018-04-01

Dissolved hydrogen measurements were made at high resolution in surface waters along a tropical north Atlantic transect between Guadeloupe and Cape Verde in 2015 (Meteor 116). Parallel water samples acquired to assess the relative abundance of the nifH gene from several types of diazotrophs, indicated that Trichodesmium and UCYN-A were dominant in this region. We show that a high degree of correlation exists between the hydrogen saturations and UCYN-A nifH abundance, and a weak correlation with Trichodesmium. The findings suggest that nitrogen fixation by UCYN-A is a major contributor to hydrogen supersaturations in this region of the ocean. The ratio of hydrogen released to nitrogen fixed has not been determined for this symbiont, but the indications are that it may be high in comparison with the small number of diazotrophs for which the ratio has been measured in laboratory cultures. We speculate that this would be consistent with the diazotroph being an exosymbiont on its haptophyte host. Our high resolution measurements of hydrogen concentrations are capable of illustrating the time and space scales of inferred activity of diazotrophs in near real-time in a way that cannot be achieved by biological sampling and rate measurements requiring incubations with 15N2. Direct measurement of high resolution spatial variability would be relatively challenging through collection and analysis of biological samples by qPCR, and extremely challenging by 15N-uptake techniques, neither of which methods yields real-time data. Nonetheless, determination of fixation rates still firmly depends on the established procedure of incubations in the presence of 15N2.

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...

Nitrogen dynamics in an Alaskan salt marsh following spring use by geese

USGS Publications Warehouse

Zacheis, Amy B.; Ruess, Roger W.; Hupp, Jerry W.

2002-01-01

Lesser snow geese (Anser caerulescens caerulescens) and Canada geese (Branta canadensis) use several salt marshes in Cook Inlet, Alaska, as stopover areas for brief periods during spring migration. We investigated the effects of geese on nitrogen cycling processes in Susitna Flats, one of the marshes. We compared net nitrogen mineralization, organic nitrogen pools and production in buried bags, nitrogen fixation by cyanobacteria, and soil and litter characteristics on grazed plots versus paired plots that had been exclosed from grazing for 3 years. Grazed areas had higher rates of net nitrogen mineralization in the spring and there was no effect of grazing on organic nitrogen availability. The increased mineralization rates in grazed plots could not be accounted for by alteration of litter quality, litter quantity, microclimate, or root biomass, which were not different between grazed and exclosed plots. In addition, fecal input was very slight in the year that we studied nitrogen cycling. We propose that trampling had two effects that could account for greater nitrogen availability in grazed areas: litter incorporation into soil, resulting in increased rates of decomposition and mineralization of litter material, and greater rates of nitrogen fixation by cyanobacteria on bare, trampled soils. A path analysis indicated that litter incorporation by trampling played a primary role in the nitrogen dynamics of the system, with nitrogen fixation secondary, and that fecal input was of little importance.

History on the biological nitrogen fixation research in graminaceous plants: special emphasis on the Brazilian experience.

PubMed

Baldani, José I; Baldani, Vera L D

2005-09-01

This review covers the history on Biological Nitrogen Fixation (BNF) in Graminaceous plants grown in Brazil, and describes research progress made over the last 40 years, most of which was coordinated by Johanna Döbereiner. One notable accomplishment during this period was the discovery of several nitrogen-fixing bacteria such as the rhizospheric (Beijerinckia fluminensis and Azotobacter paspali), associative (Azospirillum lipoferum, A. brasilense, A. amazonense) and the endophytic (Herbaspirillum seropedicae, H. rubrisubalbicans, Gluconacetobacter diazotrophicus, Burkholderia brasilensis and B. tropica). The role of these diazotrophs in association with grasses, mainly with cereal plants, has been studied and a lot of progress has been achieved in the ecological, physiological, biochemical, and genetic aspects. The mechanisms of colonization and infection of the plant tissues are better understood, and the BNF contribution to the soil/plant system has been determined. Inoculation studies with diazotrophs showed that endophytic bacteria have a much higher BNF contribution potential than associative diazotrophs. In addition, it was found that the plant genotype influences the plant/bacteria association. Recent data suggest that more studies should be conducted on the endophytic association to strengthen the BNF potential. The ongoing genome sequencing programs: RIOGENE (Gluconacetobacter diazotrophicus) and GENOPAR (Herbaspirillum seropedicae) reflect the commitment to the BNF study in Brazil and should allow the country to continue in the forefront of research related to the BNF process in Graminaceous plants.

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%.

Dicarboxylic acid transport in Bradyrhizobium japonicum: use of Rhizobium meliloti dct gene(s) to enhance nitrogen fixation.

PubMed Central

Birkenhead, K; Manian, S S; O'Gara, F

1988-01-01

A recombinant plasmid encoding Rhizobium meliloti sequences involved in dicarboxylic acid transport (plasmid pRK290:4:46) (E. Bolton, B. Higgisson, A. Harrington, and F. O'Gara, Arch. Microbiol. 144:142-146, 1986) was used to study the relationship between dicarboxylic acid transport and nitrogen fixation in Bradyrhizobium japonicum. The expression of the dct sequences on plasmid pRK290:4:46 in B. japonicum CJ1 resulted in increased growth rates in media containing dicarboxylic acids as the sole source of carbon. In addition, strain CJ1(pRK290:4:46) exhibited enhanced succinate uptake activity when grown on dicarboxylic acids under aerobic conditions. Under free-living nitrogen-fixing conditions, strain CJ1(pRK290:4:46) exhibited higher nitrogenase (acetylene reduction) activity compared with that of the wild-type strain. This increase in nitrogenase activity also correlated with an enhanced dicarboxylic acid uptake rate under these microaerobic conditions. The regulation of dicarboxylic acid transport by factors such as metabolic inhibitors and the presence of additional carbon sources was similar in both the wild-type and the engineered strains. The implications of increasing nitrogenase activity through alterations in the dicarboxylic acid transport system are discussed. PMID:3422072

Ecological occurrence of Gluconacetobacter diazotrophicus and nitrogen-fixing Acetobacteraceae members: their possible role in plant growth promotion.

PubMed

Saravanan, V S; Madhaiyan, M; Osborne, Jabez; Thangaraju, M; Sa, T M

2008-01-01

Gluconacetobacter diazotrophicus has a long-standing history of bacterial-plant interrelationship as a symbiotic endophyte capable of fixing atmospheric nitrogen. In low nitrogen fertilized sugarcane fields it plays a significant role and its occurrence was realised in most of the sugarcane growing countries. In this mini review, the association of G. diazotrophicus with sugarcane, other crop plants and with various hosts is discussed. The factors affecting survival in the rhizosphere and the putative soil mode of transmission are emphasized. In addition, other N(2)-fixing Acetobacteraceae members, including Gluconacetobacter azotocaptans, Gluconacetobacter johannae and Swaminathania salitolerans, occurring in coffee, corn and rice plants are also covered. Lastly, the plant-growth-promoting traits identified in this group of bacteria, including N(2) fixation, phytohormone synthesis, P and Zn solubilization and biocontrol, are analysed.

Biochemical and Molecular Phylogenetic Study of Agriculturally Useful Association of a Nitrogen-Fixing Cyanobacterium and Nodule Sinorhizobium with Medicago sativa L.

PubMed

Karaushu, E V; Lazebnaya, I V; Kravzova, T R; Vorobey, N A; Lazebny, O E; Kiriziy, D A; Olkhovich, O P; Taran, N Yu; Kots, S Ya; Popova, A A; Omarova, E; Koksharova, O A

2015-01-01

Seed inoculation with bacterial consortium was found to increase legume yield, providing a higher growth than the standard nitrogen treatment methods. Alfalfa plants were inoculated by mono- and binary compositions of nitrogen-fixing microorganisms. Their physiological and biochemical properties were estimated. Inoculation by microbial consortium of Sinorhizobium meliloti T17 together with a new cyanobacterial isolate Nostoc PTV was more efficient than the single-rhizobium strain inoculation. This treatment provides an intensification of the processes of biological nitrogen fixation by rhizobia bacteria in the root nodules and an intensification of plant photosynthesis. Inoculation by bacterial consortium stimulates growth of plant mass and rhizogenesis and leads to increased productivity of alfalfa and to improving the amino acid composition of plant leaves. The full nucleotide sequence of the rRNA gene cluster and partial sequence of the dinitrogenase reductase (nifH) gene of Nostoc PTV were deposited to GenBank (JQ259185.1, JQ259186.1). Comparison of these gene sequences of Nostoc PTV with all sequences present at the GenBank shows that this cyanobacterial strain does not have 100% identity with any organisms investigated previously. Phylogenetic analysis showed that this cyanobacterium clustered with high credibility values with Nostoc muscorum.

Appraisal of the nitrogen-15 natural-abundance method for quantifying dinitrogen fixation

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

Bremer, E.; van Kessel, C.

Several investigators have questioned the use of the {sup 15}N natural-abundance method of estimating N{sub 2} fixation because of variability in soil {delta}{sup 15}N and small differences between the {delta}{sup 15}N of soil N and atmospheric N. Investigations were conducted to compare the {sup 15}N natural-abundance and {sup 15}N-isotope-dilution methods for estimating N{sub 2} fixation of field-grown pea (Pisum sativum L.) and lentil (Lens culinaris Medik.). Spatial variability was assessed at three sites by determining the {delta}{sup 15}N of non-N{sub 2}-fixing plants. Seasonal variation in {delta}{sup 15}N for spring and winter wheat (Triticum aestivum L.), flax (Linum usitatissimum L.), barleymore » (Hordeum vulgare L.), rape (Brassica napus L.) and lentil was determined at one site. Comparisons between {delta}{sup 15}N and {sup 15}N-enriched isotope-dilution methods for estimating N{sub 2} fixation by lentil were conducted at several sites over a 3-yr period. Variability in {delta}{sup 15}N of the reference plant was site dependent: the {delta}{sup 15}N ranged from 2.8 to 9.3 at the first site, 3.4 to 8.8 at the second site, and 3.5 to 6.2 at the third site. The average {delta}{sup 15}N of four of the five non-N{sub 2}-fixing plants increased from 5.4 at 42 d after planting to 6.9 at the final harvest. The fifth non-N{sub 2}-fixing plant, rape, accumulated most of its N during the first 42 d after planting, and its {delta}{sup 15}N value declined from 8.1 at 42 d after planting to 7.3 at the final harvest. Estimates of N{sub 2} fixation were not significantly different in 18 out of 21 comparisons; in two comparisons in the {delta}{sup 15}N method and in one comparison the {sup 15}N-enriched method provided higher estimates of N{sub 2} fixation. Overall, both methods appeared to provide equally reliable estimates of N{sub 2} fixation for lentil.« less

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.

Consistent effects of nitrogen fertilization on soil bacterial communities in black soils for two crop seasons in China.

PubMed

Zhou, Jing; Jiang, Xin; Wei, Dan; Zhao, Baisuo; Ma, Mingchao; Chen, Sanfeng; Cao, Fengming; Shen, Delong; Guan, Dawei; Li, Jun

2017-06-12

Long-term use of inorganic nitrogen (N) fertilization has greatly influenced the bacterial community in black soil of northeast China. It is unclear how N affects the bacterial community in two successive crop seasons in the same field for this soil type. We sampled soils from a long-term fertilizer experimental field in Harbin city with three N gradients. We applied sequencing and quantitative PCR targeting at the 16S rRNA gene to examine shifts in bacterial communities and test consistent shifts and driving-factors bacterial responses to elevated N additions. N addition decreased soil pH and bacterial 16S rDNA copy numbers, and increased soil N and crop yield. N addition consistently decreased bacterial diversity and altered bacterial community composition, by increasing the relative abundance of Proteobacteria, and decreasing that of Acidobacteria and Nitrospirae in both seasons. Consistent changes in the abundant classes and genera, and the structure of the bacterial communities across both seasons were observed. Our results suggest that increases in N inputs had consistent effects on the richness, diversity and composition of soil bacterial communities across the crop seasons in two continuous years, and the N addition and the subsequent edaphic changes were important factors in shaping bacterial community structures.

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

Co-occurrence of methanogenesis and N2 fixation in oil sands tailings.

PubMed

Collins, C E Victoria; Foght, Julia M; Siddique, Tariq

2016-09-15

Oil sands tailings ponds in northern Alberta, Canada have been producing biogenic gases via microbial metabolism of hydrocarbons for decades. Persistent methanogenic activity in tailings ponds without any known replenishment of nutrients such as fixed nitrogen (N) persuaded us to investigate whether N2 fixation or polyacrylamide (PAM; used as a tailings flocculant) could serve as N sources. Cultures comprising mature fine tailings (MFT) plus methanogenic medium supplemented with or deficient in fixed N were incubated under an N2 headspace. Some cultures were further amended with citrate, which is used in oil sands processing, as a relevant carbon source, and/or with PAM. After an initial delay, N-deficient cultures with or without PAM produced methane (CH4) at the same rate as N-containing cultures, indicating a mechanism of overcoming apparent N-deficiency. Acetylene reduction and (15)N2 incorporation in all N-deficient cultures (with or without PAM) suggested active N2 fixation concurrently with methanogenesis but inability to use PAM as a N source. 16S rRNA gene pyrosequencing revealed little difference between archaeal populations regardless of N content. However, bacterial sequences in N-deficient cultures showed enrichment of Hyphomicrobiaceae and Clostridium members that might contain N2-fixing species. The results are important in understanding long-term production of biogenic greenhouse gases in oil sands tailings. Copyright © 2016 Elsevier B.V. All rights reserved.

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

Insights into functional bacterial diversity and its effects on Alpine bog ecosystem functioning.

PubMed

Bragina, Anastasia; Berg, Christian; Müller, Henry; Moser, Daniel; Berg, Gabriele

2013-01-01

Plant-associated bacteria are important for the growth and health of their host, but little is known about its functional diversity and impact on ecosystem functioning. We studied bacterial nitrogen fixation and methane oxidation from indicator Sphagnum mosses in Alpine bogs to test a hypothesis that the plant microbiome contained different functional patterns depending on their functions within the ecosystem. A high abundance and diversity of nitrogenase genes were detected, mostly specific for each Sphagnum. In contrast, methanotrophs formed highly similar patterns despite a high abundance and diversity of methane monooxygenase genes. Our hypothesis was supported by these contrasting functional patterns together with the result that the Sphagnum sporophyte contained a high proportion of specific diazotrophs (45.5%) but no potential methanotrophs. While essential for plant growth under nutrient-limited conditions, nitrogen-fixing bacteria were highly specific and transferred with the sporophyte unlike the ubiquitous methanotrophs which are important for the climate-relevant ecosystem itself.

454-Pyrosequencing Analysis of Bacterial Communities from Autotrophic Nitrogen Removal Bioreactors Utilizing Universal Primers: Effect of Annealing Temperature.

PubMed

Gonzalez-Martinez, Alejandro; Rodriguez-Sanchez, Alejandro; Rodelas, Belén; Abbas, Ben A; Martinez-Toledo, Maria Victoria; van Loosdrecht, Mark C M; Osorio, F; Gonzalez-Lopez, Jesus

2015-01-01

Identification of anaerobic ammonium oxidizing (anammox) bacteria by molecular tools aimed at the evaluation of bacterial diversity in autotrophic nitrogen removal systems is limited by the difficulty to design universal primers for the Bacteria domain able to amplify the anammox 16S rRNA genes. A metagenomic analysis (pyrosequencing) of total bacterial diversity including anammox population in five autotrophic nitrogen removal technologies, two bench-scale models (MBR and Low Temperature CANON) and three full-scale bioreactors (anammox, CANON, and DEMON), was successfully carried out by optimization of primer selection and PCR conditions (annealing temperature). The universal primer 530F was identified as the best candidate for total bacteria and anammox bacteria diversity coverage. Salt-adjusted optimum annealing temperature of primer 530F was calculated (47°C) and hence a range of annealing temperatures of 44-49°C was tested. Pyrosequencing data showed that annealing temperature of 45°C yielded the best results in terms of species richness and diversity for all bioreactors analyzed.

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

Biological nitrogen fixation in the oxygen-minimum region of the eastern tropical North Pacific ocean.

PubMed

Jayakumar, Amal; Chang, Bonnie X; Widner, Brittany; Bernhardt, Peter; Mulholland, Margaret R; Ward, Bess B

2017-10-01

Biological nitrogen fixation (BNF) was investigated above and within the oxygen-depleted waters of the oxygen-minimum zone of the Eastern Tropical North Pacific Ocean. BNF rates were estimated using an isotope tracer method that overcame the uncertainty of the conventional bubble method by directly measuring the tracer enrichment during the incubations. Highest rates of BNF (~4 nM day -1 ) occurred in coastal surface waters and lowest detectable rates (~0.2 nM day -1 ) were found in the anoxic region of offshore stations. BNF was not detectable in most samples from oxygen-depleted waters. The composition of the N 2 -fixing assemblage was investigated by sequencing of nifH genes. The diazotrophic assemblage in surface waters contained mainly Proteobacterial sequences (Cluster I nifH), while both Proteobacterial sequences and sequences with high identities to those of anaerobic microbes characterized as Clusters III and IV type nifH sequences were found in the anoxic waters. Our results indicate modest input of N through BNF in oxygen-depleted zones mainly due to the activity of proteobacterial diazotrophs.

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.

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

Acromyrmex Leaf-Cutting Ants Have Simple Gut Microbiota with Nitrogen-Fixing Potential

PubMed Central

Zhukova, Mariya; Hansen, Lars H.; Sørensen, Søren J.; Schiøtt, Morten

2015-01-01

Ants and termites have independently evolved obligate fungus-farming mutualisms, but their gardening procedures are fundamentally different, as the termites predigest their plant substrate whereas the ants deposit it directly on the fungus garden. Fungus-growing termites retained diverse gut microbiota, but bacterial gut communities in fungus-growing leaf-cutting ants have not been investigated, so it is unknown whether and how they are specialized on an exclusively fungal diet. Here we characterized the gut bacterial community of Panamanian Acromyrmex species, which are dominated by only four bacterial taxa: Wolbachia, Rhizobiales, and two Entomoplasmatales taxa. We show that the Entomoplasmatales can be both intracellular and extracellular across different gut tissues, Wolbachia is mainly but not exclusively intracellular, and the Rhizobiales species is strictly extracellular and confined to the gut lumen, where it forms biofilms along the hindgut cuticle supported by an adhesive matrix of polysaccharides. Tetracycline diets eliminated the Entomoplasmatales symbionts but hardly affected Wolbachia and only moderately reduced the Rhizobiales, suggesting that the latter are protected by the biofilm matrix. We show that the Rhizobiales symbiont produces bacterial NifH proteins that have been associated with the fixation of nitrogen, suggesting that these compartmentalized hindgut symbionts alleviate nutritional constraints emanating from an exclusive fungus garden diet reared on a substrate of leaves. PMID:26048932

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

The Sulfate Transporter SST1 Is Crucial for Symbiotic Nitrogen Fixation in Lotus japonicus Root Nodules

PubMed Central

Krusell, Lene; Krause, Katja; Ott, Thomas; Desbrosses, Guilhem; Krämer, Ute; Sato, Shusei; Nakamura, Yasukazu; Tabata, Satoshi; James, Euan K.; Sandal, Niels; Stougaard, Jens; Kawaguchi, Masayoshi; Miyamoto, Ai; Suganuma, Norio; Udvardi, Michael K.

2005-01-01

Symbiotic nitrogen fixation (SNF) by intracellular rhizobia within legume root nodules requires the exchange of nutrients between host plant cells and their resident bacteria. Little is known at the molecular level about plant transporters that mediate such exchanges. Several mutants of the model legume Lotus japonicus have been identified that develop nodules with metabolic defects that cannot fix nitrogen efficiently and exhibit retarded growth under symbiotic conditions. Map-based cloning of defective genes in two such mutants, sst1-1 and sst1-2 (for symbiotic sulfate transporter), revealed two alleles of the same gene. The gene is expressed in a nodule-specific manner and encodes a protein homologous with eukaryotic sulfate transporters. Full-length cDNA of the gene complemented a yeast mutant defective in sulfate transport. Hence, the gene was named Sst1. The sst1-1 and sst1-2 mutants exhibited normal growth and development under nonsymbiotic growth conditions, a result consistent with the nodule-specific expression of Sst1. Data from a previous proteomic study indicate that SST1 is located on the symbiosome membrane in Lotus nodules. Together, these results suggest that SST1 transports sulfate from the plant cell cytoplasm to the intracellular rhizobia, where the nutrient is essential for protein and cofactor synthesis, including nitrogenase biosynthesis. This work shows the importance of plant sulfate transport in SNF and the specialization of a eukaryotic transporter gene for this purpose. PMID:15805486

Prospective application of Leucaena leucocephala for phytoextraction of Cd and Zn and nitrogen fixation in metal polluted soils.

PubMed

Saraswat, Shweta; Rai, J P N

2011-03-01

The study deals with phytoextraction of Zn and Cd by Leucaena leucocephala grown on effluent fed and low nitrogen soils collected from S1, S2, and S3 sites, representing decreasing metal content with increasing distance from the effluent drain. Plant nitrogen fixation potential and soil micro-biochemical attributes against metal stress were also assessed. Increasing soil metal content and plant growth enhanced metal accumulation. Relatively greater amount of Zn than Cd was accumulated by L. leucocephala, which exceeded in roots with that of other parts. Remediation factor for Cd was maximum (3.6%) in S2 grown plant. Nodule numbers, their biomass, nitrogenase activity, and leghaemoglobin content were maximum in plants grown in S3 and minimum in S1 soil having maximum metals. Maximum soil organic C, total N, C(mic), and N(mic), respiration rate, ATP content, and enzymatic activities in response to phytoremediation was recorded in S3 followed by S2 and S1. Phytoremediation for a year enhanced extractable Zn and Cd by 36% and 45%, and their total removal by 20% and 30%, respectively from S2, which suggests the possible application of L. leucocephala for the remediation of metal contaminated sites and their fertility restoration by improving microbial functionalities and N-pool.

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.

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.

Autoregulation of Nodulation Interferes with Impacts of Nitrogen Fertilization Levels on the Leaf-Associated Bacterial Community in Soybeans ▿ †

PubMed Central

Ikeda, Seishi; Anda, Mizue; Inaba, Shoko; Eda, Shima; Sato, Shusei; Sasaki, Kazuhiro; Tabata, Satoshi; Mitsui, Hisayuki; Sato, Tadashi; Shinano, Takuro; Minamisawa, Kiwamu

2011-01-01

The diversities leaf-associated bacteria on nonnodulated (Nod−), wild-type nodulated (Nod+), and hypernodulated (Nod++) soybeans were evaluated by clone library analyses of the 16S rRNA gene. To analyze the impact of nitrogen fertilization on the bacterial leaf community, soybeans were treated with standard nitrogen (SN) (15 kg N ha−1) or heavy nitrogen (HN) (615 kg N ha−1) fertilization. Under SN fertilization, the relative abundance of Alphaproteobacteria was significantly higher in Nod− and Nod++ soybeans (82% to 96%) than in Nod+ soybeans (54%). The community structure of leaf-associated bacteria in Nod+ soybeans was almost unaffected by the levels of nitrogen fertilization. However, differences were visible in Nod− and Nod++ soybeans. HN fertilization drastically decreased the relative abundance of Alphaproteobacteria in Nod− and Nod++ soybeans (46% to 76%) and, conversely, increased those of Gammaproteobacteria and Firmicutes in these mutant soybeans. In the Alphaproteobacteria, cluster analyses identified two operational taxonomic units (OTUs) (Aurantimonas sp. and Methylobacterium sp.) that were especially sensitive to nodulation phenotypes under SN fertilization and to nitrogen fertilization levels. Arbuscular mycorrhizal infection was not observed on the root tissues examined, presumably due to the rotation of paddy and upland fields. These results suggest that a subpopulation of leaf-associated bacteria in wild-type Nod+ soybeans is controlled in similar ways through the systemic regulation of autoregulation of nodulation, which interferes with the impacts of N levels on the bacterial community of soybean leaves. PMID:21239540

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.

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

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

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.

Role of antimicrobial peptides in controlling symbiotic bacterial populations.

PubMed

Mergaert, P

2018-04-25

Covering: up to 2018 Antimicrobial peptides (AMPs) have been known for well over three decades as crucial mediators of the innate immune response in animals and plants, where they are involved in the killing of infecting microbes. However, AMPs have now also been found to be produced by eukaryotic hosts during symbiotic interactions with bacteria. These symbiotic AMPs target the symbionts and therefore have a more subtle biological role: not eliminating the microbial symbiont population but rather keeping it in check. The arsenal of AMPs and the symbionts' adaptations to resist them are in a careful balance, which contributes to the establishment of the host-microbe homeostasis. Although in many cases the biological roles of symbiotic AMPs remain elusive, for a number of symbiotic interactions, precise functions have been assigned or proposed to the AMPs, which are discussed here. The microbiota living on epithelia in animals, from the most primitive ones to the mammals, are challenged by a cocktail of AMPs that determine the specific composition of the bacterial community as well as its spatial organization. In the symbiosis of legume plants with nitrogen-fixing rhizobium bacteria, the host deploys an extremely large panel of AMPs - called nodule-specific cysteine-rich (NCR) peptides - that drive the bacteria into a terminally differentiated state and manipulate the symbiont physiology to maximize the benefit for the host. The NCR peptides are used as tools to enslave the bacterial symbionts, limiting their reproduction but keeping them metabolically active for nitrogen fixation. In the nutritional symbiotic interactions of insects and protists that have vertically transmitted bacterial symbionts with reduced genomes, symbiotic AMPs could facilitate the integration of the endosymbiont and host metabolism by favouring the flow of metabolites across the symbiont membrane through membrane permeabilization.

Biochemical and Molecular Phylogenetic Study of Agriculturally Useful Association of a Nitrogen-Fixing Cyanobacterium and Nodule Sinorhizobium with Medicago sativa L.

PubMed Central

Karaushu, E. V.; Kravzova, T. R.; Vorobey, N. A.; Kiriziy, D. A.; Olkhovich, O. P.; Taran, N. Yu.; Kots, S. Ya.; Omarova, E.

2015-01-01

Seed inoculation with bacterial consortium was found to increase legume yield, providing a higher growth than the standard nitrogen treatment methods. Alfalfa plants were inoculated by mono- and binary compositions of nitrogen-fixing microorganisms. Their physiological and biochemical properties were estimated. Inoculation by microbial consortium of Sinorhizobium meliloti T17 together with a new cyanobacterial isolate Nostoc PTV was more efficient than the single-rhizobium strain inoculation. This treatment provides an intensification of the processes of biological nitrogen fixation by rhizobia bacteria in the root nodules and an intensification of plant photosynthesis. Inoculation by bacterial consortium stimulates growth of plant mass and rhizogenesis and leads to increased productivity of alfalfa and to improving the amino acid composition of plant leaves. The full nucleotide sequence of the rRNA gene cluster and partial sequence of the dinitrogenase reductase (nifH) gene of Nostoc PTV were deposited to GenBank (JQ259185.1, JQ259186.1). Comparison of these gene sequences of Nostoc PTV with all sequences present at the GenBank shows that this cyanobacterial strain does not have 100% identity with any organisms investigated previously. Phylogenetic analysis showed that this cyanobacterium clustered with high credibility values with Nostoc muscorum. PMID:26114100

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

Effect of vegetation types on soil arbuscular mycorrhizal fungi and nitrogen-fixing bacterial communities in a karst region.

PubMed

Liang, Yueming; Pan, Fujing; He, Xunyang; Chen, Xiangbi; Su, Yirong

2016-09-01

Arbuscular mycorrhizal (AM) fungi and nitrogen-fixing bacteria play important roles in plant growth and recovery in degraded ecosystems. The desertification in karst regions has become more severe in recent decades. Evaluation of the fungal and bacterial diversity of such regions during vegetation restoration is required for effective protection and restoration in these regions. Therefore, we analyzed relationships among AM fungi and nitrogen-fixing bacteria abundances, plant species diversity, and soil properties in four typical ecosystems of vegetation restoration (tussock (TK), shrub (SB), secondary forest (SF), and primary forest (PF)) in a karst region of southwest China. Abundance of AM fungi and nitrogen-fixing bacteria, plant species diversity, and soil nutrient levels increased from the tussock to the primary forest. The AM fungus, nitrogen-fixing bacterium, and plant community composition differed significantly between vegetation types (p < 0.05). Plant richness and pH were linked to the community composition of fungi and nitrogen-fixing bacteria, respectively. Available phosphorus, total nitrogen, and soil organic carbon levels and plant richness were positively correlated with the abundance of AM fungi and nitrogen-fixing bacteria (p < 0.05). The results suggested that abundance of AM fungi and nitrogen-fixing bacteria increased from the tussock to the primary forest and highlight the essentiality of these communities for vegetation restoration.

454-Pyrosequencing Analysis of Bacterial Communities from Autotrophic Nitrogen Removal Bioreactors Utilizing Universal Primers: Effect of Annealing Temperature

PubMed Central

Rodriguez-Sanchez, Alejandro; Rodelas, Belén; Abbas, Ben A.; Martinez-Toledo, Maria Victoria; van Loosdrecht, Mark C. M.; Osorio, F.; Gonzalez-Lopez, Jesus

2015-01-01

Identification of anaerobic ammonium oxidizing (anammox) bacteria by molecular tools aimed at the evaluation of bacterial diversity in autotrophic nitrogen removal systems is limited by the difficulty to design universal primers for the Bacteria domain able to amplify the anammox 16S rRNA genes. A metagenomic analysis (pyrosequencing) of total bacterial diversity including anammox population in five autotrophic nitrogen removal technologies, two bench-scale models (MBR and Low Temperature CANON) and three full-scale bioreactors (anammox, CANON, and DEMON), was successfully carried out by optimization of primer selection and PCR conditions (annealing temperature). The universal primer 530F was identified as the best candidate for total bacteria and anammox bacteria diversity coverage. Salt-adjusted optimum annealing temperature of primer 530F was calculated (47°C) and hence a range of annealing temperatures of 44–49°C was tested. Pyrosequencing data showed that annealing temperature of 45°C yielded the best results in terms of species richness and diversity for all bioreactors analyzed. PMID:26421306

Investigations of the structure and function of bacterial communities associated with Sphagnum mosses.

PubMed

Opelt, Katja; Chobot, Vladimir; Hadacek, Franz; Schönmann, Susan; Eberl, Leo; Berg, Gabriele

2007-11-01

High acidity, low temperature and extremely low concentration of nutrients form Sphagnum bogs into extreme habitats for organisms. Little is known about the bacteria associated with living Sphagnum plantlets, especially about their function for the host. Therefore, we analysed the endo- and ectophytic bacterial populations associated with two widely distributed Sphagnum species, Sphagnum magellanicum and Sphagnum fallax, by a multiphasic approach. The screening of 1222 isolates for antagonistic activity resulted in 326 active isolates. The bacterial communities harboured a high proportion of antifungal (26%) but a low proportion of antibacterial isolates (0.4%). Members of the genus Burkholderia (38%) were found to be the most dominant group of antagonistic bacteria. The finding that a large proportion (89%) of the antagonistic bacteria produced antifungal compounds may provide an explanation for the well-known antimicrobial activity of certain Sphagnum species. The secondary metabolites of the Sphagnum species themselves were analysed by HPLC-PDA. The different spectra of detected compounds may not only explain the antifungal activity but also the species specificity of the microbial communities. The latter was analysed using cultivation-independent single-stranded conformation polymorphism (SSCP) analysis. Using Burkholderia-specific primers we found a high diversity of Burkholderia isolates in the endophytic and ectophytic habitats of Sphagnum. Furthermore, a high diversity of nitrogen-fixing bacteria was detected by using nifH-specific primers, especially inside Sphagnum mosses. In conclusion, this study provides evidence that both Sphagnum species were colonized by characteristic bacterial populations, which appear to be important for pathogen defence and nitrogen fixation.

Carbon dioxide fixation in the metabolism of propylene and propylene oxide by Xanthobacter strain Py2.

PubMed Central

Small, F J; Ensign, S A

1995-01-01

Evidence for a requirement for CO2 in the productive metabolism of aliphatic alkenes and epoxides by the propylene-oxidizing bacterium Xanthobacter strain Py2 is presented. In the absence of CO2, whole-cell suspensions of propylene-grown cells catalyzed the isomerization of propylene oxide (epoxypropane) to acetone. In the presence of CO2, no acetone was produced. Acetone was not metabolized by suspensions of propylene-grown cells, in either the absence or presence of CO2. The degradation of propylene and propylene oxide by propylene-grown cells supported the fixation of 14CO2 into cell material, and the time course of 14C fixation correlated with the time course of propylene and propylene oxide degradation. The degradation of glucose and propionaldehyde by propylene-grown or glucose-grown cells did not support significant 14CO2 fixation. With propylene oxide as the substrate, the concentration dependence of 14CO2 fixation exhibited saturation kinetics, and at saturation, 0.9 mol of CO2 was fixed per mol of propylene oxide consumed. Cultures grown with propylene in a nitrogen-deficient medium supplemented with NaH13CO3 specifically incorporated 13C label into the C-1 (major labeled position) and C-3 (minor labeled position) carbon atoms of the endogenous storage compound poly-beta-hydroxybutyrate. No specific label incorporation was observed when cells were cultured with glucose or n-propanol as a carbon source. The depletion of CO2 from cultures grown with propylene, but not glucose or n-propanol, inhibited bacterial growth. We propose that propylene oxide metabolism in Xanthobacter strain Py2 proceeds by terminal carboxylation of an isomerization intermediate, which, in the absence of CO2, is released as acetone. PMID:7592382

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

Plant, fungal, bacterial, and nitrogen interactions in the litter layer of a native Patagonian forest.

PubMed

Vivanco, Lucía; Rascovan, Nicolás; Austin, Amy T

2018-01-01

Plant-microbial interactions in the litter layer represent one of the most relevant interactions for biogeochemical cycling as litter decomposition is a key first step in carbon and nitrogen turnover. However, our understanding of these interactions in the litter layer remains elusive. In an old-growth mixed Nothofagus forest in Patagonia, we studied the effects of single tree species identity and the mixture of three tree species on the fungal and bacterial composition in the litter layer. We also evaluated the effects of nitrogen (N) addition on these plant-microbial interactions. In addition, we compared the magnitude of stimulation of litter decomposition due to home field advantage (HFA, decomposition occurs more rapidly when litter is placed beneath the plant species from which it had been derived than beneath a different plant species) and N addition that we previously demonstrated in this same forest, and used microbial information to interpret these results. Tree species identity had a strong and significant effect on the composition of fungal communities but not on the bacterial community of the litter layer. The microbial composition of the litter layer under the tree species mixture show an averaged contribution of each single tree species. N addition did not erase the plant species footprint on the fungal community, and neither altered the bacterial community. N addition stimulated litter decomposition as much as HFA for certain tree species, but the mechanisms behind N and HFA stimulation may have differed. Our results suggest that stimulation of decomposition from N addition might have occurred due to increased microbial activity without large changes in microbial community composition, while HFA may have resulted principally from plant species' effects on the litter fungal community. Together, our results suggest that plant-microbial interactions can be an unconsidered driver of litter decomposition in temperate forests.

Spatially robust estimates of biological nitrogen (N) fixation imply substantial human alteration of the tropical N cycle

USGS Publications Warehouse

Sullivan, Benjamin W.; Smith, William K.; Townsend, Alan R.; Nasto, Megan K.; Reed, Sasha C.; Chazdon, Robin L.; Cleveland, Cory C.

2014-01-01

Biological nitrogen fixation (BNF) is the largest natural source of exogenous nitrogen (N) to unmanaged ecosystems and also the primary baseline against which anthropogenic changes to the N cycle are measured. Rates of BNF in tropical rainforest are thought to be among the highest on Earth, but they are notoriously difficult to quantify and are based on little empirical data. We adapted a sampling strategy from community ecology to generate spatial estimates of symbiotic and free-living BNF in secondary and primary forest sites that span a typical range of tropical forest legume abundance. Although total BNF was higher in secondary than primary forest, overall rates were roughly five times lower than previous estimates for the tropical forest biome. We found strong correlations between symbiotic BNF and legume abundance, but we also show that spatially free-living BNF often exceeds symbiotic inputs. Our results suggest that BNF in tropical forest has been overestimated, and our data are consistent with a recent top-down estimate of global BNF that implied but did not measure low tropical BNF rates. Finally, comparing tropical BNF within the historical area of tropical rainforest with current anthropogenic N inputs indicates that humans have already at least doubled reactive N inputs to the tropical forest biome, a far greater change than previously thought. Because N inputs are increasing faster in the tropics than anywhere on Earth, both the proportion and the effects of human N enrichment are likely to grow in the future.

Biochar amendment decreases soil microbial biomass and increases bacterial diversity in Moso bamboo (Phyllostachys edulis) plantations under simulated nitrogen deposition

NASA Astrophysics Data System (ADS)

Li, Quan; Lei, Zhaofeng; Song, Xinzhang; Zhang, Zhiting; Ying, Yeqing; Peng, Changhui

2018-04-01

Biochar amendment has been proposed as a strategy to improve acidic soils after overuse of nitrogen fertilizers. However, little is known of the role of biochar in soil microbial biomass carbon (MBC) and bacterial community structure and diversity after soil acidification induced by nitrogen (N) deposition. Using high-throughput sequencing of the 16S rRNA gene, we determined the effects of biochar amendment (BC0, 0 t bamboo biochar ha‑1 BC20, 20 t bamboo biochar ha‑1 and BC40, 40 t bamboo biochar ha‑1) on the soil bacterial community structure and diversity in Moso bamboo plantations that had received simulated N deposition (N30, 30 kg N ha‑1 yr‑1 N60, 60 kg N ha‑1 yr‑1 N90, 90 kg N ha‑1 yr‑1 and N-free) for 21 months. After treatment of N-free plots, BC20 significantly increased soil MBC and bacterial diversity, while BC40 significantly decreased soil MBC but increased bacterial diversity. When used to amend N30 and N60 plots, biochar significantly decreased soil MBC and the reducing effect increased with biochar amendment amount. However, these significant effects were not observed in N90 plots. Under N deposition, biochar amendment largely increased soil bacterial diversity, and these effects depended on the rates of N deposition and biochar amendment. Soil bacterial diversity was significantly related to the soil C/N ratio, pH, and soil organic carbon content. These findings suggest an optimal approach for using biochar to offset the effects of N deposition in plantation soils and provide a new perspective for understanding the potential role of biochar amendments in plantation soil.

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.

Prevalence of potential nitrogen-fixing, green sulphur bacteria in the skeleton of reef-building coral Isopora palifera

NASA Astrophysics Data System (ADS)

Yang, S. H.

2016-02-01

Microbial endoliths, which inhabit interior pores of rocks, skeletons and coral, are ubiquitous in terrestrial and marine environments. In the present study, various colored layers stratified the endolithic environment within the skeleton of Isopora palifera; however, there was a distinct green-pigmented layer in the skeleton (beneath the living coral tissue). To characterize diversity of endolithic microorganisms, 16S ribosomal RNA gene amplicon pyrosequencing was used to investigate bacterial communities in the green layer of Isopora palifera coral colonies retrieved fromGreen Island, Taiwan. The dominant bacterial group in the green layer belonged to the bacterial phylum Chlorobi, green sulphur bacteria capable of anoxygenic photosynthesis and nitrogen fixation. Specifically, bacteria of the genus Prosthecochloris were dominant in this green layer. To our knowledge, this is the first study to provide a detailed profile of endolithic bacteria in coral and to determine prevalence of Prosthecochloris in the green layer. Based on our findings, we infer that these bacteria may have an important functional role in the coral holobiont in the nutrient-limited coral reef ecosystem.

A nitrogen mass balance for California

NASA Astrophysics Data System (ADS)

Liptzin, D.; Dahlgren, R. A.

2010-12-01

Human activities have greatly altered the global nitrogen cycle and these changes are apparent in water quality, air quality, ecosystem and human health. However, the relative magnitude of the sources of new reactive nitrogen and the fate of this nitrogen is not well established. Further, the biogeochemical aspects of the nitrogen cycle are often studied in isolation from the economic and social implications of all the transformations of nitrogen. The California Nitrogen Assessment is an interdisciplinary project whose aim is evaluating the current state of nitrogen science, practice, and policy in the state of California. Because of the close proximity of large population centers, highly productive and diverse agricultural lands and significant acreage of undeveloped land, California is a particularly interesting place for this analysis. One component of this assessment is developing a mass balance of nitrogen as well as identifying gaps in knowledge and quantifying uncertainty. The main inputs of new reactive nitrogen to the state are 1) synthetic nitrogen fertilizer, 2) biological nitrogen fixation, and 3) atmospheric nitrogen deposition. Permanent losses of nitrogen include 1) gaseous losses (N2, N2O, NHx, NOy), 2) riverine discharge, 3) wastewater discharge to the ocean, and 4) net groundwater recharge. A final term is the balance of food, feed, and fiber to support the human and animal populations. The largest input of new reactive nitrogen to California is nitrogen fertilizer, but both nitrogen fixation and atmospheric deposition contribute significantly. Non-fertilizer uses, such as the production of nylon and polyurethane, constitutes about 5% of the synthetic N synthesized production. The total nitrogen fixation in California is roughly equivalent on the 400,000 ha of alfalfa and the approximately 40 million ha of natural lands. In addition, even with highly productive agricultural lands, the large population of livestock, in particular dairy cows

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,...

EnviroAtlas - Biological nitrogen fixation in natural/semi-natural ecosystems by 12-digit HUC for the Conterminous United States, 2006

EPA Pesticide Factsheets

This EnviroAtlas dataset contains data on the mean biological nitrogen fixation in natural/semi-natural ecosystems per 12-digit Hydrologic Unit (HUC) in 2006. Biological N fixation (BNF) in natural/semi-natural ecosystems was estimated using a correlation with actual evapotranspiration (AET). This correlation is based on a global meta-analysis of BNF in natural/semi-natural ecosystems (Cleveland et al. 1999). AET estimates for 2006 were calculated using a regression equation describing the correlation of AET with climate (average annual daily temperature, average annual minimum daily temperature, average annual maximum daily temperature, and annual precipitation) and land use/land cover variables in the conterminous US (Sanford and Selnick 2013). Data describing annual average minimum and maximum daily temperatures and total precipitation for 2006 were acquired from the PRISM climate dataset (http://prism.oregonstate.edu). Average annual climate data were then calculated for individual 12-digit USGS Hydrologic Unit Codes (HUC12s; http://water.usgs.gov/GIS/huc.html; 22 March 2011 release) using the Zonal Statistics tool in ArcMap 10.0. AET for individual HUC12s was estimated using equations described in Sanford and Selnick (2013). BNF in natural/semi-natural ecosystems within individual HUC12s was modeled with an equation describing the statistical relationship between BNF (kg N ha-1 yr-1) and actual evapotranspiration (AET; cm yr-1) and scaled to the proportion

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.

Metabolic features involved in drought stress tolerance mechanisms in peanut nodules and their contribution to biological nitrogen fixation.

PubMed

Furlan, Ana Laura; Bianucci, Eliana; Castro, Stella; Dietz, Karl-Josef

2017-10-01

Legumes belong to the most important crops worldwide. They increase soil fertility due their ability to establish symbiotic associations with soil microorganisms, known as rhizobia, capable of fixing nitrogen from the atmosphere. However, they are frequently exposed to abiotic stress conditions in particular drought. Such adverse conditions impair the biological nitrogen fixation (BNF) and depend largely on the legume. Therefore, two peanut cultivars with contrasting tolerance to drought, namely the more tolerant EC-98 and the sensitive Granoleico, were investigated to elucidate the relative contribution of BNF to the tolerance to drought. The tolerant cultivar EC-98 sustained growth and BNF similar to the control condition despite the reduced water potential and photosynthesis, suggesting the functioning of distinct metabolic pathways that contributed to enhance the tolerance. The biochemical and metabolomics approaches revealed that nodules from the tolerant cultivar accumulated trehalose, proline and gamma-aminobutyric acid (GABA), metabolites with known function in protecting against drought stress. The amide metabolism was severely affected in nodules from the sensitive cultivar Granoleico as revealed by the low content of asparagine and glutamine in the drought stressed plants. The sensitive cultivar upon rehydration was unable to re-establish a metabolism similar to well-watered plants. This was evidenced by the low level of metabolites and, transcripts and specific activities of enzymes from the carbon (sucrose synthase) and nitrogen (glutamine synthetase) metabolism which decreased below the values of control plants. Therefore, the increased content of metabolites with protective functions under drought stress likely is crucial for the full restoration upon rehydration. Smaller changes of drought stress-related metabolites in nodule are another trait that contributes to the effective control of BNF in the tolerant peanut cultivar (EC-98). Copyright © 2017

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.

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%).

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.

Bacterial tetraethers from Tibetan hot springs: Implications for nitrogen metabolism and biological sources

NASA Astrophysics Data System (ADS)

Li, F.; Zhang, C.; Wang, S.; Klotz, M. G.; Dong, H.; Li, W.

2013-12-01

Branched glycerol dialkyl glycerol tetraethers (bGDGTs) are considered to be produced by bacteria that are predominantly found in soils and peat bogs. Recently, however, in situ production of bGDGTs is reported from a terrestrial hot spring in the Great Basin. In this study, we analyzed water chemistry, bacterial lipids, and pyrosequencing data from 37 Tibetan hot springs in order to evaluate the linkage between biological sources, metabolic processes and the distribution of bGDGTs. Analyses of absolute and relative concentrations of intact polar- and core bGDGTs (IP-bGDGTs and C-bGDGTs) suggest that the bGDGTs are predominantly produced in situ in Tibetan hot springs. Cluster analysis separated the hot spring samples into three major groups, which showed significant relationships between bGDGTs and concentrations of ammonia, nitrite or nitrate. The nirS gene abundance also correlated significantly with bGDGTs. These results indicate that the bGDGT-producing organisms may be involved in nitrogen metabolism in the Tibetan hot springs. Pyrosequencing analysis identified eight phyla of Bacteria (Acidobacteria, Bacteroidetes, Chlorobi, Firmicutes, Nitrospirae, Proteobacteria, Verrucomicrobia and Spirochetes) that may be potential sources of bGDGTs based on significant correlations of these organisms with the distribution of different bGDGTs. Representatives of these phyla have been implicated in nitrogen oxide transformations in many diverse environments including hot springs. Overall, our results suggest that bacteria producing bGDGTs may play an important role in nitrogen cycle in the Tibetan hot springs.

Cascading influence of inorganic nitrogen sources on DOM production, composition, lability and microbial community structure in the open ocean.

PubMed

Goldberg, S J; Nelson, C E; Viviani, D A; Shulse, C N; Church, M J

2017-09-01

Nitrogen frequently limits oceanic photosynthesis and the availability of inorganic nitrogen sources in the surface oceans is shifting with global change. We evaluated the potential for abrupt increases in inorganic N sources to induce cascading effects on dissolved organic matter (DOM) and microbial communities in the surface ocean. We collected water from 5 m depth in the central North Pacific and amended duplicate 20 liter polycarbonate carboys with nitrate or ammonium, tracking planktonic carbon fixation, DOM production, DOM composition and microbial community structure responses over 1 week relative to controls. Both nitrogen sources stimulated bulk phytoplankton, bacterial and DOM production and enriched Synechococcus and Flavobacteriaceae; ammonium enriched for oligotrophic Actinobacteria OM1 and Gammaproteobacteria KI89A clades while nitrate enriched Gammaproteobacteria SAR86, SAR92 and OM60 clades. DOM resulting from both N enrichments was more labile and stimulated growth of copiotrophic Gammaproteobacteria (Alteromonadaceae and Oceanospirillaceae) and Alphaproteobacteria (Rhodobacteraceae and Hyphomonadaceae) in weeklong dark incubations relative to controls. Our study illustrates how nitrogen pulses may have direct and cascading effects on DOM composition and microbial community dynamics in the open ocean. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

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

Spatially robust estimates of biological nitrogen (N) fixation imply substantial human alteration of the tropical N cycle

PubMed Central

Sullivan, Benjamin W.; Smith, W. Kolby; Townsend, Alan R.; Nasto, Megan K.; Reed, Sasha C.; Chazdon, Robin L.; Cleveland, Cory C.

2014-01-01

Biological nitrogen fixation (BNF) is the largest natural source of exogenous nitrogen (N) to unmanaged ecosystems and also the primary baseline against which anthropogenic changes to the N cycle are measured. Rates of BNF in tropical rainforest are thought to be among the highest on Earth, but they are notoriously difficult to quantify and are based on little empirical data. We adapted a sampling strategy from community ecology to generate spatial estimates of symbiotic and free-living BNF in secondary and primary forest sites that span a typical range of tropical forest legume abundance. Although total BNF was higher in secondary than primary forest, overall rates were roughly five times lower than previous estimates for the tropical forest biome. We found strong correlations between symbiotic BNF and legume abundance, but we also show that spatially free-living BNF often exceeds symbiotic inputs. Our results suggest that BNF in tropical forest has been overestimated, and our data are consistent with a recent top-down estimate of global BNF that implied but did not measure low tropical BNF rates. Finally, comparing tropical BNF within the historical area of tropical rainforest with current anthropogenic N inputs indicates that humans have already at least doubled reactive N inputs to the tropical forest biome, a far greater change than previously thought. Because N inputs are increasing faster in the tropics than anywhere on Earth, both the proportion and the effects of human N enrichment are likely to grow in the future. PMID:24843146

In Situ Denitrification and Biological Nitrogen Fixation Under Enhanced Atmospheric Reactive Nitrogen Deposition in UK Peatlands

NASA Astrophysics Data System (ADS)

Ullah, Sami; Saiz Val, Ernesto; Sgouridis, Fotis; Peichl, Matthias; Nilsson, Mats

2017-04-01

Dinitrogen (N2) and nitrous oxide (N2O) losses due to denitrification and biological N2 fixation (BNF) are the most uncertain components of the nitrogen (N) cycle in peatlands under enhanced atmospheric reactive nitrogen (Nr) deposition. This uncertainty hampers our ability to assess the contribution of denitrification to the removal of biologically fixed and/or atmospherically deposited Nr in peatlands. This uncertainty emanates from the difficulty in measuring in situ soil N2 and N2O production and consumption in peatlands. In situ denitrification and its contribution to total N2O flux was measured monthly between April 2013 and October 2014 in peatlands in two UK catchments. An adapted 15N-Gas Flux method1 with low level addition of 15N tracer (0.03 ± 0.005 kg 15N ha-1) was used to measure denitrification and its contribution to net N2O production (DN2O/TN2O). BNF was measured in situ through incubation of selected sphagnum species under 15N2 gas tracer. Denitrification2 varied temporally and averaged 8 kg N-N2 ha-1 y-1. The contribution of denitrification was about 48% to total N2O flux3 of 0.05 kg N ha-1 y-1. Soil moisture, temperature, ecosystem respiration, pH and mineral N content mainly regulated the flux of N2 and N2O. Preliminary results showed suppression of BNF, which was 1.8 to 7 times lower in peatland mosses exposed to ˜15 to 20 kg N ha-1 y-1 Nr deposition in the UK than in peatland mosses in northern Sweden with background Nr deposition. Overall, the contribution of denitrification to Nr removal in the selected peatlands was ˜50% of the annual Nr deposition rates, making these ecosystems vulnerable to chronic N saturation. These results point to a need for a more comprehensive annual BNF measurement to more accurately account for total Nr input into peatlands and its atmospheric loss due to denitrification. References Sgouridis F, Stott A & Ullah S, 2016. Application of the 15N-Gas Flux method for measuring in situ N2 and N2O fluxes due to

Tropical Dominance of N2 Fixation in the North Atlantic Ocean

NASA Astrophysics Data System (ADS)

Marconi, Dario; Sigman, Daniel M.; Casciotti, Karen L.; Campbell, Ethan C.; Alexandra Weigand, M.; Fawcett, Sarah E.; Knapp, Angela N.; Rafter, Patrick A.; Ward, Bess B.; Haug, Gerald H.

2017-10-01

To investigate the controls on N2 fixation and the role of the Atlantic in the global ocean's fixed nitrogen (N) budget, Atlantic N2 fixation is calculated by combining meridional nitrate fluxes across World Ocean Circulation Experiment sections with observed nitrate 15N/14N differences between northward and southward transported nitrate. N2 fixation inputs of 27.1 ± 4.3 Tg N/yr and 3.0 ± 0.5 Tg N/yr are estimated north of 11°S and 24°N, respectively. That is, 90% of the N2 fixation in the Atlantic north of 11°S occurs south of 24°N in a region with upwelling that imports phosphorus (P) in excess of N relative to phytoplankton requirements. This suggests that, under the modern iron-rich conditions of the equatorial and North Atlantic, N2 fixation occurs predominantly in response to P-bearing, N-poor conditions. We estimate a N2 fixation rate of 30.5 ± 4.9 Tg N/yr north of 30°S, implying only 3 Tg N/yr between 30° and 11°S, despite evidence of P-bearing, N-poor surface waters in this region as well; this is consistent with iron limitation of N2 fixation in the South Atlantic. Since the ocean flows through the Atlantic surface in <2,500 years, similar to the residence time of oceanic fixed N, Atlantic N2 fixation can stabilize the N-to-P ratio of the global ocean. However, the calculated rate of Atlantic N2 fixation is a small fraction of global ocean estimates for either N2 fixation or fixed N loss. This suggests that, in the modern ocean, an approximate balance between N loss and N2 fixation is achieved within the combined Indian and Pacific basins.

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

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.

Strong Regionality and Dominance of Anaerobic Bacterial Taxa Characterize Diazotrophic Bacterial Communities of the Arcto-Alpine Plant Species Oxyria digyna and Saxifraga oppositifolia.

PubMed

Kumar, Manoj; van Elsas, Jan Dirk; Nissinen, Riitta

2017-01-01

Arctic and alpine biomes are most often strongly nitrogen-limited, and hence biological nitrogen fixation is a strong driver of these ecosystems. Both biomes are characterized by low temperatures and short growing seasons, but they differ in seasonality of solar radiation and in soil water balance due to underlying permafrost in the Arctic. Arcto-alpine plant species are well-adapted to the low temperatures that prevail in their habitats, and plant growth is mainly limited by the availability of nutrients, in particular nitrogen, due to slow mineralization. Nitrogen fixing bacteria are likely important for plant growth in these habitats, but very little is known of these bacteria or forces shaping their communities. In this study, we characterized the potential nitrogen fixing bacterial (PNFB) communities associated with two arcto-alpine pioneer plant species, Oxyria digyna (mountain sorrel) and Saxifraga oppositifolia (blue saxifrage), in three climate regions. Both of these plants readily colonize low nutrient mineral soils. Our goal was to investigate how climate (region) and, on the other hand, host plant and plant species shape these communities. To our knowledge, this is the first comprehensive study describing PNFB communities associated with pioneer plants in different arcto-alpine biomes. Replicate samples were taken from two arctic regions, Kilpisjärvi and Ny-Ålesund, and one alpine region, Mayrhofen. In these, the PNFB communities in the bulk and rhizosphere soils and the plant endospheres were characterized by nifH -targeted PCR and massive parallel sequencing. The data revealed strong effects of climatic region on the dominating nitrogen fixers. Specifically, nifH sequences related to Geobacter (δ- Proteobacteria ) were present in high relative abundances in the nitrogen-fixing communities in the Mayrhofen and Kilpisjärvi regions, while members of the Clostridiales prevailed in the Kilpisjärvi and Ny-Ålesund regions. The bulk and rhizosphere soil

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

Fixation distance and fixation duration to vertical road signs.

PubMed

Costa, Marco; Simone, Andrea; Vignali, Valeria; Lantieri, Claudio; Palena, Nicola

2018-05-01

The distance of first-fixation to vertical road signs was assessed in 22 participants while driving a route of 8.34 km. Fixations to road signs were recorded by a mobile eye-movement-tracking device synchronized to GPS and kinematic data. The route included 75 road signs. First-fixation distance and fixation duration distributions were positively skewed. Median distance of first-fixation was 51 m. Median fixation duration was 137 ms with a modal value of 66 ms. First-fixation distance was linearly related to speed and fixation duration. Road signs were gazed at a much closer distance than their visibility distance. In a second study a staircase procedure was used to test the presentation-time threshold that lead to a 75% accuracy in road sign identification. The threshold was 35 ms, showing that short fixations to a road signs could lead to a correct identification. Copyright © 2018 Elsevier Ltd. All rights reserved.

Influence of different Sinorhizobium meliloti inocula on abundance of genes involved in nitrogen transformations in the rhizosphere of alfalfa (Medicago sativa L.).

PubMed

Babić, Katarina Huić; Schauss, Kristina; Hai, Brigitte; Sikora, Sanja; Redzepović, Sulejman; Radl, Viviane; Schloter, Michael

2008-11-01

Inoculation of leguminous seeds with selected rhizobial strains is practised in agriculture to ameliorate the plant yield by enhanced root nodulation and nitrogen uptake of the plant. However, effective symbiosis between legumes and rhizobia does not only depend on the capacity of nitrogen fixation but also on the entire nitrogen turnover in the rhizosphere. We investigated the influence of seed inoculation with two indigenous Sinorhizobium meliloti strains exhibiting different efficiency concerning plant growth promotion on nitrogen turnover processes in the rhizosphere during the growth of alfalfa. Quantification of six target genes (bacterial amoA, nirK, nirS, nosZ, nifH and archaeal amoA) within the nitrogen cycle was performed in rhizosphere samples before nodule formation, at bud development and at the late flowering stage. The results clearly demonstrated that effectiveness of rhizobial inocula is related to abundance of nifH genes in the late flowering phase of alfalfa. Moreover, other genes involved in nitrogen turnover had been affected by the inocula, e.g. higher numbers of amoA copies were observed during flowering when the more effective strain had been inoculated. However, the respective gene abundances differed overall to a greater extent between the three plant development stages than between the inoculation variants.

Nitrogen fixation in biological soil crusts from southeast Utah, USA

USGS Publications Warehouse

Belnap, Jayne

2002-01-01

Biological soil crusts can be the dominant source of N for arid land ecosystems. We measured potential N fixation rates biweekly for 2 years, using three types of soil crusts: (1) crusts whose directly counted cells were >98% Microcoleus vaginatus (light crusts); (2) crusts dominated by M. vaginatus, but with 20% or more of the directly counted cells represented by Nostoc commune and Scytonema myochrous (dark crusts); and (3) the soil lichen Collema sp. At all observation times, Collema had higher nitrogenase activity (NA) than dark crusts, which had higher NA than light crusts, indicating that species composition is critical when estimating N inputs. In addition, all three types of crusts generally responded in a similar fashion to climate conditions. Without precipitation within a week of collection, no NA was recorded, regardless of other conditions being favorable. Low (<1°C) and high (>26°C) temperatures precluded NA, even if soils were moist. If rain or snow melt had occurred 3 or less days before collection, NA levels were highly correlated with daily average temperatures of the previous 3 days (r2=0.93 for Collema crusts; r2=0.86 for dark crusts and r2=0.83 for light crusts) for temperatures between 1°C and 26°C. If a precipitation event followed a long dry period, NA levels were lower than if collection followed a time when soils were wet for extended periods (e.g., winter). Using a combination of data from a recording weather datalogger, time-domain reflectometry, manual dry-down curves, and N fixation rates at different temperatures, annual N input from the different crust types was estimated. Annual N input from dark crusts found at relatively undisturbed sites was estimated at 9 kg ha–1 year–1. With 20% cover of the N-fixing soil lichen Collema, inputs are estimated at 13 kg ha–1 year–1. N input from light crusts, generally indicating soil surface disturbance, was estimated at 1.4 kg ha–1 year–1. The rates in light crusts are

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.

Metabolic profiling of two maize (Zea mays L.) inbred lines inoculated with the nitrogen fixing plant-interacting bacteria Herbaspirillum seropedicae and Azospirillum brasilense

PubMed Central

Brusamarello-Santos, Liziane Cristina; Gilard, Françoise; Brulé, Lenaïg; Quilleré, Isabelle; Gourion, Benjamin; Ratet, Pascal; Maltempi de Souza, Emanuel; Lea, Peter J.; Hirel, Bertrand

2017-01-01

Maize roots can be colonized by free-living atmospheric nitrogen (N2)-fixing bacteria (diazotrophs). However, the agronomic potential of non-symbiotic N2-fixation in such an economically important species as maize, has still not been fully exploited. A preliminary approach to improve our understanding of the mechanisms controlling the establishment of such N2-fixing associations has been developed, using two maize inbred lines exhibiting different physiological characteristics. The bacterial-plant interaction has been characterized by means of a metabolomic approach. Two established model strains of Nif+ diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense and their Nif- couterparts defficient in nitrogenase activity, were used to evaluate the impact of the bacterial inoculation and of N2 fixation on the root and leaf metabolic profiles. The two N2-fixing bacteria have been used to inoculate two genetically distant maize lines (FV252 and FV2), already characterized for their contrasting physiological properties. Using a well-controlled gnotobiotic experimental system that allows inoculation of maize plants with the two diazotrophs in a N-free medium, we demonstrated that both maize lines were efficiently colonized by the two bacterial species. We also showed that in the early stages of plant development, both bacterial strains were able to reduce acetylene, suggesting that they contain functional nitrogenase activity and are able to efficiently fix atmospheric N2 (Fix+). The metabolomic approach allowed the identification of metabolites in the two maize lines that were representative of the N2 fixing plant-bacterial interaction, these included mannitol and to a lesser extend trehalose and isocitrate. Whilst other metabolites such as asparagine, although only exhibiting a small increase in maize roots following bacterial infection, were specific for the two Fix+ bacterial strains, in comparison to their Fix- counterparts. Moreover, a number

Metabolic profiling of two maize (Zea mays L.) inbred lines inoculated with the nitrogen fixing plant-interacting bacteria Herbaspirillum seropedicae and Azospirillum brasilense.

PubMed

Brusamarello-Santos, Liziane Cristina; Gilard, Françoise; Brulé, Lenaïg; Quilleré, Isabelle; Gourion, Benjamin; Ratet, Pascal; Maltempi de Souza, Emanuel; Lea, Peter J; Hirel, Bertrand

2017-01-01

Maize roots can be colonized by free-living atmospheric nitrogen (N2)-fixing bacteria (diazotrophs). However, the agronomic potential of non-symbiotic N2-fixation in such an economically important species as maize, has still not been fully exploited. A preliminary approach to improve our understanding of the mechanisms controlling the establishment of such N2-fixing associations has been developed, using two maize inbred lines exhibiting different physiological characteristics. The bacterial-plant interaction has been characterized by means of a metabolomic approach. Two established model strains of Nif+ diazotrophic bacteria, Herbaspirillum seropedicae and Azospirillum brasilense and their Nif- couterparts defficient in nitrogenase activity, were used to evaluate the impact of the bacterial inoculation and of N2 fixation on the root and leaf metabolic profiles. The two N2-fixing bacteria have been used to inoculate two genetically distant maize lines (FV252 and FV2), already characterized for their contrasting physiological properties. Using a well-controlled gnotobiotic experimental system that allows inoculation of maize plants with the two diazotrophs in a N-free medium, we demonstrated that both maize lines were efficiently colonized by the two bacterial species. We also showed that in the early stages of plant development, both bacterial strains were able to reduce acetylene, suggesting that they contain functional nitrogenase activity and are able to efficiently fix atmospheric N2 (Fix+). The metabolomic approach allowed the identification of metabolites in the two maize lines that were representative of the N2 fixing plant-bacterial interaction, these included mannitol and to a lesser extend trehalose and isocitrate. Whilst other metabolites such as asparagine, although only exhibiting a small increase in maize roots following bacterial infection, were specific for the two Fix+ bacterial strains, in comparison to their Fix- counterparts. Moreover, a number

RbohB, a Phaseolus vulgaris NADPH oxidase gene, enhances symbiosome number, bacteroid size, and nitrogen fixation in nodules and impairs mycorrhizal colonization.

PubMed

Arthikala, Manoj-Kumar; Sánchez-López, Rosana; Nava, Noreide; Santana, Olivia; Cárdenas, Luis; Quinto, Carmen

2014-05-01

The reactive oxygen species (ROS) generated by respiratory burst oxidative homologs (Rbohs) are involved in numerous plant cell signaling processes, and have critical roles in the symbiosis between legumes and nitrogen-fixing bacteria. Previously, down-regulation of RbohB in Phaseolus vulgaris was shown to suppress ROS production and abolish Rhizobium infection thread (IT) progression, but also to enhance arbuscular mycorrhizal fungal (AMF) colonization. Thus, Rbohs function both as positive and negative regulators. Here, we assessed the effect of enhancing ROS concentrations, by overexpressing PvRbohB, on the P. vulgaris--rhizobia and P. vulgaris--AMF symbioses. We estimated superoxide concentrations in hairy roots overexpressing PvRbohB, determined the status of early and late events of both Rhizobium and AMF interactions in symbiont-inoculated roots, and analyzed the nodule ultrastructure of transgenic plants overexpressing PvRbohB. Overexpression of PvRbohB significantly enhanced ROS production, the formation of ITs, nodule biomass, and nitrogen-fixing activity, and increased the density of symbiosomes in nodules, and the density and size of bacteroides in symbiosomes. Furthermore, PvCAT, early nodulin, PvSS1, and PvGOGAT transcript abundances were elevated in these nodules. By contrast, mycorrhizal colonization was reduced in roots that overexpressed RbohB. Overexpression of PvRbohB augmented nodule efficiency by enhancing nitrogen fixation and delaying nodule senescence, but impaired AMF colonization. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

Integrated analysis of zone-specific protein and metabolite profiles within nitrogen-fixing Medicago truncatula-Sinorhizobium medicae nodules

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

Ogden, Aaron J.; Gargouri, Mahmoud; Park, JeongJin

Symbiotic nitrogen fixation (SNF) between rhizobia and legumes requires metabolic coordination within specialized root organs called nodules. Nodules formed in the symbiosis between S. medicae and barrel medic (M. truncatula) are indeterminate, cylindrical, and contain spatially distinct developmental zones. Bacteria in the infection zone II (ZII), interzone II-III (IZ), and nitrogen fixation zone III (ZIII) represent different stages in the metabolic progression from free-living bacteria into nitrogen fixing bacteroids. To better understand the coordination of plant and bacterial metabolism within the nodule, we used liquid and gas chromatography coupled to tandem mass spectrometry (MS) to observe protein and metabolite profilesmore » representative of ZII, IZ, ZIII, whole-nodule, and primary root. Our MS-based approach confidently identified 361 S. medicae proteins and 888 M. truncatula proteins, as well as 160 metabolites from each tissue. The data are consistent with several organ- and zone-specific protein and metabolite localization patterns characterized previously. Here, we used our comprehensive dataset to demonstrate how multiple branches of primary metabolism are coordinated between symbionts and zones, including central carbon, fatty acid, and amino acid metabolism. For example, M. truncatula glycolysis enzymes accumulate from zone I to zone III within the nodule, while equivalent S. medicae enzymes decrease in abundance. We also show the localization of S. medicae's transition to dicarboxylic acid-dependent carbon metabolism within the IZ. The spatial abundance patterns of S. medicae fatty acid (FA) biosynthesis enzymes indicate an increased demand for FA production in the IZ and ZIII as compared to ZI. Our observations provide a resource for those seeking to understand coordinated physiological changes during the development of SNF.« less

Integrated analysis of zone-specific protein and metabolite profiles within nitrogen-fixing Medicago truncatula-Sinorhizobium medicae nodules

DOE PAGES

Ogden, Aaron J.; Gargouri, Mahmoud; Park, JeongJin; ...

2017-07-10

Symbiotic nitrogen fixation (SNF) between rhizobia and legumes requires metabolic coordination within specialized root organs called nodules. Nodules formed in the symbiosis between S. medicae and barrel medic (M. truncatula) are indeterminate, cylindrical, and contain spatially distinct developmental zones. Bacteria in the infection zone II (ZII), interzone II-III (IZ), and nitrogen fixation zone III (ZIII) represent different stages in the metabolic progression from free-living bacteria into nitrogen fixing bacteroids. To better understand the coordination of plant and bacterial metabolism within the nodule, we used liquid and gas chromatography coupled to tandem mass spectrometry (MS) to observe protein and metabolite profilesmore » representative of ZII, IZ, ZIII, whole-nodule, and primary root. Our MS-based approach confidently identified 361 S. medicae proteins and 888 M. truncatula proteins, as well as 160 metabolites from each tissue. The data are consistent with several organ- and zone-specific protein and metabolite localization patterns characterized previously. Here, we used our comprehensive dataset to demonstrate how multiple branches of primary metabolism are coordinated between symbionts and zones, including central carbon, fatty acid, and amino acid metabolism. For example, M. truncatula glycolysis enzymes accumulate from zone I to zone III within the nodule, while equivalent S. medicae enzymes decrease in abundance. We also show the localization of S. medicae's transition to dicarboxylic acid-dependent carbon metabolism within the IZ. The spatial abundance patterns of S. medicae fatty acid (FA) biosynthesis enzymes indicate an increased demand for FA production in the IZ and ZIII as compared to ZI. Our observations provide a resource for those seeking to understand coordinated physiological changes during the development of SNF.« less

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.

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

Importance of Nitrogen Availability on Land Carbon Sequestration in Northern Eurasia during the 21st Century

NASA Astrophysics Data System (ADS)

Kicklighter, D. W.; Melillo, J. M.; Monier, E.; Sokolov, A. P.; Lu, X.; Zhuang, Q.

2015-12-01

Atmospheric nitrogen deposition, nitrogen fixation, and the application of nitrogen fertilizers provide subsidies to land ecosystems that can increase nitrogen availability for vegetation production and thereby influence land carbon dynamics. In addition, enhanced decomposition of soil organic matter (SOM) from warming soils and permafrost degradation may also increase nitrogen availability in Northern Eurasia. Here, we examine how changes in nitrogen availability may influence land carbon dynamics in Northern Eurasia during the 21st century by comparing results for a "business as usual" scenario (the IPCC Representative Concentration Pathways or RCP 8.5) and a stabilization scenario (RCP 4.5) between a version of the Terrestrial Ecosystem Model that does not consider the effects of atmospheric nitrogen deposition, nitrogen fixation and soil thermal dynamics on land carbon dynamics (TEM 4.4) and a version that does consider these dynamics (TEM 6.0). In these simulations, atmospheric nitrogen deposition, nitrogen fixation, and fertilizer applications provide an additional 3.3 Pg N (RCP 4.5) to 3.9 Pg N (RCP 8.5) to Northern Eurasian ecosystems over the 21st century. Land ecosystems retain about 38% (RCP4.5) to 48% (RCP 8.5) of this nitrogen subsidy. Net nitrogen mineralization estimated by TEM 6.0 provide an additional 1.0 Pg N to vegetation than estimated by TEM 4.4 over the 21st century from enhanced decomposition of SOM including SOM formerly protected by permafrost. The enhanced nitrogen availability in TEM 6.0 allows Northern Eurasian ecosystems to sequester 1.8x (RCP 8.5) to 2.4x (RCP 4.5) more carbon over the 21st century than estimated by TEM 4.4. Our results indicate that consideration of nitrogen subsidies and soil thermal dynamics have a large influence on how simulated land carbon dynamics in Northern Eurasia will respond to future changes in climate, atmospheric chemistry, and disturbances.

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

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.

Carbon and nitrogen balance of leaf-eating sesarmid crabs ( Neoepisesarma versicolor) offered different food sources

NASA Astrophysics Data System (ADS)

Thongtham, Nalinee; Kristensen, Erik

2005-10-01

Carbon and nitrogen budgets for the leaf-eating crab, Neoepisesarma versicolor, were established for individuals living on pure leaf diets. Crabs were fed fresh (green), senescent (yellow) and partly degraded (brown) leaves of the mangrove tree Rhizophora apiculata. Ingestion, egestion and metabolic loss of carbon and nitrogen were determined from laboratory experiments. In addition, bacterial abundance in various compartments of the crabs' digestive tract was enumerated after dissection of live individuals. Ingestion and egestion rates (in terms of dry weight) were highest, while the assimilation efficiency was poorest for crabs fed on brown leaves. The low assimilation efficiency was more than counteracted by the high ingestion rate providing more carbon for growth than for crabs fed green and yellow leaves. In any case, the results show that all types of leaves can provide adequate carbon while nitrogen was insufficient to support both maintenance (yellow leaves) and growth (green, yellow and brown leaves). Leaf-eating crabs must therefore obtain supplementary nitrogen by other means in order to meet their nitrogen requirement. Three hypotheses were evaluated: (1) crabs supplement their diet with bacteria and benthic microalgae by ingesting own faeces and/or selective grazing at the sediment surface; (2) assimilation of symbiotic nitrogen-fixing bacteria in the crabs' own intestinal system; and (3) nitrogen storage following occasional feeding on animal tissues (e.g. meiofauna and carcasses). It appears that hypothesis 1 is of limited importance for N. versicolor since faeces and sediment can only supply a minor fraction of the missing nitrogen due to physical constraints on the amount of material the crabs can consume. Hypothesis 2 can be ruled out because tests showed no nitrogen fixation activity in the intestinal system of N. versicolor. It is therefore likely that leaf-eating crabs provide most of their nitrogen requirement from intracellular deposits

Nitrogen-doped carbon nanofibers derived from polypyrrole coated bacterial cellulose as high-performance electrode materials for supercapacitors and Li-ion batteries

DOE PAGES

Lei, Wen; Han, Lili; Xuan, Cuijuan; ...

2016-05-24

Here, nitrogen-doped carbon nanofiber (NDCN) was synthesized via carbonization of polypyrrole (PPy) coated bacterial cellulose (BC) composites, where BC serves as templates as well as precursor, and PPy serves as the nitrogen source. The synthesized NDCN was employed as electrode for both supercapacitors and Li-ion batteries. The large surface area exposed to electrolyte resulting from the 3D carbon networks leads to sufficient electrode/electrolyte interface and creates shorter transport paths of electrolyte ions and Li + ion. Besides, the three types of N dopants in NDCN improve the electronic conductivity, as well as superior electrochemical performance.

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

Origin and Evolution of Nitrogen Fixation Genes on Symbiosis Islands and Plasmid in Bradyrhizobium

PubMed Central

Okubo, Takashi; Piromyou, Pongdet; Tittabutr, Panlada; Teaumroong, Neung; Minamisawa, Kiwamu

2016-01-01

The nitrogen fixation (nif) genes of nodule-forming Bradyrhizobium strains are generally located on symbiosis islands or symbiosis plasmids, suggesting that these genes have been transferred laterally. The nif genes of rhizobial and non-rhizobial Bradyrhizobium strains were compared in order to infer the evolutionary histories of nif genes. Based on all codon positions, the phylogenetic tree of concatenated nifD and nifK sequences showed that nifDK on symbiosis islands formed a different clade from nifDK on non-symbiotic loci (located outside of symbiosis islands and plasmids) with elongated branches; however, these genes were located in close proximity, when only the 1st and 2nd codon positions were analyzed. The guanine (G) and cytosine (C) content of the 3rd codon position of nifDK on symbiosis islands was lower than that on non-symbiotic loci. These results suggest that nif genes on symbiosis islands were derived from the non-symbiotic loci of Bradyrhizobium or closely related strains and have evolved toward a lower GC content with a higher substitution rate than the ancestral state. Meanwhile, nifDK on symbiosis plasmids clustered with nifDK on non-symbiotic loci in the tree representing all codon positions, and the GC content of symbiotic and non-symbiotic loci were similar. These results suggest that nif genes on symbiosis plasmids were derived from the non-symbiotic loci of Bradyrhizobium and have evolved with a similar evolutionary pattern and rate as the ancestral state. PMID:27431195

Plant, Microbiome, and Biogeochemistry: Quantifying moss-associated N fixation in Alaska

NASA Astrophysics Data System (ADS)

Stuart, J.; Mack, M. C.; Holland Moritz, H.; Fierer, N.; McDaniels, S.; Lewis, L.

2017-12-01

The future carbon (C) sequestration potential of the Arctic and boreal zones, currently the largest terrestrial C sink globally, is linked to nitrogen (N) cycling and N availability vis-a-vis C accumulation and plant species composition. Pristine environments in Alaska have low anthropogenic N deposition (<1 kg N ha-1 yr-1), and the main source of new N to these ecosystems is through previously overlooked N-fixation from microbial communities on mosses. Despite the importance of moss associated N-fixation, the relationship between moss species, microbial communities, and fixation rates remains ambiguous. In the summer of 2016, the fixation rates of 20 moss species from sites around both Fairbanks and Toolik Lake were quantified using 15N2 incubations. Subsequently, the microbial community and moss genome of the samples were also analyzed by collaborators. The most striking result is that all sampled moss genera fixed N, including well-studied feather mosses such as Hylocomium splendens and Pleurozium schreberi as well as less common but ecologically relevant mosses such as Aulacomnium spp., Dicranum spp., Ptilium crista-castrensis, and Tomentypnum nitens. Across all samples, preliminary fixation rates ranged from 0.004-19.994 µg N g-1 moss d-1. Depending upon percent cover, moss-associated N fixation is the largest input of new N to the ecosystem. Given this, linking variation in N-fixation rates to microbial and moss community structures can be helpful in predicting future trends of C and N cycling in northern latitudes. Vegetation changes, alterations in downstream biogeochemical N processes, and anthropogenic N deposition could all interact with or alter moss associated N-fixation, thereby changing ecosystem N inputs. Further elucidation of the species level signal in N-fixation rates and microbial community will augment our knowledge of N cycling in northern latitudes, both current and future.

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 ...

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

Abundance and activity of 16S rRNA, amoA and nifH bacterial genes during assisted phytostabilization of mine tailings

PubMed Central

Nelson, Karis N.; Neilson, Julia W.; Root, Robert A.; Chorover, Jon; Maier, Raina M.

2014-01-01

Mine tailings in semiarid regions are highly susceptible to erosion and are sources of dust pollution and potential avenues of human exposure to toxic metals. One constraint to revegetation of tailings by phytostabilization is the absence of microbial communities critical for biogeochemical cycling of plant nutrients. The objective of this study was to evaluate specific genes as in situ indicators of biological soil response during phytoremediation. The abundance and activity of 16S rRNA, nifH, and amoA were monitored during a nine month phytostabilization study using buffalo grass and quailbush grown in compost-amended, metalliferous tailings. The compost amendment provided a greater than 5-log increase in bacterial abundance, and survival of this compost-inoculum was more stable in planted treatments. Despite increased abundance, the activity of the introduced community was low, and significant increases were not detected until six and nine months in quailbush, and unplanted compost and buffalo grass treatments, respectively. In addition, increased abundances of nitrogen-fixation (nifH) and ammonia-oxidizing (amoA) genes were observed in rhizospheres of buffalo grass and quailbush, respectively. Thus, plant establishment facilitated the short term stabilization of introduced bacterial biomass and supported the growth of two key nitrogen-cycling populations in compost-amended tailings. PMID:25495940

Abundance and Activity of 16S rRNA, AmoA and NifH Bacterial Genes During Assisted Phytostabilization of Mine Tailings.

PubMed

Nelson, Karis N; Neilson, Julia W; Root, Robert A; Chorover, Jon; Maier, Raina M

2015-01-01

Mine tailings in semiarid regions are highly susceptible to erosion and are sources of dust pollution and potential avenues of human exposure to toxic metals. One constraint to revegetation of tailings by phytostabilization is the absence of microbial communities critical for biogeochemical cycling of plant nutrients. The objective of this study was to evaluate specific genes as in situ indicators of biological soil response during phytoremediation. The abundance and activity of 16S rRNA, nifH, and amoA were monitored during a nine month phytostabilization study using buffalo grass and quailbush grown in compost-amended, metalliferous tailings. The compost amendment provided a greater than 5-log increase in bacterial abundance, and survival of this compost-inoculum was more stable in planted treatments. Despite increased abundance, the activity of the introduced community was low, and significant increases were not detected until six and nine months in quailbush, and unplanted compost and buffalo grass treatments, respectively. In addition, increased abundances of nitrogen-fixation (nifH) and ammonia-oxidizing (amoA) genes were observed in rhizospheres of buffalo grass and quailbush, respectively. Thus, plant establishment facilitated the short term stabilization of introduced bacterial biomass and supported the growth of two key nitrogen-cycling populations in compost-amended tailings.

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.

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.

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.

Diversity of endophytic fungal and bacterial communities in Ilex paraguariensis grown under field conditions.

PubMed

Pérez, María Laura; Collavino, Mónica Mariana; Sansberro, Pedro Alfonso; Mroginski, Luis Amado; Galdeano, Ernestina

2016-04-01

The composition and diversity of the endophytic community associated with yerba mate (Ilex paraguariensis) was investigated using culture-depending methods. Fungi were identified based on their micromorphological characteristics and internal transcribed spacer rDNA sequence analysis; for bacteria 16S rDNA sequence analysis was used. Fungal and bacterial diversity did not show significant differences between organ age. The highest fungal diversity was registered during fall season and the lowest in winter. Bacterial diversity was higher in stems and increased from summer to winter, in contrast with leaves, which decreased. The most frequently isolated fungus was Fusarium, followed by Colletotrichum; they were both present in all the sampling seasons and organ types assayed. Actinobacteria represented 57.5 % of all bacterial isolates. The most dominant bacterial taxa were Curtobacterium and Microbacterium. Other bacteria frequently found were Methylobacterium, Sphingomonas, Herbiconiux and Bacillus. Nitrogen fixation and phosphate solubilization activity, ACC deaminase production and antagonism against plant fungal pathogens were assayed in endophytic bacterial strains. In the case of fungi, strains of Trichoderma, Penicillium and Aspergillus were assayed for antagonism against pathogenic Fusarium sp. All microbial isolates assayed showed at least one growth promoting activity. Strains of Bacillus, Pantoea, Curtobacterium, Methylobacterium, Brevundimonas and Paenibacillus had at least two growth-promoting activities, and Bacillus, Paenibacillus and the three endophytic fungi showed high antagonistic activity against Fusarium sp. In this work we have made a wide study of the culturable endophytic community within yerba mate plants and found that several microbial isolates could be considered as potential inoculants useful for improving yerba mate production.

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% ...

Urea Uptake and Carbon Fixation by Marine Pelagic Bacteria and Archaea during the Arctic Summer and Winter Seasons

PubMed Central

Connelly, Tara L.; Baer, Steven E.; Cooper, Joshua T.; Bronk, Deborah A.

2014-01-01

How Arctic climate change might translate into alterations of biogeochemical cycles of carbon (C) and nitrogen (N) with respect to inorganic and organic N utilization is not well understood. This study combined 15N uptake rate measurements for ammonium, nitrate, and urea with 15N- and 13C-based DNA stable-isotope probing (SIP). The objective was to identify active bacterial and archeal plankton and their role in N and C uptake during the Arctic summer and winter seasons. We hypothesized that bacteria and archaea would successfully compete for nitrate and urea during the Arctic winter but not during the summer, when phytoplankton dominate the uptake of these nitrogen sources. Samples were collected at a coastal station near Barrow, AK, during August and January. During both seasons, ammonium uptake rates were greater than those for nitrate or urea, and nitrate uptake rates remained lower than those for ammonium or urea. SIP experiments indicated a strong seasonal shift of bacterial and archaeal N utilization from ammonium during the summer to urea during the winter but did not support a similar seasonal pattern of nitrate utilization. Analysis of 16S rRNA gene sequences obtained from each SIP fraction implicated marine group I Crenarchaeota (MGIC) as well as Betaproteobacteria, Firmicutes, SAR11, and SAR324 in N uptake from urea during the winter. Similarly, 13C SIP data suggested dark carbon fixation for MGIC, as well as for several proteobacterial lineages and the Firmicutes. These data are consistent with urea-fueled nitrification by polar archaea and bacteria, which may be advantageous under dark conditions. PMID:25063662

Light Structures Phototroph, Bacterial and Fungal Communities at the Soil Surface

PubMed Central

Davies, Lawrence O.; Schäfer, Hendrik; Marshall, Samantha; Bramke, Irene; Oliver, Robin G.; Bending, Gary D.

2013-01-01

The upper few millimeters of soil harbour photosynthetic microbial communities that are structurally distinct from those of underlying bulk soil due to the presence of light. Previous studies in arid zones have demonstrated functional importance of these communities in reducing soil erosion, and enhancing carbon and nitrogen fixation. Despite being widely distributed, comparative understanding of the biodiversity of the soil surface and underlying soil is lacking, particularly in temperate zones. We investigated the establishment of soil surface communities on pasture soil in microcosms exposed to light or dark conditions, focusing on changes in phototroph, bacterial and fungal communities at the soil surface (0–3 mm) and bulk soil (3–12 mm) using ribosomal marker gene analyses. Microbial community structure changed with time and structurally similar phototrophic communities were found at the soil surface and in bulk soil in the light exposed microcosms suggesting that light can influence phototroph community structure even in the underlying bulk soil. 454 pyrosequencing showed a significant selection for diazotrophic cyanobacteria such as Nostoc punctiforme and Anabaena spp., in addition to the green alga Scenedesmus obliquus. The soil surface also harboured distinct heterotrophic bacterial and fungal communities in the presence of light, in particular, the selection for the phylum Firmicutes. However, these light driven changes in bacterial community structure did not extend to the underlying soil suggesting a discrete zone of influence, analogous to the rhizosphere. PMID:23894406

Carbohydrate Partitioning and the Capacity of Apparent Nitrogen Fixation of Soybean Plants Grown Outdoors

PubMed Central

Millhollon, Eddie P.; Williams, Larry E.

1986-01-01

Patterns of leaf carbohydrate partitioning and nodule activity in soybean plants grown under natural conditions and the irradiance level required to produce sufficient carbohydrate to obtain maximum rates of apparent N2-fixation (acetylene reduction) were measured. Soybean plants, grown outdoors, maintained constant levels of leaf soluble sugars while leaf starch pools varied diurnally. When root temperature was kept at 25°C and shoot temperature was allowed to vary with ambient temperature, the plants maintained constant rates of apparent N2-fixation and root+nodule respiration. Results from a second experiment, in which the entire plant was kept at 25°C, were similar to those of the first experiment. Shoot carbon exchange rate of plants from the second experiment was light saturated at photosynthetic photon flux densities between 400 and 600 micromoles per square meter per second. When plants were subjected to an extended 40-hour dark period to deplete carbohydrate reserves, apparent N2-fixation was unaffected during the first 10 hours of darkness, decreased rapidly between 10 and 16 hours, and plateaued at one-third the initial level thereafter. After the extended dark period, plants were exposed to photosynthetic photon flux density from 200 to 1000 micromoles per square meter per second for 10 hours. Photosynthetic photon flux densities of 200 micromoles per square meter per second and greater resulted in maximum leaf soluble sugar content and nodule activity. Leaf starch content increased with irradiance levels up to 600 micromoles per square meter per second with no further increase at higher irradiance levels. Results presented here indicate that maximum nodule activity occurs at irradiance levels that do not saturate the plant's photosynthetic apparatus. This response would allow for maximum N2-fixation to occur in a nodulated legume during periods of inclement weather. PMID:16664789

Nitrogen fixation in transposon mutants from Bradyrhizobium japonicum USDA 110 impaired in nitrate reductase.

PubMed

Camacho, María; Burgos, Araceli; Chamber-Pérez, Manuel A

2003-04-01

Tn5 transposon mutagenesis was carried out in Bradyrhizobium japonicum strain USDA 110 to produce defective mutants. From over one thousand clones expressing low levels of nitrate reductase activity as free-living bacteria, approximately five percent had significantly different ratios of nodulation, N2 fixation or nitrate reductase activity compared to the wild strain when determined in bacteroids from soybean nodules. Tn5 insertions were checked previously and mutants were arranged into four different groups. Only one of these groups, designated AN, was less effective at N2 fixation than the wild strain, suggesting a mutation in a domain shared by nitrogenase and NR. The remaining groups of insertions successfully nodulated and were as effective at N2 fixation as the wild strain, but showed diminished ability to reduce nitrate both in nodules and in the isolated bacteroids when assayed in vitro with NADH or methyl viologen as electron donors. PCR amplification demonstrated that Tn5 insertions took place in different genes on each mutant group and the type of mutant (CC) expressing almost no nitrate reductase activity under all treatments seemed to possess transposable elements in two genes. Induction of nitrate reductase activity by nitrate was observed only in those clones expressing a low constitutive activity (AN and AE). Nitrate reductase activity in bacteroids along nodule growth decreased in all groups including the ineffective AN group, whose nodulation was highly inhibited by nitrate at 5 mmol/L N. Host-cultivar interaction seemed to influence the regulation of nitrate reductase activity in bacteroids. Total or partial repression of nitrate reductase activity in bacteroids unaffected by N2 fixation (CC, AJ and AE groups) improved nodule resistance to nitrate and N yields of shoots over those of the wild strain. These observations may suggest that some of the energy supplied to bacteroids was wasted by its constitutive NRA.

Environmental and biogeochemical controls on N2 fixation in ombrotrophic peatlands

NASA Astrophysics Data System (ADS)

Zivkovic, T.; Moore, T. R.

2017-12-01

Northern peatlands have low atmospheric nitrogen (N) inputs and acquire N mostly via biological, microbially-driven N2-fixation. Little is known about rates and controls on N2-fixation in ombrotrophic bogs. We conducted two studies to test environmental and biogeochemical controls on N2-fixation. First, we used acetylene reduction assay (ARA) calibrated with 15N2 tracer to measure N2-fixation rates in three species of Sphagnum mosses along a hydrological gradient (beaver pond, hollow and hummock in bog margin and in bog) at Mer Bleue bog from June-October 2013 and May - November 2014. We tested the following controls: moisture availability, temperature, and PAR. The largest ARA rates throughout both seasons occurred in the pond in floating Sphagnum cuspidatum mats (50.3 ± 12.9 μmol m-2 d-1 Mean ± SE), which were up to 2.5 times larger than the rates found in the driest hummock site. There was a significant seasonal peak in both years in July and early August that coincided with the peak of the air temperature. In fact, 45% of the variance of N2 fixation rates over the two field seasons was explained by rain events, water table fluctuations and the surface peat temperature (multiple regression analysis, n = 539). Our results highlight the potential impact of climate change, namely negative effects due to potential droughts and positive effect of warming, on N2 fixation patterns in ombrotrophic peatlands. Secondly, we tested stoichiometric controls (Sphagnum tissue N and phosphorous (P) ratio) of N2-fixation. In a controlled environment, we selected eight study sites along a latitudinal gradient from temperate, boreal to subarctic zone in eastern Canada. We found that decreasing N:P ratio corresponded to increasing N2-fixation. N:P explained 65% of the variance in N2-fixation in hollows but only 20% in hummocks. 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

Use of Nitrogen-15-Enriched Escherichia coli as a Bacterial Tracer in Karst Aquifers.

PubMed

Ward, James W; Warden, John G; Bandy, Ashley M; Fryar, Alan E; Brion, Gail M; Macko, Stephen A; Romanek, Christopher S; Coyne, Mark S

2016-11-01

Karst aquifers are susceptible to contamination by microorganisms, but relatively few studies have used bacteria as tracers. We demonstrate the utility of Escherichia coli enriched in the stable isotope nitrogen-15 ( 15 N) as a novel bacterial tracer. Nonpathogenic E. coli from two springs in central Kentucky were grown on 15 N-enriched media. Survival of E. coli and persistence of the isotopic signal were assessed in two sets of laboratory experiments conducted with sterilized spring water in dark microcosms at 14 °C. First, isotopically labeled bacteria survived for 130 d at concentrations within one log unit of the average initial value, and there was no significant difference in δ 15 N values from Day 1 to Day 130. Second, water samples with E. coli were inoculated with either of two different species of protozoa (Tetrahymena pyriformis or Colpoda steinii). During 7 d, δ 15 N values increased in T. pyriformis while bacterial populations decreased. In a field test, following a 2.1-cm rainfall, 15 N-labeled E. coli, solutes (rhodamine WT dye and bromide), and latex microspheres were injected into a sinkhole approximately 530 m upgradient of a spring. Breakthrough of all tracers coincided, but microspheres were remobilized by subsequent storms, unlike other tracers. Enriched E. coli exhibited more tailing than solute tracers during the initial storm-flow recession. These results indicate that 15 N-enriched E. coli is a viable tracer of bacterial transport in karst aquifers, although predation may attenuate the isotopic signal in systems that are not rapidly flushed. © 2016, National Ground Water Association.

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.

Ubiquitous Gammaproteobacteria dominate dark carbon fixation in coastal sediments.

PubMed

Dyksma, Stefan; Bischof, Kerstin; Fuchs, Bernhard M; Hoffmann, Katy; Meier, Dimitri; Meyerdierks, Anke; Pjevac, Petra; Probandt, David; Richter, Michael; Stepanauskas, Ramunas; Mußmann, Marc

2016-08-01

Marine sediments are the largest carbon sink on earth. Nearly half of dark carbon fixation in the oceans occurs in coastal sediments, but the microorganisms responsible are largely unknown. By integrating the 16S rRNA approach, single-cell genomics, metagenomics and transcriptomics with (14)C-carbon assimilation experiments, we show that uncultured Gammaproteobacteria account for 70-86% of dark carbon fixation in coastal sediments. First, we surveyed the bacterial 16S rRNA gene diversity of 13 tidal and sublittoral sediments across Europe and Australia to identify ubiquitous core groups of Gammaproteobacteria mainly affiliating with sulfur-oxidizing bacteria. These also accounted for a substantial fraction of the microbial community in anoxic, 490-cm-deep subsurface sediments. We then quantified dark carbon fixation by scintillography of specific microbial populations extracted and flow-sorted from sediments that were short-term incubated with (14)C-bicarbonate. We identified three distinct gammaproteobacterial clades covering diversity ranges on family to order level (the Acidiferrobacter, JTB255 and SSr clades) that made up >50% of dark carbon fixation in a tidal sediment. Consistent with these activity measurements, environmental transcripts of sulfur oxidation and carbon fixation genes mainly affiliated with those of sulfur-oxidizing Gammaproteobacteria. The co-localization of key genes of sulfur and hydrogen oxidation pathways and their expression in genomes of uncultured Gammaproteobacteria illustrates an unknown metabolic plasticity for sulfur oxidizers in marine sediments. Given their global distribution and high abundance, we propose that a stable assemblage of metabolically flexible Gammaproteobacteria drives important parts of marine carbon and sulfur cycles.

Evolution of Bacterial Suicide

NASA Astrophysics Data System (ADS)

Tchernookov, Martin; Nemenman, Ilya

2013-03-01

While active, controlled cellular suicide (autolysis) in bacteria is commonly observed, it has been hard to argue that autolysis can be beneficial to an individual who commits it. We propose a theoretical model that predicts that bacterial autolysis is evolutionarily advantageous to an individualand would fixate in physically structured environments for stationary phase colonies. We perform spatially resolved agent-based simulations of the model, which predict that lower mixing in the environment results in fixation of a higher autolysis rate from a single mutated cell, regardless of the colony's genetic diversity. We argue that quorum sensing will fixate as well, even if initially rare, if it is coupled to controlling the autolysis rate. The model does not predict a strong additional competitive advantage for cells where autolysis is controlled by quorum sensing systems that distinguish self from nonself. These predictions are broadly supported by recent experimental results in B. subtilisand S. pneumoniae. Research partially supported by the James S McDonnell Foundation grant No. 220020321 and by HFSP grant No. RGY0084/2011.

Temperature effect on nitrogen removal performance and bacterial community in culture of marine anammox bacteria derived from sea-based waste disposal site.

PubMed

Kawagoshi, Yasunori; Fujisaki, Koichiro; Tomoshige, Yuki; Yamashiro, Kento; Wei, Qiaoyan; Qiao, Yanwei

2012-04-01

Anaerobic ammonium oxidation (anammox) bacteria have been detected in variety of marine environment in recent years, however, there have been only a few studies on their characteristics in the culture. The aim of this study is to reveal the effect of temperature on nitrogen removal ability and bacterial community in a culture of marine anammox bacteria (MAAOB). The MAAOB were cultured from the sediment of a sea-based waste disposal site at the North Port of Osaka Bay in Japan. The maximum nitrogen removal rate (NRR) was observed at 25°C in the MAAOB culture, and it decreased both at below 20°C and over 33°C. The activation energy of the MAAOB culture was calculated to be 54.6 kJ mol(-1) in the 5°C to 30°C range. No significant change in bacterial community according with temperature (5-37°C) was confirmed in the results of polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE). Meanwhile, a number of bacteria related to the oxidation-reduction reaction of sulfur were confirmed and it is speculated that they involved in the activity of MAAOB and nitrogen removal ability in the culture. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Study of the Bioremediation of Atrazine under Variable Carbon and Nitrogen Sources by Mixed Bacterial Consortium Isolated from Corn Field Soil in Fars Province of Iran

PubMed Central

Nasseri, Simin; Hashemi, Hassan

2013-01-01

Atrazine herbicide that is widely used in corn production is frequently detected in water resources. The main objectives of this research were focused on assessing the effects of carbon and nitrogen sources on atrazine biodegradation by mixed bacterial consortium and by evaluating the feasibility of using mixed bacterial consortium in soil culture. Shiraz corn field soil with a long history of atrazine application has been explored for their potential of atrazine biodegradation. The influence of different carbon compounds and the effect of nitrogen sources and a different pH (5.5–8.5) on atrazine removal efficiency by mixed bacterial consortium in liquid culture were investigated. Sodium citrate and sucrose had the highest atrazine biodegradation rate (87.22%) among different carbon sources. Atrazine biodegradation rate decreased more quickly by the addition of urea (26.76%) compared to ammonium nitrate. Based on the data obtained in this study, pH of 7.0 is optimum for atrazine biodegradation. After 30 days of incubation, the percent of atrazine reduction rates were significantly enhanced in the inoculated soils (60.5%) as compared to uninoculated control soils (12%) at the soil moisture content of 25%. In conclusion, bioaugmentation of soil with mixed bacterial consortium may enhance the rate of atrazine degradation in a highly polluted soil. PMID:23533452

Effect of bacterial root symbiosis and urea as source of nitrogen on performance of soybean plants grown hydroponically for Bioregenerative Life Support Systems (BLSSs)

PubMed Central

Paradiso, Roberta; Buonomo, Roberta; Dixon, Mike A.; Barbieri, Giancarlo; De Pascale, Stefania

2015-01-01

Soybean is traditionally grown in soil, where root symbiosis with Bradyrhizobium japonicum can supply nitrogen (N), by means of bacterial fixation of atmospheric N2. Nitrogen fertilizers inhibit N-fixing bacteria. However, urea is profitably used in soybean cultivation in soil, where urease enzymes of telluric microbes catalyze the hydrolysis to ammonium, which has a lighter inhibitory effect compared to nitrate. Previous researches demonstrated that soybean can be grown hydroponically with recirculating complete nitrate-based nutrient solutions. In Space, urea derived from crew urine could be used as N source, with positive effects in resource procurement and waste recycling. However, whether the plants are able to use urea as the sole source of N and its effect on root symbiosis with B. japonicum is still unclear in hydroponics. We compared the effect of two N sources, nitrate and urea, on plant growth and physiology, and seed yield and quality of soybean grown in closed-loop Nutrient Film Technique (NFT) in growth chamber, with or without inoculation with B. japonicum. Urea limited plant growth and seed yield compared to nitrate by determining nutrient deficiency, due to its low utilization efficiency in the early developmental stages, and reduced nutrients uptake (K, Ca, and Mg) throughout the whole growing cycle. Root inoculation with B. japonicum did not improve plant performance, regardless of the N source. Specifically, nodulation increased under fertigation with urea compared to nitrate, but this effect did not result in higher leaf N content and better biomass and seed production. Urea was not suitable as sole N source for soybean in closed-loop NFT. However, the ability to use urea increased from young to adult plants, suggesting the possibility to apply it during reproductive phase or in combination with nitrate in earlier developmental stages. Root symbiosis did not contribute significantly to N nutrition and did not enhance the plant ability to use

Bacterial Diversity and Nitrogen Utilization Strategies in the Upper Layer of the Northwestern Pacific Ocean.

PubMed

Li, Yuan-Yuan; Chen, Xiao-Huang; Xie, Zhang-Xian; Li, Dong-Xu; Wu, Peng-Fei; Kong, Ling-Fen; Lin, Lin; Kao, Shuh-Ji; Wang, Da-Zhi

2018-01-01

Nitrogen (N) is a primary limiting nutrient for bacterial growth and productivity in the ocean. To better understand bacterial community and their N utilization strategy in different N regimes of the ocean, we examined bacterial diversity, diazotrophic diversity, and N utilization gene expressions in the northwestern Pacific Ocean (NWPO) using a combination of high-throughput sequencing and real-time qPCR methods. 521 and 204 different operational taxonomic units (OTUs) were identified in the 16s rRNA and nifH libraries from nine surface samples. Of the 16s rRNA gene OTUs, 11.9% were observed in all samples while 3.5 and 15.9% were detected only in N-sufficient and N-deficient samples. Proteobacteria, Cyanobacteria and Bacteroidetes dominated the bacterial community. Prochlorococcus and Pseudoalteromonas were the most abundant at the genus level in N-deficient regimes, while SAR86, Synechococcus and SAR92 were predominant in the Kuroshio-Oyashio confluence region. The distribution of the nifH gene presented great divergence among sampling stations: Cyanobacterium_UCYN-A dominated the N-deficient stations, while clusters related to the Alpha-, Beta- , and Gamma-Proteobacteria were abundant in other stations. Temperature was the main factor that determined bacterial community structure and diversity while concentration of NO X -N was significantly correlated with structure and distribution of N 2 -fixing microorganisms. Expression of the ammonium transporter was much higher than that of urea transporter subunit A ( urtA ) and ferredoxin-nitrate reductase , while urtA had an increased expression in N-deficient surface water. The predicted ammonium transporter and ammonium assimilation enzymes were most abundant in surface samples while urease and nitrogenase were more abundant in the N-deficient regions. These findings underscore the fact that marine bacteria have evolved diverse N utilization strategies to adapt to different N habitats, and that urea metabolism is of

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.

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...

Responses of Ammonia-Oxidizing Bacterial and Archaeal Populations to Organic Nitrogen Amendments in Low-Nutrient Groundwater ▿

PubMed Central

Reed, David W.; Smith, Jason M.; Francis, Christopher A.; Fujita, Yoshiko

2010-01-01

To evaluate the potential for organic nitrogen addition to stimulate the in situ growth of ammonia oxidizers during a field scale bioremediation trial, samples collected from the Eastern Snake River Plain Aquifer in Idaho before, during, and after the addition of molasses and urea were subjected to PCR analysis of ammonia monooxygenase subunit A (amoA) genes. Ammonia-oxidizing bacteria (AOB) and archaea (AOA) were present in all of the samples tested, with AOA amoA genes outnumbering AOB amoA genes in all of the samples. Following urea addition, nitrate levels rose and bacterial amoA copy numbers increased dramatically, suggesting that urea hydrolysis stimulated nitrification. Bacterial amoA diversity was limited to two Nitrosomonas phylotypes, whereas archaeal amoA analyses revealed 20 distinct operational taxonomic units, including several that were markedly different from all previously reported sequences. Results from this study demonstrate the likelihood of stimulating ammonia-oxidizing communities during field scale manipulation of groundwater conditions to promote urea hydrolysis. PMID:20190081

Persistence of biological nitrogen fixation in high latitude grass-clover grasslands under different management practices

NASA Astrophysics Data System (ADS)

Tzanakakis, Vasileios; Sturite, Ievina; Dörsch, Peter

2016-04-01

Biological nitrogen fixation (BNF) can substantially contribute to N supply in permanent grasslands, improving N yield and forage quality, while reducing inorganic N inputs. Among the factors critical to the performance of BNF in grass-legume mixtures are selected grass and legume species, proportion of legumes, the soil-climatic conditions, in particular winter conditions, and management practices (e.g. fertilization and compaction). In high latitude grasslands, low temperatures can reduce the performance of BNF by hampering the legumés growth and by suppressing N2 fixation. Estimation of BNF in field experiments is not straightforward. Different methods have been developed providing different results. In the present study, we evaluated the performance of BNF, in a newly established field experiment in North Norway over four years. The grassland consisted of white clover (Trifolium repens L.) and red clover (Trifolium pretense L.) sawn in three proportions (0, 15 and 30% in total) together with timothy (Pheum pretense L.) and meadow fescue (Festuca pratensis L.). Three levels of compaction were applied each year (no tractor, light tractor, heavy tractor) together with two different N rates (110 kg N/ha as cattle slurry or 170 kg N/ha as cattle slurry and inorganic N fertilizer). We applied two different methods, the 15N natural abundance and the difference method, to estimate BNF in the first harvest of each year. Overall, the difference method overestimated BNF relative to the 15N natural abundance method. BNF in the first harvest was compared to winter survival of red and white clover plants, which decreased with increasing age of the grassland. However, winter conditions did not seem to affect the grassland's ability to fix N in spring. The fraction of N derived from the atmosphere (NdfA) in white and red clover was close to 100% in each spring, indicating no suppression of BNF. BNF increased the total N yield of the grasslands by up to 75%, mainly due to high

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...

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.

Chemoautotrophic carbon fixation rates and active bacterial communities in intertidal marine sediments.

PubMed

Boschker, Henricus T S; Vasquez-Cardenas, Diana; Bolhuis, Henk; Moerdijk-Poortvliet, Tanja W C; Moodley, Leon

2014-01-01

Chemoautotrophy has been little studied in typical coastal marine sediments, but may be an important component of carbon recycling as intense anaerobic mineralization processes in these sediments lead to accumulation of high amounts of reduced compounds, such as sulfides and ammonium. We studied chemoautotrophy by measuring dark-fixation of 13C-bicarbonate into phospholipid derived fatty acid (PLFA) biomarkers at two coastal sediment sites with contrasting sulfur chemistry in the Eastern Scheldt estuary, The Netherlands. At one site where free sulfide accumulated in the pore water right to the top of the sediment, PLFA labeling was restricted to compounds typically found in sulfur and ammonium oxidizing bacteria. At the other site, with no detectable free sulfide in the pore water, a very different PLFA labeling pattern was found with high amounts of label in branched i- and a-PLFA besides the typical compounds for sulfur and ammonium oxidizing bacteria. This suggests that other types of chemoautotrophic bacteria were also active, most likely Deltaproteobacteria related to sulfate reducers. Maximum rates of chemoautotrophy were detected in first 1 to 2 centimeters of both sediments and chemosynthetic biomass production was high ranging from 3 to 36 mmol C m(-2) d(-1). Average dark carbon fixation to sediment oxygen uptake ratios were 0.22±0.07 mol C (mol O2)(-1), which is in the range of the maximum growth yields reported for sulfur oxidizing bacteria indicating highly efficient growth. Chemoautotrophic biomass production was similar to carbon mineralization rates in the top of the free sulfide site, suggesting that chemoautotrophic bacteria could play a crucial role in the microbial food web and labeling in eukaryotic poly-unsaturated PLFA was indeed detectable. Our study shows that dark carbon fixation by chemoautotrophic bacteria is a major process in the carbon cycle of coastal sediments, and should therefore receive more attention in future studies on

N2-fixation dynamics during ecosystem recovery in longleaf pine savannas

NASA Astrophysics Data System (ADS)

Tierney, J. A.

2016-12-01

Biological nitrogen fixation (BNF) can alleviate nitrogen (N) deficiencies that inhibit ecosystem recovery. BNF may be particularly important in ecosystems recovering from land-use change and perturbations from fire, as these disturbances can exacerbate N limitation. Here, we investigated how BNF dynamics change throughout ecosystem development in restored longleaf pine savannas, and how BNF responds to fire. We conducted this study in 59 1-ha plots of longleaf pine distributed across gradients of stand age and fire frequency at two sites in the southeastern US. We determined BNF contributions by three functional groups of N2-fixers (herbaceous legumes, biological soil crusts, and asymbiotic N2-fixing bacteria) by quantifying their abundances, assessing nitrogenase activity, and scaling these estimates up to the plot-level. To determine aboveground N demands, we measured tree growth using diameter increments and allometric equations paired with tissue-specific N concentrations. We fit linear models to evaluate the effects of stand age and time since fire on BNF and N demands throughout stand development, and performed separate analyses on mature stands to determine how fire return interval affects BNF. We observed distinct temporal patterns of N2-fixation across stand development among the three groups of N2 fixers. N2-fixation by legumes and asymbiotic bacteria remained low until stands reached maturity, while N2-fixation by biological soil crusts (BSCs) was high in juvenile stands and decreased with stand age. These patterns suggest a compensatory shift in the importance of these functional groups throughout stand development such that contributions from BSCs are critical for meeting N demands when disturbances may hinder the establishment of legumes and asymbiotic bacteria. N2-fixation by BSCs and asymbiotic bacteria throughout stand development was not affected by time since fire, but legume abundance increased the year following fire, suggesting a recovery

Different nitrogen sources change the transcriptome of welan gum-producing strain Sphingomonas sp. ATCC 31555.

PubMed

Xu, Xiaopeng; Nie, Zuoming; Zheng, Zhiyong; Zhu, Li; Zhang, Hongtao; Zhan, Xiaobei

2017-09-01

To reveal effects of different nitrogen sources on the expressions and functions of genes in Sphingomonas sp. ATCC 31555, it was cultivated in medium containing inorganic nitrogen (IN), organic nitrogen (ON), or inorganic-organic combined nitrogen (CN). Welan gum production and bacterial biomass were determined, and RNA sequencing (RNA-seq) was performed. Differentially expressed genes (DEGs) between the different ATCC 31555 groups were identified, and their functions were analyzed. Welan gum production and bacterial biomass were significantly higher in the ON and CN groups compared with those in the IN group. RNA-seq produced 660 unigenes, among which 488, 731, and 844 DEGs were identified between the IN vs. ON, IN vs. CN, and ON vs. CN groups, respectively. All the DEGs were related significantly to metabolic process and signal transduction. DEGs between the IN vs. CN and ON vs. CN groups were potentially associated with bacterial chemotaxis. Real-time PCR confirmed the expressions of selected DEGs. Organic nitrogen led to higher bacterial biomass and welan gum production than inorganic nitrogen, which might reflect differences in gene expression associated with metabolic process, signal transduction, and bacterial chemotaxis induced by different nitrogen sources.

Assessing carbon and nitrogen removal in a novel anoxic-aerobic cyanobacterial-bacterial photobioreactor configuration with enhanced biomass sedimentation.

PubMed

de Godos, I; Vargas, V A; Guzmán, H O; Soto, R; García, B; García, P A; Muñoz, R

2014-09-15

The carbon and nitrogen removal potential of an innovative anoxic-aerobic photobioreactor configuration operated with both internal and external recyclings was evaluated under different cyanobacterial-bacterial sludge residence times (9-31 days) during the treatment of wastewaters with low C/N ratios. Under optimal operating conditions, the two-stage photobioreactor was capable of providing organic carbon and nitrogen removals over 95% and 90%, respectively. The continuous biomass recycling from the settler resulted in the enrichment and predominance of rapidly-settling cyanobacterial-bacterial flocs and effluent suspended solid concentrations lower than 35 mg VSS L(-1). These flocs exhibited sedimentation rates of 0.28-0.42 m h(-1) but sludge volumetric indexes of 333-430 ml/g. The decoupling between the hydraulic retention time and sludge retention time mediated by the external recycling also avoided the washout of nitrifying bacteria and supported process operation at biomass concentrations of 1000-1500 mg VSS L(-1). The addition of additional NaHCO3 to the process overcame the CO2 limitation resulting from the intense competition for inorganic carbon between cyanobacteria and nitrifying bacteria in the photobioreactor, which supported the successful implementation of a nitrification-denitrification process. Unexpectedly, this nitrification-denitrification process occurred both simultaneously in the photobioreactor alone (as a result of the negligible dissolved oxygen concentrations) and sequentially in the two-stage anoxic-aerobic configuration with internal NO3(-)/NO2(-) recycling. Copyright © 2014 Elsevier Ltd. All rights reserved.