Science.gov

Sample records for nitrogen fixation studies

  1. The Fixation of Nitrogen.

    ERIC Educational Resources Information Center

    Andrew, S. P. S.

    1978-01-01

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

  2. Update: Biological Nitrogen Fixation.

    ERIC Educational Resources Information Center

    Wiseman, Alan; And Others

    1985-01-01

    Updates knowledge on nitrogen fixation, indicating that investigation of free-living nitrogen-fixing organisms is proving useful in understanding bacterial partners and is expected to lead to development of more effective symbioses. Specific areas considered include biochemistry/genetics, synthesis control, proteins and enzymes, symbiotic systems,…

  3. Molecular Biology of Nitrogen Fixation

    ERIC Educational Resources Information Center

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

    1975-01-01

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

  4. Nitrogen fixation and CO/sub 2/ metabolism: proceedings

    SciTech Connect

    Ludden, P.W.; Burris, J.E.

    1985-01-01

    Photosynthesis and nitrogen fixation are key metabolic processes which lead to the production of reduced carbon and nitrogen compounds. These compounds are essential for the maintenance and continuation of life on earth. In this volume many recent advances in the study of nitrogen fixation and photosynthetic carbon dioxide fixation are presented. The papers were presented in seven sessions. These sessions were the biochemistry of the legume nodule, genetics and molecular biology of nitrogen fixation, enzymes and cofactors involved in inorganic nitrogen reductions, aspects of nitrogen fixation by associations and symbioses, physiology of free-living nitrogen fixers, interactions between carbon metabolism and nitrogen fixation, photorespiration in plants, and photosynthetic carbon fixation. (DT)

  5. 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.

  6. 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.

  7. Eighth international congress on nitrogen fixation

    SciTech Connect

    Not Available

    1990-01-01

    This volume contains the proceedings of the Eighth International Congress on Nitrogen Fixation held May 20--26, 1990 in Knoxville, Tennessee. The volume contains abstracts of individual presentations. Sessions were entitled Recent Advances in the Chemistry of Nitrogen Fixation, Plant-microbe Interactions, Limiting Factors of Nitrogen Fixation, Nitrogen Fixation and the Environment, Bacterial Systems, Nitrogen Fixation in Agriculture and Industry, Plant Function, and Nitrogen Fixation and Evolution.

  8. Understanding Nitrogen Fixation

    SciTech Connect

    Paul J. Chirik

    2012-05-25

    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

  9. 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

  10. Missing nitrogen fixation in the Benguela region

    NASA Astrophysics Data System (ADS)

    Wasmund, Norbert; Struck, Ulrich; Hansen, Anja; Flohr, Anita; Nausch, Günther; Grüttmüller, Annett; Voss, Maren

    2015-12-01

    Opposing opinions on the importance of nitrogen fixation in the northern Benguela upwelling region provoked us to investigate the magnitude of nitrogen fixation in front of northern Namibia and southern Angola. Measurements of nitrogen fixation rates using the 15N method at 66 stations during seven cruises from 2008 to 2014 showed that, in general, the 15N content in the biomass did not increase after tracer incubation with 15N2, indicating that no nitrogen fixation occurred. Correspondingly, the filamentous nitrogen-fixing cyanobacterium Trichodesmium was almost not present. The abundant picocyanobacteria did obviously not perform nitrogen fixation to a significant degree. The artificial improvement of conditions for nitrogen fixation in mesocosm experiments, including phosphate and iron additions and a warmer temperature, failed to induce nitrogen fixation. A plausible explanation of these findings is a lack of conditioned cells for nitrogen fixation in the Benguela region.

  11. Traffic lights in trichodesmium. Regulation of photosynthesis for nitrogen fixation studied by chlorophyll fluorescence kinetic microscopy.

    PubMed

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

    2004-08-01

    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 (F(0)), 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 F(0) 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 F(0) compared to the normal F(0) in the dark phase and a PSII activity, measured as variable fluorescence (F(v) = F(m) - F(0)), 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 F(0) 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

  12. Trichodesmium and nitrogen fixation in the Kuroshio

    NASA Astrophysics Data System (ADS)

    Shiozaki, T.; Takeda, S.; Itoh, S.; Kodama, T.; Liu, X.; Hashihama, F.; Furuya, K.

    2015-07-01

    Nitrogen fixation in the Kuroshio influences nitrogen balance in the North Pacific Ocean. The genus Trichodesmium is recognized as a major diazotroph in the Kuroshio. Although its abundance is higher in the Kuroshio than in adjacent waters, the reason for this difference remains unclear. The present study investigated the abundance of Trichodesmium spp. and nitrogen fixation together with concentrations of dissolved iron and phosphate, whose availabilities potentially control diazotrophy, in the Kuroshio and its marginal seas. We performed the observations near the Miyako Islands, which form part of the Ryukyu Islands, situated along the Kuroshio, since satellite analysis suggested that material transport could occur from the islands to the Kuroshio. Trichodesmium spp. bloomed (> 20 000 filaments L-1) near the Miyako Islands, and the abundance was high in the Kuroshio and the Kuroshio bifurcation region of the East China Sea, but was low in the Philippine Sea. The abundance of Trichodesmium spp. was significantly correlated with the total nitrogen fixation activity. The surface concentrations of dissolved iron (0.19-0.89 nM) and phosphate (< 3-36 nM) were similar for all of the study areas, indicating that the nutrient distribution could not explain the spatial differences in Trichodesmium spp. abundance and nitrogen fixation. We used a numerical model to simulate the transportation of water around the Ryukyu Islands to the Kuroshio. Our results indicate that Trichodesmium growing around the islands situated along the Kuroshio is potentially important for determining diazotrophy in this region.

  13. Biochemical Approaches to Improved Nitrogen Fixation

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  14. 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.

  15. 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.

  16. Methanotrophy Induces Nitrogen Fixation in Boreal Mosses

    NASA Astrophysics Data System (ADS)

    Tiirola, M. A.

    2014-12-01

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

  17. Nitrogen fixation and nitrogen transformations in marine symbioses.

    PubMed

    Fiore, Cara L; Jarett, Jessica K; Olson, Nathan D; Lesser, Michael P

    2010-10-01

    Many marine organisms have coevolved symbiotic relationships with nitrogen-fixing bacteria in nitrogen limited environments such as coral reefs. In addition, some of these organisms also harbor microbes that carry out nitrification and denitrification. Prokaryotes involved in nitrogen fixation and other nitrogen transformations are symbionts in a range of eukaryotic hosts in the marine environment including shipworms, diatoms, corals and sponges. Molecular genetic approaches, and other analytical techniques, have provided exciting new insights into symbiont diversity and the relationship between host and symbiont. We review the current state of knowledge of these symbioses and highlight important avenues for future studies.

  18. Nitrogen fixation and nitrogen transformations in marine symbioses.

    PubMed

    Fiore, Cara L; Jarett, Jessica K; Olson, Nathan D; Lesser, Michael P

    2010-10-01

    Many marine organisms have coevolved symbiotic relationships with nitrogen-fixing bacteria in nitrogen limited environments such as coral reefs. In addition, some of these organisms also harbor microbes that carry out nitrification and denitrification. Prokaryotes involved in nitrogen fixation and other nitrogen transformations are symbionts in a range of eukaryotic hosts in the marine environment including shipworms, diatoms, corals and sponges. Molecular genetic approaches, and other analytical techniques, have provided exciting new insights into symbiont diversity and the relationship between host and symbiont. We review the current state of knowledge of these symbioses and highlight important avenues for future studies. PMID:20674366

  19. 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

  20. Variable Nitrogen Fixation in Wild Populus.

    PubMed

    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

  1. Variable Nitrogen Fixation in Wild Populus.

    PubMed

    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.

  2. Cultivar effects on nitrogen fixation in peas and lentils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Increasing nitrogen fixation in legume crops could increase cropping productivity and reduce nitrogen fertilizer use. Studies have found that crop genotype, rhizobial strain, and occasionally genotype-specific interactions affect N fixation, but this knowledge has not yet been used to evaluate or br...

  3. Eighth international congress on nitrogen fixation. Final program

    SciTech Connect

    Not Available

    1990-12-31

    This volume contains the proceedings of the Eighth International Congress on Nitrogen Fixation held May 20--26, 1990 in Knoxville, Tennessee. The volume contains abstracts of individual presentations. Sessions were entitled Recent Advances in the Chemistry of Nitrogen Fixation, Plant-microbe Interactions, Limiting Factors of Nitrogen Fixation, Nitrogen Fixation and the Environment, Bacterial Systems, Nitrogen Fixation in Agriculture and Industry, Plant Function, and Nitrogen Fixation and Evolution.

  4. Bacterial alternative nitrogen fixation systems.

    PubMed

    Joerger, R D; Bishop, P E

    1988-01-01

    The introduction briefly reviews some of the salient features of the well-characterized conventional molybdo-enzyme system for N2 fixation. This is followed by a brief account of the discovery of an alternative N2 fixation system that does not require molybdenum in the N2-fixing bacterum Azotobacter vinelandii. The next section cites observations from the early literature on N2 fixation suggesting may not always require molybdenum. Next, recent evidence for an alternative N2 fixation system in A. vinelandii is discussed. A brief description of our discovery of an alternative nitrogenase which is not a molybdenum or vanadium enzyme is presented, followed by a summary of recent papers describing an alternative vanadium-containing nitrogenase. Available information on the genetics and regulation of alternative N2 fixation systems is discussed. Finally, the possible/probable presence of alternative N2 fixation systems in bacteria other than Azotobacter species is covered.

  5. 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.

  6. 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.

  7. Nitrogen Inputs via Nitrogen Fixation in Northern Plants and Soils

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

    Dominated by cold and often acidic water logged environments, mineralization of organic matter is slow in the majority of northern ecosystems. Measures of extractable ammonium and nitrate are generally low and can be undetectable in peat pore waters. Despite this apparent nitrogen limitation, many of these environments produce deep deposits of soil organic matter. Biological nitrogen fixation carried out by autotrophic and heterotrophic diazotrophs associated with cryptograms provides the majority of known nitrogen inputs in these northern ecosystems. Nitrogen fixation was assessed in a variety of northern soils within rhizospheres of dominant plant communities. We investigated the availability of this newly fixed nitrogen to the vascular plant community in nitrogen limited northern plant communities. We tracked nitrogen flow from 15N2 gas fixed in Sphagnum mosses into tissues of two native vascular plant species, boreal cranberry (Vaccinium oxycoccus) and black spruce (Picea mariana). 15N-labeled Sphagnum microcosms were grown within variable mesh size exclusion/inclusion fabrics in a nitrogen addition experiment in situ in order to investigate the role of mycorrhizal fungi in the uptake of newly fixed nitrogen. Up to 24% of daily fixed 15N label was transferred to vascular plant tissues during 2 months. Nitrogen addition resulted in decreased N2 fixation rates; however, with higher nitrogen availability there was a higher rate of 15N label uptake into the vascular plants, likely the result of increased production of dissolved organic nitrogen. Reliance on mycorrhizal networks for nitrogen acquisition was indicated by nitrogen isotope fractionation patterns. Moreover, N2 fixation activities in mosses were stimulated when vascular plants were grown in moss microcosms versus "moss only" treatments. Results indicate that bog vascular plants may derive considerable nitrogen from atmospheric N2 biologically fixed within Sphagnum mosses. This work demonstrates that

  8. Changes in North Atlantic nitrogen fixation controlled by ocean circulation.

    PubMed

    Straub, Marietta; Sigman, Daniel M; Ren, Haojia; Martínez-García, Alfredo; Meckler, A Nele; Hain, Mathis P; Haug, Gerald H

    2013-09-12

    In the ocean, the chemical forms of nitrogen that are readily available for biological use (known collectively as 'fixed' nitrogen) fuel the global phytoplankton productivity that exports carbon to the deep ocean. Accordingly, variation in the oceanic fixed nitrogen reservoir has been proposed as a cause of glacial-interglacial changes in atmospheric carbon dioxide concentration. Marine nitrogen fixation, which produces most of the ocean's fixed nitrogen, is thought to be affected by multiple factors, including ocean temperature and the availability of iron and phosphorus. Here we reconstruct changes in North Atlantic nitrogen fixation over the past 160,000 years from the shell-bound nitrogen isotope ratio ((15)N/(14)N) of planktonic foraminifera in Caribbean Sea sediments. The observed changes cannot be explained by reconstructed changes in temperature, the supply of (iron-bearing) dust or water column denitrification. We identify a strong, roughly 23,000-year cycle in nitrogen fixation and suggest that it is a response to orbitally driven changes in equatorial Atlantic upwelling, which imports 'excess' phosphorus (phosphorus in stoichiometric excess of fixed nitrogen) into the tropical North Atlantic surface. In addition, we find that nitrogen fixation was reduced during glacial stages 6 and 4, when North Atlantic Deep Water had shoaled to become glacial North Atlantic intermediate water, which isolated the Atlantic thermocline from excess phosphorus-rich mid-depth waters that today enter from the Southern Ocean. Although modern studies have yielded diverse views of the controls on nitrogen fixation, our palaeobiogeochemical data suggest that excess phosphorus is the master variable in the North Atlantic Ocean and indicate that the variations in its supply over the most recent glacial cycle were dominated by the response of regional ocean circulation to the orbital cycles.

  9. Nitrogen Fixation and Hydrogen Metabolism in Cyanobacteria

    PubMed Central

    Bothe, Hermann; Schmitz, Oliver; Yates, M. Geoffrey; Newton, William E.

    2010-01-01

    Summary: 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 N2 fixation and/or H2 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 H2 as a source of combustible energy. To enhance the rates of H2 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 H2 formation and so improve the chances of utilizing cyanobacteria as a source for the generation of clean energy. PMID:21119016

  10. Nitrogen Fixation: An Interdisciplinary Frontier

    ERIC Educational Resources Information Center

    Schneller, Stewart W.

    1972-01-01

    Research has progressed from three major directions--biological, chemical, and non-enzymatic. Hopefully all three will converge and will explain not only the process, but also introduce new means for modifying the molecule to meet the nutritional needs of man in areas now incapable of self support without fixed nitrogen. (DF)

  11. Applying reversible mutations of nodulation and nitrogen-fixation genes to study social cheating in Rhizobium etli-legume interaction.

    PubMed

    Ling, Jun; Zheng, Huiming; Katzianer, David S; Wang, Hui; Zhong, Zengtao; Zhu, Jun

    2013-01-01

    Mutualisms are common in nature, though these symbioses can be quite permeable to cheaters in situations where one individual parasitizes the other by discontinuing cooperation yet still exploits the benefits of the partnership. In the Rhizobium-legume system, there are two separate contexts, namely nodulation and nitrogen fixation processes, by which resident Rhizobium individuals can benefit by cheating. Here, we constructed reversible and irreversible mutations in key nodulation and nitrogen-fixation pathways of Rhizobium etli and compared their interaction with plant hosts Phaseolus vulgaris to that of wild type. We show that R. etli reversible mutants deficient in nodulation factor production are capable of intra-specific cheating, wherein mutants exploit other Rhizobium individuals capable of producing these factors. Similarly, we show that R. etli mutants are also capable of cheating inter-specifically, colonizing the host legume yet contributing nothing to the partnership in terms of nitrogen fixation. Our findings indicate that cheating is possible in both of these frameworks, seemingly without damaging the stability of the mutualism itself. These results may potentially help explain observations suggesting that legume plants are commonly infected by multiple bacterial lineages during the nodulation process.

  12. Applying Reversible Mutations of Nodulation and Nitrogen-Fixation Genes to Study Social Cheating in Rhizobium etli-Legume Interaction

    PubMed Central

    Wang, Hui; Zhong, Zengtao; Zhu, Jun

    2013-01-01

    Mutualisms are common in nature, though these symbioses can be quite permeable to cheaters in situations where one individual parasitizes the other by discontinuing cooperation yet still exploits the benefits of the partnership. In the Rhizobium-legume system, there are two separate contexts, namely nodulation and nitrogen fixation processes, by which resident Rhizobium individuals can benefit by cheating. Here, we constructed reversible and irreversible mutations in key nodulation and nitrogen-fixation pathways of Rhizobium etli and compared their interaction with plant hosts Phaseolus vulgaris to that of wild type. We show that R. etli reversible mutants deficient in nodulation factor production are capable of intra-specific cheating, wherein mutants exploit other Rhizobium individuals capable of producing these factors. Similarly, we show that R. etli mutants are also capable of cheating inter-specifically, colonizing the host legume yet contributing nothing to the partnership in terms of nitrogen fixation. Our findings indicate that cheating is possible in both of these frameworks, seemingly without damaging the stability of the mutualism itself. These results may potentially help explain observations suggesting that legume plants are commonly infected by multiple bacterial lineages during the nodulation process. PMID:23922937

  13. 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.

  14. Effect of butachlor on growth and nitrogen fixation by Anabaena sphaerica.

    PubMed

    Suseela, M R

    2001-07-01

    Present study was carried out to examine the effect of Butachlor on growth and nitrogen fixation by Anabaena sphaerica. The increased concentration of the pesticide did not have any adverse effect on the alga. Rather it accelerated the algal contribution in terms of biomass and nitrogen fixation.

  15. High rates of nitrogen fixation in equatorial upwelling region

    NASA Astrophysics Data System (ADS)

    Balcerak, Ernie

    2013-05-01

    Surface waters in upwelling regions of the ocean are generally rich in nutrients. Scientists had thought that these areas would have low rates of nitrogen fixation because diazotrophs—microbes that convert nitrogen gas from the atmosphere into usable forms, such as ammonia—could use the nutrients in the water directly instead of having to fix nitrogen gas. However, researchers recently recorded high rates of nitrogen fixation in an upwelling region in the equatorial Atlantic.

  16. Nitrogen Fixation and Molecular Oxygen: Comparative Genomic Reconstruction of Transcription Regulation in Alphaproteobacteria

    PubMed Central

    Tsoy, Olga V.; Ravcheev, Dmitry A.; Čuklina, Jelena; Gelfand, Mikhail S.

    2016-01-01

    Biological nitrogen fixation plays a crucial role in the nitrogen cycle. An ability to fix atmospheric nitrogen, reducing it to ammonium, was described for multiple species of Bacteria and Archaea. The transcriptional regulatory network for nitrogen fixation was extensively studied in several representatives of the class Alphaproteobacteria. This regulatory network includes the activator of nitrogen fixation NifA, working in tandem with the alternative sigma-factor RpoN as well as oxygen-responsive regulatory systems, one-component regulators FnrN/FixK and two-component system FixLJ. Here we used a comparative genomics approach for in silico study of the transcriptional regulatory network in 50 genomes of Alphaproteobacteria. We extended the known regulons and proposed the scenario for the evolution of the nitrogen fixation transcriptional network. The reconstructed network substantially expands the existing knowledge of transcriptional regulation in nitrogen-fixing microorganisms and can be used for genetic experiments, metabolic reconstruction, and evolutionary analysis.

  17. Nitrogen Fixation and Molecular Oxygen: Comparative Genomic Reconstruction of Transcription Regulation in Alphaproteobacteria.

    PubMed

    Tsoy, Olga V; Ravcheev, Dmitry A; Čuklina, Jelena; Gelfand, Mikhail S

    2016-01-01

    Biological nitrogen fixation plays a crucial role in the nitrogen cycle. An ability to fix atmospheric nitrogen, reducing it to ammonium, was described for multiple species of Bacteria and Archaea. The transcriptional regulatory network for nitrogen fixation was extensively studied in several representatives of the class Alphaproteobacteria. This regulatory network includes the activator of nitrogen fixation NifA, working in tandem with the alternative sigma-factor RpoN as well as oxygen-responsive regulatory systems, one-component regulators FnrN/FixK and two-component system FixLJ. Here we used a comparative genomics approach for in silico study of the transcriptional regulatory network in 50 genomes of Alphaproteobacteria. We extended the known regulons and proposed the scenario for the evolution of the nitrogen fixation transcriptional network. The reconstructed network substantially expands the existing knowledge of transcriptional regulation in nitrogen-fixing microorganisms and can be used for genetic experiments, metabolic reconstruction, and evolutionary analysis.

  18. Significant nonsymbiotic nitrogen fixation in Patagonian ombrotrophic bogs.

    PubMed

    Knorr, Klaus-Holger; Horn, Marcus A; Borken, Werner

    2015-06-01

    Nitrogen (N) nutrition in pristine peatlands relies on the natural input of inorganic N through atmospheric deposition or biological dinitrogen (N2 ) fixation. However, N2 fixation and its significance for N cycling, plant productivity, and peat buildup are mostly associated with the presence of Sphagnum mosses. Here, we report high nonsymbiotic N2 -fixation rates in two pristine Patagonian bogs with diversified vegetation and natural N deposition. Nonsymbiotic N2 fixation was measured in samples from 0 to 10, 10 to 20, and 40 to 50 cm depth using the (15) N2 assay as well as the acetylene reduction assay (ARA). The ARA considerably underestimated N2 fixation and can thus not be recommended for peatland studies. Based on the (15) N2 assay, high nonsymbiotic N2 -fixation rates of 0.3-1.4 μmol N2  g(-1)  day(-1) were found down to 50 cm under micro-oxic conditions (2 vol.%) in samples from plots covered by Sphagnum magellanicum or by vascular cushion plants, latter characterized by dense and deep aerenchyma roots. Peat N concentrations point to greater potential of nonsymbiotic N2 fixation under cushion plants, likely because of the availability of easily decomposable organic compounds and oxic conditions in the rhizosphere. In the Sphagnum plots, high N2 fixation below 10 cm depth rather reflects the potential during dry periods or low water level when oxygen penetrates the top peat layer and triggers peat mineralization. Natural abundance of the (15) N isotope of live Sphagnum (5.6 δ‰) from 0 to 10 cm points to solely N uptake from atmospheric deposition and nonsymbiotic N2 fixation. A mean (15) N signature of -0.7 δ‰ of peat from the cushion plant plots indicates additional N supply from N mineralization. Our findings suggest that nonsymbiotic N2 fixation overcomes N deficiency in different vegetation communities and has great significance for N cycling and peat accumulation in pristine peatlands.

  19. Evaluation of kidney bean yield and nitrogen fixation under low soil nitrogen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    U.S. Kidney bean cultivars as a group have limited genetic diversity. They also require more intensive crop management than most other bean market classes. The objective of this study was to evaluate biological nitrogen fixation and seed yield in a diverse group of kidney lines. A 250 line Andean be...

  20. 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

  1. New insights into the evolutionary history of biological nitrogen fixation.

    PubMed

    Boyd, Eric S; Peters, John W

    2013-01-01

    Nitrogenase, which catalyzes the ATP-dependent reduction of dinitrogen (N2) to ammonia (NH3), accounts for roughly half of the bioavailable nitrogen supporting extant life. The fundamental requirement for fixed forms of nitrogen for life on Earth, both at present and in the past, has led to broad and significant interest in the origin and evolution of biological N2 fixation. One key question is whether the limited availability of fixed nitrogen was a factor in life's origin or whether there were ample sources of fixed nitrogen produced by abiotic processes or delivered through the weathering of bolide impact materials to support this early life. If the latter, the key questions become what were the characteristics of the environment that precipitated the evolution of this oxygen sensitive process, when did this occur, and how was its subsequent evolutionary history impacted by the advent of oxygenic photosynthesis and the rise of oxygen in the Earth's biosphere. Since the availability of fixed sources of nitrogen capable of supporting early life is difficult to glean from the geologic record, there are limited means to get direct insights into these questions. Indirect insights, however, can be gained through phylogenetic studies of nitrogenase structural gene products and additional gene products involved in the biosynthesis of the complex metal-containing prosthetic groups associated with this enzyme complex. Insights gained from such studies, as reviewed herein, challenge traditional models for the evolution of biological nitrogen fixation and provide the basis for the development of new conceptual models that explain the stepwise evolution of this highly complex life sustaining process.

  2. Diel variation of nitrogen fixation in Lake Valencia, Venezuela

    SciTech Connect

    Levine, S.N.; Lewis, W.M. Jr.

    1984-07-01

    During 1981 the authors examined the diel variations of nitrogen fixation in Lake Valencia, Venezuela. Four species of heterocyst-bearing blue-green algae were common but subdominant in the phytoplankton. In samples taken from and incubated at 0.5 m, the rate of nitrogen fixation per unit volume of water was lowest at night, increased from dawn until early afternoon, and then diminished between late afternoon and the first hour of darkness. This pattern was caused partly by diel changes in light intensity and partly by diel migrations of heterocystous blue-green algae. The heterocyst-specific nitrogen fixation rates at 0.5 m were much less variable than the nitrogen fixation rates per unit volume of water. Heterocyst-specific rates rose rapidly in early morning and fell slowly in the evening, but were almost constant over much of the day. Heterocyst-specific nitrogen fixation rates were very close to those predicted by a model based on the light dependency of nitrogen fixation.

  3. Novel regulatory cascades controlling expression of nitrogen-fixation genes in Geobacter sulfurreducens

    PubMed Central

    Ueki, Toshiyuki; Lovley, Derek R.

    2010-01-01

    Geobacter species often play an important role in bioremediation of environments contaminated with metals or organics and show promise for harvesting electricity from waste organic matter in microbial fuel cells. The ability of Geobacter species to fix atmospheric nitrogen is an important metabolic feature for these applications. We identified novel regulatory cascades controlling nitrogen-fixation gene expression in Geobacter sulfurreducens. Unlike the regulatory mechanisms known in other nitrogen-fixing microorganisms, nitrogen-fixation gene regulation in G. sulfurreducens is controlled by two two-component His–Asp phosphorelay systems. One of these systems appears to be the master regulatory system that activates transcription of the majority of nitrogen-fixation genes and represses a gene encoding glutamate dehydrogenase during nitrogen fixation. The other system whose expression is directly activated by the master regulatory system appears to control by antitermination the expression of a subset of the nitrogen-fixation genes whose transcription is activated by the master regulatory system and whose promoter contains transcription termination signals. This study provides a new paradigm for nitrogen-fixation gene regulation. PMID:20660485

  4. Evolution of photosynthesis and biospheric oxygenation contingent upon nitrogen fixation?

    NASA Astrophysics Data System (ADS)

    Grula, John W.

    2005-10-01

    How photosynthesis by Precambrian cyanobacteria oxygenated Earth's biosphere remains incompletely understood. Here it is argued that the oxic transition, which took place between approximately 2.3 and 0.5 Gyr ago, required a great proliferation of cyanobacteria, and this in turn depended on their ability to fix nitrogen via the nitrogenase enzyme system. However, the ability to fix nitrogen was not a panacea, and the rate of biospheric oxygenation may still have been affected by nitrogen constraints on cyanobacterial expansion. Evidence is presented for why cyanobacteria probably have a greater need for fixed nitrogen than other prokaryotes, underscoring the importance of their ability to fix nitrogen. The connection between nitrogen fixation and the evolution of photosynthesis is demonstrated by the similarities between nitrogenase and enzymes critical for the biosynthesis of (bacterio)chlorophyll. It is hypothesized that biospheric oxygenation would not have occurred if the emergence of cyanobacteria had not been preceded by the evolution of nitrogen fixation, and if these organisms had not also acquired the ability to fix nitrogen at the beginning of or very early in their history. The evolution of nitrogen fixation also appears to have been a precondition for the evolution of (bacterio)chlorophyll-based photosynthesis. Given that some form of chlorophyll is obligatory for true photosynthesis, and its light absorption and chemical properties make it a ‘universal pigment’, it may be predicted that the evolution of nitrogen fixation and photosynthesis are also closely linked on other Earth-like planets.

  5. Oxygen relations of nitrogen fixation in cyanobacteria.

    PubMed Central

    Fay, P

    1992-01-01

    The enigmatic coexistence of O2-sensitive nitrogenase and O2-evolving photosynthesis in diazotrophic cyanobacteria has fascinated researchers for over two decades. Research efforts in the past 10 years have revealed a range of O2 sensitivity of nitrogenase in different strains of cyanobacteria and a variety of adaptations for the protection of nitrogenase from damage by both atmospheric and photosynthetic sources of O2. The most complex and apparently most efficient mechanisms for the protection of nitrogenase are incorporated in the heterocysts, the N2-fixing cells of cyanobacteria. Genetic studies indicate that the controls of heterocyst development and nitrogenase synthesis are closely interrelated and that the expression of N2 fixation (nif) genes is regulated by pO2. Images PMID:1620069

  6. Light response of nitrogen fixation in Lake Valencia, Venezuela

    SciTech Connect

    Lewis, W.M. Jr.; Levine, S.N.

    1984-07-01

    Water samples from six depths in the upper water column of Lake Valencia, Venezuela, were incubated in situ with acetylene at several depths corresponding to different light exposures. On five dates there was sufficient nitrogen fixation to define the light-response curve for samples taken at 2 and 100 cm. The light response of nitrogen fixation was modeled successfully with an equation previously developed for the light response of photosynthesis. All parameter values of the response curves for samples originating at a given depth were relatively stable across dates, but differed between depths. Rates of increase of N fixation with irradiance in the subsaturation range (..cap alpha..) were much lower than is typical of photosynthesis. While the general shape of nitrogen fixation curves was similar to that of photosynthesis curves, critical parameters were more sensitive to the light history (depth of origin) of cells than would be expected for photosynthesis.

  7. Nitrogen fixation and the diazotroph community in the temperate coastal region of the northwestern North Pacific

    NASA Astrophysics Data System (ADS)

    Shiozaki, T.; Nagata, T.; Ijichi, M.; Furuya, K.

    2015-08-01

    Nitrogen fixation in temperate oceans is a potentially important, but poorly understood process that may influence the marine nitrogen budget. This study determined seasonal variations in nitrogen fixation and the diazotroph community within the euphotic zone in the temperate coastal region of the northwestern North Pacific. Nitrogen fixation as high as 13.6 nmol N L-1 d-1 was measured from early summer to fall when the surface temperature exceeded 14.2 °C (but was lower than 24.3 °C) and the surface nitrate concentration was low (≤ 0.30 μM), although we also detected nitrogen fixation in subsurface layers (42-62 m) where nitrate concentrations were high (> 1 μM). Clone library analysis results indicated that nifH gene sequences were omnipresent throughout the investigation period. During the period when nitrogen fixation was detected (early summer to fall), the genes affiliated with UCYN-A, Trichodesmium, and γ-proteobacterial phylotype γ-24774A11 were frequently recovered. In contrast, when nitrogen fixation was undetectable (winter to spring), many sequences affiliated with Cluster III diazotrophs (putative anaerobic bacteria) were recovered. Quantitative PCR analysis revealed that UCYN-A was relatively abundant from early to late summer compared with Trichodesmium and γ-24774A11, whereas Trichodesmium abundance was the highest among the three groups during fall.

  8. Variability for Biological Nitrogen Fixation Capacity in Beans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As legumes, common beans have the capacity to form a symbiotic relationship with soil bacteria called rhizobia and fix nitrogen from the atmosphere. Common beans however are considered to be poor nitrogen fixers as compared to other legumes. Identification of genetic variability for N fixation capac...

  9. Low Carbon Costs of Nitrogen Fixation in Tropical Dry Forests

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

    Legume tree species with the ability to fix nitrogen (N) are highly diverse and widespread across tropical forests but in particular in the dry tropics. Their ecological success in lower latitudes has been called a "paradox": soil N in the tropics is thought to be high, while acquiring N through fixation incurs high energetic costs. However, the long held assumptions that N fixation is limited by photosynthate and that N fixation penalizes plant productivity have rarely been tested, particularly in legume tree species. We show results from three different experiments where we grew eleven species of tropical dry forest legumes. We quantified plant biomass and N fixation using nodulation and the 15N natural isotope abundance (Ndfa or nitrogen derived from fixation). These data show little evidence for costs of N fixation in seedlings grown under different soil fertility, light regimes, and with different microbial communities. Seedling productivity did not incur major costs because of N fixation: indeed, the average slope between Ndfa and biomass was positive (range in slopes: -0.03 to 0.3). Moreover, foliar N, which varied among species, was tightly constrained and not correlated with Ndfa. This finding implies that legume species have a target N that does not change depending on N acquisition strategies. The process of N fixation in tropical legumes may be more carbon efficient than previously thought. This view is more consistent with the hyperabundance of members of this family in tropical ecosystems.

  10. 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.

  11. 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

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

    PubMed

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

    2016-04-19

    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 N2fixation, the primary source of nitrogen to the ocean. Here, we present results from two cruises to the ETSP where rates of N2fixation and its contribution to export production were determined with a suite of geochemical and biological measurements. N2fixation 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 N2fixation 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 N2fixation 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 N2fixation at all six stations, studies of nitrogenase gene abundance and expression from the same cruises align with the geochemical data and together indicate that N2fixation 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 N2fixation in the ETSP. PMID:26976587

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

    PubMed

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

    2016-04-19

    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 N2fixation, the primary source of nitrogen to the ocean. Here, we present results from two cruises to the ETSP where rates of N2fixation and its contribution to export production were determined with a suite of geochemical and biological measurements. N2fixation 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 N2fixation 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 N2fixation 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 N2fixation at all six stations, studies of nitrogenase gene abundance and expression from the same cruises align with the geochemical data and together indicate that N2fixation 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 N2fixation in the ETSP.

  14. Problems and Promises of Assaying the Genetic Potential for Nitrogen Fixation in the Marine Environment

    PubMed

    Zehr; Capone

    1996-11-01

    Nitrogen fixation in the sea has attracted the attention of ecologists for decades. Much is known about the habitats in which it occurs and some of the factors that limit N2 fixation activity in different environments, but we still know little about the organisms that fix nitrogen, and what limits the growth and distribution of these organisms in marine environments. Molecular biology technological developments have provided tools for detecting and characterizing N2-fixing organisms in the environment. These techniques hold great promise for unraveling the mysteries and paradoxes of N2 fixation in the sea. In this review, we address the theoretical basis for the use of a molecular approach to N2 fixation, highlight the strengths and weaknesses of the approach, and provide case studies that demonstrate the potential contribution of molecular biology approaches to studies of N2 fixation in the sea.

  15. Regulation of Development and Nitrogen Fixation in Anabaena

    SciTech Connect

    James W Golden

    2004-08-05

    The nitrogen-fixing filamentous cyanobacterium Anabaena sp. strain PCC 7120 is being used as a simple model of microbial development and pattern formation in a multicellular prokaryotic organism. Anabaena reduces atmospheric nitrogen to ammonia in highly specialized, terminally differentiated cells called heterocysts. Anabaena is an important model system because of the multicellular growth pattern, the suspected antiquity of heterocyst development, and the contribution of fixed nitrogen to the environment. We are especially interested in understanding the molecular signaling pathways and genetic regulation that control heterocyst development. In the presence of an external source of reduced nitrogen, the differentiation of heterocysts is inhibited. When Anabaena is grown on dinitrogen, a one-dimensional developmental pattern of single heterocysts separated by approximately ten vegetative cells is established to form a multicellular organism composed of two interdependent cell types. The goal of this project is to understand the signaling and regulatory pathways that commit a vegetative cell to terminally differentiate into a nitrogen-fixing heterocyst. Several genes identified by us and by others were chosen as entry points into the regulatory network. Our research, which was initially focused on transcriptional regulation by group 2 sigma factors, was expanded to include group 3 sigma factors and their regulators after the complete Anabaena genome sequence became available. Surprisingly, no individual sigma factor is essential for heterocyst development. We have used the isolation of extragenic suppressors to study genetic interactions between key regulatory genes such as patS, hetR, and hetC in signaling and developmental pathways. We identified a hetR R223W mutation as a bypass suppressor of patS overexpression. Strains containing the hetR R223W allele fail to respond to pattern formation signals and overexpression of this allele results in a lethal phenotype

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

    PubMed

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

    2015-11-23

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

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

    PubMed

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

    2015-03-01

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

  18. Nitrogen fixation by corona discharge on the early precambrian Earth.

    PubMed

    Nna-Mvondo, Delphine; Navarro-González, Rafael; Raulin, François; Coll, Patrice

    2005-10-01

    We report the first experimental study of nitrogen fixation by corona discharge on the anoxic primitive Earth. The energy yields of nitric oxide (NO) and nitrous oxide (N(2)O) were experimentally determined over a wide range of CO(2)-N(2) mixtures simulating the evolution of the Earth's atmosphere during the Hadean and Archean eras (from 4.5 ba to 2.5 ba). NO, the principal form of fixed nitrogen in lightning and coronal discharge in early Earth, is produced ten times less efficiently in the latter type of electrical discharge with an estimated maximum annual production rate of the order of 10(10) g yr(-1). For N(2)O the maximum production rate was estimated to be approximately 10(9) g yr(-1). These low rates of syntheses indicate that corona discharges as point discharges on the clouds and ground did not play a significant role in the overall pool of reactive nitrogen needed for the emergence and sustainability of life.

  19. Molecular evidence for sediment nitrogen fixation in a temperate New England estuary

    PubMed Central

    Newell, Silvia E.; Pritchard, Kaitlyn R.; Foster, Sarah Q.

    2016-01-01

    Primary production in coastal waters is generally nitrogen (N) limited with denitrification outpacing nitrogen fixation (N2-fixation). However, recent work suggests that we have potentially underestimated the importance of heterotrophic sediment N2-fixation in marine ecosystems. We used clone libraries to examine transcript diversity of nifH (a gene associated with N2-fixation) in sediments at three sites in a temperate New England estuary (Waquoit Bay, Massachusetts, USA) and compared our results to net sediment N2 fluxes previously measured at these sites. We observed nifH expression at all sites, including a site heavily impacted by anthropogenic N. At this N impacted site, we also observed mean net sediment N2-fixation, linking the geochemical rate measurement with nifH expression. This same site also had the lowest diversity (non-parametric Shannon = 2.75). At the two other sites, we also detected nifH transcripts, however, the mean N2 flux indicated net denitrification. These results suggest that N2-fixation and denitrification co-occur in these sediments. Of the unique sequences in this study, 67% were most closely related to uncultured bacteria from various marine environments, 17% to Cluster III, 15% to Cluster I, and only 1% to Cluster II. These data add to the growing body of literature that sediment heterotrophic N2-fixation, even under high inorganic nitrogen concentrations, may be an important yet overlooked source of N in coastal systems. PMID:26977375

  20. Molecular evidence for sediment nitrogen fixation in a temperate New England estuary.

    PubMed

    Newell, Silvia E; Pritchard, Kaitlyn R; Foster, Sarah Q; Fulweiler, Robinson W

    2016-01-01

    Primary production in coastal waters is generally nitrogen (N) limited with denitrification outpacing nitrogen fixation (N2-fixation). However, recent work suggests that we have potentially underestimated the importance of heterotrophic sediment N2-fixation in marine ecosystems. We used clone libraries to examine transcript diversity of nifH (a gene associated with N2-fixation) in sediments at three sites in a temperate New England estuary (Waquoit Bay, Massachusetts, USA) and compared our results to net sediment N2 fluxes previously measured at these sites. We observed nifH expression at all sites, including a site heavily impacted by anthropogenic N. At this N impacted site, we also observed mean net sediment N2-fixation, linking the geochemical rate measurement with nifH expression. This same site also had the lowest diversity (non-parametric Shannon = 2.75). At the two other sites, we also detected nifH transcripts, however, the mean N2 flux indicated net denitrification. These results suggest that N2-fixation and denitrification co-occur in these sediments. Of the unique sequences in this study, 67% were most closely related to uncultured bacteria from various marine environments, 17% to Cluster III, 15% to Cluster I, and only 1% to Cluster II. These data add to the growing body of literature that sediment heterotrophic N2-fixation, even under high inorganic nitrogen concentrations, may be an important yet overlooked source of N in coastal systems.

  1. Nitrogen-fixation catalyst based on graphene: every part counts.

    PubMed

    Le, Yuan-Qi; Gu, Jia; Tian, Wei Quan

    2014-11-11

    The catalytic profile and function of each component of a molybdenum-graphene based catalyst (Mo/N-doped graphene) for nitrogen fixation, which combines the merits of these two components, is evaluated computationally. The Mo/N part acts as an active centre for N2 bond breaking and the graphene part works as an electron transmitter and electron reservoir.

  2. A new perspective on environmental controls of marine nitrogen fixation

    NASA Astrophysics Data System (ADS)

    Landolfi, A.; Koeve, W.; Dietze, H.; Kähler, P.; Oschlies, A.

    2015-06-01

    Growing slowly, marine N2 fixers are generally expected to be competitive only where nitrogen (N) supply is low relative to that of phosphorus (P) with respect to the cellular N:P ratio (R) of nonfixing phytoplankton. This is at odds with observed high N2 fixation rates in the oligotrophic North Atlantic where the ratio of nutrients supplied to the surface is elevated in N relative to the average R (16:1). In this study, we investigate several mechanisms to solve this puzzle: iron limitation, phosphorus enhancement by preferential remineralization or stoichiometric diversity of phytoplankton, and dissolved organic phosphorus (DOP) utilization. Combining resource competition theory and a global coupled ecosystem-circulation model, we find that the additional N and energy investments required for exoenzymatic breakdown of DOP give N2 fixers a competitive advantage in oligotrophic P-starved regions. Accounting for this mechanism expands the ecological niche of N2 fixers also to regions where the nutrient supply is high in N relative to R, yielding, in our model, a pattern consistent with the observed high N2 fixation rates in the oligotrophic North Atlantic.

  3. Nitrogen Fixation and Molecular Oxygen: Comparative Genomic Reconstruction of Transcription Regulation in Alphaproteobacteria.

    PubMed

    Tsoy, Olga V; Ravcheev, Dmitry A; Čuklina, Jelena; Gelfand, Mikhail S

    2016-01-01

    Biological nitrogen fixation plays a crucial role in the nitrogen cycle. An ability to fix atmospheric nitrogen, reducing it to ammonium, was described for multiple species of Bacteria and Archaea. The transcriptional regulatory network for nitrogen fixation was extensively studied in several representatives of the class Alphaproteobacteria. This regulatory network includes the activator of nitrogen fixation NifA, working in tandem with the alternative sigma-factor RpoN as well as oxygen-responsive regulatory systems, one-component regulators FnrN/FixK and two-component system FixLJ. Here we used a comparative genomics approach for in silico study of the transcriptional regulatory network in 50 genomes of Alphaproteobacteria. We extended the known regulons and proposed the scenario for the evolution of the nitrogen fixation transcriptional network. The reconstructed network substantially expands the existing knowledge of transcriptional regulation in nitrogen-fixing microorganisms and can be used for genetic experiments, metabolic reconstruction, and evolutionary analysis. PMID:27617010

  4. Nitrogen Fixation and Molecular Oxygen: Comparative Genomic Reconstruction of Transcription Regulation in Alphaproteobacteria

    PubMed Central

    Tsoy, Olga V.; Ravcheev, Dmitry A.; Čuklina, Jelena; Gelfand, Mikhail S.

    2016-01-01

    Biological nitrogen fixation plays a crucial role in the nitrogen cycle. An ability to fix atmospheric nitrogen, reducing it to ammonium, was described for multiple species of Bacteria and Archaea. The transcriptional regulatory network for nitrogen fixation was extensively studied in several representatives of the class Alphaproteobacteria. This regulatory network includes the activator of nitrogen fixation NifA, working in tandem with the alternative sigma-factor RpoN as well as oxygen-responsive regulatory systems, one-component regulators FnrN/FixK and two-component system FixLJ. Here we used a comparative genomics approach for in silico study of the transcriptional regulatory network in 50 genomes of Alphaproteobacteria. We extended the known regulons and proposed the scenario for the evolution of the nitrogen fixation transcriptional network. The reconstructed network substantially expands the existing knowledge of transcriptional regulation in nitrogen-fixing microorganisms and can be used for genetic experiments, metabolic reconstruction, and evolutionary analysis. PMID:27617010

  5. Fixation of nitrogen in the prebiotic atmosphere

    NASA Technical Reports Server (NTRS)

    Yung, Y. L.; Mcelroy, M. B.

    1979-01-01

    Reactions between nitrogen and water in the air surrounding lightning discharges can provide an important source of nitric oxide even under conditions where oxygen is a minor atmospheric constituent. Estimates are given for the associated source of soluble nitrite and nitrate. It is shown that lightning and subsequent atmospheric chemistry can provide a source of nitrate for the primitive ocean as large as one million tons of nitrogen per year, sufficient to fill the ocean to its present level of nitrate in less than one million years.

  6. Horizontal heterogeneity of nitrogen fixation in Lake Valencia, Venezuela

    SciTech Connect

    Levine, S.N.; Lewis, W.M. Jr.

    1985-11-01

    Spatial and temporal variability of nitrogen fixation in Lake Valencia, Venezuela, were quantified on the basis of duplicate water samples collected from a depth of 0.5 m at 16 sites on 10 dates. The concentration of heterocysts in samples were determined and the samples were incubated with acetylene in situ. Two-way ANOVA was used to separate the variance associated with site (fixed spatial patchiness), date (temporal variation), the interaction between site and date (ephemeral spatial patchiness), and sampling error. The nitrogen fixers in Lake Valencia are arranged in large (40-200 km/sup 2/), ephemeral patches with distinctive fixation rates per heterocyst. Both variability in fixation per heterocyst and variability in heterocyst concentration contribute significantly to variation in fixation per unit volume of lake water, but the variability attributable to heterocyst abundance is greater. Spatial variation in fixation and heterocyst concentration exceeds temporal variation in these parameters, and the ephemeral component of patchiness is much greater than the fixed component.

  7. 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.

  8. A possible nitrogen crisis for Archaean life due to reduced nitrogen fixation by lightning

    NASA Astrophysics Data System (ADS)

    Navarro-González, Rafael; McKay, Christopher P.; Mvondo, Delphine Nna

    2001-07-01

    Nitrogen is an essential element for life and is often the limiting nutrient for terrestrial ecosystems. As most nitrogen is locked in the kinetically stable form, N2, in the Earth's atmosphere, processes that can fix N2 into biologically available forms-such as nitrate and ammonia-control the supply of nitrogen for organisms. On the early Earth, nitrogen is thought to have been fixed abiotically, as nitric oxide formed during lightning discharge. The advent of biological nitrogen fixation suggests that at some point the demand for fixed nitrogen exceeded the supply from abiotic sources, but the timing and causes of the onset of biological nitrogen fixation remain unclear. Here we report an experimental simulation of nitrogen fixation by lightning over a range of Hadean (4.5-3.8Gyr ago) and Archaean (3.8-2.5Gyr ago) atmospheric compositions, from predominantly carbon dioxide to predominantly dinitrogen (but always without oxygen). We infer that, as atmospheric CO2 decreased over the Archaean period, the production of nitric oxide from lightning discharge decreased by two orders of magnitude until about 2.2Gyr. After this time, the rise in oxygen (or methane) concentrations probably initiated other abiotic sources of nitrogen. Although the temporary reduction in nitric oxide production may have lasted for only 100Myr or less, this was potentially long enough to cause an ecological crisis that triggered the development of biological nitrogen fixation.

  9. Nitrogen Fixation (Acetylene Reduction) in a Salt Marsh Amended with Sewage Sludge and Organic Carbon and Nitrogen Compounds 1

    PubMed Central

    Hanson, Roger B.

    1977-01-01

    Seasonal distribution of nitrogen fixation by Spartina alterniflora epiphytes and in surface and soil samples was investigated in a Georgia salt marsh which was amended with sewage sludge or with glucose and/or ammonium nitrate. There was no significant difference between the rates of fixation in the unamended and sewage sludge plots. Additional perturbation experiments suggested that nitrogen addition indirectly stimulates nitrogen fixation by enhancing Spartina production and root exudation. Glucose additions, on the other hand, suppressed nitrogen fixation on a long-term basis. It is suggested that the microbial population in the soil out-competed the plants for the available nitrogen and in turn suppressed plant production and possibly root exudation. A comparison of nitrogen fixation in clipped and unclipped Spartina plots substantiated the suggestion that root exudation probably supports nitrogen fixation. Fixation in the clipped plots was significantly lower (P < 0.05) than the rates in the unclipped plots. PMID:16345239

  10. 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

  11. Equatorial upwelling enhances nitrogen fixation in the Atlantic Ocean

    NASA Astrophysics Data System (ADS)

    Subramaniam, Ajit; Mahaffey, Claire; Johns, William; Mahowald, Natalie

    2013-05-01

    Surface waters in upwelling regions are thought to be nutrient rich and hence inhibit nitrogen fixation (diazotrophy) because diazotrophs can preferentially assimilate nitrate and ammonia instead of expending energy to fix dinitrogen. We found average nitrogen fixation rates to be two to seven times higher in the surface waters of the upwelling region of the eastern equatorial Atlantic than typically measured here during non-upwelling periods. We posit that in this region, low nitrate-phosphate ratio waters are upwelled, and an initial bloom of non-diazotrophic phytoplankton removes recently upwelled nitrate. Thereby, diazotrophy is fuelled by residual phosphate and by a combination of aeolian and upwelled sources of iron. Annually, we estimate that approximately 47 Gmol of new nitrogen is introduced by diazotrophy in upwelled waters alone and 195 Gmol N is fixed in the equatorial Atlantic region. Our findings challenge the paradigm that the highest nitrogen fixation rates occur in oligotrophic gyres and instead provide evidence of its importance in upwelling regimes where phosphate- and iron-rich waters rich are upwelled.

  12. 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

  13. Physiological limitations and the genetic improvement of symbiotic nitrogen fixation

    SciTech Connect

    Gara, F.O.; Manian, S. ); Drevon, J.J. )

    1988-01-01

    The rhizobium legume symbiosis continues to be of strategic importance particularly in the context of food production. As the world population grows, it is necessary that new developments take place in crop improvement. The development and application of new technologies in biological sciences over past years has made the entire area of plant-microbial interaction an exciting and challenging research area to be involved in. In view of the importance of symbiotic nitrogen fixation, it is not surprising that it still represents one of the priority areas for commercial development in agricultural biotechnology. Since this symbiosis involves an association between procaryotic and eucaryhotic partners, it requires of necessity a coordinated and interdisciplinary approach. This book focuses on physiological limitations affecting symbiotic nitrogen fixation and the potential for overcoming such limitations by using genetic technologies.

  14. [Nitrogen fixation potential of biological soil crusts in southeast edge of Tengger Desert, Northwest China].

    PubMed

    Zhang, Peng; Li, Xin-Rong; Zhang, Zhi-Shan; Pan, Yan-Xia; Liu, Yan-Mei; Su, Jie-Qiong

    2012-08-01

    Taking three typical types of biological soil crusts (BSCs), i.e., cyanobacterial-algal crust, lichen crust, and moss crust, in the southeast fringe of Tengger Desert as test objects, this paper studied their nitrogen fixation potential, seasonal fluctuation, and responses to the environmental factors from June 2010 to May 2011. During the whole study period, the nitrogenase activity (NA) of the cyanobacterial-algal, lichen, and moss crusts had significant difference, being 14-133, 20-101, and 4-28 micromol x m(-2) x h(-1), respectively, which indicated the critical role of the species composition of BSCs in nitrogen fixation. The NA of the three crust types had similar response characteristics to environmental factors. The NA had less correlation with the precipitation during the study period, but was positively correlated to the < 3 mm precipitation in the former 3 days of the experiment, indicating that the three types of BSCs could reach the maximum rate of nitrogen fixation under 3 mm precipitation. The NA of the three crust types had a significant quadratic functional relationship with air temperature, i.e., decreased after an initial increase. The NA of cyanobacterial-algal and lichen crusts declined rapidly when the temperature exceeded 30 degrees C, while the NA of moss crust began to decline when the temperature exceeded 25 degrees C, suggesting that different types of BSCs had different optimal temperature range of nitrogen fixation. The seasonal change of the NA of the three crust types was in the order of autumn > spring > summer > winter. The high air temperature in summer and the low air temperature (< 0 degrees C) in winter inhibited the NA of the crusts, while the suitable water and heat conditions in late spring and early autumn promoted the NA, which implied that the seasonal fluctuation of the NA was mainly controlled by the joint effect of the water and heat conditions. Under humid condition, the BSCs in the temperate desert zone had nitrogen

  15. Enhanced symbiotic nitrogen fixation with P. syringae pv tabaci

    SciTech Connect

    Langston-Unkefer, P.J.; Knight, T.J. New Mexico State Univ., Las Cruces ); Sengupta-Gopalan, C. )

    1989-04-01

    Infestation of legumes such as alfalfa and soybeans with the plant pathogen Pseudomonas syringae pv. tabaci is accompanied by increased plant growth, nodulation, overall nitrogen fixation, and total assimilated nitrogen. These effects are observed only in plants infested with Tox{sup +} pathogen; the toxin is tabtoxinine-{beta}-lactam, an active site-directed irreversible inhibitor of glutamine synthetase. The key to the legumes survival of this treatment is the insensitivity of the nodule-specific form of glutamine synthetase to the toxin. As expected, significant changes are observed in ammonia assimilation in these plants. The biochemical and molecular biological consequences of this treatment are being investigated.

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

    PubMed

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

    2016-08-01

    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.

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

    PubMed

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

    2016-08-01

    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

  18. EFFECT OF NITROGEN AND METAL ADDITIONS ON NITROGEN FIXATION ACTIVITY IN BIOLOGICAL SOIL CRUSTS

    NASA Astrophysics Data System (ADS)

    Alexander, K.; Lui, D.; Anbar, A. D.; Garcia-Pichel, F.; Hartnett, H. E.

    2009-12-01

    Biological soil crusts (BSCs) are diverse consortia of microorganisms that live in intimate association with soils in arid environments. Also called cryptogamic or microbiotic crusts, these communities can include cyanobacteria, algae, heterotrophic bacteria, fungi, lichens, and mosses. Together, these organisms provide many services to their surrounding ecosystems, including reduction of water runoff, promotion of water infiltration, and prevention of soil erosion. The cyanobacteria and algae also provide fixed carbon (C) to the soil through photosynthesis, and because atmospheric deposition of nitrogen (N) in arid environments is low, the major input of biologically available N comes from cyanobacteria capable of converting nitrogen gas (N2) to ammonium (NH4+). Biological soil crusts are easily destroyed by livestock grazing, motor vehicle travel, and many forms of recreational and agricultural land use. Loss of BSC cover can leave the soil vulnerable to intense erosion that can remove the nutrients necessary to sustain plant and animal life, thus accelerating the process of desertification. In order to preserve existing crusts and encourage the development of new crusts, it is crucial to understand the nutrient requirements of metabolism and growth in these microbial communities. This study investigated the affect of nitrogen and metal additions on N2-fixation activity in cyanobacterially-dominated crusts from the Colorado Plateau near Moab, Utah. Although N2-fixation has been studied in this system before, the affect of nutrient additions on N2-fixation activity has not been documented. The goal of this work was to understand how N and metal supplementation affects crust N metabolism. Three experiments were conducted to observe how N2-fixation activity changed with the addition of N, molybdenum (Mo), and vanadium (V). Molybdenum and vanadium were chosen because they are most commonly found at the active site of the enzyme nitrogenase, the molecule responsible

  19. The sensitivity of marine N2 fixation to dissolved inorganic nitrogen

    PubMed Central

    Knapp, Angela N.

    2012-01-01

    The dominant process adding nitrogen (N) to the ocean, di-nitrogen (N2) fixation, is mediated by prokaryotes (diazotrophs) sensitive to a variety of environmental factors. In particular, it is often assumed that consequential rates of marine N2 fixation do not occur where concentrations of nitrate (NO−3) and/or ammonium (NH+4) exceed 1μM because of the additional energetic cost associated with assimilating N2 gas relative to NO−3 or NH+4. However, an examination of culturing studies and in situ N2 fixation rate measurements from marine euphotic, mesopelagic, and benthic environments indicates that while elevated concentrations of NO−3 and/or NH+4 can depress N2 fixation rates, the process can continue at substantial rates in the presence of as much as 30μM NO−3 and/or 200μM NH+4. These findings challenge expectations of the degree to which inorganic N inhibits this process. The high rates of N2 fixation measured in some benthic environments suggest that certain benthic diazotrophs may be less sensitive to prolonged exposure to NO−3 and/or NH+4 than cyanobacterial diazotrophs. Additionally, recent work indicates that cyanobacterial diazotrophs may have mechanisms for mitigating NO−3 inhibition of N2 fixation. In particular, it has been recently shown that increasing phosphorus (P) availability increases diazotroph abundance, thus compensating for lower per-cell rates of N2 fixation that result from NO−3 inhibition. Consequently, low ambient surface ocean N:P ratios such as those generated by the increasing rates of N loss thought to occur during the last glacial to interglacial transition may create conditions favorable for N2 fixation and thus help to stabilize the marine N inventory on relevant time scales. These findings suggest that restricting measurements of marine N2 fixation to oligotrophic surface waters may underestimate global rates of this process and contribute to uncertainties in the marine N budget. PMID:23091472

  20. Nitrogen fixation rates in algal turf communities of a degraded versus less degraded coral reef

    NASA Astrophysics Data System (ADS)

    den Haan, Joost; Visser, Petra M.; Ganase, Anjani E.; Gooren, Elfi E.; Stal, Lucas J.; van Duyl, Fleur C.; Vermeij, Mark J. A.; Huisman, Jef

    2014-12-01

    Algal turf communities are ubiquitous on coral reefs in the Caribbean and are often dominated by N2-fixing cyanobacteria. However, it is largely unknown (1) how much N2 is actually fixed by turf communities and (2) which factors affect their N2 fixation rates. Therefore, we compared N2 fixation activity by turf communities at different depths and during day and night-time on a degraded versus a less degraded coral reef site on the island of Curaçao. N2 fixation rates measured with the acetylene reduction assay were slightly higher in shallow (5-10-m depth) than in deep turf communities (30-m depth), and N2 fixation rates during the daytime significantly exceeded those during the night. N2 fixation rates by the turf communities did not differ between the degraded and less degraded reef. Both our study and a literature survey of earlier studies indicated that turf communities tend to have lower N2 fixation rates than cyanobacterial mats. However, at least in our study area, turf communities were more abundant than cyanobacterial mats. Our results therefore suggest that turf communities play an important role in the nitrogen cycle of coral reefs. N2 fixation by turfs may contribute to an undesirable positive feedback that promotes the proliferation of algal turf communities while accelerating coral reef degradation.

  1. Nitrogen Fixation Tolerance to Soil Water Deficit Among Commercial Cultivars and breeding Lines of Peanut

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanut is often grown in the U.S. on sandy soil with limited water holding capacity. Since nitrogen fixation activity of other grain legume species, and some peanut cultivars, has been found to be especially sensitive to soil drying, this study was undertaken to examine a possible limitation in U.S...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    NASA Astrophysics Data System (ADS)

    Dekas, A. E.; Orphan, V.

    2011-12-01

    Biological nitrogen fixation (the conversion of N2 to NH3) is a critical process in the oceans, counteracting the production of N2 gas by dissimilatory bacterial metabolisms and providing a source of bioavailable nitrogen to many nitrogen-limited ecosystems. Although current measurements of N2 production and consumption in the oceans indicate that the nitrogen cycle is not balanced, recent findings on the limits of nitrogen fixation suggest that the perceived imbalance is an artifact of an incomplete assessment of marine diazotrophy. One currently poorly studied and potentially underappreciated habitat for diazotrophic organisms is the sediments of the deep-sea. In the present study we investigate the distribution and magnitude of benthic marine diazotrophy at several active deep-sea methane seeps (Mound 12, Costa Rica; Eel River Basin, CA, USA; Hydrate Ridge, OR, USA; and Monterey Canyon, CA, USA). Using 15N2 and 15NH4 sediment incubation experiments followed by single-cell (FISH-NanoSIMS) and bulk isotopic analysis (EA-IRMS), we observed total protein synthesis (15N uptake from 15NH4) and nitrogen fixation (15N update from 15N2). The highest rates of nitrogen fixation observed in the methane seep sediment incubation experiments were over an order of magnitude greater than those previously published from non-seep deep-sea sediments (Hartwig and Stanley, Deep-Sea Research, 1978, 25:411-417). However, methane seep diazotrophy appears to be highly spatially variable, with sediments exhibiting no nitrogen fixation originating only centimeters away from sediments actively incorporating 15N from 15N2. The greatest spatial variability in diazotrophy was observed with depth in the sediment, and corresponded to steep gradients in sulfate and methane. The maximum rates of nitrogen fixation were observed within the methane-sulfate transition zone, where organisms mediating the anaerobic oxidation of methane are typically in high abundance. Additionally, incubation

  4. Screening Prosopis (mesquite) germplasm for biomass production and nitrogen fixation

    SciTech Connect

    Felker, P.; Cannell, G.H.; Clark, P.R.; Osborn, J.F.

    1980-01-01

    The nitrogen-fixing trees of the genus Prosopis (mesquite or algaroba) are well adapted to the semi-arid and often saline regions of the world. These trees may produce firewood or pods for livestock food, they may stabilize sand dunes and they may enrich the soil by production of leaf litter supported by nitrogen fixation. A collection of nearly 500 Prosopis accessions representing North and South American and African germplasm has been established. Seventy of these accessions representing 14 taxa are being grown under field conditions where a 30-fold range in biomass productivity among accessions has been estimated. In a greehouse experiment, 13 Prosopis taxa grew on nitrogen-free medium nodulated, and had a 10-fold difference in nitrogen fixation (acetylene reduction). When Prosopis is propagated by seed the resulting trees are extremely variable in growth rate and presence or absence of thorns. Propagation of 6 Prosopis taxa by stem cuttings has been achieved with low success (1 to 10%) in field-grown plants and with higher success (50 to 100%) with young actively growing greenhouse plants.

  5. Can we trust current estimates for biological nitrogen fixation?

    NASA Astrophysics Data System (ADS)

    Bellenger, Jean-Philippe; Kraepiel, Anne

    2016-04-01

    Biological nitrogen fixation (BNF) consists on the reduction of atmospheric dinitrogen (N2) into bioavailable ammonium. This reaction accounts for up to 97% of nitrogen (N) input in unmanaged terrestrial ecosystems. Closing the N budget is a long standing challenge in many ecosystems. Recent studies have highlighted that current methods used to assess BNF are affected by critical biases. These findings challenge our confidence in many N budgets and call for a profound reconsideration of our methodological approaches. Beside these methodological issues, our ability to properly assess BNF might be further altered as a result of a misconception regarding the importance of BNF enzymatic diversity in nature. BNF is catalyzed by the enzyme nitrogenase (Nase) for which three isoforms have been identified so far; the molybdenum (Mo), vanadium (V) and iron-only (Fe) isoforms. Currently BNF is mostly considered to primarily depend on the Mo isoform. The contribution of the alternative Nases (V and Fe isoforms) to BNF in natural habitats has been mostly overlooked. However, recent findings have challenged this traditional view of the Nases hierarchy (Mo isoform predominance) with deep implications for BNF assessment in the field. Here, I will present an overview of recent findings, provided by various research groups, challenging current methods used to assess BNF. I will also present a summary of recent studies highlighting the importance of alternative Nases in nature. I will finally illustrate how altering our view on the Mo-Nase predominance can deeply affect our confidence in current BNF estimates. I will conclude by presenting new methodological approaches that will contribute to significantly improve our ability to understand and estimate BNF in the field by improving our capacity to access BNF spatio-temporal variability and enzymatic diversity.

  6. Local versus basin-scale limitation of marine nitrogen fixation.

    PubMed

    Weber, Thomas; Deutsch, Curtis

    2014-06-17

    Nitrogen (N) fixation by diazotrophic plankton is the primary source of this crucial nutrient to the ocean, but the factors limiting its rate and distribution are controversial. According to one view, the ecological niche of diazotrophs is primarily controlled by the ocean through internally generated N deficits that suppress the growth of their competitors. A second view posits an overriding limit from the atmosphere, which restricts diazotrophs to regions where dust deposition satisfies their high iron (Fe) requirement, thus separating N sources from sinks at a global scale. Here we use multiple geochemical signatures of N2 fixation to show that the Fe limitation of diazotrophs is strong enough to modulate the regional distribution of N2 fixation within ocean basins--particularly the Fe-poor Pacific--but not strong enough to influence its partition between basins, which is instead governed by rates of N loss. This scale-dependent limitation of N2 fixation reconciles local observations of Fe stress in diazotroph communities with an inferred spatial coupling of N sources and sinks. Within this regime of intermediate Fe control, the oceanic N reservoir would respond only weakly to enhanced dust fluxes during glacial climates, but strongly to the reduced fluxes hypothesized under anthropogenic climate warming. PMID:24889607

  7. Local versus basin-scale limitation of marine nitrogen fixation.

    PubMed

    Weber, Thomas; Deutsch, Curtis

    2014-06-17

    Nitrogen (N) fixation by diazotrophic plankton is the primary source of this crucial nutrient to the ocean, but the factors limiting its rate and distribution are controversial. According to one view, the ecological niche of diazotrophs is primarily controlled by the ocean through internally generated N deficits that suppress the growth of their competitors. A second view posits an overriding limit from the atmosphere, which restricts diazotrophs to regions where dust deposition satisfies their high iron (Fe) requirement, thus separating N sources from sinks at a global scale. Here we use multiple geochemical signatures of N2 fixation to show that the Fe limitation of diazotrophs is strong enough to modulate the regional distribution of N2 fixation within ocean basins--particularly the Fe-poor Pacific--but not strong enough to influence its partition between basins, which is instead governed by rates of N loss. This scale-dependent limitation of N2 fixation reconciles local observations of Fe stress in diazotroph communities with an inferred spatial coupling of N sources and sinks. Within this regime of intermediate Fe control, the oceanic N reservoir would respond only weakly to enhanced dust fluxes during glacial climates, but strongly to the reduced fluxes hypothesized under anthropogenic climate warming.

  8. Functional ecology of free-living nitrogen fixation: A contemporary perspective

    USGS Publications Warehouse

    Reed, Sasha C.; Cleveland, Cory C.; Townsend, Alan R.

    2011-01-01

    Nitrogen (N) availability is thought to frequently limit terrestrial ecosystem processes, and explicit consideration of N biogeochemistry, including biological N2 fixation, is central to understanding ecosystem responses to environmental change. Yet, the importance of free-living N2 fixation—a process that occurs on a wide variety of substrates, is nearly ubiquitous in terrestrial ecosystems, and may often represent the dominant pathway for acquiring newly available N—is often underappreciated. Here, we draw from studies that investigate free-living N2 fixation from functional, physiological, genetic, and ecological perspectives. We show that recent research and analytical advances have generated a wealth of new information that provides novel insight into the ecology of N2 fixation as well as raises new questions and priorities for future work. These priorities include a need to better integrate free-living N2 fixation into conceptual and analytical evaluations of the N cycle's role in a variety of global change scenarios.

  9. 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.

  10. 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

  11. 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.).

  12. Nitrogen Fixation on Early Mars and Other Terrestrial Planets: Experimental Demonstration of Abiotic Fixation Reactions to Nitrite and Nitrate

    NASA Astrophysics Data System (ADS)

    Summers, David P.; Khare, Bishun

    2007-05-01

    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 NO2. 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 NO2 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 NO2 reaction with ice, adsorbed water, etc.).

  13. An Alternative Path for the Evolution of Biological Nitrogen Fixation

    PubMed Central

    Boyd, Eric S.; Hamilton, Trinity L.; Peters, John W.

    2011-01-01

    Nitrogenase catalyzed nitrogen fixation is the process by which life converts dinitrogen gas into fixed nitrogen in the form of bioavailable ammonia. The most common form of nitrogenase today requires a complex metal cluster containing molybdenum (Mo), although alternative forms exist which contain vanadium (V) or only iron (Fe). It has been suggested that Mo-independent forms of nitrogenase (V and Fe) were responsible for N2 fixation on early Earth because oceans were Mo-depleted and Fe-rich. Phylogenetic- and structure-based examinations of multiple nitrogenase proteins suggest that such an evolutionary path is unlikely. Rather, our results indicate an evolutionary path whereby Mo-dependent nitrogenase emerged within the methanogenic archaea and then gave rise to the alternative forms suggesting that they arose later, perhaps in response to local Mo limitation. Structural inferences of nitrogenase proteins and related paralogs suggest that the ancestor of all nitrogenases had an open cavity capable of binding metal clusters which conferred reactivity. The evolution of the nitrogenase ancestor and its associated bound metal cluster was controlled by the availability of fixed nitrogen in combination with local environmental factors that influenced metal availability until a point in Earth’s geologic history where the most desirable metal, Mo, became sufficiently bioavailable to bring about and refine the solution (Mo-nitrogenase) we see perpetuated in extant biology. PMID:22065963

  14. Nitrogen fixation and respiratory electron transport in the cyanobacterium Cyanothece under different light/dark cycles.

    PubMed

    Rabouille, Sophie; Van de Waal, Dedmer B; Matthijs, Hans C P; Huisman, Jef

    2014-03-01

    Incompatibility of nitrogen fixation and oxygen production compels unicellular diazotrophic cyanobacteria to perform photosynthesis during daytime and restrict nitrogen fixation to nighttime. The marine diazotroph Cyanothece BG 043511 was grown in continuous culture under three light/dark regimes (16L : 8D, 12L : 12D, and 8L : 16D h); we monitored nitrogen fixation and potential photosynthetic efficiency simultaneously online to reveal how their temporal separation is affected by different LD regimes. An increase in nitrogen fixation rate at night coincided with a rise in pulse-amplitude modulated fluorescence, indicating that the enhanced respiratory electron transport to fuel diazotrophy affects the oxidation state of the plastoquinone pool. This may offer an alternative approach to assess instantaneous nitrogen fixation activity. Regardless of photoperiod, the maximum rate of nitrogen fixation was conserved at about 20 h after the onset of the light. Consequently, nitrogen fixation rates peaked at different moments in the dark: relatively early in the 16L : 8D cycle, at midnight in 12L : 12D, and relatively late in 8L : 16D. Under 16L : 8D, nitrogen fixation extended into the light, demonstrating the functional plasticity of nitrogen fixation in Cyanothece. Highest daily amounts of nitrogen fixed were obtained in 12L : 12D, which is consistent with the natural LD cycle of subtropical latitudes in which Cyanothece thrives.

  15. Nitrogen fixation in distinct microbial niches within a chemoautotrophy-driven cave ecosystem

    PubMed Central

    Desai, Mahesh S; Assig, Karoline; Dattagupta, Sharmishtha

    2013-01-01

    Microbial sulfur and carbon cycles in ecosystems driven by chemoautotrophy—present at deep-sea hydrothermal vents, cold seeps and sulfidic caves—have been studied to some extent, yet little is known about nitrogen fixation in these systems. Using a comprehensive approach comprising of 15N2 isotope labeling, acetylene reduction assay and nitrogenase gene expression analyses, we investigated nitrogen fixation in the sulfide-rich, chemoautotrophy-based Frasassi cave ecosystem (Italy). Nitrogen fixation was examined in three different microbial niches within the cave waters: (1) symbiotic bacterial community of Niphargus amphipods, (2) Beggiatoa-dominated biofilms, which occur at the sulfide–oxygen interface, and (3) sulfidic sediment. We found evidence for nitrogen fixation in all the three niches, and the nitrogenase gene (homologs of nifH) expression data clearly show niche differentiation of diazotrophic Proteobacteria within the water streams. The nifH transcript originated from the symbiotic community of Niphargus amphipods might belong to the Thiothrix ectosymbionts. Two abundantly expressed nifH genes in the Beggiatoa-dominated biofilms are closely related to those from Beggiatoa- and Desulfovibrio-related bacteria. These two diazotrophs were consistently found in Beggiatoa-dominated biofilms collected at various time points, thus illustrating species-specific associations of the diazotrophs in biofilm formation, and micron-scale niche partitioning of sulfur-oxidizing and sulfate-reducing bacteria driven by steep redox gradients within the biofilm. Finally, putative heterotrophs (Geobacter, Azoarcus and Desulfovibrio related) were the active diazotrophs in the sulfidic sediment. Our study is the first to shed light on nitrogen fixation in permanently dark caves and suggests that diazotrophy may be widespread in chemosynthetic communities. PMID:23924780

  16. 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.

  17. 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

  18. Diversity of nitrogen fixation strategies in Mediterranean legumes.

    PubMed

    Menge, Duncan N L; Wolf, Amelia A; Funk, Jennifer L

    2015-06-01

    Symbiotic N2 fixation (SNF) brings nitrogen into ecosystems, fuelling much of the world's agriculture(1) and sustaining carbon storage(2,3). However, it can also cause nitrogen saturation, exacerbating eutrophication and greenhouse warming(4-7). The balance of these effects depends on the degree to which N2-fixing plants adjust how much N2 they fix based on their needs (their SNF 'strategies')(5,6). Genetic, biochemical and physiological details of SNF are well known for certain economically important species(8,9), but the diversity of N2-fixing plants(10) and bacteria(11) is enormous, and little is known about most N2-fixing symbioses in natural ecosystems(12). Here, we show that co-occurring, closely related herbs exhibit diverse SNF strategies. In response to a nitrogen supply gradient, four species fixed less N2 than they needed (over-regulation), two fixed what they needed (facultative) and two did not downregulate SNF (obligate). No species downregulated but fixed more N2 than it needed (under-regulation or incomplete downregulation), but some species under-regulated or incompletely downregulated structural allocation to SNF. In fact, most species maintained nodules (the root structures that house symbionts) when they did not fix N2, suggesting decoupling of SNF activity and structure. Simulations showed that over-regulation of SNF activity is more adaptive than under-regulation or incomplete downregulation, and that different strategies have wildly different effects on ecosystem-level nitrogen cycling.

  19. Nitrogen fixation strategies can explain the latitudinal shift in nitrogen-fixing tree abundance.

    PubMed

    Menge, Duncan N L; Lichstein, Jeremy W; Angeles-Pérez, Gregorio

    2014-08-01

    The rarity of symbiotic nitrogen-fixing trees in higher-latitude compared to lower-latitude forests is paradoxical because higher-latitude soils are relatively N poor. Using national-scale forest inventories from the United States and Mexico, we show that the latitudinal abundance distribution of N-fixing trees (more than 10 times less abundant poleward of 35 degrees N) coincides with a latitudinal transition in symbiotic N-fixation type: rhizobial N-fixing trees (which are typically facultative, regulating fixation to meet nutritional demand) dominate equatorward of 35 degrees N, whereas actinorhizal N-fixing trees (typically obligate, maintaining fixation regardless of soil nutrition) dominate to the north. We then use theoretical and statistical models to show that a latitudinal shift in N-fixation strategy (facultative vs. obligate) near 35 degrees N can explain the observed change in N-fixing tree abundance, even if N availability is lower at higher latitudes, because facultative fixation leads to much higher landscape-scale N-fixing tree abundance than obligate fixation.

  20. Nitrogen fixation rate and chlorophyll content of the lichen Peltigera canina exposed to sulfur dioxide

    SciTech Connect

    Henriksson, E.; Pearson, L.C.

    1981-01-01

    In general, the rate of nitrogen fixation decreased when the lichen Peltigera canina (L.) Willd. was exposed to sulfur dioxide gas at levels from 0.1 to 500 ppm; at 30 ppm, however, nitrogen fixation was stimulated. The chlorophyll content decreased as the level of sulfur dioxide increased.

  1. 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

  2. The Sensitivity of Moss-Associated Nitrogen Fixation towards Repeated Nitrogen Input.

    PubMed

    Rousk, Kathrin; Michelsen, Anders

    2016-01-01

    Nitrogen (N2) fixation is a major source of available N in ecosystems that receive low amounts of atmospheric N deposition. In boreal forest and subarctic tundra, the feather moss Hylocomium splendens is colonized by N2 fixing cyanobacteria that could contribute fundamentally to increase the N pool in these ecosystems. However, N2 fixation in mosses is inhibited by N input. Although this has been shown previously, the ability of N2 fixation to grow less sensitive towards repeated, increased N inputs remains unknown. Here, we tested if N2 fixation in H. splendens can recover from increased N input depending on the N load (0, 5, 20, 80, 320 kg N ha(-1) yr(-1)) after a period of N deprivation, and if sensitivity towards increased N input can decrease after repeated N additions. Nitrogen fixation in the moss was inhibited by the highest N addition, but was promoted by adding 5 kg N ha(-1) yr(-1), and increased in all treatments during a short period of N deprivation. The sensitivity of N2 fixation towards repeated N additions seem to decrease in the 20 and 80 kg N additions, but increased in the highest N addition (320 kg N ha(-1) yr(-1)). Recovery of N in leachate samples increased with increasing N loads, suggesting low retention capabilities of mosses if N input is above 5 kg N ha(-1) yr(-1). Our results demonstrate that the sensitivity towards repeated N additions is likely to decrease if N input does not exceed a certain threshold. PMID:26731691

  3. The Sensitivity of Moss-Associated Nitrogen Fixation towards Repeated Nitrogen Input

    PubMed Central

    Rousk, Kathrin; Michelsen, Anders

    2016-01-01

    Nitrogen (N2) fixation is a major source of available N in ecosystems that receive low amounts of atmospheric N deposition. In boreal forest and subarctic tundra, the feather moss Hylocomium splendens is colonized by N2 fixing cyanobacteria that could contribute fundamentally to increase the N pool in these ecosystems. However, N2 fixation in mosses is inhibited by N input. Although this has been shown previously, the ability of N2 fixation to grow less sensitive towards repeated, increased N inputs remains unknown. Here, we tested if N2 fixation in H. splendens can recover from increased N input depending on the N load (0, 5, 20, 80, 320 kg N ha-1 yr-1) after a period of N deprivation, and if sensitivity towards increased N input can decrease after repeated N additions. Nitrogen fixation in the moss was inhibited by the highest N addition, but was promoted by adding 5 kg N ha-1 yr-1, and increased in all treatments during a short period of N deprivation. The sensitivity of N2 fixation towards repeated N additions seem to decrease in the 20 and 80 kg N additions, but increased in the highest N addition (320 kg N ha-1 yr-1). Recovery of N in leachate samples increased with increasing N loads, suggesting low retention capabilities of mosses if N input is above 5 kg N ha-1 yr-1. Our results demonstrate that the sensitivity towards repeated N additions is likely to decrease if N input does not exceed a certain threshold. PMID:26731691

  4. The Sensitivity of Moss-Associated Nitrogen Fixation towards Repeated Nitrogen Input.

    PubMed

    Rousk, Kathrin; Michelsen, Anders

    2016-01-01

    Nitrogen (N2) fixation is a major source of available N in ecosystems that receive low amounts of atmospheric N deposition. In boreal forest and subarctic tundra, the feather moss Hylocomium splendens is colonized by N2 fixing cyanobacteria that could contribute fundamentally to increase the N pool in these ecosystems. However, N2 fixation in mosses is inhibited by N input. Although this has been shown previously, the ability of N2 fixation to grow less sensitive towards repeated, increased N inputs remains unknown. Here, we tested if N2 fixation in H. splendens can recover from increased N input depending on the N load (0, 5, 20, 80, 320 kg N ha(-1) yr(-1)) after a period of N deprivation, and if sensitivity towards increased N input can decrease after repeated N additions. Nitrogen fixation in the moss was inhibited by the highest N addition, but was promoted by adding 5 kg N ha(-1) yr(-1), and increased in all treatments during a short period of N deprivation. The sensitivity of N2 fixation towards repeated N additions seem to decrease in the 20 and 80 kg N additions, but increased in the highest N addition (320 kg N ha(-1) yr(-1)). Recovery of N in leachate samples increased with increasing N loads, suggesting low retention capabilities of mosses if N input is above 5 kg N ha(-1) yr(-1). Our results demonstrate that the sensitivity towards repeated N additions is likely to decrease if N input does not exceed a certain threshold.

  5. Growth, photosynthesis and nitrogen fixation of Anabaena doliolum exposed to Assam crude extract

    SciTech Connect

    Gaur, J.P.; Singh, A.K. )

    1990-03-01

    Petroleum contaminants pose serious threat to the structure and functioning of aquatic ecosystems. Although their effects on various algal species have been studied before, heterocystous blue-green algae (cyanobacteria) have been little explored in this regard. Inadequate information concerning the effects of petroleum oils on them, especially on their nitrogen fixing system, prompted the authors to undertake this study. In this communication the authors report the influence of Assam crude on growth, photosynthesis ({sup 14}C incorporation), nitrogen fixation (nitrogenase activity) and heterocyst differentiation in Anabaena doliolum. This species and other heterocystous cyanobacteria occur widely in soil and aquatic ecosystems.

  6. Nitrogen fixation by cyanobacteria stimulates production in Baltic food webs.

    PubMed

    Karlson, Agnes M L; Duberg, Jon; Motwani, Nisha H; Hogfors, Hedvig; Klawonn, Isabell; Ploug, Helle; Barthel Svedén, Jennie; Garbaras, Andrius; Sundelin, Brita; Hajdu, Susanna; Larsson, Ulf; Elmgren, Ragnar; Gorokhova, Elena

    2015-06-01

    Filamentous, nitrogen-fixing cyanobacteria form extensive summer blooms in the Baltic Sea. Their ability to fix dissolved N2 allows cyanobacteria to circumvent the general summer nitrogen limitation, while also generating a supply of novel bioavailable nitrogen for the food web. However, the fate of the nitrogen fixed by cyanobacteria remains unresolved, as does its importance for secondary production in the Baltic Sea. Here, we synthesize recent experimental and field studies providing strong empirical evidence that cyanobacterial nitrogen is efficiently assimilated and transferred in Baltic food webs via two major pathways: directly by grazing on fresh or decaying cyanobacteria and indirectly through the uptake by other phytoplankton and microbes of bioavailable nitrogen exuded from cyanobacterial cells. This information is an essential step toward guiding nutrient management to minimize noxious blooms without overly reducing secondary production, and ultimately most probably fish production in the Baltic Sea.

  7. 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

  8. 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.

  9. 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

  10. Symbiotic Nitrogen Fixation and the Challenges to Its Extension to Nonlegumes.

    PubMed

    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; Ané, Jean-Michel; Peters, John W

    2016-07-01

    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

  11. Symbiotic Nitrogen Fixation and the Challenges to Its Extension to Nonlegumes.

    PubMed

    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; Ané, Jean-Michel; Peters, John W

    2016-07-01

    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.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  13. 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).

  14. 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

  15. 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.

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

    PubMed

    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.

  17. Regulation of Development and Nitrogen Fixation in Anabaena

    SciTech Connect

    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-term 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 rpoN (sigma 54

  18. 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.

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

    SciTech Connect

    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 found 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.

  20. Chloroplast gene sequence data suggest a single origin of the predisposition for symbiotic nitrogen fixation in angiosperms.

    PubMed Central

    Soltis, D E; Soltis, P S; Morgan, D R; Swensen, S M; Mullin, B C; Dowd, J M; Martin, P G

    1995-01-01

    Of the approximately 380 families of angiosperms, representatives of only 10 are known to form symbiotic associations with nitrogen-fixing bacteria in root nodules. The morphologically based classification schemes proposed by taxonomists suggest that many of these 10 families of plants are only distantly related, engendering the hypothesis that the capacity to fix nitrogen evolved independently several, if not many, times. This has in turn influenced attitudes toward the likelihood of transferring genes responsible for symbiotic nitrogen fixation to crop species lacking this ability. Phylogenetic analysis of DNA sequences for the chloroplast gene rbcL indicates, however, that representatives of all 10 families with nitrogen-fixing symbioses occur together, with several families lacking this association, in a single clade. This study therefore indicates that only one lineage of closely related taxa achieved the underlying genetic architecture necessary for symbiotic nitrogen fixation in root nodules. PMID:7708699

  1. 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.

  2. Systems biology of bacterial nitrogen fixation: High-throughput technology and its integrative description with constraint-based modeling

    PubMed Central

    2011-01-01

    fixation, all of these in qualitative agreement with observations made in R. etli and other Rhizobiaceas. Conclusions In this work we present a genome scale study of the metabolic activity in bacterial nitrogen fixation. This approach leads us to construct a computational model that serves as a guide for 1) integrating high-throughput data, 2) describing and predicting metabolic activity, and 3) designing experiments to explore the genotype-phenotype relationship in bacterial nitrogen fixation. PMID:21801415

  3. 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.

  4. Nitrogen fixation and nitrogenase activities in members of the family Rhodospirillaceae.

    PubMed Central

    Madigan, M; Cox, S S; Stegeman, R A

    1984-01-01

    Strains of all 18 species of the family Rhodospirillaceae (nonsulfur photosynthetic bacteria) were studied for their comparative nitrogen-fixing abilities. All species, with the exception of Rhodocyclus purpureus, were capable of growth with N2 as the sole nitrogen source under photosynthetic (anaerobic) conditions. Most rapid growth on N2 was observed in strains of Rhodopseudomonas capsulata. Within the genus Rhodopseudomonas, the species R. capsulata, R. sphaeroides, R. viridis, R. gelatinosa, and R. blastica consistently showed the highest in vivo nitrogenase rates (with the acetylene reduction technique); nitrogenase rates in other species of Rhodopseudomonas and in most species of Rhodospirillum were notably lower. Chemotrophic (dark microaerobic) nitrogen fixation occurred in all species with the exception of one strain of Rhodospirillum fulvum; oxygen requirements for dark N2 fixation varied considerably among species and even within strains of the same species. We conclude that the capacity to fix molecular nitrogen is virtually universal among members of the Rhodospirillaceae but that the efficacy of the process varies considerably among species. PMID:6581158

  5. Responses of soil nitrogen fixation to Spartina alterniflora invasion and nitrogen addition in a Chinese salt marsh.

    PubMed

    Huang, Jingxin; Xu, Xiao; Wang, Min; Nie, Ming; Qiu, Shiyun; Wang, Qing; Quan, Zhexue; Xiao, Ming; Li, Bo

    2016-02-12

    Biological nitrogen fixation (BNF) is the major natural process of nitrogen (N) input to ecosystems. To understand how plant invasion and N enrichment affect BNF, we compared soil N-fixation rates and N-fixing microbes (NFM) of an invasive Spartina alterniflora community and a native Phragmites australis community in the Yangtze River estuary, with and without N addition. Our results indicated that plant invasion relative to N enrichment had a greater influence on BNF. At each N level, the S. alterniflora community had a higher soil N-fixation rate but a lower diversity of the nifH gene in comparison with the native community. The S. alterniflora community with N addition had the highest soil N-fixation rate and the nifH gene abundance across all treatments. Our results suggest that S. alterniflora invasion can increase soil N fixation in the high N-loading estuarine ecosystem, and thus may further mediate soil N availability.

  6. Responses of soil nitrogen fixation to Spartina alterniflora invasion and nitrogen addition in a Chinese salt marsh

    PubMed Central

    Huang, Jingxin; Xu, Xiao; Wang, Min; Nie, Ming; Qiu, Shiyun; Wang, Qing; Quan, Zhexue; Xiao, Ming; Li, Bo

    2016-01-01

    Biological nitrogen fixation (BNF) is the major natural process of nitrogen (N) input to ecosystems. To understand how plant invasion and N enrichment affect BNF, we compared soil N-fixation rates and N-fixing microbes (NFM) of an invasive Spartina alterniflora community and a native Phragmites australis community in the Yangtze River estuary, with and without N addition. Our results indicated that plant invasion relative to N enrichment had a greater influence on BNF. At each N level, the S. alterniflora community had a higher soil N-fixation rate but a lower diversity of the nifH gene in comparison with the native community. The S. alterniflora community with N addition had the highest soil N-fixation rate and the nifH gene abundance across all treatments. Our results suggest that S. alterniflora invasion can increase soil N fixation in the high N-loading estuarine ecosystem, and thus may further mediate soil N availability. PMID:26869197

  7. Effects of drought on characters related to nitrogen fixation in peanut

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Twelve peanut genotypes were tested under three water regimes in two greenhouses to investigate the effects of drought on biomass production and N2 fixation. Drought reduced biomass production from 36.5 to 56.0% and reduced nitrogen fixation from 26.8 to 68.8%. Most genotypes with high biomass produ...

  8. Inferring the evolutionary history of Mo-dependent nitrogen fixation from phylogenetic studies of nifK and nifDK.

    PubMed

    Hartmann, Linda S; Barnum, Susan R

    2010-07-01

    The ability to fix nitrogen is widely, but sporadically distributed among the Bacteria and Archaea suggesting either a vertically inherited, ancient function with widespread loss across genera or an adaptive feature transferred laterally between co-inhabitants of nitrogen-poor environments. As previous phylogenetic studies of nifH and nifD have not completely resolved the evolutionary history of nitrogenase, sixty nifD, nifK, and combined nifDK genes were analyzed using Bayesian, maximum likelihood, and parsimony algorithms to determine whether the individual and combined datasets could provide additional information. The results show congruence between the 16S and nifDK phylogenies at the phyla level and generally support vertical descent with loss. However, statistically significant differences between tree topographies suggest a complex evolutionary history with the underlying pattern of vertical descent obscured by recurring lateral transfer events and different patterns of evolution between the genes. Results support inheritance from the Last Common ancestor or an ancient lateral transfer of the nif genes between Bacteria and Archaea, ongoing gene transfer between cohabitants of similar biogeographic regions, acquisition of nitrogen-fixing capability via symbiosis islands, possible xenologous displacement of one gene in the operon, and possible retention of ancestral genes in heterocystous cyanobacteria. Analyses support the monophyly of the Cyanobacteria, alphabetagamma-Proteobacteria, and Actinobacteria (Frankia) and provide strong support for the placement of Frankia nif genes at the base of combined the Cyanobacteria/Proteobacteria clades.

  9. Genetic regulation of nitrogen fixation in Rhizobium meliloti.

    PubMed

    Cebolla, A; Palomares, A J

    1994-12-01

    The soil bacterium Rhizobium meliloti fixes dinitrogen when associated with root nodules formed on its plant host, Medicago sativa (alfalfa). The expression of most of the known genes required for nitrogen fixation (nif and fix genes), including the structural genes for nitrogenase, is induced in response to a decrease in oxygen concentration. Induction of nif and fix gene expression by low oxygen is physiologically relevant because a low-oxygen environment is maintained in root nodules to prevent inactivation of the highly oxygen-sensitive nitrogenase enzyme. The genes responsible for sensing and transducing the low oxygen signal, fixL and fixJ, encode proteins (FixL and FixJ, respectively) that are homologous to a large family of bacterial proteins involved in signal transduction, the two component regulatory system proteins. The two components consist of a sensor protein, to which FixL is homologous, and a response regulator protein, to which FixJ is homologous. The sensor protein respond to an activating signal by autophosphorylating and then transferring the phosphate to its cognate response regulator protein. The phosphorylated response regulator, which is often a transcriptional activator, is then able to activate its target. A cascade model of nif and fix gene regulation in R. meliloti has been proposed, whereby FixL acts as an oxygen sensor as the initial event in the cascade and transmits this information to FixJ. FixJ, which possesses a putative helix-turn-helix DNA-binding motif, then activates transcription of the nifA and fixK genes. The nifA and fixK gene products, are transcriptional activators of at least 14 other nif and fix genes.

  10. Influence of Co and B12 on the growth and nitrogen fixation of Trichodesmium

    PubMed Central

    Rodriguez, Irene B.; Ho, Tung-Yuan

    2015-01-01

    We investigated the influence of varying cobalt (Co) and B12 concentrations to growth and nitrogen fixation of Trichodesmium, a major diazotroph in the tropical and subtropical oligotrophic ocean. Here we show that sufficient inorganic Co, 20 pmol L-1, sustains the growth of Trichodesmium either with or without an additional B12 supply. We also found that in these culture conditions, nitrogen levels fixed by Trichodesmium were higher in treatments with insufficient B12 than in treatments with higher B12 availability. Under limited inorganic Co availability, ranging from 0.2 to 2 pmol L-1, Trichodesmium growth was significantly compromised in cultures without B12. In these low Co concentrations, addition of 400 pmol L-1 of B12 supported phytoplankton growth indicating that B12 supply augmented for the low Co concentrations. Our study demonstrates that Trichodesmium has an absolute Co requirement, which is not replaceable with Zn, and that B12 supply alleviates stress in cases where Co is limiting. These results show that the interlocking availabilities of Co and B12 may influence the growth and nitrogen fixation of Trichodesmium in the ocean. PMID:26150813

  11. 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.

  12. MALDI mass spectrometry-assisted molecular imaging of metabolites during nitrogen fixation in the Medicago truncatula-Sinorhizobium meliloti symbiosis.

    PubMed

    Ye, Hui; Gemperline, Erin; Venkateshwaran, Muthusubramanian; Chen, Ruibing; Delaux, Pierre-Marc; Howes-Podoll, Maegen; Ané, Jean-Michel; Li, Lingjun

    2013-07-01

    Symbiotic associations between leguminous plants and nitrogen-fixing rhizobia culminate in the formation of specialized organs called root nodules, in which the rhizobia fix atmospheric nitrogen and transfer it to the plant. Efficient biological nitrogen fixation depends on metabolites produced by and exchanged between both partners. The Medicago truncatula-Sinorhizobium meliloti association is an excellent model for dissecting this nitrogen-fixing symbiosis because of the availability of genetic information for both symbiotic partners. Here, we employed a powerful imaging technique - matrix-assisted laser desorption/ionization (MALDI)/mass spectrometric imaging (MSI) - to study metabolite distribution in roots and root nodules of M. truncatula during nitrogen fixation. The combination of an efficient, novel MALDI matrix [1,8-bis(dimethyl-amino) naphthalene, DMAN] with a conventional matrix 2,5-dihydroxybenzoic acid (DHB) allowed detection of a large array of organic acids, amino acids, sugars, lipids, flavonoids and their conjugates with improved coverage. Ion density maps of representative metabolites are presented and correlated with the nitrogen fixation process. We demonstrate differences in metabolite distribution between roots and nodules, and also between fixing and non-fixing nodules produced by plant and bacterial mutants. Our study highlights the benefits of using MSI for detecting differences in metabolite distributions in plant biology.

  13. 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...

  14. Nitrogen fixation. (Latest citations from the Selected Water Resources Abstracts database). Published Search

    SciTech Connect

    Not Available

    1993-01-01

    The bibliography contains citations concerning nitrogen fixation in terrestrial and aquatic ecosystems and its impact on future resources. The biochemistry and biophysics of the nitrogen-fixing bacteria, algae, and blue-green algae are discussed. (Contains a minimum of 97 citations and includes a subject term index and title list.)

  15. A minimal nitrogen fixation gene cluster from Paenibacillus sp. WLY78 enables expression of active nitrogenase in Escherichia coli.

    PubMed

    Wang, Liying; Zhang, Lihong; Liu, Zhanzhi; Liu, Zhangzhi; 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 σ(70) (σ(A))-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.

  16. 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

  17. 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

  18. Symbiotic nitrogen fixation in the fungus gardens of leaf-cutter ants.

    PubMed

    Pinto-Tomás, Adrián A; Anderson, Mark A; Suen, Garret; Stevenson, David M; Chu, Fiona S T; Cleland, W Wallace; Weimer, Paul J; Currie, Cameron R

    2009-11-20

    Bacteria-mediated acquisition of atmospheric N2 serves as a critical source of nitrogen in terrestrial ecosystems. Here we reveal that symbiotic nitrogen fixation facilitates the cultivation of specialized fungal crops by leaf-cutter ants. By using acetylene reduction and stable isotope experiments, we demonstrated that N2 fixation occurred in the fungus gardens of eight leaf-cutter ant species and, further, that this fixed nitrogen was incorporated into ant biomass. Symbiotic N2-fixing bacteria were consistently isolated from the fungus gardens of 80 leaf-cutter ant colonies collected in Argentina, Costa Rica, and Panama. The discovery of N2 fixation within the leaf-cutter ant-microbe symbiosis reveals a previously unrecognized nitrogen source in neotropical ecosystems.

  19. Direct nitrogen fixation at the edges of graphene nanoplatelets as efficient electrocatalysts for energy conversion

    PubMed Central

    Jeon, In-Yup; Choi, Hyun-Jung; Ju, Myung Jong; Choi, In Taek; Lim, Kimin; Ko, Jaejung; Kim, Hwan Kyu; Kim, Jae Cheon; Lee, Jae-Joon; Shin, Dongbin; Jung, Sun-Min; Seo, Jeong-Min; Kim, Min-Jung; Park, Noejung; Dai, Liming; Baek, Jong-Beom

    2013-01-01

    Nitrogen fixation is essential for the synthesis of many important chemicals (e.g., fertilizers, explosives) and basic building blocks for all forms of life (e.g., nucleotides for DNA and RNA, amino acids for proteins). However, direct nitrogen fixation is challenging as nitrogen (N2) does not easily react with other chemicals. By dry ball-milling graphite with N2, we have discovered a simple, but versatile, scalable and eco-friendly, approach to direct fixation of N2 at the edges of graphene nanoplatelets (GnPs). The mechanochemical cracking of graphitic C−C bonds generated active carbon species that react directly with N2 to form five- and six-membered aromatic rings at the broken edges, leading to solution-processable edge-nitrogenated graphene nanoplatelets (NGnPs) with superb catalytic performance in both dye-sensitized solar cells and fuel cells to replace conventional Pt-based catalysts for energy conversion. PMID:23877200

  20. Direct nitrogen fixation at the edges of graphene nanoplatelets as efficient electrocatalysts for energy conversion

    NASA Astrophysics Data System (ADS)

    Jeon, In-Yup; Choi, Hyun-Jung; Ju, Myung Jong; Choi, In Taek; Lim, Kimin; Ko, Jaejung; Kim, Hwan Kyu; Kim, Jae Cheon; Lee, Jae-Joon; Shin, Dongbin; Jung, Sun-Min; Seo, Jeong-Min; Kim, Min-Jung; Park, Noejung; Dai, Liming; Baek, Jong-Beom

    2013-07-01

    Nitrogen fixation is essential for the synthesis of many important chemicals (e.g., fertilizers, explosives) and basic building blocks for all forms of life (e.g., nucleotides for DNA and RNA, amino acids for proteins). However, direct nitrogen fixation is challenging as nitrogen (N2) does not easily react with other chemicals. By dry ball-milling graphite with N2, we have discovered a simple, but versatile, scalable and eco-friendly, approach to direct fixation of N2 at the edges of graphene nanoplatelets (GnPs). The mechanochemical cracking of graphitic C-C bonds generated active carbon species that react directly with N2 to form five- and six-membered aromatic rings at the broken edges, leading to solution-processable edge-nitrogenated graphene nanoplatelets (NGnPs) with superb catalytic performance in both dye-sensitized solar cells and fuel cells to replace conventional Pt-based catalysts for energy conversion.

  1. Direct nitrogen fixation at the edges of graphene nanoplatelets as efficient electrocatalysts for energy conversion.

    PubMed

    Jeon, In-Yup; Choi, Hyun-Jung; Ju, Myung Jong; Choi, In Taek; Lim, Kimin; Ko, Jaejung; Kim, Hwan Kyu; Kim, Jae Cheon; Lee, Jae-Joon; Shin, Dongbin; Jung, Sun-Min; Seo, Jeong-Min; Kim, Min-Jung; Park, Noejung; Dai, Liming; Baek, Jong-Beom

    2013-01-01

    Nitrogen fixation is essential for the synthesis of many important chemicals (e.g., fertilizers, explosives) and basic building blocks for all forms of life (e.g., nucleotides for DNA and RNA, amino acids for proteins). However, direct nitrogen fixation is challenging as nitrogen (N₂) does not easily react with other chemicals. By dry ball-milling graphite with N₂, we have discovered a simple, but versatile, scalable and eco-friendly, approach to direct fixation of N₂ at the edges of graphene nanoplatelets (GnPs). The mechanochemical cracking of graphitic C--C bonds generated active carbon species that react directly with N₂ to form five- and six-membered aromatic rings at the broken edges, leading to solution-processable edge-nitrogenated graphene nanoplatelets (NGnPs) with superb catalytic performance in both dye-sensitized solar cells and fuel cells to replace conventional Pt-based catalysts for energy conversion.

  2. The transcriptional activator NrpA is crucial for inducing nitrogen fixation in Methanosarcina mazei Gö1 under nitrogen-limited conditions.

    PubMed

    Weidenbach, Katrin; Ehlers, Claudia; Schmitz, Ruth A

    2014-08-01

    With the aim of unraveling their potential involvement in the regulation of nitrogen metabolism in Methanosarcina mazei strain Gö1, we characterized five genes that are differentially transcribed in response to changing nitrogen availability and encoding putative transcriptional regulators. Study of the respective mutant strains under nitrogen-limited conditions revealed a growth delay for M. mazei MM0444::pac and MM1708::pac, and strongly reduced diazotrophic growth for MM0872::pac, whereas the absence of MM2441 or MM2525 did not affect growth behaviour. Transcriptome analyses further demonstrated that only MM1708 - encoding a CxxCG zinc finger protein - plays a regulatory role in nitrogen metabolism, most likely by specifically enhancing transcription of the N2 fixation (nif) operon under nitrogen-limited conditions. In agreement with this, a palindromic binding motif was predicted in silico in the nifH promoter region, nine nucleotides upstream of the BRE box, and confirmed to bind purified maltose-binding protein-MM1708 by electromobility shift assays. As MM1708 itself is under the control of the global nitrogen repressor NrpR, this adds a secondary level to the transcriptional regulation of the nif genes, and is most likely crucial for maximal nif induction under nitrogen-limited conditions. This is in accordance with the finding that protein expression of NifH is highly reduced in the absence of MM1708 under nitrogen-limited conditions. On the basis of our findings, we hypothesize that, in M. mazei, nitrogen fixation is controlled by a hierarchical network of two transcriptional regulators, the global nitrogen repressor NrpR, and the newly identified activator NrpA (MM1708), thereby providing tight control of N2 fixation.

  3. 15N discrimination and the sensitivity of nitrogen fixation to changes in dietary nitrogen in Reticulitermes flavipes (Isoptera: Rhinotermitidae).

    PubMed

    Meuti, Megan E; Jones, Susan C; Curtis, Peter S

    2010-12-01

    Xylophagous termites possess symbiotic bacteria that fix atmospheric nitrogen (N(2)). Although symbiotic N(2) fixation is central to termite nutrition and ecologically important, it is energetically costly. Using stable isotopes, we tested the hypothesis that symbiotic N(2) fixation would decrease in workers of the eastern subterranean termite, Reticulitermes flavipes Kollar, which were exposed to high nitrogen diets. To calculate the isotope discrimination factor occurring as a result of digestion, Δ(dig), and which occurs as the result of N(2) fixation, Δ(fix), symbiotic N(2) fixation was inhibited via force feeding termites the antibiotic kanamycin. Antibiotic-treated termites and control (N(2)-fixing) termites were exposed to different concentrations of dietary N (0, 0.21, and 0.94% N), their (15)N signatures were obtained, and the percent nitrogen derived from the atmosphere within termite samples was calculated. As we hypothesized, symbiotic N(2) fixation rates were negatively correlated with dietary N, suggesting that high concentrations of dietary N suppressed symbiotic N(2) fixation in R. flavipes. A comparison of the (15)N isotope signatures of antibiotic-treated termites with their food sources demonstrated that Δ(dig) = 2.284‰, and a comparison of the (15)N isotope signatures of antibiotic-treated termites with control termites indicated that Δ(fix) = -1.238‰. These are the first estimates of Δ(dig) for R. flavipes, and the first estimate of Δ(fix) for any N(2)-fixing termite species.

  4. Nitrogen fixation by Klebsiella pneumoniae is inhibited by certain multicopy hybrid nif plasmids.

    PubMed

    Riedel, G E; Brown, S E; Ausubel, F M

    1983-01-01

    In our studies of nif gene regulation, we have observed that certain hybrid nif plasmids drastically inhibit the expression of the chromosomal nif genes of Klebsiella pneumonia. Wild-type (Nif+) K. pneumoniae strains that acquire certain hybrid nif plasmids also acquire the Nif- phenotype; these strains lose 90 to 99% of all detectable nitrogen fixation activity and grow poorly (or not at all) on solid media with N2 as the sole nitrogen source. We describe experiments which defined this inhibition of the Nif+ phenotype by hybrid nif plasmids and identify and characterize four nif DNA regions associated with this inhibition. We show that plasmids carrying these nif regions could recombine with, but not complement, nif chromosomal mutations. Our results suggest that inhibition of the Nif+ phenotype will provide a useful bioassay for some of the factors that mediate nif gene expression.

  5. Nitrogen fixation in an oligotrophic, saline desert lake: Pyramid Lake, Nevada

    SciTech Connect

    Horne, A.J.; Galat, D.L.

    1985-11-01

    High rates of nitrogen fixation by a short-lived but dense unialgal bloom of the planktonic blue-green Nodularia spumigena provided 99.5% of the alga's needs and 81% of Pyramid Lake's annual total combined nitrogen input in 1979. The bloom was spatially very heterogeneous. Bloom size, duration, and presumably N/sub 2/ fixation vary from year to year, but in 1979 about 900 t of nitrogenwere fixed in 2 months in this large deep lake. The annual rate of N/sub 2/ fixation was about 2 g m/sup -2/. In this year of low inflow the Truckee River provided 54 t of inorganic nitrogen and 83 t of organic nitrogen. Planktonic N/sub 2/ fixation has not been measured during high inflow years and may have been small relative to river input. Lakewide average heterocyst to vegetative cell (h:c) ratios followed seasonal trends in N/sub 2/ fixation, but synoptic samples showed only a weak relation between h:c and N/sub 2/ fixation. N/sub 2/ fixation was induced by low epilimnetic levels of inorganic nitrogen and ended before lake overturn in the fall. High rates of N/sub 2/ fixation were confined to the upper 5% of the epilimnetic volume and thus occurred only in calm weather when Nodularia colonies floated to the lake surface. Access to freshly dissolved atmospheric CO/sub 2/ may account for the near-surface dependence, since the lake pH is normally about 9.2. Nodularia will not show the same degree of near-surface dependence in near-neutral lakes or in the ocean.

  6. Regulation of symbiotic nitrogen fixation in root nodules of alfalfa (Medicago sativa) infected with Rhizobium meliloti.

    PubMed

    Kamberger, W

    1977-10-24

    Symbiotic nitrogen fixation of Rhizobium meliloti bacteroids in Medicago sativa root nodules was suppressed by several inorganic nitrogen sources. Amino acids like glutamine, glutamic acid and aspartic acid, which can serve as sole nitrogen sources for the unnodulated plant did not influence nitrogenase activity of effective nodules, even at high concetrations. Ammonia and nitrate suppressed symbiotic nitrogen fixation in vivo only at concentrations much higher than those needed for suppression of nitrogenase activity in free living nitrogen fixing bacteria. The kinetics of suppression were slow compared with that of free living nitrogen fixing bacteria. On the other hand, nitrite, which acts as a direct inhibitor of nitrogenase, suppressed very quickly and at low concentrations. Glutamic acid and glutamine enhanced the effect of ammonia dramatically, while the suppression by nitrate was enhanced only slightly.

  7. Nitrogen fixation and identification of potential diazotrophs in the Canadian Arctic

    NASA Astrophysics Data System (ADS)

    Blais, Marjolaine; Tremblay, Jean-Éric; Jungblut, Anne D.; Gagnon, Jonathan; Martin, Johannie; Thaler, Mary; Lovejoy, Connie

    2012-09-01

    Global gaseous nitrogen (N2) fixation rates may be underestimated and data is lacking from many regions without conspicuous diazotrophic cyanobacteria, such as cold oceans. We estimated N2 fixation rates at diverse sites in the Canadian Arctic, including the mouth of the Mackenzie River, the offshore Beaufort Sea, Lancaster Sound, Baffin Bay and a river influenced fjord. We also identified potential diazotrophic communities using a targeted survey of the nifH gene. Nitrogen fixation rates ranged from 0.02 nmol N L-1 d-1 in Baffin Bay to 4.45 nmol N L-1 d-1 in the Mackenzie River plume. Sequences recovered from the nifH gene survey belonged mainly to Cluster III, a group of nifH sequences associated with diverse microorganisms, with some α- andγ-proteobacterianifH genes at most sites. Cyanobacteria nifH genes with best matches to Nostocales, which are common in Arctic freshwaters, were recovered from the marine Beaufort Sea. The geographic pattern of N2 fixation rates and nifHgene identities suggest that the Mackenzie River is the source of a diazotrophic community that contributes new nitrogen to the nitrogen-depleted surface waters of the Beaufort Sea. This first record of N2 fixation at high latitudes refines our understanding of the global nitrogen budget.

  8. Using synthetic biology to distinguish and overcome regulatory and functional barriers related to nitrogen fixation.

    PubMed

    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 σ(54)-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 σ(54)-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.

  9. Nitrogen fixation island and rhizosphere competence traits in the genome of root-associated Pseudomonas stutzeri A1501

    PubMed Central

    Yan, Yongliang; Yang, Jian; Dou, Yuetan; Chen, Ming; Ping, Shuzhen; Peng, Junping; Lu, Wei; Zhang, Wei; Yao, Ziying; Li, Hongquan; Liu, Wei; He, Sheng; Geng, Lizhao; Zhang, Xiaobing; Yang, Fan; Yu, Haiying; Zhan, Yuhua; Li, Danhua; Lin, Zhanglin; Wang, Yiping; Elmerich, Claudine; Lin, Min; Jin, Qi

    2008-01-01

    The capacity to fix nitrogen is widely distributed in phyla of Bacteria and Archaea but has long been considered to be absent from the Pseudomonas genus. We report here the complete genome sequencing of nitrogen-fixing root-associated Pseudomonas stutzeri A1501. The genome consists of a single circular chromosome with 4,567,418 bp. Comparative genomics revealed that, among 4,146 protein-encoding genes, 1,977 have orthologs in each of the five other Pseudomonas representative species sequenced to date. The genome contains genes involved in broad utilization of carbon sources, nitrogen fixation, denitrification, degradation of aromatic compounds, biosynthesis of polyhydroxybutyrate, multiple pathways of protection against environmental stress, and other functions that presumably give A1501 an advantage in root colonization. Genetic information on synthesis, maturation, and functioning of nitrogenase is clustered in a 49-kb island, suggesting that this property was acquired by lateral gene transfer. New genes required for the nitrogen fixation process have been identified within the nif island. The genome sequence offers the genetic basis for further study of the evolution of the nitrogen fixation property and identification of rhizosphere competence traits required in the interaction with host plants; moreover, it opens up new perspectives for wider application of root-associated diazotrophs in sustainable agriculture. PMID:18495935

  10. High cell-specific rates of nitrogen and carbon fixation by the cyanobacterium Aphanizomenon sp. at low temperatures in the Baltic Sea.

    PubMed

    Svedén, Jennie B; Adam, Birgit; Walve, Jakob; Nahar, Nurun; Musat, Niculina; Lavik, Gaute; Whitehouse, Martin J; Kuypers, Marcel M M; Ploug, Helle

    2015-12-01

    Aphanizomenon is a widespread genus of nitrogen (N2)-fixing cyanobacteria in lakes and estuaries, accounting for a large fraction of the summer N2-fixation in the Baltic Sea. However, information about its cell-specific carbon (C)- and N2-fixation rates in the early growth season has not previously been reported. We combined various methods to study N2-fixation, photosynthesis and respiration in field-sampled Baltic Sea Aphanizomenon sp. during early summer at 10°C. Stable isotope incubations at in situ light intensities during 24 h combined with cell-specific secondary ion mass spectrometry showed an average net N2-fixation rate of 55 fmol N cell(-1) day(-1). Dark net N2-fixation rates over a course of 12 h were 20% of those measured in light. C-fixation, but not N2-fixation, was inhibited by high ambient light intensities during daytime. Consequently, the C:N fixation ratio varied substantially over the diel cycle. C- and N2-fixation rates were comparable to those reported for Aphanizomenon sp. in August at 19°C, using the same methods. High respiration rates (23% of gross photosynthesis) were measured with (14)C-incubations and O2-microsensors, and presumably reflect the energy needed for high N2-fixation rates. Hence, Aphanizomenon sp. is an important contributor to N2-fixation at low in situ temperatures in the early growth season.

  11. High cell-specific rates of nitrogen and carbon fixation by the cyanobacterium Aphanizomenon sp. at low temperatures in the Baltic Sea.

    PubMed

    Svedén, Jennie B; Adam, Birgit; Walve, Jakob; Nahar, Nurun; Musat, Niculina; Lavik, Gaute; Whitehouse, Martin J; Kuypers, Marcel M M; Ploug, Helle

    2015-12-01

    Aphanizomenon is a widespread genus of nitrogen (N2)-fixing cyanobacteria in lakes and estuaries, accounting for a large fraction of the summer N2-fixation in the Baltic Sea. However, information about its cell-specific carbon (C)- and N2-fixation rates in the early growth season has not previously been reported. We combined various methods to study N2-fixation, photosynthesis and respiration in field-sampled Baltic Sea Aphanizomenon sp. during early summer at 10°C. Stable isotope incubations at in situ light intensities during 24 h combined with cell-specific secondary ion mass spectrometry showed an average net N2-fixation rate of 55 fmol N cell(-1) day(-1). Dark net N2-fixation rates over a course of 12 h were 20% of those measured in light. C-fixation, but not N2-fixation, was inhibited by high ambient light intensities during daytime. Consequently, the C:N fixation ratio varied substantially over the diel cycle. C- and N2-fixation rates were comparable to those reported for Aphanizomenon sp. in August at 19°C, using the same methods. High respiration rates (23% of gross photosynthesis) were measured with (14)C-incubations and O2-microsensors, and presumably reflect the energy needed for high N2-fixation rates. Hence, Aphanizomenon sp. is an important contributor to N2-fixation at low in situ temperatures in the early growth season. PMID:26511856

  12. Biological nitrogen fixation in acidic high-temperature geothermal springs in Yellowstone National Park, Wyoming.

    PubMed

    Hamilton, Trinity L; Lange, Rachel K; Boyd, Eric S; Peters, John W

    2011-08-01

    The near ubiquitous distribution of nifH genes in sediments sampled from 14 high-temperature (48.0-89.0°C) and acidic (pH 1.90-5.02) geothermal springs in Yellowstone National Park suggested a role for the biological reduction of dinitrogen (N(2)) to ammonia (NH(3)) (e.g. nitrogen fixation or diazotrophy) in these environments. nifH genes from these environments formed three unique phylotypes that were distantly related to acidiphilic, mesophilic diazotrophs. Acetylene reduction assays and (15) N(2) tracer studies in microcosms containing sediments sampled from acidic and high-temperature environments where nifH genes were detected confirmed the potential for biological N(2) reduction in these environments. Rates of acetylene reduction by sediment-associated populations were positively correlated with the concentration of NH(4)(+), suggesting a potential relationship between NH(4)(+) consumption and N(2) fixation activity. Amendment of microcosms with NH(4)(+) resulted in increased lag times in acetylene reduction assays. Manipulation of incubation temperature and pH in acetylene reduction assays indicated that diazotrophic populations are specifically adapted to local conditions. Incubation of sediments in the presence of a N(2) headspace yielded a highly enriched culture containing a single nifH phylotype. This phylotype was detected in all 14 geothermal spring sediments examined and its abundance ranged from ≈ 780 to ≈ 6800 copies (g dry weight sediment)(-1), suggesting that this organism may contribute N to the ecosystems. Collectively, these results for the first time demonstrate thermoacidiphilic N(2) fixation in the natural environment and extend the upper temperature for biological N(2) fixation in terrestrial systems.

  13. Biotic nitrogen fixation in the bryosphere is inhibited more by drought than warming.

    PubMed

    Whiteley, Jonathan A; Gonzalez, Andrew

    2016-08-01

    The boreal forest is of particular interest to climate change research due to its large circumpolar distribution and accumulated soil carbon pool. Carbon uptake in this ecosystem is nitrogen (N)-limited, therefore factors affecting carbon or nitrogen dynamics in the boreal forest can have consequences for global climate. We used a 2-year field experiment to investigate the response of biotic nitrogen fixation by cyanobacteria associated with boreal forest bryophytes, in a factorial experiment combining simulated climate change with habitat fragmentation treatments. We simulated climate change conditions using open-top greenhouse chambers in the field, which increased mean and maximum temperatures, and created a precipitation gradient from ambient levels in the center to extreme drought conditions at the periphery of the chamber. The dry patches near the chamber walls exhibited almost no N-fixation, despite having similar densities of cyanobacteria (predominantly Stigonema sp.) as other patches. Rates of N-fixation were best explained by a model containing moisture, fragmentation, cyanobacteria density and time; warming was not a significant variable affecting N-fixation. There was no significant interaction between warming and fragmentation. These results suggest that cyanobacteria responded physiologically to drought by reducing N-fixation activity long before any changes in density. Ecosystem processes, such as N-fixation, can respond in the short term to environmental change much more rapidly than changes in the underlying community structure. Such rapid physiological responses may occur faster than demographic insurance effects of biodiversity. PMID:27098528

  14. Biotic nitrogen fixation in the bryosphere is inhibited more by drought than warming.

    PubMed

    Whiteley, Jonathan A; Gonzalez, Andrew

    2016-08-01

    The boreal forest is of particular interest to climate change research due to its large circumpolar distribution and accumulated soil carbon pool. Carbon uptake in this ecosystem is nitrogen (N)-limited, therefore factors affecting carbon or nitrogen dynamics in the boreal forest can have consequences for global climate. We used a 2-year field experiment to investigate the response of biotic nitrogen fixation by cyanobacteria associated with boreal forest bryophytes, in a factorial experiment combining simulated climate change with habitat fragmentation treatments. We simulated climate change conditions using open-top greenhouse chambers in the field, which increased mean and maximum temperatures, and created a precipitation gradient from ambient levels in the center to extreme drought conditions at the periphery of the chamber. The dry patches near the chamber walls exhibited almost no N-fixation, despite having similar densities of cyanobacteria (predominantly Stigonema sp.) as other patches. Rates of N-fixation were best explained by a model containing moisture, fragmentation, cyanobacteria density and time; warming was not a significant variable affecting N-fixation. There was no significant interaction between warming and fragmentation. These results suggest that cyanobacteria responded physiologically to drought by reducing N-fixation activity long before any changes in density. Ecosystem processes, such as N-fixation, can respond in the short term to environmental change much more rapidly than changes in the underlying community structure. Such rapid physiological responses may occur faster than demographic insurance effects of biodiversity.

  15. 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)).

  16. Effect of insecticides and phenolics on nitrogen fixation by Nostoc linckia

    SciTech Connect

    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, cypermethrin 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.

  17. Nitrogen fixation in lichens is important for improved rock weathering.

    PubMed

    Seneviratne, Gamini; Indrasena, I K

    2006-12-01

    It is known that cyanobacteria in cyanolichens fix nitrogen for their nutrition.However, specific uses of the fixed nitrogen have not been examined. The present study shows experimentally that a mutualistic interaction between a heterotrophic N2 fixer and lichen fungi in the presence of a carbon source can contribute to enhanced release of organic acids, leading to improved solubilization of the mineral substrate. Three lichen fungi were isolated from Xanthoparmelia mexicana, a foliose lichen, and they were cultured separately or with a heterotrophic N2 fixer in nutrient broth media in the presence of a mineral substrate. Cells of the N2-fixing bacteria attached to the mycelial mats of all fungi, forming biofilms. All biofilms showed higher solubilizations of the substrate than cultures of their fungi alone. This finding has bearing on the significance of the origin and existence of N2-fixing activity in the evolution of lichen symbiosis. Further, our results may explain why there are N2-fixing photobionts even in the presence of non- fixing photobionts (green algae) in some remarkable lichens such as Placopsis gelida. Our study sheds doubt on the idea that the establishment of terrestrial eukaryotes was possible only through the association between a fungus and a phototroph.

  18. Enzymology of biological nitrogen fixation. Final report, May 1, 1987--April 30, 1996

    SciTech Connect

    1997-05-01

    Biological nitrogen fixation is of central importance in the earth`s nitrogen economy. Fixation of nitrogen is accomplished by a variety of microorganisms, all of them procaryotic. Some operate independently and some function symbiotically or associatively with photosynthesizing plants. Biological nitrogen fixation is accomplished via the reaction: N{sub 2} + 8H{sup +} + 8e{sup {minus}} {yields} 2NH{sub 3} + H{sub 2}. This reaction requires a minimum of 16 ATP under ideal laboratory conditions, so it is obvious that the energy demand of the reaction is very high. When certain nitrogen-fixing organisms are supplied fixed nitrogen (e.g., ammonium) the organisms use the fixed nitrogen and turn off their nitrogenase system, thus conserving energy. When the fixed nitrogen is exhausted, the organism reactivates its nitrogenase. The system is turned off by dinitrogenase reductase ADP-ribosyl transferase (DRAT) and turned back on by dinitrogenase reductase-activating glycohydrolase (DRAG). The authors have investigated the details of how DRAT and DRAG are formed, how they function, and the genetics of their formation and operation.

  19. 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.

  20. 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

  1. Identifying Regulatory Changes to Facilitate Nitrogen Fixation in the Nondiazotroph Synechocystis sp. PCC 6803.

    PubMed

    Mueller, Thomas J; Welsh, Eric A; Pakrasi, Himadri B; Maranas, Costas D

    2016-03-18

    The incorporation of biological nitrogen fixation into a nondiazotrophic photosynthetic organism provides a promising solution to the increasing fixed nitrogen demand, but is accompanied by a number of challenges for accommodating two incompatible processes within the same organism. Here we present regulatory influence networks for two cyanobacteria, Synechocystis PCC 6803 and Cyanothece ATCC 51142, and evaluate them to co-opt native transcription factors that may be used to control the nif gene cluster once it is transferred to Synechocystis. These networks were further examined to identify candidate transcription factors for other metabolic processes necessary for temporal separation of photosynthesis and nitrogen fixation, glycogen catabolism and cyanophycin synthesis. Two transcription factors native to Synechocystis, LexA and Rcp1, were identified as promising candidates for the control of the nif gene cluster and other pertinent metabolic processes, respectively. Lessons learned in the incorporation of nitrogen fixation into a nondiazotrophic prokaryote may be leveraged to further progress the incorporation of nitrogen fixation in plants.

  2. Nitrogen fixation (Acetylene Reduction) by annual winter legumes on a coal surface mine

    SciTech Connect

    Gabrielson, F.C.

    1982-01-01

    The winter annuals, crimson clover, rose clover, subterranean clover and hairy vetch, were evaluated for nitrogen fixing capacity on coal surface mine substrates by measuring their ability to reduce acetylene to ethylene. The effects of fertilizer, Abruzzi rye, Kentucky 31 fescue grass and a phytotoxic plant Chenopodium album on nitrogen fixation were also assessed. Crimson clover was recommended as the best legume to use on topsoil and shale in the south. Hairy vetch gave good results on shale and subterranean clover did well on topsoil. The use of these species for revegetation is discussed. Overall, no correlation between substrate pH and ethylene levels was found and effects of substrate depended upon the legume species. Super phosphate fertilizer supported less nitrogen fixation than 13-13-13. Abruzzi rye in some unknown way inhibited plant density and nitrogen fixation by legumes but not by free living substrate micro-organisms. Shale from under dead Chenopodium plants in both field and greehouse experiments did not inhibit nitrogen fixation. 7 tables.

  3. Nitrogen fixation (acetylene reduction) by annual winter legumes on a coal surface mine

    SciTech Connect

    Gabrielson, F.C.

    1982-01-01

    The winter annuals, crimson clover, rose clover, subterranean clover and hairy vetch, were evaluated for their ability to fix nitrogen on coal surface mine substrates by measuring their ability to reduce acetylene to ethylene. The effects of fertilizer, Abruzzi ryegrass, Kentucky 31 fescue grass and a phytotoxic plant Chenopodium album on nitrogen fixation was also assessed. Crimson clover was recommended as the best legume to use on topsoil and shale in the South. Hairy vetch gave good results on shale and subterranean clover did well on topsoil. The use of these species for revegetation is discussed. Overall, no correlation between substrate pH and ethylene levels was found and effects of substrate depended upon the legume species. Super phosphate fertilizer supported less nitrogen fixation than 13-13-13. Abruzzi ryegrass in some unknown way inhibited plant density and nitrogen fixation by legumes but not by free living substrate micro-organisms. Shale from under dead Chenopodium plants in both field and greenhouse experiments did not inhibit nitrogen fixation. 11 references, 7 tables.

  4. Quantitative trait locus analysis of symbiotic nitrogen fixation activity in the model legume Lotus japonicus.

    PubMed

    Tominaga, Akiyoshi; Gondo, Takahiro; Akashi, Ryo; Zheng, Shao-Hui; Arima, Susumu; Suzuki, Akihiro

    2012-05-01

    Many legumes form nitrogen-fixing root nodules. An elevation of nitrogen fixation in such legumes would have significant implications for plant growth and biomass production in agriculture. To identify the genetic basis for the regulation of nitrogen fixation, quantitative trait locus (QTL) analysis was conducted with recombinant inbred lines derived from the cross Miyakojima MG-20 × Gifu B-129 in the model legume Lotus japonicus. This population was inoculated with Mesorhizobium loti MAFF303099 and grown for 14 days in pods containing vermiculite. Phenotypic data were collected for acetylene reduction activity (ARA) per plant (ARA/P), ARA per nodule weight (ARA/NW), ARA per nodule number (ARA/NN), NN per plant, NW per plant, stem length (SL), SL without inoculation (SLbac-), shoot dry weight without inoculation (SWbac-), root length without inoculation (RLbac-), and root dry weight (RWbac-), and finally 34 QTLs were identified. ARA/P, ARA/NN, NW, and SL showed strong correlations and QTL co-localization, suggesting that several plant characteristics important for symbiotic nitrogen fixation are controlled by the same locus. QTLs for ARA/P, ARA/NN, NW, and SL, co-localized around marker TM0832 on chromosome 4, were also co-localized with previously reported QTLs for seed mass. This is the first report of QTL analysis for symbiotic nitrogen fixation activity traits.

  5. OxyR-regulated catalase activity is critical for oxidative stress resistance, nodulation and nitrogen fixation in Azorhizobium caulinodans.

    PubMed

    Zhao, Yue; Nickels, Logan M; Wang, Hui; Ling, Jun; Zhong, Zengtao; Zhu, Jun

    2016-07-01

    The legume-rhizobial interaction results in the formation of symbiotic nodules in which rhizobia fix nitrogen. During the process of symbiosis, reactive oxygen species (ROS) are generated. Thus, the response of rhizobia to ROS is important for successful nodulation and nitrogen fixation. In this study, we investigated how Azorhizobium caulinodans, a rhizobium that forms both root and stem nodules on its host plant, regulates ROS resistance. We found that in-frame deletions of a gene encoding the putative catalase-peroxidase katG or a gene encoding a LysR-family regulatory protein, oxyR, exhibited increased sensitivity to H2O2 We then showed that OxyR positively regulated katG expression in an H2O2-independent fashion. Furthermore, we found that deletion of katG or oxyR led to significant reduction in the number of stem nodules and decrease of nitrogen fixation capacities in symbiosis. Our results revealed that KatG and OxyR are not only critical for antioxidant defense in vitro, but also important for nodule formation and nitrogen fixation during interaction with plant hosts. PMID:27190162

  6. OxyR-regulated catalase activity is critical for oxidative stress resistance, nodulation and nitrogen fixation in Azorhizobium caulinodans.

    PubMed

    Zhao, Yue; Nickels, Logan M; Wang, Hui; Ling, Jun; Zhong, Zengtao; Zhu, Jun

    2016-07-01

    The legume-rhizobial interaction results in the formation of symbiotic nodules in which rhizobia fix nitrogen. During the process of symbiosis, reactive oxygen species (ROS) are generated. Thus, the response of rhizobia to ROS is important for successful nodulation and nitrogen fixation. In this study, we investigated how Azorhizobium caulinodans, a rhizobium that forms both root and stem nodules on its host plant, regulates ROS resistance. We found that in-frame deletions of a gene encoding the putative catalase-peroxidase katG or a gene encoding a LysR-family regulatory protein, oxyR, exhibited increased sensitivity to H2O2 We then showed that OxyR positively regulated katG expression in an H2O2-independent fashion. Furthermore, we found that deletion of katG or oxyR led to significant reduction in the number of stem nodules and decrease of nitrogen fixation capacities in symbiosis. Our results revealed that KatG and OxyR are not only critical for antioxidant defense in vitro, but also important for nodule formation and nitrogen fixation during interaction with plant hosts.

  7. Nitrogen fixation in the Gulf of California and the Eastern Tropical North Pacific

    NASA Astrophysics Data System (ADS)

    White, Angelicque E.; Foster, Rachel A.; Benitez-Nelson, Claudia R.; Masqué, Pere; Verdeny, Elisabet; Popp, Brian N.; Arthur, Karen E.; Prahl, Fredrick G.

    2013-02-01

    Di-nitrogen (N2) fixation plays a well-recognized role in the enhancement of primary production and arguably particle export in oligotrophic regions of the subtropical and tropical oceans. However, recent evidence suggests that N2 fixation may also be significant in regions of the surface ocean proximate to or overlying zones of intense subsurface denitrification. In this study, we present results from a series of research cruises in the Gulf of California (GoCal) and adjacent waters of the Eastern Tropical North Pacific (ETNP). Measurements include microscopy, genomic analyses, incubations, stable isotopic measurements, and sediment traps coupled with 238U:234Th disequilibria. Combined, these results suggest that N2 fixing microorganisms are present and active throughout the region, with larger sized Richelia and Trichodesmium spp. recorded within the warmer waters at the entrance to and within the GoCal, and smaller, unicellular diazotrophs observed in the cooler waters of the northern ETNP. N2 fixation rates in the summer varied from 15-70 μmol N m-2 d-1, with episodic blooms contributing as much as 795 μmol N m-2 d-1. While the estimated contribution of N2 fixation to particle export was highly variable, blooms of diatom-Richelia symbioses accounted for as much as ∼44% of the measured summer carbon flux at 100 m. Alternately, evaluation of the N isotopic composition of sinking material and the magnitude of measured N2 fixation rates indicate negligible to small enhancements of new production when blooms of either colonial Trichodesmium spp. or unicellular diazotrophs were encountered. Consistent with previous research, we also found that while fluxes of C to sediment traps are similar in winter and summer months, the efficiency of C export (export/surface productivity) in the GoCal region is elevated during summer relative to the more productive diatom-dominated winter phase of the seasonal cycle. The episodic and variable nature of N2 fixation recorded in

  8. Transcriptomic Analysis of Sinorhizobium meliloti and Medicago truncatula Symbiosis Using Nitrogen Fixation-Deficient Nodules.

    PubMed

    Lang, Claus; Long, Sharon R

    2015-08-01

    The bacterium Sinorhizobium meliloti interacts symbiotically with legume plant hosts such as Medicago truncatula to form nitrogen-fixing root nodules. During symbiosis, plant and bacterial cells differentiate in a coordinated manner, resulting in specialized plant cells that contain nitrogen-fixing bacteroids. Both plant and bacterial genes are required at each developmental stage of symbiosis. We analyzed gene expression in nodules formed by wild-type bacteria on six plant mutants with defects in nitrogen fixation. We observed differential expression of 482 S. meliloti genes with functions in cell envelope homeostasis, cell division, stress response, energy metabolism, and nitrogen fixation. We simultaneously analyzed gene expression in M. truncatula and observed differential regulation of host processes that may trigger bacteroid differentiation and control bacterial infection. Our analyses of developmentally arrested plant mutants indicate that plants use distinct means to control bacterial infection during early and late symbiotic stages.

  9. Increasing Nitrogen Fixation and Seed Development in Soybean Requires Complex Adjustments of Nodule Nitrogen Metabolism and Partitioning Processes.

    PubMed

    Carter, Amanda M; Tegeder, Mechthild

    2016-08-01

    Legumes are able to access atmospheric di-nitrogen (N2) through a symbiotic relationship with rhizobia that reside within root nodules. In soybean, following N2 fixation by the bacteroids, ammonia is finally reduced in uninfected cells to allantoin and allantoic acid [1]. These ureides present the primary long-distance transport forms of nitrogen (N), and are exported from nodules via the xylem for shoot N supply. Transport of allantoin and allantoic acid out of nodules requires the function of ureide permeases (UPS1) located in cells adjacent to the vasculature [2, 3]. We expressed a common bean UPS1 transporter in cortex and endodermis cells of soybean nodules and found that delivery of N from nodules to shoot, as well as seed set, was significantly increased. In addition, the number of transgenic nodules was increased and symbiotic N2 fixation per nodule was elevated, indicating that transporter function in nodule N export is a limiting step in bacterial N acquisition. Further, the transgenic nodules showed considerable increases in nodule N assimilation, ureide synthesis, and metabolite levels. This suggests complex adjustments of nodule N metabolism and partitioning processes in support of symbiotic N2 fixation. We propose that the transgenic UPS1 plants display metabolic and allocation plasticity to overcome N2 fixation and seed yield limitations. Overall, it is demonstrated that transporter function in N export from nodules is a key step for enhancing atmospheric N2 fixation and nodule function and for improving shoot N nutrition and seed development in legumes.

  10. Selection and breeding of corn to enhance associative bacterial nitrogen fixation

    SciTech Connect

    Ela, S.W.; Anderson, M.A.; Brill, W.J.

    1980-01-01

    We have increased, through screening and breeding, the ability of corn (maize, Zea mays L.) to support bacterial nitrogen fixation in or on corn roots. Isotopic N fixed from /sup 15/N/sub 2/ was found on the roots. Even though the nitrogen-fixing association depends on germ plasm from tropical corn, the activity can be bred into corn currently used in midwestern United States agriculture.

  11. Nitrogen fixation and nifH diversity in human gut microbiota.

    PubMed

    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 (15)N2 incorporation assay showed significant enrichment of (15)N 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

  12. Nitrogen fixation and nifH diversity in human gut microbiota.

    PubMed

    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

    2016-08-24

    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 (15)N2 incorporation assay showed significant enrichment of (15)N 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.

  13. 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

  14. Nitrogen fixation in sediments along a depth transect through the Peruvian oxygen minimum zone

    NASA Astrophysics Data System (ADS)

    Gier, Jessica; Sommer, Stefan; Löscher, Carolin R.; Dale, Andrew W.; Schmitz, Ruth A.; Treude, Tina

    2016-07-01

    The potential coupling of nitrogen (N2) fixation and sulfate reduction (SR) was explored in sediments of the Peruvian oxygen minimum zone (OMZ). Sediment samples were retrieved by a multiple corer at six stations along a depth transect (70-1025 m water depth) at 12° S, covering anoxic and hypoxic bottom water conditions. Benthic N2 fixation, determined by the acetylene reduction assay, was detected at all sites, with highest rates between 70 and 253 m and lower rates at greater depth. SR rates decreased with increasing water depth. N2 fixation and SR overlapped in sediments, suggesting a potential coupling of both processes. However, a weak positive correlation of their activity distribution was detected by principle component analysis. A potential link between N2 fixation and sulfate-reducing bacteria was indicated by the molecular analysis of nifH genes. Detected nifH sequences clustered with the sulfate-reducing bacteria Desulfonema limicola at the 253 m station. However, nifH sequences of other stations clustered with uncultured organisms, Gammaproteobacteria, and Firmicutes (Clostridia) rather than with known sulfate reducers. The principle component analysis revealed that benthic N2 fixation in the Peruvian OMZ is controlled by organic matter (positive) and free sulfide (negative). No correlation was found between N2 fixation and ammonium concentrations (even at levels > 2022 µM). N2 fixation rates in the Peruvian OMZ sediments were in the same range as those measured in other organic-rich sediments.

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

    SciTech Connect

    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 biomass 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.

  16. Soil spatial variability and symbiotic nitrogen fixation by legumes

    SciTech Connect

    Reichardt, K. )

    1990-09-01

    The isotope dilution method for the estimation of N{sub 2} fixation by legumes is analyzed, comparing the application of {sup 15}N-enriched fertilizer with {sup 15}N-labeled soil. Soil variability of other dynamic processes in the soil are discussed in light of the distribution of the {sup 15}N label in the system. Field data were collected along six transects, 45 m long, with 30 plots (replicates) each. The legume (Vicia faba L.) was used as a fixing crop, barley (Hordeum vulgare L.) and oil radish (Raphinus sativus L.) as nonfixing standard crops. Isotope methods were also compared with the yield difference method. Results show that isotope methods were very sensitive to the distribution of the label in the soil and that dynamic processes involving N can significantly affect this distribution over a whole field. Best results were obtained with {sup 15}N-labeled soil. The particular site used, having been farmed for more than 20 years with {sup 15}N trials, showed a homogeneous residual {sup 15}N label that made it possible to estimate N{sub 2} fixation without the application of extra label. Estimates of N{sub 2} fixation with the isotope method were well correlated with the yield difference method when fertilizer use efficiency of the fixing and nonfixing crops were similar. Results also indicate that a good reference crop for one method might not be the best for the other method, and one reason for this is the variability of soil parameters and of dynamic processes occurring in the soil.

  17. Application of plant genomics for improved symbiotic nitrogen fixation in plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Because genome sequencing, transcript profiling, proteome analysis, metabolite profiling, mutant analysis, and comparative genomics have progressed at a logarithmic pace, we know more about the plant genes involved in symbiotic nitrogen fixation (SNF) than could have been imagined a decade ago. Howe...

  18. Nitrogen Fixation by Vibrio parahaemolyticus and Its Implications for a New Ecological Niche▿

    PubMed Central

    Criminger, J. D.; Hazen, T. H.; Sobecky, P. A.; Lovell, C. R.

    2007-01-01

    A Vibrio parahaemolyticus strain isolated from the rhizosphere of the ecosystem dominant estuarine grass, Spartina alterniflora, was characterized and shown to carry nifH, the gene encoding the nitrogenase iron protein, and to fix N2. Nitrogen fixation may contribute substantially to the adaptability, niche breadth, and ecological significance of V. parahaemolyticus. PMID:17675440

  19. Effect of polynuclear hydrocarbons on algal nitrogen fixation (acetylene reduction)

    SciTech Connect

    Bastian, M.V.; Toetz, D.W.

    1985-08-01

    The objective of this research was to determine the effects of polynuclear aromatic hydrocarbons (PAH) on N/sub 2/ fixation by the alga, Anabaena flos-aquea. The reduction of acetylene (C/sub 2/H/sub 2/) to ethylene (C/sub 2/H/sub 4/) was measured as a measure of the capacity of an organism to fix atmospheric N/sub 2/ and reduce it to an assimilable form. The primary advantage of this assay is its speed since chemical exposure and quantitative chromatographic analysis can be completed in a few hours.

  20. Denitrification and Nitrogen Fixation Dynamics in the Area Surrounding an Individual Ghost Shrimp (Neotrypaea californiensis) Burrow System

    PubMed Central

    Bertics, Victoria J.; Sohm, Jill A.; Magnabosco, Cara

    2012-01-01

    Bioturbated sediments are thought of as areas of increased denitrification or fixed-nitrogen (N) loss; however, recent studies have suggested that not all N may be lost from these environments, with some N returning to the system via microbial dinitrogen (N2) fixation. We investigated denitrification and N2 fixation in an intertidal lagoon (Catalina Harbor, CA), an environment characterized by bioturbation by thalassinidean shrimp (Neotrypaea californiensis). Field studies were combined with detailed measurements of denitrification and N2 fixation surrounding a single ghost shrimp burrow system in a narrow aquarium (15 cm by 20 cm by 5 cm). Simultaneous measurements of both activities were performed on samples taken within a 1.5-cm grid for a two-dimensional illustration of their intensity and distribution. These findings were then compared with rate measurements performed on bulk environmental sediment samples collected from the lagoon. Results for the aquarium indicated that both denitrification and N2 fixation have a patchy distribution surrounding the burrow, with no clear correlation to each other, sediment depth, or distance from the burrow. Field denitrification rates were, on average, lower in a bioturbated region than in a seemingly nonbioturbated region; however, replicates showed very high variability. A comparison of denitrification field results with previously reported N2 fixation rates from the same lagoon showed that in the nonbioturbated region, depth-integrated (10 cm) denitrification rates were higher than integrated N2 fixation rates (∼9 to 50 times). In contrast, in the bioturbated sediments, depending on the year and bioturbation intensity, some (∼6.2%) to all of the N lost via denitrification might be accounted for via N2 fixation. PMID:22447588

  1. 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. PMID:24595111

  2. Seasonally dependent iron limitation of nitrogen fixation in tropical forests of karst landscapes

    NASA Astrophysics Data System (ADS)

    Winbourne, J. B.; Brewer, S.; Houlton, B. Z.

    2015-12-01

    Limestone tropical forests in karst topography are one of the most poorly studied ecosystems on Earth, and has been substantially cleared by human activities throughout much of Central America. This ecosystem is noted for its high level of plant productivity, biomass, endemism and biological diversity compared to nearby neighboring tropical forests on volcanic rock substrates (Brewer et al. 2002). A question remains as to how limestone tropical forests are able to maintain the high nutrient demands of plant photosynthesis and tree biomass growth. Here, we demonstrate that rates of nitrogen (N) fixation are higher in limestone versus volcanic soil substrates, with direct evidence for the emergence of seasonally dependent iron limitation of N fixation in limestone tropical forest. N fixation rates showed a three-fold increase in response to iron additions, especially during the wet season when N demands of the forest trees are highest. In contrast, adjacent forests growing on the more classical acidic volcanic soils showed no response to iron or other nutrient additions. Biologically available pools of iron were exceedingly low in the limestone forest site, consistent with the complexation of iron under high pH conditions. Biological acquisition of iron, as measured by the concentration of iron chelating compounds (i.e. siderophores), provided additional evidence for iron limitation of microbial processes in limestone tropical forests, where concentrations were six times higher than those at the volcanic site. Our results suggest that the functioning of limestone tropical forest is strongly regulated by interactions between iron, soil pH, and N cycling.

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

    SciTech Connect

    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 particularly 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.

  4. Regional constraints to biological nitrogen fixation in post-fire forest communities.

    PubMed

    Yelenik, Stephanie; Perakis, Steven; Hibbs, David

    2013-03-01

    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 x ha(-1) x 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.

  5. 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.

  6. The effect of nutrients on carbon and nitrogen fixation by the UCYN-A-haptophyte symbiosis.

    PubMed

    Krupke, Andreas; Mohr, Wiebke; LaRoche, Julie; Fuchs, Bernhard M; Amann, Rudolf I; Kuypers, Marcel M M

    2015-07-01

    Symbiotic relationships between phytoplankton and N2-fixing microorganisms play a crucial role in marine ecosystems. The abundant and widespread unicellular cyanobacteria group A (UCYN-A) has recently been found to live symbiotically with a haptophyte. Here, we investigated the effect of nitrogen (N), phosphorus (P), iron (Fe) and Saharan dust additions on nitrogen (N2) fixation and primary production by the UCYN-A-haptophyte association in the subtropical eastern North Atlantic Ocean using nifH expression analysis and stable isotope incubations combined with single-cell measurements. N2 fixation by UCYN-A was stimulated by the addition of Fe and Saharan dust, although this was not reflected in the nifH expression. CO2 fixation by the haptophyte was stimulated by the addition of ammonium nitrate as well as Fe and Saharan dust. Intriguingly, the single-cell analysis using nanometer scale secondary ion mass spectrometry indicates that the increased CO2 fixation by the haptophyte in treatments without added fixed N is likely an indirect result of the positive effect of Fe and/or P on UCYN-A N2 fixation and the transfer of N2-derived N to the haptophyte. Our results reveal a direct linkage between the marine carbon and nitrogen cycles that is fuelled by the atmospheric deposition of dust. The comparison of single-cell rates suggests a tight coupling of nitrogen and carbon transfer that stays balanced even under changing nutrient regimes. However, it appears that the transfer of carbon from the haptophyte to UCYN-A requires a transfer of nitrogen from UCYN-A. This tight coupling indicates an obligate symbiosis of this globally important diazotrophic association. PMID:25535939

  7. The effect of nutrients on carbon and nitrogen fixation by the UCYN-A–haptophyte symbiosis

    PubMed Central

    Krupke, Andreas; Mohr, Wiebke; LaRoche, Julie; Fuchs, Bernhard M; Amann, Rudolf I; Kuypers, Marcel MM

    2015-01-01

    Symbiotic relationships between phytoplankton and N2-fixing microorganisms play a crucial role in marine ecosystems. The abundant and widespread unicellular cyanobacteria group A (UCYN-A) has recently been found to live symbiotically with a haptophyte. Here, we investigated the effect of nitrogen (N), phosphorus (P), iron (Fe) and Saharan dust additions on nitrogen (N2) fixation and primary production by the UCYN-A–haptophyte association in the subtropical eastern North Atlantic Ocean using nifH expression analysis and stable isotope incubations combined with single-cell measurements. N2 fixation by UCYN-A was stimulated by the addition of Fe and Saharan dust, although this was not reflected in the nifH expression. CO2 fixation by the haptophyte was stimulated by the addition of ammonium nitrate as well as Fe and Saharan dust. Intriguingly, the single-cell analysis using nanometer scale secondary ion mass spectrometry indicates that the increased CO2 fixation by the haptophyte in treatments without added fixed N is likely an indirect result of the positive effect of Fe and/or P on UCYN-A N2 fixation and the transfer of N2-derived N to the haptophyte. Our results reveal a direct linkage between the marine carbon and nitrogen cycles that is fuelled by the atmospheric deposition of dust. The comparison of single-cell rates suggests a tight coupling of nitrogen and carbon transfer that stays balanced even under changing nutrient regimes. However, it appears that the transfer of carbon from the haptophyte to UCYN-A requires a transfer of nitrogen from UCYN-A. This tight coupling indicates an obligate symbiosis of this globally important diazotrophic association. PMID:25535939

  8. The effect of nutrients on carbon and nitrogen fixation by the UCYN-A-haptophyte symbiosis.

    PubMed

    Krupke, Andreas; Mohr, Wiebke; LaRoche, Julie; Fuchs, Bernhard M; Amann, Rudolf I; Kuypers, Marcel M M

    2015-07-01

    Symbiotic relationships between phytoplankton and N2-fixing microorganisms play a crucial role in marine ecosystems. The abundant and widespread unicellular cyanobacteria group A (UCYN-A) has recently been found to live symbiotically with a haptophyte. Here, we investigated the effect of nitrogen (N), phosphorus (P), iron (Fe) and Saharan dust additions on nitrogen (N2) fixation and primary production by the UCYN-A-haptophyte association in the subtropical eastern North Atlantic Ocean using nifH expression analysis and stable isotope incubations combined with single-cell measurements. N2 fixation by UCYN-A was stimulated by the addition of Fe and Saharan dust, although this was not reflected in the nifH expression. CO2 fixation by the haptophyte was stimulated by the addition of ammonium nitrate as well as Fe and Saharan dust. Intriguingly, the single-cell analysis using nanometer scale secondary ion mass spectrometry indicates that the increased CO2 fixation by the haptophyte in treatments without added fixed N is likely an indirect result of the positive effect of Fe and/or P on UCYN-A N2 fixation and the transfer of N2-derived N to the haptophyte. Our results reveal a direct linkage between the marine carbon and nitrogen cycles that is fuelled by the atmospheric deposition of dust. The comparison of single-cell rates suggests a tight coupling of nitrogen and carbon transfer that stays balanced even under changing nutrient regimes. However, it appears that the transfer of carbon from the haptophyte to UCYN-A requires a transfer of nitrogen from UCYN-A. This tight coupling indicates an obligate symbiosis of this globally important diazotrophic association.

  9. 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. PMID:22283408

  10. 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.

  11. Approaches for enhancement of N₂ fixation efficiency of chickpea (Cicer arietinum L.) under limiting nitrogen conditions.

    PubMed

    Esfahani, Maryam Nasr; Sulieman, Saad; Schulze, Joachim; Yamaguchi-Shinozaki, Kazuko; Shinozaki, Kazuo; Tran, Lam-Son

    2014-04-01

    Chickpea (Cicer arietinum) is an important pulse crop in many countries in the world. The symbioses between chickpea and Mesorhizobia, which fix N₂ inside the root nodules, are of particular importance for chickpea's productivity. With the aim of enhancing symbiotic efficiency in chickpea, we compared the symbiotic efficiency of C-15, Ch-191 and CP-36 strains of Mesorhizobium ciceri in association with the local elite chickpea cultivar 'Bivanij' as well as studied the mechanism underlying the improvement of N₂ fixation efficiency. Our data revealed that C-15 strain manifested the most efficient N₂ fixation in comparison with Ch-191 or CP-36. This finding was supported by higher plant productivity and expression levels of the nifHDK genes in C-15 nodules. Nodule specific activity was significantly higher in C-15 combination, partially as a result of higher electron allocation to N₂ versus H⁺. Interestingly, a striking difference in nodule carbon and nitrogen composition was observed. Sucrose cleavage enzymes displayed comparatively lower activity in nodules established by either Ch-191 or CP-36. Organic acid formation, particularly that of malate, was remarkably higher in nodules induced by C-15 strain. As a result, the best symbiotic efficiency observed with C-15-induced nodules was reflected in a higher concentration of the total and several major amino metabolites, namely asparagine, glutamine, glutamate and aspartate. Collectively, our findings demonstrated that the improved efficiency in chickpea symbiotic system, established with C-15, was associated with the enhanced capacity of organic acid formation and the activities of the key enzymes connected to the nodule carbon and nitrogen metabolism.

  12. Nitrogen fixation in biological soil crusts from southeast Utah, USA

    USGS Publications Warehouse

    Belnap, J.

    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 (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 expected to be highly variable, as

  13. 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

  14. The role of nitrogen fixation in cyanobacterial bloom toxicity in a temperate, eutrophic lake.

    PubMed

    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 N(2) 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, N(2) 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 N(2) fixation rates were observed. Then, following large early summer N(2) 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 N(2) fixation rates and Aphanizomenon abundance increased before the

  15. 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

    2014-01-01

    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. PMID:24336216

  16. 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

    2014-01-01

    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.

  17. 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.

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

    PubMed Central

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

    2016-01-01

    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. PMID:27476439

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

    PubMed

    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.

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

    PubMed

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

    2016-01-01

    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. PMID:27476439

  1. Nitrogen fixation in sediments along a depth transect through the Peruvian oxygen minimum zone

    NASA Astrophysics Data System (ADS)

    Gier, J.; Sommer, S.; Löscher, C. R.; Dale, A. W.; Schmitz, R. A.; Treude, T.

    2015-09-01

    Benthic nitrogen (N2) fixation and sulfate reduction (SR) were investigated in the Peruvian oxygen minimum zone (OMZ). Sediment samples, retrieved by a multiple corer were taken at six stations (70-1025 m) along a depth transect at 12° S, covering anoxic and hypoxic bottom water conditions. Benthic N2 fixation was detected at all sites, with high rates measured in OMZ mid-waters between the 70 and 253 m and lowest N2 fixation rates below 253 m down to 1025 m water depth. SR rates were decreasing with increasing water depth, with highest rates at the shallow site. Benthic N2 fixation depth profiles largely overlapped with SR depth profiles, suggesting that both processes are coupled. The potential of N2 fixation by SR bacteria was verified by the molecular analysis of nifH genes. Detected nifH sequences clustered with SR bacteria that have been demonstrated to fix N2 in other benthic environments. Depth-integrated rates of N2 fixation and SR showed no direct correlation along the 12° S transect, suggesting that the benthic diazotrophs in the Peruvian OMZ are being controlled by additional various environmental factors. The organic matter availability and the presence of sulfide appear to be major drivers for benthic diazotrophy. It was further found that N2 fixation was not inhibited by high ammonium concentrations. N2 fixation rates in OMZ sediments were similar to rates measured in other organic-rich sediments. Overall, this work improves our knowledge on N sources in marine sediments and contributes to a better understanding of N cycling in OMZ sediments.

  2. Nitrogen fixation by immobilized NIF derepressed Klebsiella pneumoniae cells

    SciTech Connect

    Venkatasubramanian, K.; Toda, Y.

    1980-01-01

    In vitro production of ammonia through biological means poses a number of challenges. The organisms should be able to accumulate considerable concentrations of ammonia in the medium. Secondly, nonphotosynthetic organisms must be supplied with high-energy substrates to carry out the fixation reaction. Thirdly, the organisms must be kept in a viable state to produce ammonia over long periods of time. In this article, preliminary results on the production of ammonia by a mutant strain of Klebsiella pneumoniae in continuous reactor systems are discussed. Continuous production of ammonia becomes feasible through the immobilization of the whole microbial cells and then through the use of the resulting catalyst system in a flow-through reactor. The rationale for immobilizing microbial cells and the advantages of such an approach over traditional fermentation processes are briefly described as they relate to the microbial production of ammonia. The microbial cells can be immobilized in such a way that their viability is still maintained in the immobilized state. This, in turn, obviates addition of cofactors, which is often an expensive step associated with immobilized multi-enzyme systems. Reconstituted bovine-hide collagen as the carrier matrix for fixing the cells was the carrier of choice for our work on immobilized Klebsiella cells. Polyacrylamide gels were examined as an alternate carrier matrix but results from this were found to be inferior to those collagen immobilized cell system.

  3. 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

  4. Effect of Arsenic on Nodulation and Nitrogen Fixation of Blackgram (Vigna mungo).

    PubMed

    Mandal, Santi M; Gouri, Samiran S; De, Debasis; Das, Bidus K; Mondal, Keshab C; Pati, Bikas R

    2011-01-01

    Rhizobium-legume symbiotic interaction is an efficient model system for soil remediation and reclamation. We earlier isolated an arsenic (As) (2.8 mM arsenate) tolerant and symbiotically effective Rhizobium strain, VMA301 from Vigna mungo and in this study we further characterized its efficacy for arsenic removal from the soil and its nitrogen fixation capacity. Although nodule formation is delayed in plants with As-treated composite when the inoculum was prepared without arsenic in culture medium, whereas it attains the significant number of nodules compare to plant grown in As-free soil when the inoculum was prepared with arsenic supplemented medium. Arsenic accumulation was higher in roots than root nodules. Nitrogenase activity is reduced to almost 2 fold in plants with As-treated soil but not abolished. These results suggest that this strain, VMA301, has been able to establish an effective symbiotic interaction in V. mungo in As-contaminated soil and can perform dual role of arsenic bioremediation as well as soil nitrogen improvement.

  5. Biological invasion by Myrica faya in Hawaii: Plant demography, nitrogen fixation, ecosystem effects

    SciTech Connect

    Vitousek, P.M.; Walker, L.R. )

    1989-09-01

    Myrica faya, an introduced actinorhizal nitrogen fixer, in invading young volcanic sites in Hawaii Volcanoes National Park. We examined the population biology of the invader and ecosystem-level consequences of its invasion in open-canopied forests resulting from volcanic cinder-fall. Although Myrica faya is nominally dioecious, both males and females produce large amounts of fruit that are utilized by a number of exotic and native birds, particularly the exotic Zosterops japonica. In areas of active colonization, Myrica seed rain under perch trees of the dominant native Metrosideros polymorpha ranged from 6 to 60 seeds m{sup {minus}2} yr{sup {minus}1}; no seeds were captured in the open. Planted seeds of Myrica also germinated an established better under isolated individuals of Metrosideros than in the open. Diameter growth of Myrica is > 15-fold greater than that of Metrosideros, and the Myrica population is increasing rapidly. Rates of nitrogen fixation were measured using the acetylene reduction assay calibrated with {sup 15}N. Myrica nodules reduced acetylene at between 5 and 20 {mu}mol g{sup {minus}1} h{sup {minus}1}, a rate that extrapolated to nitrogen fixation of 18 kg ha{sup {minus}1} in a densely colonized site. By comparison, all native sources of nitrogen fixation summed to 0.2 kg ha{sup {minus}1} yr{sup {minus}1}, and precipitation added < 4 kg ha{sup {minus}1} yr{sup {minus}1}. Measurements of litter decomposition and nitrogen release, soil nitrogen mineralization, and plant growth in bioassays all demonstrated that nitrogen fixed by Myrica becomes available to other organisms as well. We conclude that biological invasion by Myrica faya alters ecosystem-level properties in this young volcanic area; at least in this case, the demography and physiology of one species controls characteristics of a whole ecosystem.

  6. Nitrogen fixation in sediments along a depth transect through the Eastern Boundary Upwelling Systems off Peru and Mauritania

    NASA Astrophysics Data System (ADS)

    Gier, J.; Sommer, S.; Löscher, C. R.; Dale, A.; Schmitz, R. A.; Treude, T.

    2015-12-01

    The distribution of benthic nitrogen (N2) fixation and its relevance for N cycling in the Eastern Boundary Upwelling Systems (EBUS) are still unknown. Recent studies confirm that benthic N2 fixation can be coupled to sulfate reduction (SR) and that several species of sulfate reducing bacteria have the genetic ability to fix N due to the presence of the gene encoding for the nitrogenase enzyme. We investigated benthic N2 fixation and SR in the Peruvian oxygen minimum zone at 12°S and in the Mauritanian upwelling system at 18°N along a depth transect. Sediments were retrieved by a multicorer and a benthic lander at six stations in both regions. Benthic N2 fixation occurred throughout the sediment in both EBUS. Off Peru the highest integrated (0-20 cm) N2 fixation rate of 0.4 mmol N/m2/d was measured inside the core of the OMZ at 253 m water depth. Off Mauritania the highest integrated (0-20 cm) N2 fixation rate of 0.15 mmol N/m2/d was measured at 90 m, coinciding with a low bottom water oxygen concentration (30 μM). N2 fixation depth profiles often overlapped with SR activity. Moreover, sequencing data yielded insights into the composition and diversity of the nifH gene pool in EBUS sediments. Interestingly, detected sequences in both EBUS clustered with SR bacteria, such as Desulfovibrio vulgaris and several of the novel detected clades belonged to uncultured diazotrophs. Our results suggest that N2 fixation and SR were coupled to a large extent in both regions. However, potential environmental factors controlling benthic diazotrophs in the EBUS appear to be the availability of sulfide and organic matter. Additionally, no inhibition of N2 fixation at high ammonium concentrations was found, which highlights gaps in our knowledge regards the interaction between ammonium availability and diazotrophy. Our results contribute to a better understanding of N cycling in EBUS sediments and sources of fixed N.

  7. Environmental Forcing of Nitrogen Fixation in the Eastern Tropical and Sub-Tropical North Atlantic Ocean

    PubMed Central

    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 (N2) 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 106 L−1 nifH gene copies, unicellular group A cyanobacteria with up to 105 L−1 nifH gene copies and gamma A proteobacteria with up to 104 L−1 nifH gene copies. N2 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). CO2-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, N2 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 N2 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 N2 fixation rates. However, the balance between a decreased supply of DIP and DFe from deep waters as a result of more pronounced stratification and an enhanced supply of these nutrients with a predicted increase in deposition of Saharan dust may ultimately determine the consequences of climate

  8. 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

  9. Increasing Nitrogen Fixation and Seed Development in Soybean Requires Complex Adjustments of Nodule Nitrogen Metabolism and Partitioning Processes.

    PubMed

    Carter, Amanda M; Tegeder, Mechthild

    2016-08-01

    Legumes are able to access atmospheric di-nitrogen (N2) through a symbiotic relationship with rhizobia that reside within root nodules. In soybean, following N2 fixation by the bacteroids, ammonia is finally reduced in uninfected cells to allantoin and allantoic acid [1]. These ureides present the primary long-distance transport forms of nitrogen (N), and are exported from nodules via the xylem for shoot N supply. Transport of allantoin and allantoic acid out of nodules requires the function of ureide permeases (UPS1) located in cells adjacent to the vasculature [2, 3]. We expressed a common bean UPS1 transporter in cortex and endodermis cells of soybean nodules and found that delivery of N from nodules to shoot, as well as seed set, was significantly increased. In addition, the number of transgenic nodules was increased and symbiotic N2 fixation per nodule was elevated, indicating that transporter function in nodule N export is a limiting step in bacterial N acquisition. Further, the transgenic nodules showed considerable increases in nodule N assimilation, ureide synthesis, and metabolite levels. This suggests complex adjustments of nodule N metabolism and partitioning processes in support of symbiotic N2 fixation. We propose that the transgenic UPS1 plants display metabolic and allocation plasticity to overcome N2 fixation and seed yield limitations. Overall, it is demonstrated that transporter function in N export from nodules is a key step for enhancing atmospheric N2 fixation and nodule function and for improving shoot N nutrition and seed development in legumes. PMID:27451897

  10. [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

  11. [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

  12. Growth, nitrogen fixation, yield and kernel quality of peanut in response to lime, organic and inorganic fertilizer levels.

    PubMed

    Basu, Manisha; Bhadoria, P B S; Mahapatra, S C

    2008-07-01

    The aim of this investigation was to study the effect of different levels of chemical fertilizers alone and in combination with farmyard manure and lime on growth, nitrogen fixation, yield and kernel quality of peanut in an acid lateritic soil. Five fertilization levels viz., no chemical fertilizer (CF) (F0), CF @ 20:40:30 (F1), CF @ 40:80:60 (F2) kg ha(-1) NPK, F1 +2.5 t ha(-1) FYM (F3) and F2 +5 t ha(-1) FYM (F4) with and without liming (2 t ha(-1)) were tested. Results revealed that integrated application of FYM+CF at F3 level significantly (P0.05) improved the nitrogen content of nodules (12.4%), kernel yield (19.3%), mineral composition, oil content (4.8%), protein content (28.2%) and hydration coefficient (11.6%) of kernels over sole CF at F1 level. Maximum level of CF or FYM+CF though improved the population of symbiotic nitrogen fixing bacteria in the peanut rhizosphere, however, could not improve nitrogen fixation, yield and kernel quality. PMID:17993273

  13. Nitrogen fixation by photosynthetic bacteria in lowland rice culture.

    PubMed

    Habte, M; Alexander, M

    1980-02-01

    Propanil (3',4'-dichloropropionanilide) was a potent inhibitor of the nitrogenase activity of blue-green algae (cyanobacteria) in flooded soil, but the herbicide at comparable concentrations was not toxic to rice, protozoa, and nitrogen-fixing bacteria. Ethanol-amended flooded soils treated with propanil exhibited higher rates of nitrogenase activity than those not treated with the herbicide. The enhanced nitrogenase activity in propanil-treated soils was associated with a rise in the population of purple sulfur bacteria, especially of cells resembling Chromatium and Thiospirillum. By employing propanil and a means of excluding light from the floodwater to prevent the development of phototrophs during rice growth under lowland conditions, the relative activities of blue-green algae, photosynthetic bacteria, and the rhizosphere microflora were determined. The results suggest that the potential contribution of photosynthetic bacteria may be quite high. PMID:16345507

  14. Nitrogen Fixation by Photosynthetic Bacteria in Lowland Rice Culture

    PubMed Central

    Habte, M.; Alexander, M.

    1980-01-01

    Propanil (3′,4′-dichloropropionanilide) was a potent inhibitor of the nitrogenase activity of blue-green algae (cyanobacteria) in flooded soil, but the herbicide at comparable concentrations was not toxic to rice, protozoa, and nitrogen-fixing bacteria. Ethanol-amended flooded soils treated with propanil exhibited higher rates of nitrogenase activity than those not treated with the herbicide. The enhanced nitrogenase activity in propanil-treated soils was associated with a rise in the population of purple sulfur bacteria, especially of cells resembling Chromatium and Thiospirillum. By employing propanil and a means of excluding light from the floodwater to prevent the development of phototrophs during rice growth under lowland conditions, the relative activities of blue-green algae, photosynthetic bacteria, and the rhizosphere microflora were determined. The results suggest that the potential contribution of photosynthetic bacteria may be quite high. PMID:16345507

  15. [Effects of low molecular organic acids on nitrogen accumulation, nodulation, and nitrogen fixation of soybean (Glycine max L.) under phosphorus deficiency stress].

    PubMed

    Wang, Shu-Qi; Han, Xiao-Zeng; Qiao, Yun-Fa; Yan, Jun; Li, Xiao-Hui

    2009-05-01

    A greenhouse sand culture experiment was conducted to study the effects of citric acid, oxalic acid, malic acid, and their mixture on the nitrogen accumulation, nodulation, and nitrogen fixation of soybean. After the application of test low molecular weight organic acids, the nitrogen accumulation in the aboveground part of soybean decreased by 17.6%-44.9% at seedling stage, 29.8%-88.4% at flowering stage, 9.18%-69.6% at podding stage, and 2.21%-41.7% at maturing stage). In the meanwhile, the nodule number, nitrogenase activity, and leghemoglobin content decreased by 11.4%-59.6%, 80.5%-91.7%, and 11.9%-59.9%, respectively, resulting in a significant decrease (9.71%-64.5%) of nitrogen fixation of soybean, compared with the control. The inhibitory effect of test low molecular weight organic acids increased with their increasing concentration. Oxalic acid had a higher inhibitory effect than citric acid and malic acid, and the mixture of the three organic acids had an enhanced inhibitory effect.

  16. Correlation between nitrogen fixation rate and alginate productivity of an indigenous Azotobacter vinelandii from Iran

    PubMed Central

    Nosrati, R; Owlia, P; Saderi, H; Olamaee, M; Rasooli, I; Akhavian, Tehrani A

    2012-01-01

    Background and Objectives Azotobacter vinelandii, a gamma-proteobacterium, is an obligate aerobic free-living gram-negative soil bacterium capable of fixing nitrogen. Oxygen transfer rate into the cell is reduced by the increase of alginate concentrations during the course of A. vinelandii cultivation. This phenomenon provides a low intracellular oxygen concentration needed for nitrogenase activity. The aim of this study was to design a simple strategy to explain the alginate production, cell growth and nitrogenase activity correlation in A. vinelandii under aerobic conditions. Material and Methods Thirty-five different soil samples were taken from the rhizosphere of agricultural crops of Iran. Enrichment and isolation strategies were employed for microbial isolation. Physiological and biochemical characteristics were determined. Molecular identification was performed using selective nifH-g1 primers. Alginate production and nitrogenase activity assay by each isolate of Azotobacter were carried out. Bacterial growth, alginate production and Nitrogenase activity were conducted by time-coursed quantitative measurements. Results Total of 26 isolates were selected after enrichment, isolation, and screening. The isolate was identified by molecular tests as A. vinelandii. The highest alginate productions of 1.02 g/l and 0.91g/l were noted after 4 days in 8 isolates, cell biomass of which were estimated 4.88-5.26 g/l. Six of 8 isolates were able to fix atmospheric N2 on nitrogen-free medium. Rates obtained in isolates were in the range of 12.1 to 326.4 nmol C2H4 h-1 vial-1. Conclusions Nitrogen fixation and alginate production yielded significant and positive Pearson's correlation coefficient of R2 = 0.760, p ∼ 0.02. Finally association between bacterial growth, alginate production and nitrogenase activity almost noticeable yielded significant and positive Pearson's correlation coefficient R2= 0.723, p ∼ 0.04. PMID:23066492

  17. Bedrock nitrogen inputs support litter nitrogen fixation and temperate forest ecosystem fertility

    NASA Astrophysics Data System (ADS)

    Dynarski, K. A.; Mitchell, S. A.; Morford, S.; Houlton, B. Z.

    2015-12-01

    Nitrogen (N) is one of the most frequently limiting nutrients to terrestrial ecosystem productivity worldwide. As atmospheric carbon dioxide concentrations continue to rise, progressive N limitation is expected to constrain the ability of terrestrial ecosystems to store additional C, making an understanding of N inputs to terrestrial ecosystems increasingly important. In temperate forests, rock reservoirs and biological N fixation (BNF) represent two significant, but poorly characterized, inputs of bioavailable N. Recent research has demonstrated that bedrock can provide a substantial amount of ecosystem-available N in moderate-to-high relief areas with N-rich sedimentary bedrock. In these same ecosystems, asymbiotic BNF performed by heterotrophic microbes in plant litter can provide an additional N input of up to ~2 kg N ha-1 yr-1. Here, we tested the hypothesis that rock N inputs support increased litter BNF via enhanced ecosystem N fertility. We measured rates of BNF along with rock, soil, foliage, and litter chemistry across sites varying substantially in rock N concentrations (from 32 to 800 ppm N). The sites are dominated by Douglas fir and share similar climates and landscape positions (eroding slopes), yet display marked increases in foliar and soil N content as a function of rock N concentrations (foliar: R2=0.18, p<0.001, soil: R2=0.50, p=0.001). We found a significant positive correlation between rock N content and litter BNF rates (R2=0.11, p=0.0035), with rates of BNF at sites with greater than 400 ppm N in bedrock more than double rates of BNF at sites with lower than 400 ppm N in bedrock (p<0.001). These patterns could not be explained by increases in other rock-derived nutrients such as phosphorus or molybdenum, as neither of these known BNF controls increased over the rock N gradient. We found declining foliar lignin:N ratios with increased rock N, suggesting that rock N inputs can increase litter quality, supporting greater microbial activity

  18. Large-scale impact of the island mass effect through nitrogen fixation in the western South Pacific Ocean

    NASA Astrophysics Data System (ADS)

    Shiozaki, Takuhei; Kodama, Taketoshi; Furuya, Ken

    2014-04-01

    We describe a new mechanism for the island mass effect fueled by nitrogen fixation. The nitrogen fixation activities and δ15N of suspended particles in the surface water in the South Pacific were examined. Active nitrogen fixation and abundant Trichodesmium spp. were observed near islands in the western subtropical region, which was attributable to the material supplied by land runoff. High primary production was extensively centered around the islands and was characterized by low δ15N of suspended particles and a reduction in phosphate concentrations at the surface compared with the subtropical gyre and eastern equatorial upwelling. This suggested that Trichodesmium spp. were advected to areas remote from these islands, and consequently, the elevated primary production fueled by nitrogen fixation extended over a large area around them. Because the proposed island mass effect is triggered by a terrigenous nutrient supply, this ecosystem is potentially vulnerable to human activity on small islands.

  19. On the influence of "non-Redfield" dissolved organic nutrient dynamics on the spatial distribution of N2 fixation and the size of the marine fixed nitrogen inventory

    NASA Astrophysics Data System (ADS)

    Somes, Christopher J.; Oschlies, Andreas

    2015-07-01

    Dissolved organic nitrogen (DON) and phosphorus (DOP) represent the most abundant form of their respective nutrient pool in the surface layer of the oligotrophic oceans and play an important role in nutrient cycling and productivity. Since DOP is generally more labile than DON, it provides additional P that may stimulate growth of nitrogen-fixing diazotrophs that supply fixed nitrogen to balance denitrification in the ocean. In this study, we introduce semirecalcitrant components of DON and DOP as state variables in an existing global ocean-atmosphere-sea ice-biogeochemistry model of intermediate complexity to assess their impact on the spatial distribution of nitrogen fixation and the size of the marine fixed nitrogen inventory. Large-scale surface data sets of global DON and Atlantic Ocean DOP are used to constrain the model. Our simulations suggest that both preferential DOP remineralization and phytoplankton DOP uptake are important "non-Redfield" processes (i.e., deviate from molar N:P = 16) that need to be accounted for to explain the observed patterns of DOP. Additional non-Redfield DOP sensitivity experiments testing dissolved organic matter (DOM) production rate uncertainties that best reproduce the observed spatial patterns of DON and DOP stimulate additional nitrogen fixation that increases the size of the global marine fixed nitrogen inventory by 4.7 ± 1.7% compared to the simulation assuming Redfield DOM stoichiometry that underestimates the observed nitrogen inventory. The extra 8 Tg yr-1 of nitrogen fixation stimulated in the Atlantic Ocean is mainly responsible for this increase due to its large spatial separation from water column denitrification, which buffers any potential nitrogen surplus in the Pacific Ocean. Our study suggests that the marine fixed nitrogen budget is sensitive to non-Redfield DOP dynamics because access to the relatively labile DOP pool expands the ecological niche for nitrogen-fixing diazotrophs.

  20. Dinitrogen fixation and dissolved organic nitrogen fueled primary production and particulate export during the VAHINE mesocosm experiment (New Caledonia lagoon)

    NASA Astrophysics Data System (ADS)

    Berthelot, H.; Moutin, T.; L'Helguen, S.; Leblanc, K.; Hélias, S.; Grosso, O.; Leblond, N.; Charrière, B.; Bonnet, S.

    2015-07-01

    In the oligotrophic ocean characterized by nitrate (NO3-) depletion in surface waters, dinitrogen (N2) fixation and dissolved organic nitrogen (DON) can represent significant nitrogen (N) sources for the ecosystem. In this study, we deployed large in situ mesocosms in New Caledonia in order to investigate (1) the contribution of N2 fixation and DON use to primary production (PP) and particle export and (2) the fate of the freshly produced particulate organic N (PON), i.e., whether it is preferentially accumulated and recycled in the water column or exported out of the system. The mesocosms were fertilized with phosphate (PO43-) in order to prevent phosphorus (P) limitation and promote N2 fixation. The diazotrophic community was dominated by diatom-diazotroph associations (DDAs) during the first part of the experiment for 10 days (P1) followed by the unicellular N2-fixing cyanobacteria UCYN-C for the last 9 days (P2) of the experiment. N2 fixation rates averaged 9.8 ± 4.0 and 27.7 ± 8.6 nmol L-1 d-1 during P1 and P2, respectively. NO3- concentrations (< 0.04 μmol L-1) in the mesocosms were a negligible source of N, indicating that N2 fixation was the main driver of new production throughout the experiment. The contribution of N2 fixation to PP was not significantly different (p > 0.05) during P1 (9.0 ± 3.3 %) and P2 (12.6 ± 6.1 %). However, the e ratio that quantifies the efficiency of a system to export particulate organic carbon (POCexport) compared to PP (e ratio = POCexport/PP) was significantly higher (p < 0.05) during P2 (39.7 ± 24.9 %) than during P1 (23.9 ± 20.2 %), indicating that the production sustained by UCYN-C was more efficient at promoting C export than the production sustained by DDAs. During P1, PON was stable and the total amount of N provided by N2 fixation (0.10 ± 0.02 μmol L-1) was not significantly different (p > 0.05) from the total amount of PON exported (0.10 ± 0.04 μmol L-1), suggesting a rapid and probably direct export of the

  1. Interspecies Transfer and Regulation of Pseudomonas stutzeri A1501 Nitrogen Fixation Island in Escherichia coli.

    PubMed

    Han, Yunlei; Lu, Na; Chen, Qinghua; Zhan, Yuhua; Liu, Wei; Lu, Wei; Zhu, Baoli; Lin, Min; Yang, Zhirong; Yan, Yongliang

    2015-08-01

    Until now, considerable effort has been made to engineer novel nitrogen-fixing organisms through the transfer of nif genes from various diazotrophs to non-nitrogen fixers; however, regulatory coupling of the heterologous nif genes with the regulatory system of the new host is still not well understood. In this work, a 49 kb nitrogen fixation island from P. stutzeri A1501 was transferred into E. coli using a novel and efficient transformation strategy, and a series of recombinant nitrogen-fixing E. coli strains were obtained. We found that the nitrogenase activity of the recombinant E. coli strain EN-01, similar to the parent strain P. stutzeri A1501, was dependent on external ammonia concentration, oxygen tension, and temperature. We further found that there existed a regulatory coupling between the E. coli general nitrogen regulatory system and the heterologous P. stutzeri nif island in the recombinant E. coli strain. We also provided evidence that the E. coli general nitrogen regulator GlnG protein was involved in the activation of the nif-specific regulator NifA via a direct interaction with the NifA promoter. To the best of our knowledge, this work plays a groundbreaking role in increasing understanding of the regulatory coupling of the heterologous nitrogen fixation system with the regulatory system of the recipient host. Furthermore, it will shed light on the structure and functional integrity of the nif island and will be useful for the construction of novel and more robust nitrogen-fixing organisms through biosynthetic engineering.

  2. 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.

  3. Comparative sequence analysis of nitrogen fixation-related genes in six legumes.

    PubMed

    Kim, Dong Hyun; Parupalli, Swathi; Azam, Sarwar; Lee, Suk-Ha; Varshney, Rajeev K

    2013-01-01

    Legumes play an important role as food and forage crops in international agriculture especially in developing countries. Legumes have a unique biological process called nitrogen fixation (NF) by which they convert atmospheric nitrogen to ammonia. Although legume genomes have undergone polyploidization, duplication and divergence, NF-related genes, because of their essential functional role for legumes, might have remained conserved. To understand the relationship of divergence and evolutionary processes in legumes, this study analyzes orthologs and paralogs for selected 20 NF-related genes by using comparative genomic approaches in six legumes i.e., Medicago truncatula (Mt), Cicer arietinum, Lotus japonicus, Cajanus cajan (Cc), Phaseolus vulgaris (Pv), and Glycine max (Gm). Subsequently, sequence distances, numbers of synonymous substitutions per synonymous site (Ks) and non-synonymous substitutions per non-synonymous site (Ka) between orthologs and paralogs were calculated and compared across legumes. These analyses suggest the closest relationship between Gm and Cc and the highest distance between Mt and Pv in six legumes. Ks proportional plots clearly showed ancient genome duplication in all legumes, whole genome duplication event in Gm and also speciation pattern in different legumes. This study also reports some interesting observations e.g., no peak at Ks 0.4 in Gm-Gm, location of two independent genes next to each other in Mt and low Ks values for outparalogs for three genes as compared to other 12 genes. In summary, this study underlines the importance of NF-related genes and provides important insights in genome organization and evolutionary aspects of six legume species analyzed. PMID:23986765

  4. Climbing Nitrogenase: Towards a Mechanism of Enzymatic Nitrogen Fixation

    PubMed Central

    Dean, Dennis R.; Seefeldt, Lance C.

    2009-01-01

    Conspectus “Nitrogen fixation”—the reduction of dinitrogen (N2) to two ammonia (NH3) molecules—by the Mo-dependent nitrogenase is essential for all life. Despite four decades of research, a daunting number of unanswered questions about the mechanism of nitrogenase make it the ‘Everest of enzymes’. This Account describes our efforts to climb one “face” of this mountain by meeting two interdependent challenges central to determining the mechanism of biological N2 reduction. The first challenge is to determine the reaction pathway: the composition and structure of each of the substrate-derived moieties bound to the catalytic FeMocofactor (FeMo-co) of the molybdenum-iron (MoFe) protein of nitrogenase. To overcome this challenge, we need to discriminate between the two classes of potential reaction pathways: 1) a “distal” (D) pathway, in which H atoms add sequentially at a single N or 2) an “alternating” (A) pathway, in which H atoms add alternately to the two N atoms of N2. Secondly, we need to characterize the dynamics of conversion among intermediates within the accepted Lowe-Thorneley kinetic scheme for N2 reduction. That goal requires us to experimentally determine both the number of electrons/protons delivered to the MoFe protein and their “inventory”—a partition into those residing on each of the reaction components and released as H2 or NH3. The principal obstacle to this “climb” has been the inability to generate N2 reduction intermediates for characterization. A combination of genetic, biochemical, and spectroscopic approaches recently overcame this obstacle. These experiments identified one of the four-iron Fe-S faces of the active-site FeMo-cofactor as the specific site of reactivity, indicated that the sidechain of residue α70V controls access to this face, and supported the involvement of the sidechain of residue α195H in proton delivery. We can now freeze-quench trap N2 reduction pathway intermediates and use ENDOR

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

    PubMed

    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 NH(4) (+). The reported fluid chemistry included N(2), NH(4) (+) (5-112 μM), NO(2) (-) (27-48 μM), and NO(3) (-) (17-72 μM). In this study, the correlation between low NH(4) (+) concentrations in dominantly meteoric fluids and higher NH(4) (+) in rock-reacted fluids is used to hypothesize that NH(4) (+) is sourced from NH(4) (+)-bearing biotite. However, biotite samples from the host rocks and ore-body minerals were analyzed by Fourier transform infrared (FTIR) microscopy and none-contained NH(4) (+). However, the nitrogenase-encoding gene nifH was successfully amplified from DNA of the fluid sample with high NH(4) (+), suggesting that subsurface microbes have the capability to fix N(2). If so, unregulated nitrogen fixation may account for the relatively high NH(4) (+) concentrations in the fluids. Additionally, the amoA and nxrB genes for archaeal ammonium monooxygenase and nitrite oxidoreductase, respectively, were amplified from the high NH(4) (+) 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 NH(4) (+) 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

  6. 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

  7. Primary productivity and nitrogen fixation by Trichodesmium spp. in the Arabian Sea

    NASA Astrophysics Data System (ADS)

    Parab, Sushma G.; Matondkar, S. G. P.

    2012-12-01

    Trichodesmium was studied with the purpose of understanding its distribution, organic production and nitrogen fixation in the Arabian Sea. Out of the 143 stations sampled, a total of 93 stations showed the presence of Trichodesmium filaments. Two species of Trichodesmium namely, T. thiebautii and T. erythraeum were found. These were distributed on the basis of the physico-chemical conditions of the Arabian Sea. This was the first time that we managed to detect and record the presence of Trichodesmium thiebautii bloom in the Arabian Sea at depths as deep as 60-70 m. Total counts of Trichodesmium varied between 0 and 400737 filamentsL- 1. T. thiebautii developed in offshore waters during the fall intermonsoon, when the water temperature was around 28 °C and nitrate content was as low as 0.34 μM. After the northeast monsoon, Trichodesmium erythraeum developed in the offshore area and then spread to coastal waters. Both species of Trichodesmium together produced a total of 0.263 TgCyear- 1 and fixed a total of 0.2976 TgNyear- 1 in the Arabian Sea. The study revealed that Trichodesmium was a major contributor to the organic matter productivity of the Arabian Sea during the period from November to April. The seasonality of the blooms of Trichodesmium is discussed with the help of Ocean Color Monitoring (OCM) data and biogeochemical implication of these findings in the Arabian Sea.

  8. Quantifying the effect of fire disturbance on free-living nitrogen fixation in tropical ecosystems

    NASA Astrophysics Data System (ADS)

    De Oliveira Bomfim, B.; Silva, L. C. R.; Marimon-Junior, B. H.; Marimon, B.; Horwath, W. R.; Neves, L.

    2015-12-01

    Tropical forests and savannas are among the most important biomes on Earth, supporting more than half of all plant and animal species on the planet. Despite growing interest in biogeochemical processes that affect tropical forest dynamics, many, including biological nitrogen fixation (BNF), are still poorly understood. Free-living N-fixers are thought to play a key role in tropical ecosystems, alleviating N and P limitation, supporting above and below ground biomass production, as well as carbon storage in plants and soil, but this influence has yet to be quantified. Of particular interest, the spatial distribution and identity of free-living BNF under disturbance regimes that commonly lead to the conversion of forests to savannas is currently unknown. To address this critical gap in knowledge, we measured free-living BNF quantifying rates of N fixation under contrasting fire regimes in the Amazon-Cerrado transition of central Brazil. Samples were collected in 4 ha of floodable forests affected by fire and 1 ha of unburned (seasonally flooded) forest located at the Araguaia State Park, Mato Grosso State, Brazil. Free-living N-fixation rates were measured by both 15N2 (98 atom% 15N) and acethylene reduction assay (ARA). Samples were incubated in the field and left in the dark at room temperature for 12 hours. In the next few weeks we will quantify N fixation rates that will be presented in the upcoming AGU meeting.

  9. 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.

  10. Habitability of the early earth: Clues from the physiology of nitrogen fixation and photosynthesis

    NASA Astrophysics Data System (ADS)

    Towe, Kenneth M.

    1985-12-01

    In the absence of direct evidence concerning the nature of the early Earth environments, it is acceptable under the uniformitarian principle to attempt to define primitive habitats from modern procaryotic physiology. Combining the rock and fossil record with present phylogenetic reconstuctions, application of this paleoecological approach to the evolutionary biochemistry and physiology of nitrogen fixation and photosynthesis leads to several inferences about the nature of Archean environments: 1. To stimulate nitrogenase evolution and avoid its repression, the activity of the NH{4/+} ion was less than 10-3, and probably lower. 2. To be consistent with a moderately protective ozone screen, while not also repressing nitrogenase activity, incursions of abiotic dissolved oxygen at levels in the range 10-1.2-10-3.5 PAL would have been acceptable. 3. To induce the formation and activity of RuBP carboxylase, the pCO2 was less than 100 PAL. 4. To support Photosystem I activity, sulfide concentrations of at least 10-4 M were present in the photic zone. 5. To avoid a too-rapid oxidation of sulfide, the pH was probably between 6 7, where H2S exceeds HS-. Evolutionary ‘pressure’ to stimulate the later development of oxygenic photosynthesis (Photosystem II), would require several subsequent habitat modifications: 1. Lowering the sulfide to < 10-4 M to inhibit Photosystem I. 2. Raising the pH above neutral (HS- > H2S), to mediate more rapid oxidation of HS-. 3. Maintaining either an illumination below 300 400 lux (to avoid photosynthetic O2 self-repression of nitrogen fixation), or an adequate local source of combined nitrogen (aNH{4/+} > 10-4) to repress nitrogen fixation entirely.

  11. Nitrogen Fixation Associated with Rinsed Roots and Rhizomes of the Eelgrass Zostera marina.

    PubMed

    Capone, D G; Budin, J M

    1982-12-01

    Nitrogen fixation was associated with the rinsed roots and rhizomes of the seagrass, Zostera marina L. Nitrogenase activity (acetylene reduction) was greater on rhizomes compared to roots, and on older roots and rhizomes relative to younger tissue. Compared to aerobic assays, anaerobic or microaerobic conditions enhanced the rate of acetylene reduction by rhizomes with attached roots, with the highest activity (100 nanomoles per gram dry weight per hour) occurring at pO(2) = 0.01 atmosphere. Addition of glucose, sucrose, or succinate also increased the rate of acetylene reduction under anaerobic conditions, with glucose providing the most stimulation. In one experiment, comparison of acetylene reduction assays with (15)N(2) incorporation yielded a ratio of about 2.6:1. Seagrass communities are thought to be limited by the availability of nitrogen and, therefore, nitrogenase activity directly associated with their roots and rhizomes suggests the possibility of a N(2)-fixing flora which may subsidize their nutritional demand for nitrogen.

  12. 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

  13. The abiotic fixation of nitrogen on mars and other terrestrial planets: conversion of nitrogen, through NO, into nitrate, nitrite, ammonia, and nitrous oxide.

    NASA Astrophysics Data System (ADS)

    Summers, David; Basa, Ranor; Khare, Bishun; Rodoni, David

    The abiotic fixation of nitrogen is critical to understanding habitability, planetary evolution and the potential origin of life on terrestrial planets such as Mars. A non-biological source of biochemically accessible nitrogen is necessary for the origin and early evolution of life. The Martian surface has become uninhabitable, in part due to loss of atmospheric gases, such as nitrogen, resulting in an incapacity to sustain liquid surface water. Chemical sequestration in the crust is one possible mechanism for such loss. The products of nitrogen fixation also impact the climate and geochemistry of the planet. Shock heating of a non-reducing atmosphere will produce NO. This process has been well studied. We have been experimentally studying the pathways possible from NO to more stable forms in the atmosphere and crust. Our work has observed that there are multiple pathways for the fixation. One pathway observed is consistent with the theoretically predicted route via photochemical formation of HNO. Inter-estingly, this pathway is coupled to the formation of formaldehyde from CO. With liquid water, this pathway leads to nitrate and nitrite. In the presence of just water vapor, HNO appears to mostly dimerize to form N2 O. A second pathway involves the formation of NO2 from CO2 and NO. This pathway becomes more dominant without water, but the reaction of NO2 with any form of water, even just adsorbed water, can lead to nitric acid. Finally, with FeS suspended in liquid water, the direct reduction of NO to ammonia is observed. This last pathway represents the most efficient way to reduced nitrogen, with product yields in excess of 50 % in a single step. In conjunction with the reduction of NO, there is also a catalytic disproportionation at the mineral surface, converting NO to NO2 and N2 O, providing an abiotic source of nitrous oxide. This chemistry has implications for a number epochs in Martian history. For example, chemistry in the presence of water is relevant to

  14. Foraminiferal isotope evidence of reduced nitrogen fixation in the ice age Atlantic Ocean.

    PubMed

    Ren, H; Sigman, D M; Meckler, A N; Plessen, B; Robinson, R S; Rosenthal, Y; Haug, G H

    2009-01-01

    Fixed nitrogen (N) is a limiting nutrient for algae in the low-latitude ocean, and its oceanic inventory may have been higher during ice ages, thus helping to lower atmospheric CO2 during those intervals. In organic matter within planktonic foraminifera shells in Caribbean Sea sediments, we found that the 15N/14N ratio from the last ice age is higher than that from the current interglacial, indicating a higher nitrate 15N/14N ratio in the Caribbean thermocline. This change and other species-specific differences are best explained by less N fixation in the Atlantic during the last ice age. The fixation decrease was most likely a response to a known ice age reduction in ocean N loss, and it would have worked to balance the ocean N budget and to curb ice age-interglacial change in the N inventory.

  15. Effect of simulated acid precipitation on algal fixation of nitrogen and carbon dioxide in forest soils

    SciTech Connect

    Chang, F.H.; Alexander, M.

    1983-01-01

    Three forest soils from areas exposed to acid precipitation were incubated for 21 days in the light to enhance the development of indigenous algae. The rates of nitrogen fixation (acetylene reduction) in the light and dark were significantly less if the soils were treated with simulated rain at pH 3.5 than at pH 5.6. The inhibition increased with increasing amounts of simulated rain at pH 3.5. The fixation of CO/sub 2/ in the light was significantly less in the three soils following their exposure to simulated precipitation at pH 3.5 than to the same solutions at pH 5.6, and the extent of suppression rose with increasing amounts of synthetic rain. It is suggested that algae in terrestrial ecosystems may be especially susceptible to acid precipitation.

  16. 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

  17. Multifaceted Investigation of Metabolites During Nitrogen Fixation in Medicago via High Resolution MALDI-MS Imaging and ESI-MS

    NASA Astrophysics Data System (ADS)

    Gemperline, Erin; Jayaraman, Dhileepkumar; Maeda, Junko; Ané, Jean-Michel; Li, Lingjun

    2015-01-01

    Legumes have developed the unique ability to establish a symbiotic relationship with soil bacteria known as rhizobia. This interaction results in the formation of root nodules in which rhizobia thrive and reduce atmospheric dinitrogen into plant-usable ammonium through biological nitrogen fixation (BNF). Owing to the availability of genetic information for both of the symbiotic partners, the Medicago truncatula- Sinorhizobium meliloti association is an excellent model for examining the BNF process. Although metabolites are important in this symbiotic association, few studies have investigated the array of metabolites that influence this process. Of these studies, most target only a few specific metabolites, the roles of which are either well known or are part of a well-characterized metabolic pathway. Here, we used a multifaceted mass spectrometric (MS) approach to detect and identify the key metabolites that are present during BNF using the Medicago truncatula- Sinorhizobium meliloti association as the model system. High mass accuracy and high resolution matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) Orbitrap instruments were used in this study and provide complementary results for more in-depth characterization of the nitrogen-fixation process. We used well-characterized plant and bacterial mutants to highlight differences between the metabolites that are present in functional versus nonfunctional nodules. Our study highlights the benefits of using a combination of mass spectrometric techniques to detect differences in metabolite composition and the distributions of these metabolites in plant biology.

  18. The value of biodiversity in legume symbiotic nitrogen fixation and nodulation for biofuel and food production.

    PubMed

    Gresshoff, Peter M; Hayashi, Satomi; Biswas, Bandana; Mirzaei, Saeid; Indrasumunar, Arief; Reid, Dugald; Samuel, Sharon; Tollenaere, Alina; van Hameren, Bethany; Hastwell, April; Scott, Paul; Ferguson, Brett J

    2015-01-01

    Much of modern agriculture is based on immense populations of genetically identical or near-identical varieties, called cultivars. However, advancement of knowledge, and thus experimental utility, is found through biodiversity, whether naturally-found or induced by the experimenter. Globally we are confronted by ever-growing food and energy challenges. Here we demonstrate how such biodiversity from the food legume crop soybean (Glycine max L. Merr) and the bioenergy legume tree Pongamia (Millettia) pinnata is a great value. Legume plants are diverse and are represented by over 18,000 species on this planet. Some, such as soybean, pea and medics are used as food and animal feed crops. Others serve as ornamental (e.g., wisteria), timber (e.g., acacia/wattle) or biofuel (e.g., Pongamia pinnata) resources. Most legumes develop root organs (nodules) after microsymbiont induction that serve as their habitat for biological nitrogen fixation. Through this, nitrogen fertiliser demand is reduced by the efficient symbiosis between soil Rhizobium-type bacteria and the appropriate legume partner. Mechanistic research into the genetics, biochemistry and physiology of legumes is thus strategically essential for future global agriculture. Here we demonstrate how molecular plant science analysis of the genetics of an established food crop (soybean) and an emerging biofuel P. pinnata feedstock contributes to their utility by sustainable production aided by symbiotic nitrogen fixation. PMID:25240795

  19. 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

  20. Discovery of symbiotic nitrogen fixation and chemoautotrophy in cold-water corals.

    PubMed

    Middelburg, Jack J; Mueller, Christina E; Veuger, Bart; Larsson, Ann I; Form, Armin; van Oevelen, Dick

    2015-12-08

    Cold-water corals (CWC) are widely distributed around the world forming extensive reefs at par with tropical coral reefs. They are hotspots of biodiversity and organic matter processing in the world's deep oceans. Living in the dark they lack photosynthetic symbionts and are therefore considered to depend entirely on the limited flux of organic resources from the surface ocean. While symbiotic relations in tropical corals are known to be key to their survival in oligotrophic conditions, the full metabolic capacity of CWC has yet to be revealed. Here we report isotope tracer evidence for efficient nitrogen recycling, including nitrogen assimilation, regeneration, nitrification and denitrification. Moreover, we also discovered chemoautotrophy and nitrogen fixation in CWC and transfer of fixed nitrogen and inorganic carbon into bulk coral tissue and tissue compounds (fatty acids and amino acids). This unrecognized yet versatile metabolic machinery of CWC conserves precious limiting resources and provides access to new nitrogen and organic carbon resources that may be essential for CWC to survive in the resource-depleted dark ocean.

  1. Discovery of symbiotic nitrogen fixation and chemoautotrophy in cold-water corals.

    PubMed

    Middelburg, Jack J; Mueller, Christina E; Veuger, Bart; Larsson, Ann I; Form, Armin; van Oevelen, Dick

    2015-01-01

    Cold-water corals (CWC) are widely distributed around the world forming extensive reefs at par with tropical coral reefs. They are hotspots of biodiversity and organic matter processing in the world's deep oceans. Living in the dark they lack photosynthetic symbionts and are therefore considered to depend entirely on the limited flux of organic resources from the surface ocean. While symbiotic relations in tropical corals are known to be key to their survival in oligotrophic conditions, the full metabolic capacity of CWC has yet to be revealed. Here we report isotope tracer evidence for efficient nitrogen recycling, including nitrogen assimilation, regeneration, nitrification and denitrification. Moreover, we also discovered chemoautotrophy and nitrogen fixation in CWC and transfer of fixed nitrogen and inorganic carbon into bulk coral tissue and tissue compounds (fatty acids and amino acids). This unrecognized yet versatile metabolic machinery of CWC conserves precious limiting resources and provides access to new nitrogen and organic carbon resources that may be essential for CWC to survive in the resource-depleted dark ocean. PMID:26644069

  2. Discovery of symbiotic nitrogen fixation and chemoautotrophy in cold-water corals

    PubMed Central

    Middelburg, Jack J.; Mueller, Christina E.; Veuger, Bart; Larsson, Ann I.; Form, Armin; Oevelen, Dick van

    2015-01-01

    Cold-water corals (CWC) are widely distributed around the world forming extensive reefs at par with tropical coral reefs. They are hotspots of biodiversity and organic matter processing in the world’s deep oceans. Living in the dark they lack photosynthetic symbionts and are therefore considered to depend entirely on the limited flux of organic resources from the surface ocean. While symbiotic relations in tropical corals are known to be key to their survival in oligotrophic conditions, the full metabolic capacity of CWC has yet to be revealed. Here we report isotope tracer evidence for efficient nitrogen recycling, including nitrogen assimilation, regeneration, nitrification and denitrification. Moreover, we also discovered chemoautotrophy and nitrogen fixation in CWC and transfer of fixed nitrogen and inorganic carbon into bulk coral tissue and tissue compounds (fatty acids and amino acids). This unrecognized yet versatile metabolic machinery of CWC conserves precious limiting resources and provides access to new nitrogen and organic carbon resources that may be essential for CWC to survive in the resource-depleted dark ocean. PMID:26644069

  3. Simultaneous Measurement of Nitrogen Fixation Estimated by Acetylene-Ethylene Assay and Nitrate Absorption by Soybeans 1

    PubMed Central

    Wych, Robert D.; Rains, D. William

    1978-01-01

    An apparatus was designed for simultaneous measurement of rates of N2 fixation estimated by C2H2-C2H4 assay (N2[C2H2] fixation) and NO3− absorption by roots of intact, nodulated soybeans (Glycine max [L.] Merr.). The principal design features include: (a) a gas-tight mist chamber in which nodulated roots can be exposed simultaneously to C2H2 in the gas phase and to a liquid phase containing NO3− sprayed in a fine mist; and (b) provision for sampling the gas phase for C2H4 determination, and the liquid phase for NO3− determination. We studied NO3− absorption by soybeans as affected by nodulation, NO3− concentration during assay, and previous N nutrition during growth in nutrient solution culture in controlled environment chambers. It was established that 0.5 mm NO3− nearly saturated the NO3− absorption system of both nodulated and unnodulated soybeans when the concentration dependence of NO3− absorption rate was measured just after flowering began. Nitrate absorption rates were measured after development of N stress in unnodulated plants, and during recovery from N stress in nodulated plants. The results suggested that the lower capacity for NO3− absorption of nodulated plants was a consequence of N stress during the period of nodule growth and development. Nitrogen [C2H2] fixation rates were compared in intact plants assayed in the mist chamber and in excised roots assayed in both the mist chamber and in glass jars. Excised roots had a lower N2[C2H2] fixation rate than intact plants. The decline observed during the first hour after shoot removal was more pronounced for glass jar-assayed excised roots than for mist chamber-assayed excised roots. We discuss the advantages of our method for assessing the capability of a nodulated legume to acquire nitrogen through both N2 fixation and absorption and assimilation of NO3−. PMID:16660534

  4. The effect of cutting, mulching and applications of farmyard manure on nitrogen fixation in a red clover/grass sward.

    PubMed

    Hatch, D J; Goodlass, G; Joynes, A; Shepherd, M A

    2007-12-01

    In organic farming, maximising the amount of nitrogen (N) which is fixed and retained within the soil is of paramount importance for the yield of the following crop. The aim of this study was to establish the extent to which increased soil fertility, farmyard manure (FYM) applications and/or mulching, could adversely affect fixation. At two sites, situated in the South West (SW) and North East (NE) of England, N(2) fixation was estimated in 'organically' managed red clover/grass plots, both with and without green manure (i.e. surface mulched) and/or the addition of FYM. The FYM was incorporated into the seedbeds at both sites in autumn 2002 at the rate of 170 kg total Nha(-1), as either well-composted (SW site), or not actively-composted (NE site) manures. The same FYM application rate was repeated as top-dressings to both sites in autumn 2003. The plots were cut three or four times each year over two growing seasons. In the first harvest year (2003), incorporation of FYM had beneficial effects of increasing dry matter and N yields significantly at the first cut, but there were no significant differences in subsequent cuts. The same pattern was found in the second harvest year (2004) after the top dressings of FYM, suggesting that most of the N in both types of FYM was in recalcitrant forms. Over the two growing seasons, mulching did not affect red clover/grass dry matter or N yields, but did reduce the proportion of N(2) fixed, by up to 60 kg Nha(-1) when compared with plots from which the clover/grass herbage was cut and removed. Thus, the gain in N from FYM or green manure tended to be offset by a similar reduction in N(2) fixation. These results demonstrate the close association between the availability of soil N and the feed-back system which operates on N(2) fixation by red clover.

  5. Nitrogen fixation and nitrogenase (nifH) expression in tropical waters of the eastern north atlantic.

    SciTech Connect

    Turk, K. A.; Rees, A. P.; Zehr, J. P.; Pereira, N.; Swift, P.; Shelley, R.; Lohan, M.; Woodward, E. M. S.; Gilbert, J.

    2011-01-01

    Expression of nifH in 28 surface water samples collected during fall 2007 from six stations in the vicinity of the Cape Verde Islands (north-east Atlantic) was examined using reverse transcription-polymerase chain reaction (RT-PCR)-based clone libraries and quantitative RT-PCR (RT-qPCR) analysis of seven diazotrophic phylotypes. Biological nitrogen fixation (BNF) rates and nutrient concentrations were determined for these stations, which were selected based on a range in surface chlorophyll concentrations to target a gradient of primary productivity. BNF rates greater than 6 nmolN I{sup -1} h{sup -1} were measured at two of the near-shore stations where high concentrations of Fe and PO{sub 4}{sup 3-} were also measured. Six hundred and five nifH transcripts were amplified by RT-PCR, of which 76% are described by six operational taxonomic units, including Trichodesmium and the uncultivated UCYN-A, and four non-cyanobacterial diazotrophs that clustered with uncultivated Proteobacteria. Although all five cyanobacterial phylotypes quantified in RT-qPCR assays were detected at different stations in this study, UCYN-A contributed most significantly to the pool of nifH transcripts in both coastal and oligotrophic waters. A comparison of results from RT-PCR clone libraries and RT-qPCR indicated that a {gamma}-proteobacterial phylotype was preferentially amplified in clone libraries, which underscores the need to use caution interpreting clone-library-based nifH studies, especially when considering the importance of uncultivated proteobacterial diazotrophs.

  6. Efficient Inactivation of Symbiotic Nitrogen Fixation Related Genes in Lotus japonicus Using CRISPR-Cas9

    PubMed Central

    Wang, Longxiang; Wang, Longlong; Tan, Qian; Fan, Qiuling; Zhu, Hui; Hong, Zonglie; Zhang, Zhongming; Duanmu, Deqiang

    2016-01-01

    The targeted genome editing technique, CRISPR/Cas9 system, has been widely used to modify genes of interest in a predictable and precise manner. In this study, we describe the CRISPR/Cas9-mediated efficient editing of representative SNF (symbiotic nitrogen fixation) related genes in the model legume Lotus japonicus via Agrobacterium-mediated stable or hairy root transformation. We first predicted nine endogenous U6 genes in Lotus and then demonstrated the efficacy of the LjU6-1 gene promoter in driving expression of single guide RNAs (sgRNAs) by using a split yellow fluorescence protein (YFP) reporter system to restore the fluorescence in Arabidopsis protoplasts. Next, we chose a customized sgRNA targeting SYMRK (symbiosis receptor-like kinase) loci and achieved ~35% mutagenic efficiency in 20 T0 transgenic plants, two of them containing biallelic homozygous mutations with a 2-bp deletion near the PAM region. We further designed two sgRNAs targeting three homologous leghemoglobin loci (LjLb1, LjLb2, LjLb3) for testing the possibility of generating multi-gene knockouts. 20 out of 70 hairy root transgenic plants exhibited white nodules, with at least two LjLbs disrupted in each plant. Compared with the constitutively active CaMV 35S promoter, the nodule-specific LjLb2 promoter was also effective in gene editing in nodules by hairy root transformation. Triple mutant knockout of LjLbs was also obtained by stable transformation using two sgRNAs. Collectively, these studies demonstrate that the CRISPR/Cas9 system should greatly facilitate functional analyses of SNF related genes in Lotus japonicus.

  7. Efficient Inactivation of Symbiotic Nitrogen Fixation Related Genes in Lotus japonicus Using CRISPR-Cas9

    PubMed Central

    Wang, Longxiang; Wang, Longlong; Tan, Qian; Fan, Qiuling; Zhu, Hui; Hong, Zonglie; Zhang, Zhongming; Duanmu, Deqiang

    2016-01-01

    The targeted genome editing technique, CRISPR/Cas9 system, has been widely used to modify genes of interest in a predictable and precise manner. In this study, we describe the CRISPR/Cas9-mediated efficient editing of representative SNF (symbiotic nitrogen fixation) related genes in the model legume Lotus japonicus via Agrobacterium-mediated stable or hairy root transformation. We first predicted nine endogenous U6 genes in Lotus and then demonstrated the efficacy of the LjU6-1 gene promoter in driving expression of single guide RNAs (sgRNAs) by using a split yellow fluorescence protein (YFP) reporter system to restore the fluorescence in Arabidopsis protoplasts. Next, we chose a customized sgRNA targeting SYMRK (symbiosis receptor-like kinase) loci and achieved ~35% mutagenic efficiency in 20 T0 transgenic plants, two of them containing biallelic homozygous mutations with a 2-bp deletion near the PAM region. We further designed two sgRNAs targeting three homologous leghemoglobin loci (LjLb1, LjLb2, LjLb3) for testing the possibility of generating multi-gene knockouts. 20 out of 70 hairy root transgenic plants exhibited white nodules, with at least two LjLbs disrupted in each plant. Compared with the constitutively active CaMV 35S promoter, the nodule-specific LjLb2 promoter was also effective in gene editing in nodules by hairy root transformation. Triple mutant knockout of LjLbs was also obtained by stable transformation using two sgRNAs. Collectively, these studies demonstrate that the CRISPR/Cas9 system should greatly facilitate functional analyses of SNF related genes in Lotus japonicus. PMID:27630657

  8. Impact of the ahas transgene for herbicides resistance on biological nitrogen fixation and yield of soybean.

    PubMed

    Hungria, Mariangela; Nakatani, André Shigueyoshi; Souza, Rosinei Aparecida; Sei, Fernando Bonafé; de Oliveira Chueire, Ligia Maria; Arias, Carlos Arrabal

    2015-02-01

    Studies on the effects of transgenes in soybean [Glycine max (L.) Merr.] and the associated use of specific herbicides on biological nitrogen fixation (BNF) are still few, although it is important to ensure minimal impacts on benefits provided by the root-nodule symbiosis. Cultivance CV127 transgenic soybean is a cultivar containing the ahas gene, which confers resistance to herbicides of the imidazolinone group. The aim of this study was to assess the effects of the ahas transgene and of imidazolinone herbicide on BNF parameters and soybean yield. A large-scale set of field experiments was conducted, for three cropping seasons, at nine sites in Brazil, with a total of 20 trials. The experiment was designed as a completely randomized block with four replicates and the following treatments: (T1) near isogenic transgenic soybean (Cultivance CV127) + herbicide of the imidazolinone group (imazapyr); (T2) near isogenic transgenic soybean + conventional herbicides; and (T3) parental conventional soybean (Conquista) + conventional herbicides; in addition, two commercial cultivars were included, Monsoy 8001 (M-SOY 8001) (T4), and Coodetec 217 (CD 217) (T5). At the R2 growth stage, plants were collected and BNF parameters evaluated. In general, there were no effects on BNF parameters due to the transgenic trait or associated with the specific herbicide. Similarly, at the final harvest, no grain-yield effects were detected related to the ahas gene or to the specific herbicide. However, clear effects on BNF and grain yield were attributed to location and cropping season.

  9. Efficient Inactivation of Symbiotic Nitrogen Fixation Related Genes in Lotus japonicus Using CRISPR-Cas9.

    PubMed

    Wang, Longxiang; Wang, Longlong; Tan, Qian; Fan, Qiuling; Zhu, Hui; Hong, Zonglie; Zhang, Zhongming; Duanmu, Deqiang

    2016-01-01

    The targeted genome editing technique, CRISPR/Cas9 system, has been widely used to modify genes of interest in a predictable and precise manner. In this study, we describe the CRISPR/Cas9-mediated efficient editing of representative SNF (symbiotic nitrogen fixation) related genes in the model legume Lotus japonicus via Agrobacterium-mediated stable or hairy root transformation. We first predicted nine endogenous U6 genes in Lotus and then demonstrated the efficacy of the LjU6-1 gene promoter in driving expression of single guide RNAs (sgRNAs) by using a split yellow fluorescence protein (YFP) reporter system to restore the fluorescence in Arabidopsis protoplasts. Next, we chose a customized sgRNA targeting SYMRK (symbiosis receptor-like kinase) loci and achieved ~35% mutagenic efficiency in 20 T0 transgenic plants, two of them containing biallelic homozygous mutations with a 2-bp deletion near the PAM region. We further designed two sgRNAs targeting three homologous leghemoglobin loci (LjLb1, LjLb2, LjLb3) for testing the possibility of generating multi-gene knockouts. 20 out of 70 hairy root transgenic plants exhibited white nodules, with at least two LjLbs disrupted in each plant. Compared with the constitutively active CaMV 35S promoter, the nodule-specific LjLb2 promoter was also effective in gene editing in nodules by hairy root transformation. Triple mutant knockout of LjLbs was also obtained by stable transformation using two sgRNAs. Collectively, these studies demonstrate that the CRISPR/Cas9 system should greatly facilitate functional analyses of SNF related genes in Lotus japonicus. PMID:27630657

  10. Environmental and genotypic effects on the respiration associated with symbiotic nitrogen fixation in peas.

    PubMed

    Mahon, J D

    1979-05-01

    Estimated values for the respiration associated with symbiotic nitrogen fixation in Pisum sativum L. were independent of irradiance, temperature, plant age, and CO(2) concentration, despite large variation in the total rates of C(2)H(2) reduction and root + nodule respiration. Similar values were also found in Phaseolus vulgaris L., Vicia faba L. and Glycine max (L.) Merr. Among all combinations of four Pisum cultivars with four Rhizobium leguminosarum inoculants only the plant genotype significantly affected the fixation-linked respiration, although both plant and bacterial types significantly influenced the total rate of C(2)H(2) reduction. On the basis of measured rates of H(2) evolution and C(2)H(2) reduction, or total nitrogen gain in the same system, the least respiration per unit of ammonia produced symbiotically was estimated as 4.8 to 6.9 moles CO(2) (mole NH(3))(-1) in Laxton's Progress and the greatest as 9.3 to 13.3 moles CO(2) (mole NH(3))(-1) in an Indian cultivar, as compared to a theoretical minimum respiration requirement of 4.7 moles CO(2) (mole NH(3))(-1) in peas.

  11. 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.

  12. Distribution and evolution of nitrogen fixation genes in the phylum Bacteroidetes.

    PubMed

    Inoue, Jun-ichi; Oshima, Kenshiro; Suda, Wataru; Sakamoto, Mitsuo; Iino, Takao; Noda, Satoko; Hongoh, Yuichi; Hattori, Masahira; Ohkuma, Moriya

    2015-01-01

    Diazotrophs had not previously been identified among bacterial species in the phylum Bacteroidetes until the rapid expansion of bacterial genome sequences, which revealed the presence of nitrogen fixation (nif) genes in this phylum. We herein determined the draft genome sequences of Bacteroides graminisolvens JCM 15093(T) and Geofilum rubicundum JCM 15548(T). In addition to these and previously reported 'Candidatus Azobacteroides pseudotrichonymphae' and Paludibacter propionicigenes, an extensive survey of the genome sequences of diverse Bacteroidetes members revealed the presence of a set of nif genes (nifHDKENB) in strains of Dysgonomonas gadei, Dysgonomonas capnocytophagoides, Saccharicrinis fermentans, and Alkaliflexus imshenetskii. These eight species belonged to and were distributed sporadically within the order Bacteroidales. Acetylene reduction activity was detected in the five species examined, strongly suggesting their diazotrophic nature. Phylogenetic analyses showed monophyletic clustering of the six Nif protein sequences in the eight Bacteroidales species, implying that nitrogen fixation is ancestral to Bacteroidales and has been retained in these species, but lost in many other lineages. The identification of nif genes in Bacteroidales facilitates the prediction of the organismal origins of related sequences directly obtained from various environments.

  13. Distribution and Evolution of Nitrogen Fixation Genes in the Phylum Bacteroidetes

    PubMed Central

    Inoue, Jun-ichi; Oshima, Kenshiro; Suda, Wataru; Sakamoto, Mitsuo; Iino, Takao; Noda, Satoko; Hongoh, Yuichi; Hattori, Masahira; Ohkuma, Moriya

    2015-01-01

    Diazotrophs had not previously been identified among bacterial species in the phylum Bacteroidetes until the rapid expansion of bacterial genome sequences, which revealed the presence of nitrogen fixation (nif) genes in this phylum. We herein determined the draft genome sequences of Bacteroides graminisolvens JCM 15093T and Geofilum rubicundum JCM 15548T. In addition to these and previously reported ‘Candidatus Azobacteroides pseudotrichonymphae’ and Paludibacter propionicigenes, an extensive survey of the genome sequences of diverse Bacteroidetes members revealed the presence of a set of nif genes (nifHDKENB) in strains of Dysgonomonas gadei, Dysgonomonas capnocytophagoides, Saccharicrinis fermentans, and Alkaliflexus imshenetskii. These eight species belonged to and were distributed sporadically within the order Bacteroidales. Acetylene reduction activity was detected in the five species examined, strongly suggesting their diazotrophic nature. Phylogenetic analyses showed monophyletic clustering of the six Nif protein sequences in the eight Bacteroidales species, implying that nitrogen fixation is ancestral to Bacteroidales and has been retained in these species, but lost in many other lineages. The identification of nif genes in Bacteroidales facilitates the prediction of the organismal origins of related sequences directly obtained from various environments. PMID:25736980

  14. Environmental and Genotypic Effects on the Respiration Associated with Symbiotic Nitrogen Fixation in Peas 1

    PubMed Central

    Mahon, John D.

    1979-01-01

    Estimated values for the respiration associated with symbiotic nitrogen fixation in Pisum sativum L. were independent of irradiance, temperature, plant age, and CO2 concentration, despite large variation in the total rates of C2H2 reduction and root + nodule respiration. Similar values were also found in Phaseolus vulgaris L., Vicia faba L. and Glycine max (L.) Merr. Among all combinations of four Pisum cultivars with four Rhizobium leguminosarum inoculants only the plant genotype significantly affected the fixation-linked respiration, although both plant and bacterial types significantly influenced the total rate of C2H2 reduction. On the basis of measured rates of H2 evolution and C2H2 reduction, or total nitrogen gain in the same system, the least respiration per unit of ammonia produced symbiotically was estimated as 4.8 to 6.9 moles CO2 (mole NH3)−1 in Laxton's Progress and the greatest as 9.3 to 13.3 moles CO2 (mole NH3)−1 in an Indian cultivar, as compared to a theoretical minimum respiration requirement of 4.7 moles CO2 (mole NH3)−1 in peas. PMID:16660833

  15. 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. PMID:24972068

  16. Aerosol Deposition of Molybdenum: A Control on Nitrogen-Fixation and Tropical Forest Function

    NASA Astrophysics Data System (ADS)

    Wong, M.; Howarth, R. W.; Marino, R. M.; Mahowald, N. M.; Williams, E. R.

    2015-12-01

    Nitrogen fixation, the primary source of new nitrogen (N) to tropical forests, is exclusively catalyzed by the nitrogenase enzyme, which almost always requires molybdenum (Mo). Increasing evidence in recent years suggests that Mo availability may be low in highly weathered soils and can constrain N-fixation rates. Mo is generally either present in a highly soluble form (MoO42-) that is susceptible to leaching or tightly bound in minerals unavailable for biological uptake. To address how Mo is retained in highly weathered tropical systems to support N-fixation, atmospheric transport through dust and sea-salt aerosol spray were examined. Using a global atmospheric transport model computed from modeled meteorological fields, extrapolated dust and sea-salt aerosol Mo sources were used to calculate global distribution of Mo deposition. Dust deposition occurs across the entirety of some tropical forests, particularly the world's largest tropical forest in the Amazon Basin. The model indicates that the Amazon Basin receives substantial inputs of dust, especially the entire northern Amazon Basin, while the southern half receives less. Most of the dust reaching the Amazon originates from the Sahara Desert, and about half of this dust originates from one part of the Sahara, the Bodélé Depression. Mo in dust from the Bodélé Depression was measured with an average concentration of 1.14 ± 0.05 μg/g, similar to the crustal abundance. The model predicts Mo inputs from sea-salt aerosols in coastal regions up to 0.002 mg m-2yr-1. Significant sea-salt deposition occurs up to 300 km inland. Mo from fossil fuel combustion and biomass burning were also evaluated to determine the potential influence of anthropogenic emissions on releasing Mo into the environment.

  17. Asparagine: an amide of particular distinction in the regulation of symbiotic nitrogen fixation of legumes.

    PubMed

    Sulieman, Saad; Tran, Lam-Son Phan

    2013-09-01

    Symbiotic nitrogen fixation is tightly regulated by a range of fine processes at the nodule level, over which the host plant has overall control through the whole life of the plant. The operation of this control at the nodule level is not yet fully understood, but greater knowledge will ultimately lead to a better improvement of N2 fixation through the use of crop legumes and genetic engineering of crop plants for higher performance. It has been suggested that, nodule responses to the nutritional complexity of the rhizosphere environment involve a great deal of coordination of sensing and signal transduction. This regulation can be achieved through several mechanisms, including changes in carbon metabolism, oxygen supply and/or overproduction of reactive oxygen and nitrogen species. Recently, the cycling of amino acids observed between the plant and bacteroid fractions suggests a new and important regulatory mechanism involved in nodule responses. Most of the recent transcriptional findings are consistent with the earlier biochemical and physiological reports. Current research revealed unique advances for nodule metabolism, especially on the regulation of asparagine synthetase gene expression and the control of asparagine (ASN) to N2 fixing activity. A large amount of ASN is found accumulating in the root nodules of the symbiotic plants under restricted environments, such as drought, salinity and nutrient deficiency. Exceptionally, ASN phloem feeding has resulted in an increased concentration of the ASN amide in nodules followed by a remarkable decrease in nodule activity. In this review, recent progress concerning the possible role of ASN in whole-plant-based down-regulation of symbiotic N2 fixation will be reviewed.

  18. Nitrite fixation by humic substances: Nitrogen-15 nuclear magnetic resonance evidence for potential intermediates in chemodenitrification

    USGS Publications Warehouse

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

    2000-01-01

    Studies have suggested that NO2/-, produced during nitrification and denitrification, can become incorporated into soil organic matter and, in one of the processes associated with chemodenitrification, react with organic matter to form trace N gases, including N2O. To gain an understanding of the nitrosation chemistry on a molecular level, soil and aquatic humic substances were reacted with 15N-labeled NaNO2, and analyzed by liquid phase 15N and 13C nuclear magnetic resonance (NMR). The International Humic Substances Society (IHSS) Pahokee peat and peat humic acid were also reacted with Na15NO2 and analyzed by solid-state 15N NMR. In Suwannee River, Armadale, and Laurentian fulvic acids, phenolic rings and activated methylene groups underwent nitrosation to form nitrosophenols (quinone monoximes) and ketoximes, respectively. The oximes underwent Beckmann rearrangements to 2??amides, and Beckmann fragmentations to nitriles. The nitriles in turn underwent hydrolysis to 1??amides. Peaks tentatively identified as imine, indophenol, or azoxybenzene nitrogens were clearly present in spectra of samples nitrosated at pH 6 but diminished at pH 3. The 15N NMR spectrum of the peat humic acid exhibited peaks corresponding with N-nitroso groups in addition to nitrosophenols, ketoximes, and secondary Beckmann reaction products. Formation of N-nitroso groups was more significant in the whole peat compared with the peat humic acid. Carbon-13 NMR analyses also indicated the occurrence of nitrosative demethoxylation in peat and soil humic acids. Reaction of 15N-NH3 fixated fulvic acid with unlabeled NO2/- resulted in nitrosative deamination of aminohydroquinone N, suggesting a previously unrecognized pathway for production of N2 gas in soils fertilized with NH3.Studies have suggested that NO2-, produced during nitrification and denitrification, can become incorporated into soil organic matter and, in one of the processes associated with chemodenitrification, react with organic

  19. Physical forcing of nitrogen fixation and diazotroph community structure in the North Pacific subtropical gyre

    NASA Astrophysics Data System (ADS)

    Church, Matthew J.; Mahaffey, Claire; Letelier, Ricardo M.; Lukas, Roger; Zehr, Jonathan P.; Karl, David M.

    2009-06-01

    Dinitrogen (N2) fixing microorganisms (termed diazotrophs) exert important control on the ocean carbon cycle. However, despite increased awareness on the roles of these microorganisms in ocean biogeochemistry and ecology, the processes controlling variability in diazotroph distributions, abundances, and activities remain largely unknown. In this study, we examine 3 years (2004-2007) of approximately monthly measurements of upper ocean diazotroph community structure and rates of N2 fixation at Station ALOHA (22°45'N, 158°W), the field site for the Hawaii Ocean Time-series program in the central North Pacific subtropical gyre (NPSG). The structure of the N2-fixing microorganism assemblage varied widely in time with unicellular N2-fixing microorganisms frequently dominating diazotroph abundances in the late winter and early spring, while filamentous microorganisms (specifically various heterocyst-forming cyanobacteria and Trichodesmium spp.) fluctuated episodically during the summer. On average, a large fraction (˜80%) of the daily N2 fixation was partitioned into the biomass of <10 μm microorganisms. Rates of N2 fixation were variable in time, with peak N2 fixation frequently coinciding with periods when heterocystous N2-fixing cyanobacteria were abundant. During the summer months when sea surface temperatures exceeded 25.2°C and concentrations of nitrate plus nitrite were at their annual minimum, rates of N2 fixation often increased during periods of positive sea surface height anomalies, as reflected in satellite altimetry. Our results suggest mesoscale physical forcing may comprise an important control on variability in N2 fixation and diazotroph community structure in the NPSG.

  20. Genetic Variability in Nodulation and Root Growth Affects Nitrogen Fixation and Accumulation in Pea

    PubMed Central

    Bourion, Virginie; Laguerre, Gisele; Depret, Geraldine; Voisin, Anne-Sophie; Salon, Christophe; Duc, Gerard

    2007-01-01

    Background and Aims Legume nitrogen is derived from two different sources, symbiotically fixed atmospheric N2 and soil N. The effect of genetic variability of root and nodule establishment on N acquisition and seed protein yield was investigated under field conditions in pea (Pisum sativum). In addition, these parameters were related to the variability in preference for rhizobial genotypes. Methods Five different spring pea lines (two hypernodulating mutants and three cultivars), previously identified in artificial conditions as contrasted for both root and nodule development, were characterized under field conditions. Root and nodule establishment was examined from the four-leaf stage up to the beginning of seed filling and was related to the patterns of shoot dry matter and nitrogen accumulation. The genetic structure of rhizobial populations associated with the pea lines was obtained by analysis of nodule samples. The fraction of nitrogen derived from symbiotic fixation was estimated at the beginning of seed filling and at physiological maturity, when seed protein content and yield were determined. Key Results The hypernodulating mutants established nodules earlier and maintained them longer than was the case for the three cultivars, whereas their root development and nitrogen accumulation were lower. The seed protein yield was higher in ‘Athos’ and ‘Austin’, the two cultivars with increased root development, consistent with their higher N absorption during seed filling. Conclusion The hypernodulating mutants did not accumulate more nitrogen, probably due to the C cost for nodulation being higher than for root development. Enhancing exogenous nitrogen supply at the end of the growth cycle, by increasing the potential for root N uptake from soil, seems a good option for improving pea seed filling. PMID:17670753

  1. Constitutive expression of nitrogen fixation (nif) genes of Klebsiella pneumoniae due to a DNA duplication.

    PubMed

    Sibold, L; Elmerich, C

    1982-01-01

    A spontaneous mutant of Klebsiella pneumoniae exhibiting nitrogen fixing activity in the presence of ammonia was isolated from a nifL ::Mu mutant. The main features of the nif constitutive mutation, designated nif-8388, were as follows: (i) neither ammonia nor bases repressed, but amino acids partially repressed, nitrogen fixation; (ii) the mutation caused an escape from the regulatory effect of glnA and glnG mutations of K. pneumoniae but not that of a glnF mutation; (iii) it enabled the activation of the nifH -lac fusion in the presence of oxygen with or without ammonia and a nifL -lac fusion in the presence of ammonia without oxygen; (iv) the mutation allowed nitrogen fixation at 37 degrees C when plasmid-borne. Restriction analysis and Southern hybridization using Mu DNA and the 8.1-kb nifQBALF EcoRI fragment as probes demonstrated that the nif-8388 mutation was a tandem duplication of 10 kb in the nifL region in which no Mu DNA was present. This duplication led to an operon fusion between nifLA and his since Nifc expression was shown to be increased with a specific inducer of the his operon. These results provide further evidence that the nifA product is a nif-specific activator, and that the nifL product is involved in oxygen repression and temperature control. In addition, they suggest that there is an autoactivation of nifLA transcription by the nifA product and that glnF could act in nif regulation by a mechanism other than the glnG-mediated control of nifLA transcription. PMID:6327278

  2. Dissolved hydrogen and nitrogen fixation in the oligotrophic North Pacific Subtropical Gyre.

    PubMed

    Wilson, Samuel T; del Valle, Daniela A; Robidart, Julie C; Zehr, Jonathan P; Karl, David M

    2013-10-01

    The production of hydrogen (H2) is an inherent component of biological dinitrogen (N2) fixation, and there have been several studies quantifying H2 production relative to N2 fixation in cultures of diazotrophs. However, conducting the relevant measurements for a field population is more complex as shown by this study of N2 fixation, H2 consumption and dissolved H2 concentrations in the oligotrophic North Pacific Ocean. Measurements of H2 oxidation revealed microbial consumption of H2 was equivalent to 1-7% of ethylene produced during the acetylene reduction assay and 11-63% of (15)N2 assimilation on a molar scale. Varying abundances of Crocosphaera and Trichodesmium as revealed by nifH gene abundances broadly corresponded with diel changes observed in both N2 fixation and H2 oxidation. However, no corresponding changes were observed in the dissolved H2 concentrations which remained consistently supersaturated (147-560%) relative to atmospheric equilibrium. The results from this field study allow the efficiency of H2 cycling by natural populations of diazotrophs to be compared to cultured representatives. The findings indicate that dissolved H2 concentrations may depend not only on the community composition of diazotrophs but also upon relevant environmental parameters such as light intensity or the presence of other H2-metabolizing microorganisms. PMID:24115620

  3. Dissolved hydrogen and nitrogen fixation in the oligotrophic North Pacific Subtropical Gyre.

    PubMed

    Wilson, Samuel T; del Valle, Daniela A; Robidart, Julie C; Zehr, Jonathan P; Karl, David M

    2013-10-01

    The production of hydrogen (H2) is an inherent component of biological dinitrogen (N2) fixation, and there have been several studies quantifying H2 production relative to N2 fixation in cultures of diazotrophs. However, conducting the relevant measurements for a field population is more complex as shown by this study of N2 fixation, H2 consumption and dissolved H2 concentrations in the oligotrophic North Pacific Ocean. Measurements of H2 oxidation revealed microbial consumption of H2 was equivalent to 1-7% of ethylene produced during the acetylene reduction assay and 11-63% of (15)N2 assimilation on a molar scale. Varying abundances of Crocosphaera and Trichodesmium as revealed by nifH gene abundances broadly corresponded with diel changes observed in both N2 fixation and H2 oxidation. However, no corresponding changes were observed in the dissolved H2 concentrations which remained consistently supersaturated (147-560%) relative to atmospheric equilibrium. The results from this field study allow the efficiency of H2 cycling by natural populations of diazotrophs to be compared to cultured representatives. The findings indicate that dissolved H2 concentrations may depend not only on the community composition of diazotrophs but also upon relevant environmental parameters such as light intensity or the presence of other H2-metabolizing microorganisms.

  4. 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

  5. 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.

  6. 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.

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

    PubMed

    Moyes, Andrew B; Kueppers, Lara M; Pett-Ridge, Jennifer; Carper, Dana L; Vandehey, Nick; O'Neil, James; Frank, A Carolin

    2016-04-01

    Coniferous forest nitrogen (N) budgets indicate unknown sources of N. A consistent association between limber pine (Pinus flexilis) and potential N2 -fixing acetic acid bacteria (AAB) indicates that native foliar endophytes may supply subalpine forests with N. To assess whether the P. flexilis-AAB association is consistent across years, we re-sampled P. flexilis twigs at Niwot Ridge, CO and characterized needle endophyte communities via 16S rRNA Illumina sequencing. To investigate whether endophytes have access to foliar N2 , we incubated twigs with (13) N2 -enriched air and imaged radioisotope distribution in needles, the first experiment of its kind using (13) N. We used the acetylene reduction assay to test for nitrogenase activity within P. flexilis twigs four times from June to September. We found evidence for N2 fixation in P. flexilis foliage. N2 diffused readily into needles and nitrogenase activity was positive across sampling dates. We estimate that this association could provide 6.8-13.6 μg N m(-2)  d(-1) to P. flexilis stands. AAB dominated the P. flexilis needle endophyte community. We propose that foliar endophytes represent a low-cost, evolutionarily stable N2 -fixing strategy for long-lived conifers. This novel source of biological N2 fixation has fundamental implications for understanding forest N budgets.

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

    PubMed

    Moyes, Andrew B; Kueppers, Lara M; Pett-Ridge, Jennifer; Carper, Dana L; Vandehey, Nick; O'Neil, James; Frank, A Carolin

    2016-04-01

    Coniferous forest nitrogen (N) budgets indicate unknown sources of N. A consistent association between limber pine (Pinus flexilis) and potential N2 -fixing acetic acid bacteria (AAB) indicates that native foliar endophytes may supply subalpine forests with N. To assess whether the P. flexilis-AAB association is consistent across years, we re-sampled P. flexilis twigs at Niwot Ridge, CO and characterized needle endophyte communities via 16S rRNA Illumina sequencing. To investigate whether endophytes have access to foliar N2 , we incubated twigs with (13) N2 -enriched air and imaged radioisotope distribution in needles, the first experiment of its kind using (13) N. We used the acetylene reduction assay to test for nitrogenase activity within P. flexilis twigs four times from June to September. We found evidence for N2 fixation in P. flexilis foliage. N2 diffused readily into needles and nitrogenase activity was positive across sampling dates. We estimate that this association could provide 6.8-13.6 μg N m(-2)  d(-1) to P. flexilis stands. AAB dominated the P. flexilis needle endophyte community. We propose that foliar endophytes represent a low-cost, evolutionarily stable N2 -fixing strategy for long-lived conifers. This novel source of biological N2 fixation has fundamental implications for understanding forest N budgets. PMID:27000956

  9. Biological nitrogen fixation: rates, patterns and ecological controls in terrestrial ecosystems.

    PubMed

    Vitousek, Peter M; Menge, Duncan N L; Reed, Sasha C; Cleveland, Cory C

    2013-07-01

    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 (15)N 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.

  10. Biological nitrogen fixation: rates, patterns and ecological controls in terrestrial ecosystems

    PubMed Central

    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. PMID:23713117

  11. Partner choice in nitrogen-fixation mutualisms of legumes and rhizobia.

    PubMed

    Simms, Ellen L; Taylor, D Lee

    2002-04-01

    Mutualistic interactions are widespread and obligatory for many organisms, yet their evolutionary persistence in the face of cheating is theoretically puzzling. Nutrient-acquisition symbioses between plants and soil microbes are critically important to plant evolution and ecosystem function, yet we know almost nothing about the evolutionary dynamics and mechanisms of persistence of these ancient mutualisms. Partner-choice and partner-fidelity are mechanisms for dealing with cheaters, and can theoretically allow mutualisms to persist despite cheaters.Many models of cooperative behavior assume pairwise interactions, while most plant-microbe nutrient-acquisition symbioses involve a single plant interacting with numerous microbes. Market models, in contrast, are well suited to mutualisms in which single plants attempt to conduct mutually beneficial resource exchange with multiple individuals. Market models assume that one partner chooses to trade with a subset of individuals selected from a market of potential partners. Hence, determining whether partner-choice occurs in plant-microbe mutualisms is critical to understanding the evolutionary persistence and dynamics of these symbioses. The nitrogen-fixation/carbon-fixation mutualism between leguminous plants and rhizobial bacteria is widespread, ancient, and important for ecosystem function and human nutrition. It also involves single plants interacting simultaneously with several to many bacterial partners, including ineffective ("cheating") strains. We review the existing literature and find that this mutualism displays several elements of partner-choice, and may match the requirements of the market paradigm. We conclude by identifying profitable questions for future research.

  12. Comparison of two Cellulomonas strains and their interaction with Azospirillum brasilense in degradation of wheat straw and associated nitrogen fixation

    SciTech Connect

    Halsall, D.M.; Gibson, A.H.

    1986-04-01

    A mutant strain of Cellulomonas sp. CS1-17 was compared with Cellulomonas gelida 2480 as the cellulolytic component of a mixed culture which was responsible for the breakdown of wheat straw to support asymbiotic nitrogen fixation by Azospirillum brasilense Sp7 (ATCC 29145). Cellulomonas sp. strain CS1-17 was more efficient than was C. gelida in cellulose breakdown at lower oxygen concentrations and, in mixed culture with A. brasilense, it supported higher nitrogenase activity(C/sub 2/H/sub 2/ reduction) and nitrogen fixation with straw as the carbon source. Based on gravimetric determinations of straw breakdown and total N determinations, the efficiency of nitrogen fixation was 72 and 63 mg of N per g of straw utilized for the mixtures containing Cellulomonas sp. and C. gelida, respectively. Both Cellulomonas spp. and Azospirillum spp. exhibited a wide range of pH tolerance. When introduced into sterilized soil, the Cellulomonas sp.-Azospirillum brasilense association was more effective in nitrogen fixation at a pH of 7.0 than at the native soil pH (5.6). This was also true of the indigenous diazotrophic microflora of this soil. The potential implications of this work to the field situation are discussed. 16 references.

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

    SciTech Connect

    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 range 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.

  14. Identification of a cis-acting element in nitrogen fixation genes recognized by CnfR in the nonheterocystous nitrogen-fixing cyanobacterium Leptolyngbya boryana.

    PubMed

    Tsujimoto, Ryoma; Kamiya, Narumi; Fujita, Yuichi

    2016-08-01

    The filamentous cyanobacterium Leptolyngbya boryana has the ability to fix nitrogen without any heterocysts under microoxic conditions. Previously, we identified the cnfR gene for a master transcriptional activator for nitrogen fixation (nif) genes in a 50-kb gene cluster containing nif and nif-related genes in L. boryana. We showed that CnfR activates the transcription of nif genes in response to low oxygen conditions, which allows the oxygen-vulnerable enzyme nitrogenase to function. However, the regulatory mechanism that underlies regulation by CnfR remains unknown. In this study, we identified a conserved cis-acting element that is recognized by CnfR. We established a reporter system in the non-diazotrophic cyanobacterium Synechocystis sp. PCC 6803 using luciferase genes (luxAB). Reporter analysis was performed with a series of truncated and modified upstream regulatory regions of nifB and nifP. The cis-element can be divided into nine motifs I-IX, and it is located 76 bp upstream of the transcriptional start sites of nifB and nifP. Six motifs of them are essential for transcriptional activation by CnfR. This cis-acting element is conserved in the upstream regions of nif genes in all diazotrophic cyanobacteria, including Anabaena and Cyanothece, thereby suggesting that the transcriptional regulation by CnfR is widespread in nitrogen-fixing cyanobacteria.

  15. Inputs of Nitrogen to Bogs of Alberta, Canada: the Importance of Biological Nitrogen Fixation VS. Atmospheric Deposition from Oil Sands Mining

    NASA Astrophysics Data System (ADS)

    Prsa, T.; Vile, M. A.; Wieder, R.; Vitt, D. H.

    2010-12-01

    Bogs of Alberta, Canada are peatlands that are both Sphagnum-moss dominated and nutrient limited. Due to their ombrotrophic nature, nitrogen (N) is deposited only via atmospheric deposition (wet/dry) and biological N2 fixation. Historically, bogs of Alberta are unpolluted with low rates of atmospheric N deposition (< 1 kg ha-1 yr-1), as opposed to eastern Canada and western Europe where rates are considerably higher (>15 kg ha-1 yr-1). Due to the extensive rich bitumen deposits under northern Alberta, however, the Oil Sands Mining (OSM) industry has been growing exponentially since the late 1960’s. Bogs situated near OSM, therefore, are likely to experience increased N deposition and the consequences and impacts of such a phenomenon are as yet, unknown. Additional N inputs into these N-limited ecosystems may cause an imbalance in the N-cycle, specifically, biological N2 fixation. Our goal was to quantify inputs of N to the system from both rates of biological N2 fixation and bulk atmospheric deposition. In summer 2010, we used acetylene reduction assay (ARA) to indirectly measure N2 fixation rates in the four most abundant moss species: Sphagnum fuscum, S. capillifolium, S. angustifolium and Pleurozium schreberi at three bog sites varying in proximity to OSM: McMurray, McKay and Utikuma bog (51, 24 and 300 km, respectively) throughout the growing season (May-August comprising 6 sampling efforts). We measured atmospheric N deposition with ion exchange resin columns (10 per site). An ANOVA and subsequent ad hoc test indicated that Utikuma had significantly lower atmospheric N deposition rates (0.130 ± 0.19 mg m-2 d-1; µ ± SE) than both McMurray and McKay (0.337 ± 0.03 and 0.262 ± 0.03 mg m-2 da-1, respectively; F2,24 = 9.04, p<0.0012), demonstrating that sites closest to the OSM region do exhibit higher rates of atmospheric N deposition. Alternatively, for inputs of N via N2 fixation, we found that McMurray (700.6 ± 144.7 µmol m-2 da-1) had significantly

  16. Azide resistance in Rhizobium ciceri linked with superior symbiotic nitrogen fixation.

    PubMed

    Bhaskar, V Vijay

    2004-12-01

    Isolated azide resistant (AzR) native R. ciceri strain 18-7 was resistant to sodium azide at 10 microg/ml. To find if nif-reiteration is responsible for azide resistance and linked to superior symbiotic nitrogen fixation, transposon (Tn5) induced azide sensitive mutants were generated. Using 4 kb nif-reiterated Sinorhizobium meliloti DNA, a clone C4 that complemented azide sensitivity was isolated by DNA hybridization from genomic library of chickpea Rhizobium strain Rcd301. EcoRI restriction mapping revealed the presence of 7 recognition sites with a total insert size of 19.17 kb. Restriction analysis of C4 clone and nif-reiterated DNA (pRK 290.7) with EcoRI and XhoI revealed similar banding pattern. Wild type strain 18-7, mutant M126 and complemented mutant M126(C4) were characterized for symbiotic properties (viz., acetylene reduction assay, total nitrogen content, nodule number and fresh and dry weight of the infected plants) and explanta nitrogenase activity. Our results suggested that azide resistance, nif-reiteration, and superior symbiotic effectiveness were interlinked with no correlation between ex-planta nitrogenase activity and azide resistance in R. ciceri.

  17. Comparative Shotgun Proteomic Analysis of Wastewater-Cultured Microalgae: Nitrogen Sensing and Carbon Fixation for Growth and Nutrient Removal in Chlamydomonas reinhardtii.

    PubMed

    Patel, Anil K; Huang, Eric L; Low-Décarie, Etienne; Lefsrud, Mark G

    2015-08-01

    Chlamydomonas reinhardtii was batch-cultured for 12 days under continuous illumination to investigate nitrogen uptake and metabolic responses to wastewater processing. Our approach compared two conditions: (1) artificial wastewater containing nitrate and ammonia and (2) nutrient-sufficient control containing nitrate as sole form of nitrogen. Treatments did not differ in final biomass; however, comparison of group proteomes revealed significant differences. Label-free shotgun proteomic analysis identified 2358 proteins, of which 92 were significantly differentially abundant. Wastewater cells showed higher relative abundances of photosynthetic antenna proteins, enzymes related to carbon fixation, and biosynthesis of amino acids and secondary metabolites. Control cells showed higher abundances of enzymes and proteins related to nitrogen metabolism and assimilation, synthesis and utilization of starch, amino acid recycling, evidence of oxidative stress, and little lipid biosynthesis. This study of the eukaryotic microalgal proteome response to nitrogen source, availability, and switching highlights tightly controlled pathways essential to the maintenance of culture health and productivity in concert with light absorption and carbon assimilation. Enriched pathways in artificial wastewater, notably, photosynthetic carbon fixation and biosynthesis of plant hormones, and those in nitrate only control, most notably, nitrogen, amino acid, and starch metabolism, represent potential targets for genetic improvement requiring targeted elucidation. PMID:25997359

  18. Comparative Shotgun Proteomic Analysis of Wastewater-Cultured Microalgae: Nitrogen Sensing and Carbon Fixation for Growth and Nutrient Removal in Chlamydomonas reinhardtii.

    PubMed

    Patel, Anil K; Huang, Eric L; Low-Décarie, Etienne; Lefsrud, Mark G

    2015-08-01

    Chlamydomonas reinhardtii was batch-cultured for 12 days under continuous illumination to investigate nitrogen uptake and metabolic responses to wastewater processing. Our approach compared two conditions: (1) artificial wastewater containing nitrate and ammonia and (2) nutrient-sufficient control containing nitrate as sole form of nitrogen. Treatments did not differ in final biomass; however, comparison of group proteomes revealed significant differences. Label-free shotgun proteomic analysis identified 2358 proteins, of which 92 were significantly differentially abundant. Wastewater cells showed higher relative abundances of photosynthetic antenna proteins, enzymes related to carbon fixation, and biosynthesis of amino acids and secondary metabolites. Control cells showed higher abundances of enzymes and proteins related to nitrogen metabolism and assimilation, synthesis and utilization of starch, amino acid recycling, evidence of oxidative stress, and little lipid biosynthesis. This study of the eukaryotic microalgal proteome response to nitrogen source, availability, and switching highlights tightly controlled pathways essential to the maintenance of culture health and productivity in concert with light absorption and carbon assimilation. Enriched pathways in artificial wastewater, notably, photosynthetic carbon fixation and biosynthesis of plant hormones, and those in nitrate only control, most notably, nitrogen, amino acid, and starch metabolism, represent potential targets for genetic improvement requiring targeted elucidation.

  19. Evolutionary dynamics of nitrogen fixation in the legume-rhizobia symbiosis.

    PubMed

    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

  20. The prospect function of terrestrial nitrogen-fixing blue-green algae on the fixation of desert

    NASA Astrophysics Data System (ADS)

    Yang, Yusuo; Lei, Jiaqiang

    2003-07-01

    The Terrestrial Nitrogen-fixing Blue-green Algae, which are possessed of both photosynthesis and nitrogen fixation, are the leading organisms in the adverse circumstances. With their typical cell structures and physiological abilities, they are strongly resistant to drought, infertility etc. The growth of Terrestrial Nitrogen-fixing Blue-green Algae can rich the soils in nitrogen and organic compounds, which are benefit to other microbes and plants. Terrestrial Nitrogen-fixing Blue-green Algae are widely distributed in Gurbantunggut Desert. It was estimated that about 40% of the surface of the desert are covered by the "Black Crust". "Black Crust" is mainly occupied by Terrestrial Nitrogen-fixing Blue-green Algae. It is Terrestrial Nitrogen-fixing Blue-green Algae that construct the mechanical crust with a little other algae and fungi through biological, chemical and physical actions. So Terrestrial Nitrogen-fixing Blue-green Algae play an important part in desert fixation. It was analyzed that there are three species of the blue-greens in the "Black Crust": Microcoleus vaginatus(Vauch)Gom.,Scytonema ocellatum Lynbye and Schizothrix mella Gardner. We had isolated Microcoleus vaginatus(Vauch)Gom. and Scytonema ocellatum Lynbye. Some tests had been made to prove the feasibility of the desert fixation of the Blue-greens. Under experiment conditions, the blue-greens grown on the surface of sand, covered the sand quickly after the inoculation, and formed a mechanical fixed surface layer (7 days for Microcoleus vaginatus, 15-21 days for Scytonema ocellatum).

  1. Comparative Assessment of Nitrogen Fixation Methodologies, Conducted in the Oligotrophic North Pacific Ocean

    PubMed Central

    Böttjer, Daniela; Church, Matthew J.; Karl, David M.

    2012-01-01

    Resolution of the nitrogen (N) cycle in the marine environment requires an accurate assessment of dinitrogen (N2) fixation. We present here an update on progress in conducting field measurements of acetylene reduction (AR) and 15N2 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 (C2H4) with a reducing compound photodetector. The rates of C2H4 production were measurable for nonconcentrated seawater samples after an incubation period of 3 to 4 h. The 15N2 tracer measurements compared the addition of 15N2 as a gas bubble and dissolved as 15N2 enriched seawater. On all sampling occasions and at all depths, a 2- to 6-fold increase in the rate of 15N2 assimilation was measured when 15N2-enriched seawater was added to the seawater sample compared to the addition of 15N2 as a gas bubble. In addition, we show that the 15N2-enriched seawater can be prepared prior to its use with no detectable loss (<1.7%) of dissolved 15N2 during 4 weeks of storage, facilitating its use in the field. The ratio of C2H4 production to 15N2 assimilation varied from 7 to 27 when measured simultaneously in surface seawater samples. Collectively, the modifications to the AR assay and the 15N2 assimilation technique present opportunities for more accurate and high frequency measurements (e.g., diel scale) of N2 fixation, providing further insight into the contribution of different groups of diazotrophs to the input of N in the global oceans. PMID:22773638

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

    PubMed Central

    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 15N2 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. PMID:23985746

  3. 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. PMID:23985746

  4. 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. PMID:22773638

  5. Spatial and Temporal Variation in Feather Moss Associated Nitrogen Fixation in Coniferous and Deciduous Dominated Alaskan Boreal Forests

    NASA Astrophysics Data System (ADS)

    Jean, M.; Mack, M. C.; Johnstone, J. F.

    2015-12-01

    Dominant canopy tree species have strong effects on the composition and function of understory species. In boreal forests, forest floor bryophytes and their associated microbes are a primary source of ecosystem nitrogen (N) inputs, and thus an important process regulating ecosystem productivity. Bryophyte composition and abundance varies with forest composition, yet how such changes can affect ecosystem processes such as N fixation is still poorly understood. Our goal is to investigate how cyanobacteria-based N fixation occurring in the two most common feather mosses in the Alaskan boreal forest (Pleurozium schreberi and Hylocomium splendens) varies among coniferous and deciduous forest types, over the growing season, and across a nutrient availability gradient. Twelve patches of H. splendens and P. schreberi were identified in three pairs (blocks) of adjacent stands of paper birch (Betula neoalaskana) and black spruce (Picea mariana) near Fairbanks, interior Alaska. Sampling occurred in one block in June, July, August, and September 2014, and in the three blocks once in August 2014. Moss leaf area, moisture and weight, as well as environmental variables such as air temperature and canopy cover were recorded. Fixation rates were consistently higher for P. schreberi than for H. splendens. Overall, N fixation rates were lower in birch than in spruce stands and peaked in August, or July for P. schreberi in birch stands. Moreover, fixation rates varied along the nutrient availability gradient, with fixation rates higher where nutrient availability was lower. This difference was especially clear in spruce stands. Our preliminary results suggest that moss species, canopy type, and environmental factors all influence N fixation rates in Alaskan boreal forests. Our results will enhance the knowledge of the processes that drive N fixation in boreal forests, which is important for predicting ecosystem consequences of changing forest composition.

  6. CO2-adapted legumes ameliorate but do not prevent the negative effect of elevated CO2 on nitrogen fixation

    NASA Astrophysics Data System (ADS)

    Newton, P.; Bowatte, S.; Lieffering, M.; Li, F.

    2015-12-01

    The response of biological nitrogen fixation (BNF) to climate and elevated CO2 (eCO2) is a key uncertainty in modelling C cycle projections. In addition, as BNF provides 50% of the nitrogen (N) input to agricultural production and as ecosystem responses to eCO2 are strongly influenced by N availability then the eCO2 impact on BNF is central to modelling legume-based system responses to climate change. Greater photoassimilate production under eCO2 should lead to enhanced BNF and this response is a feature of ecosystem models thus providing the N inputs necessary to provide continuing stimulation of NPP. FACE experiments provide a 'realistic' environment for eCO2 studies; however, even if run for multiple years, they still may not capture adaptation to eCO2 particularly in ecosystems dominated by perennial species. We tested the effect of eCO2 on BNF and the potential importance of adaption by growing legumes that had been exposed to high or ambient CO2 concentrations at a natural CO2 spring in a long-running (16 year) FACE experiment on grassland. BNF was significantly lower under eCO2 but the reduction was less marked where plants had originated in a high CO2 environment. An ecosystem model run with reduced BNF proved a better fit to the experimental data for the FACE experiment than where BNF was enhanced or unchanged under eCO2.

  7. Isotopic evidence for biological nitrogen fixation by molybdenum-nitrogenase from 3.2 Gyr.

    PubMed

    Stüeken, Eva E; Buick, Roger; Guy, Bradley M; Koehler, Matthew C

    2015-04-30

    Nitrogen is an essential nutrient for all organisms that must have been available since the origin of life. Abiotic processes including hydrothermal reduction, photochemical reactions, or lightning discharge could have converted atmospheric N2 into assimilable NH4(+), HCN, or NOx species, collectively termed fixed nitrogen. But these sources may have been small on the early Earth, severely limiting the size of the primordial biosphere. The evolution of the nitrogen-fixing enzyme nitrogenase, which reduces atmospheric N2 to organic NH4(+), thus represented a major breakthrough in the radiation of life, but its timing is uncertain. Here we present nitrogen isotope ratios with a mean of 0.0 ± 1.2‰ from marine and fluvial sedimentary rocks of prehnite-pumpellyite to greenschist metamorphic grade between 3.2 and 2.75 billion years ago. These data cannot readily be explained by abiotic processes and therefore suggest biological nitrogen fixation, most probably using molybdenum-based nitrogenase as opposed to other variants that impart significant negative fractionations. Our data place a minimum age constraint of 3.2 billion years on the origin of biological nitrogen fixation and suggest that molybdenum was bioavailable in the mid-Archaean ocean long before the Great Oxidation Event.

  8. Isotopic evidence for biological nitrogen fixation by molybdenum-nitrogenase from 3.2 Gyr.

    PubMed

    Stüeken, Eva E; Buick, Roger; Guy, Bradley M; Koehler, Matthew C

    2015-04-30

    Nitrogen is an essential nutrient for all organisms that must have been available since the origin of life. Abiotic processes including hydrothermal reduction, photochemical reactions, or lightning discharge could have converted atmospheric N2 into assimilable NH4(+), HCN, or NOx species, collectively termed fixed nitrogen. But these sources may have been small on the early Earth, severely limiting the size of the primordial biosphere. The evolution of the nitrogen-fixing enzyme nitrogenase, which reduces atmospheric N2 to organic NH4(+), thus represented a major breakthrough in the radiation of life, but its timing is uncertain. Here we present nitrogen isotope ratios with a mean of 0.0 ± 1.2‰ from marine and fluvial sedimentary rocks of prehnite-pumpellyite to greenschist metamorphic grade between 3.2 and 2.75 billion years ago. These data cannot readily be explained by abiotic processes and therefore suggest biological nitrogen fixation, most probably using molybdenum-based nitrogenase as opposed to other variants that impart significant negative fractionations. Our data place a minimum age constraint of 3.2 billion years on the origin of biological nitrogen fixation and suggest that molybdenum was bioavailable in the mid-Archaean ocean long before the Great Oxidation Event. PMID:25686600

  9. 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

  10. 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

  11. The reallocation of carbon in P deficient lupins affects biological nitrogen fixation.

    PubMed

    Kleinert, Aleysia; Venter, Mauritz; Kossmann, Jens; Valentine, Alexander

    2014-11-01

    It is not known how phosphate (P) deficiency affects the allocation of carbon (C) to biological nitrogen fixation (BNF) in legumes. The alteration of the respiratory and photosynthetic C costs of BNF was investigated under P deficiency. Although BNF can impose considerable sink stimulation on host respiratory and photosynthetic C, it is not known how the change in the C and energy allocation during P deficiency may affect BNF. Nodulated Lupinus luteus plants were grown in sand culture, using a modified Long Ashton nutrient solution containing no nitrogen (N) for ca. four weeks, after which one set was exposed to a P-deficient nutrient medium, while the other set continued growing on a P-sufficient nutrient medium. Phosphorus stress was measured at 20 days after onset of P-starvation. During P stress the decline in nodular P levels was associated with lower BNF and nodule growth. There was also a shift in the balance of photosynthetic and respiratory C toward a loss of C during P stress. Below-ground respiration declined under limiting P conditions. However, during this decline there was also a shift in the proportion of respiratory energy from maintenance toward growth respiration. Under P stress, there was an increased allocation of C toward root growth, thereby decreasing the amount of C available for maintenance respiration. It is therefore possible that the decline in BNF under P deficiency may be due to this change in resource allocation away from respiration associated with direct nutrient uptake, but rather toward a long term nutrient acquisition strategy of increased root growth.

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

    SciTech Connect

    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, 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.

  13. Nitrogen Fixation in Peanut Nodules during Dark Periods and Detopped Conditions with Special Reference to Lipid Bodies 1

    PubMed Central

    Siddique, Abu-baker M.; Bal, Arya K.

    1991-01-01

    The peanut plant (Arachis hypogaea L.), unlike other known legumes, can sustain nitrogen fixation when prolonged periods of darkness or detopping curtail the supply of photosynthate to the nodule. This ability to withstand photosynthate stress is attributed to the presence of lipid bodies in infected nodule cells. In both dark-treated and detopped plants, the lipid bodies show a gradual decrease in numbers, suggesting their utilization as a source of energy and carbon for nitrogen fixation. Lipolytic activity can be localized in the lipid bodies, and the existence of β-oxidation pathway and, glyoxylate cycle is shown by the release of 14CO2 from 14C lineoleoyl coenzyme A by the nodule homogenate. Images Figure 2 Figure 4 PMID:16668069

  14. Nitrogen fixation in peanut nodules during dark periods and detopped conditions with special reference to lipid bodies

    SciTech Connect

    Siddique, A.M.; Bal, A.K. )

    1991-03-01

    The peanut plant (Arachis hypogaea L.), unlike other known legumes, can sustain nitrogen fixation when prolonged periods of darkness or detopping curtail the supply of photosynthate to the nodule. This ability to withstand photosynthate stress is attributed to the presence of lipid bodies in infected nodule cells. In both dark-treated and detopped plants, the lipid bodies show a gradual decrease in numbers, suggesting their utilization as a source of energy and carbon for nitrogen fixation. Lipolytic activity can be localized in the lipid bodies, and the existence of {beta}-oxidation pathway and glyoxylate cycle is shown by the release of {sup 14}CO{sub 2} from {sup 14}C lineoleoyl coenzyme A by the nodule homogenate.

  15. Growth condition study of algae function in ecosystem for CO2 bio-fixation.

    PubMed

    Tsai, David Dah-Wei; Ramaraj, Rameshprabu; Chen, Paris Honglay

    2012-02-01

    Algae niche play a crucial role on carbon cycle and have great potential for CO(2) sequestration. This study was to investigate the CO(2) bio-fixation by the high rate pond (HRP) to mimic the algae function of nature. All the reactors can keep CO(2) consumption efficiencies over 100%. The statistical analyses proved HRPs were close to the natural system from all the growth conditions. The HRP could show the "natural optimization as nature" to perform as well as the artificial reactor of continuously stirred tank reactor (CSTR). In the nutrition study, the carbon mass balance indicated CO(2) was the main carbon source. Accordingly, the HRPs can keep a neutral pH range to provide dissolved oxygen (DO), to promote total nitrogen (TN)/total phosphorous (TP) removal efficiencies and to demonstrate self-purification process. Furthermore, the observations of different nitrogen species in the reactors demonstrated that the major nitrogen source was decided by pH. This finding logically explained the complex nitrogen uptake by algae in nature. Consequently, this study took advantage of HRP to explore the processes of efficient CO(2) uptake with the corresponding growth condition in the ecosystem. Those results contributed the further understanding of the role of CO(2) bio-fixation in nature and demonstrated HRP could be a potential ecological engineering alternative.

  16. Growth condition study of algae function in ecosystem for CO2 bio-fixation.

    PubMed

    Tsai, David Dah-Wei; Ramaraj, Rameshprabu; Chen, Paris Honglay

    2012-02-01

    Algae niche play a crucial role on carbon cycle and have great potential for CO(2) sequestration. This study was to investigate the CO(2) bio-fixation by the high rate pond (HRP) to mimic the algae function of nature. All the reactors can keep CO(2) consumption efficiencies over 100%. The statistical analyses proved HRPs were close to the natural system from all the growth conditions. The HRP could show the "natural optimization as nature" to perform as well as the artificial reactor of continuously stirred tank reactor (CSTR). In the nutrition study, the carbon mass balance indicated CO(2) was the main carbon source. Accordingly, the HRPs can keep a neutral pH range to provide dissolved oxygen (DO), to promote total nitrogen (TN)/total phosphorous (TP) removal efficiencies and to demonstrate self-purification process. Furthermore, the observations of different nitrogen species in the reactors demonstrated that the major nitrogen source was decided by pH. This finding logically explained the complex nitrogen uptake by algae in nature. Consequently, this study took advantage of HRP to explore the processes of efficient CO(2) uptake with the corresponding growth condition in the ecosystem. Those results contributed the further understanding of the role of CO(2) bio-fixation in nature and demonstrated HRP could be a potential ecological engineering alternative. PMID:22196805

  17. Local inhibition of nitrogen fixation and nodule metabolism in drought-stressed soybean

    PubMed Central

    Gil-Quintana, Erena; Larrainzar, Estíbaliz; Seminario, Amaia; Díaz-Leal, Juan Luis; Alamillo, Josefa M.; Pineda, Manuel; Arrese-Igor, Cesar; Wienkoop, Stefanie; González, Esther M.

    2013-01-01

    Drought stress is a major factor limiting symbiotic nitrogen fixation (NF) in soybean crop production. However, the regulatory mechanisms involved in this inhibition are still controversial. Soybean plants were symbiotically grown in a split-root system (SRS), which allowed for half of the root system to be irrigated at field capacity while the other half remained water deprived. NF declined in the water-deprived root system while nitrogenase activity was maintained at control values in the well-watered half. Concomitantly, amino acids and ureides accumulated in the water-deprived belowground organs regardless of transpiration rates. Ureide accumulation was found to be related to the decline in their degradation activities rather than increased biosynthesis. Finally, proteomic analysis suggests that plant carbon metabolism, protein synthesis, amino acid metabolism, and cell growth are among the processes most altered in soybean nodules under drought stress. Results presented here support the hypothesis of a local regulation of NF taking place in soybean and downplay the role of ureides in the inhibition of NF. PMID:23580751

  18. Limitation of Acetylene Reduction (Nitrogen Fixation) by Photosynthesis in Soybean Having Low Water Potentials 1

    PubMed Central

    Huang, Chi-Ying; Boyer, John S.; Vanderhoef, Larry N.

    1975-01-01

    The role of photosynthesis and transpiration in the desiccation-induced inhibition of acetylene reduction (nitrogen fixation) was investigated in soybean (Glycine max [L.] Merr. var. Beeson) using an apparatus that permitted simultaneous measurements of acetylene reduction, net photosynthesis, and transpiration. The inhibition of acetylene reduction caused by low water potentials and their aftereffects could be reproduced by depriving shoots of atmospheric CO2 even though the soil remained at water potentials that should have favored rapid acetylene reduction. The inhibition of acetylene reduction at low water potentials could be partially reversed by exposing the shoots to high CO2 concentrations. When transpiration was varied independently of photosynthesis and dark respiration in plants having high water potentials, no effects on acetylene reduction could be observed. There was no correlation between transpiration and acetylene reduction in the CO2 experiments. Therefore, the correlation that was observed between transpiration and acetylene reduction during desiccation was fortuitous. We conclude that the inhibition of shoot photosynthesis accounted for the inhibition of nodule acetylene reduction at low water potentials. PMID:16659277

  19. Interactions among nitrogen fixation and soil phosphorus acquisition strategies in lowland tropical rain forests.

    PubMed

    Nasto, Megan K; Alvarez-Clare, Silvia; Lekberg, Ylva; Sullivan, Benjamin W; Townsend, Alan R; Cleveland, Cory C

    2014-10-01

    Paradoxically, symbiotic dinitrogen (N2 ) fixers are abundant in nitrogen (N)-rich, phosphorus (P)-poor lowland tropical rain forests. One hypothesis to explain this pattern states that N2 fixers have an advantage in acquiring soil P by producing more N-rich enzymes (phosphatases) that mineralise organic P than non-N2 fixers. We assessed soil and root phosphatase activity between fixers and non-fixers in two lowland tropical rain forest sites, but also addressed the hypothesis that arbuscular mycorrhizal (AM) colonisation (another P acquisition strategy) is greater on fixers than non-fixers. Root phosphatase activity and AM colonisation were higher for fixers than non-fixers, and strong correlations between AM colonisation and N2 fixation at both sites suggest that the N-P interactions mediated by fixers may generally apply across tropical forests. We suggest that phosphatase enzymes and AM fungi enhance the capacity of N2 fixers to acquire soil P, thus contributing to their high abundance in tropical forests.

  20. Purification and binding analysis of the nitrogen fixation regulatory NifA protein from Azospirillum brasilense.

    PubMed

    Passaglia, L M; Van Soom, C; Schrank, A; Schrank, I S

    1998-11-01

    NifA protein activates transcription of nitrogen fixation operons by the alternative sigma 54 holoenzyme form of RNA polymerase. This protein binds to a well-defined upstream activator sequence (UAS) located at the -200/-100 position of nif promoters with the consensus motif TGT-N10-ACA. NifA of Azospirillum brasilense was purified in the form of a glutathione-S-transferase (GST)-NifA fusion protein and proteolytic release of GST yielded inactive and partially soluble NifA. However, the purified NifA was able to induce the production of specific anti-A. brasilense NifA-antiserum that recognized NifA from A. brasilense but not from K. pneumoniae. Both GST-NifA and NifA expressed from the E. coli tac promoter are able to activate transcription from the nifHDK promoter but only in an A. brasilense background. In order to investigate the mechanism that regulates NifA binding capacity we have used E. coli total protein extracts expressing A. brasilense nifA in mobility shift assays. DNA fragments carrying the two overlapping, wild-type or mutated UAS motifs present in the nifH promoter region revealed a retarded band of related size. These data show that the binding activity present in the C-terminal domain of A. brasilense NifA protein is still functional even in the presence of oxygen.

  1. A purple acid phosphatase plays a role in nodule formation and nitrogen fixation in Astragalus sinicus.

    PubMed

    Wang, Jianyun; Si, Zaiyong; Li, Fang; Xiong, Xiaobo; Lei, Lei; Xie, Fuli; Chen, Dasong; Li, Yixing; Li, Youguo

    2015-08-01

    The AsPPD1 gene from Astragalus sinicus encodes a purple acid phosphatase. To address the functions of AsPPD1 in legume-rhizobium symbiosis, its expression patterns, enzyme activity, subcellular localization, and phenotypes associated with its over-expression and RNA interference (RNAi) were investigated. The expression of AsPPD1 was up-regulated in roots and nodules after inoculation with rhizobia. Phosphate starvation reduced the levels of AsPPD1 transcripts in roots while increased those levels in nodules. We confirmed the acid phosphatase and phosphodiesterase activities of recombinant AsPPD1 purified from Pichia pastoris, and demonstrated its ability to hydrolyze ADP and ATP in vitro. Subcellular localization showed that AsPPD1 located on the plasma membranes in hairy roots and on the symbiosomes membranes in root nodules. Over-expression of AsPPD1 in hairy roots inhibited nodulation, while its silencing resulted in nodules early senescence and significantly decreased nitrogenase activity. Furthermore, HPLC measurement showed that AsPPD1 overexpression affects the ADP levels in the infected roots and nodules, AsPPD1 silencing affects the ratio of ATP/ADP and the energy charge in nodules, and quantitative observation demonstrated the changes of AsPPD1 transcripts level affected nodule primordia formation. Taken together, it is speculated that AsPPD1 contributes to symbiotic ADP levels and energy charge control, and this is required for effective nodule organogenesis and nitrogen fixation.

  2. Crystallization of a flavodoxin involved in nitrogen fixation in Rhodobacter capsulatus

    SciTech Connect

    Pérez-Dorado, Inmaculada; Bortolotti, Ana; Cortez, Néstor; Hermoso, Juan A.

    2008-05-01

    The flavodoxin NifF from R. capsulatus, a candidate for nitrogenase reduction during nitrogen fixation, has been crystallized using the hanging-drop vapour-diffusion method. Preliminary X-ray data processing at 2.17 Å resolution allowed determination of the crystal system and unit-cell parameters. Flavodoxins are small electron-transfer proteins that contain one molecule of noncovalently bound flavin mononucleotide (FMN). The flavodoxin NifF from the photosynthetic bacterium Rhodobacter capsulatus is reduced by one electron from ferredoxin/flavodoxin:NADP(H) reductase and was postulated to be an electron donor to nitrogenase in vivo. NifF was cloned and overexpressed in Escherichia coli, purified and concentrated for crystallization using the hanging-drop vapour-diffusion method at 291 K. Crystals grew from a mixture of PEG 3350 and PEG 400 at pH 5.5 and belong to the tetragonal space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 66.49, c = 121.32 Å. X-ray data sets have been collected to 2.17 Å resolution.

  3. Short-term nitrate (nitrite) inhibition of nitrogen fixation in Azotobacter chroococcum

    SciTech Connect

    Cejudo, F.J.; Paneque, A.

    1986-01-01

    Nitrate-grown Azotobacter chroococcum ATCC 4412 cells lack the ability to fix N/sub 2/. Nitrogenase activity developed after the cells were suspended in a combined nitrogen-free medium and was paralleled by a concomitant decrease in nitrate assimilation capacity. In such treated cells exhibiting transitory nitrate assimilation and N/sub 2/-fixation capacity, nitrate or nitrite caused a short-term inhibitory effect on nitrogenase activity which ceased once the anion was exhausted from the medium. The glutamate analog L-methionine-DL-sulfoximine, an inhibitor of glutamine synthetase, prevented inhibition of nitrogenase activity by nitrate or nitrite without affecting the uptake of these anions, which were reduced and stoichiometrically released into the external medium as ammonium. Inhibition of nitrogenase by nitrate (nitrite) did not take place in A. chroococcum MCD1, which is unable to assimilate either. The authors conclude that the short-term inhibitory effect of nitrate (nitrite) on nitrogenase activity is due to some organic product(s) formed during the assimilation of the ammonium resulting from nitrate (nitrite) reduction.

  4. 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.

  5. Nitrogen fixation capacity of Azotobacter spp. strains isolated from soils in different ecosystems and relationship between them and the microbiological properties of soils.

    PubMed

    Kizilkaya, Ridvan

    2009-01-01

    The objectives of this study were to count and culture Azotobacter spp. in sampled soils, to determine the nitrogen (N) fixing capacity byAzotobacter spp. in pure culture and different soils, and to explore the relationships between N fixation capacity of Azotobacter spp. and microbiological properties of soils in Northern Anatolia, Turkey. Statistically significant relationships were found between the population of Azotobacter spp. in soils and microbial biomass C (Cmic), dehydrogenase (DHA), beta-glucosidase (GA), alkaline phosphatase (APA) and arylsulphatase (ASA) activities. However, relationships between the population of Azotobacter spp. and basal soil respiration (BSR), urease (UA) and catalase (CA) activities were insignificant. The N fixation capacities of native 3 day old Azotobacter chroococcum strains added to Ashby Media varied from 3.50 to 29.35 microg N ml(-1) on average 10.24. In addition, N fixation capacities of Azotobacter spp. strains inoculated with clayey soil, loam soil, and sandy clay loam soil during eight week incubation period were 4.78-15.91 microg N g(-1), 9.03-13.47 microg N g(-1) and 6.51-16.60 microg N g(-1), respectively. It was concluded that the most N fixation by Azotobacter spp. was in sandy clay loam soils.

  6. 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

  7. Ureide assay for measuring nitrogen fixation by nodulated soybean calibrated by sup 15 N methods. [Glycine max

    SciTech Connect

    Herridge, D.F. ); Peoples, M.B. )

    1990-06-01

    We report experiments to quantify the relationships between the relative abundance of ureide-N in root-bleeding sap, vacuum-extracted sap, and hot water extracts of stems and petioles of nodulated soybean (Glycine max (L.) Merrill cv Bragg) and the proportion of plant N derived from nitrogen fixation. Additional experiments examined the effects of plant genotype and strain of rhizobia on these relationships. In each of the five experiments reported, plants of cv Bragg (experiment 1), cv Lincoln (experiments 3, 4, 5), or six cultivars/genotypes (experiment 2) were grown in a sand:vermiculite mixture in large pots in a naturally lit, temperature-controlled glasshouse during summer. Pots were inoculated at sowing with effective Bradyrhizobium japonicum CB 1809 (USDA 136) or with one of 21 different strains of rhizobia. The proportions of plant N derived from nitrogen fixation were determined using {sup 15}N dilution. Results show that assessment of nitrogen fixation by soybean using the ureide technique should now be possible with the standard curves presented, irrespective of genotype or strain of rhizobia occupying the nodules.

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

    PubMed

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

    2014-11-01

    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.

  9. Nodulation and nitrogen fixation by Mimosa spp. in the Cerrado and Caatinga biomes of Brazil.

    PubMed

    dos Reis, Fábio Bueno; Simon, Marcelo F; Gross, Eduardo; Boddey, Robert M; Elliott, Geoffrey N; Neto, Nicolau E; Loureiro, M de Fatima; de Queiroz, Luciano P; Scotti, Maria Rita; Chen, Wen-Ming; Norén, Agneta; Rubio, Maria C; de Faria, Sergio M; Bontemps, Cyril; Goi, Silvia R; Young, J Peter W; Sprent, Janet I; James, Euan K

    2010-06-01

    *An extensive survey of nodulation in the legume genus Mimosa was undertaken in two major biomes in Brazil, the Cerrado and the Caatinga, in both of which there are high degrees of endemicity of the genus. *Nodules were collected from 67 of the 70 Mimosa spp. found. Thirteen of the species were newly reported as nodulating. Nodules were examined by light and electron microscopy, and all except for M. gatesiae had a structure typical of effective Mimosa nodules. The endosymbiotic bacteria in nodules from all of the Mimosa spp. were identified as Burkholderia via immunolabelling with an antibody against Burkholderia phymatum STM815. *Twenty of the 23 Mimosa nodules tested were shown to contain nitrogenase by immunolabelling with an antibody to the nitrogenase Fe- (nifH) protein, and using the delta(15)N ((15)N natural abundance) technique, contributions by biological N(2) fixation of up to 60% of total plant N were calculated for Caatinga Mimosa spp. *It is concluded that nodulation in Mimosa is a generic character, and that the preferred symbionts of Brazilian species are Burkholderia. This is the first study to demonstrate N(2) fixation by beta-rhizobial symbioses in the field.

  10. Proteome Profiling of the Rhodobacter capsulatus Molybdenum Response Reveals a Role of IscN in Nitrogen Fixation by Fe-Nitrogenase

    PubMed Central

    Hoffmann, Marie-Christine; Wagner, Eva; Langklotz, Sina; Pfänder, Yvonne; Hött, Sina; Bandow, Julia E.

    2015-01-01

    ABSTRACT Rhodobacter capsulatus is capable of synthesizing two nitrogenases, a molybdenum-dependent nitrogenase and an alternative Mo-free iron-only nitrogenase, enabling this diazotroph to grow with molecular dinitrogen (N2) as the sole nitrogen source. Here, the Mo responses of the wild type and of a mutant lacking ModABC, the high-affinity molybdate transporter, were examined by proteome profiling, Western analysis, epitope tagging, and lacZ reporter fusions. Many Mo-controlled proteins identified in this study have documented or presumed roles in nitrogen fixation, demonstrating the relevance of Mo control in this highly ATP-demanding process. The levels of Mo-nitrogenase, NifHDK, and the Mo storage protein, Mop, increased with increasing Mo concentrations. In contrast, Fe-nitrogenase, AnfHDGK, and ModABC, the Mo transporter, were expressed only under Mo-limiting conditions. IscN was identified as a novel Mo-repressed protein. Mo control of Mop, AnfHDGK, and ModABC corresponded to transcriptional regulation of their genes by the Mo-responsive regulators MopA and MopB. Mo control of NifHDK and IscN appeared to be more complex, involving different posttranscriptional mechanisms. In line with the simultaneous control of IscN and Fe-nitrogenase by Mo, IscN was found to be important for Fe-nitrogenase-dependent diazotrophic growth. The possible role of IscN as an A-type carrier providing Fe-nitrogenase with Fe-S clusters is discussed. IMPORTANCE Biological nitrogen fixation is a central process in the global nitrogen cycle by which the abundant but chemically inert dinitrogen (N2) is reduced to ammonia (NH3), a bioavailable form of nitrogen. Nitrogen reduction is catalyzed by nitrogenases found in diazotrophic bacteria and archaea but not in eukaryotes. All diazotrophs synthesize molybdenum-dependent nitrogenases. In addition, some diazotrophs, including Rhodobacter capsulatus, possess catalytically less efficient alternative Mo-free nitrogenases, whose expression

  11. High levels of heterogeneity in diazotroph diversity and activity within a putative hotspot for marine nitrogen fixation.

    PubMed

    Messer, Lauren F; Mahaffey, Claire; M Robinson, Charlotte; Jeffries, Thomas C; Baker, Kirralee G; Bibiloni Isaksson, Jaime; Ostrowski, Martin; Doblin, Martina A; Brown, Mark V; Seymour, Justin R

    2016-06-01

    Australia's tropical waters represent predicted 'hotspots' for nitrogen (N2) fixation based on empirical and modelled data. However, the identity, activity and ecology of diazotrophs within this region are virtually unknown. By coupling DNA and cDNA sequencing of nitrogenase genes (nifH) with size-fractionated N2 fixation rate measurements, we elucidated diazotroph dynamics across the shelf region of the Arafura and Timor Seas (ATS) and oceanic Coral Sea during Austral spring and winter. During spring, Trichodesmium dominated ATS assemblages, comprising 60% of nifH DNA sequences, while Candidatus Atelocyanobacterium thalassa (UCYN-A) comprised 42% in the Coral Sea. In contrast, during winter the relative abundance of heterotrophic unicellular diazotrophs (δ-proteobacteria and γ-24774A11) increased in both regions, concomitant with a marked decline in UCYN-A sequences, whereby this clade effectively disappeared in the Coral Sea. Conservative estimates of N2 fixation rates ranged from <1 to 91 nmol l(-1) day(-1), and size fractionation indicated that unicellular organisms dominated N2 fixation during both spring and winter, but average unicellular rates were up to 10-fold higher in winter than in spring. Relative abundances of UCYN-A1 and γ-24774A11 nifH transcripts negatively correlated to silicate and phosphate, suggesting an affinity for oligotrophy. Our results indicate that Australia's tropical waters are indeed hotspots for N2 fixation and that regional physicochemical characteristics drive differential contributions of cyanobacterial and heterotrophic phylotypes to N2 fixation.

  12. 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

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

    PubMed

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

    2016-09-29

    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.

  14. Isolation of a Rhizobium phaseoli cytochrome mutant with enhanced respiration and symbiotic nitrogen fixation.

    PubMed Central

    Soberón, M; Williams, H D; Poole, R K; Escamilla, E

    1989-01-01

    Cultured cells of a Rhizobium phaseoli wild-type strain (CE2) possess b-type and c-type cytochromes and two terminal oxidases: cytochromes o and aa3. Cytochrome aa3 was partially expressed when CE2 cells were grown on minimal medium, during symbiosis, and in well-aerated liquid cultures in a complex medium (PY2). Two cytochrome mutants of R. phaseoli were obtained and characterized. A Tn5-mob-induced mutant, CFN4201, expressed diminished amounts of b-type and c-type cytochromes, showed an enhanced expression of cytochrome oxidases, and had reduced levels of N,N,N',N'-tetramethyl-p-phenylenediamine, succinate, and NADH oxidase activities. Nodules formed by this strain had no N2 fixation activity. The other mutant, CFN4205, which was isolated by nitrosoguanidine mutagenesis, had reduced levels of cytochrome o and higher succinate oxidase activity but similar NADH and N,N,N',N'-tetramethyl-p-phenylenediamine oxidase activities when compared with the wild-type strain. Strain CFN4205 expressed a fourfold-higher cytochrome aa3 content when cultured on minimal and complex media and had twofold-higher cytochrome aa3 levels during symbiosis when compared with the wild-type strain. Nodules formed by strain CFN4205 fixed 33% more N2 than did nodules formed by the wild-type strain, as judged by the total nitrogen content found in plants nodulated by these strains. Finally, low-temperature photodissociation spectra of whole cells from strains CE2 and CFN4205 reveal cytochromes o and aa3. Both cytochromes react with O2 at -180 degrees C to give a light-insensitive compound. These experiments identify cytochromes o and aa3 as functional terminal oxidases in R. phaseoli. PMID:2644201

  15. Organization and regulation of the genes for nitrogen fixation in Rhodopseudomonas capsulata

    SciTech Connect

    Haselkorn, R.

    1989-05-01

    The research carried out over the past two years with DOE support has led to the elucidation of the complete nucleotide sequence of three genes required for the transcription of the genes for nitrogen fixation in Rhodobacter capsulatus. These genes are called nifR1, nifR2 and nifR4 in R. capsulatus. They were found to be homologous to ntrC, ntrB and ntrA in Klebsiella, respectively. Genetic analysis of ammonia-constitutive (NiF{sup c}) mutants of R. capsulatus, isolated using a nifH::lac fusion, showed that ammonia repression involves components of the adenylylation cascade (e.g., the glnB gene) as in Klebsiella. Oxygen control of nif gene expression was linked to the degree of DNA supercoiling by showing that inhibitors of DNA gyrase prevent nif gene expression. Attempts to relate the degree of supercoiling or of gyrase activity in R. capsulatus to the oxygen level have been inconclusive. The gene encoding the DNA gyrase B subunit of R. capsulatus was cloned. The genes encoding the two large subunits of R. Capsulatus RNA polymerase were cloned, mapped and partially sequenced. The nif genes and the glnA gene are being physically mapped on the R. capsulatus chromosome using pulsed field electrophoresis to separate large segments of the bacterial chromosome. A start was made on the purification of R. capsulatus RNA polymerase. A system for expressing genes in R. capsulatus was applied to nifR4; analogous constructions will be made for nifR1, nifR2 and nifA.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    • Plant productivity is predicted to increase in northern latitudes due to climate warming; however, this may depend on whether biological N-fixation also increases. We evaluated how variation in temperature and light affect N-fixation by two boreal feather mosses, Pleurozium schreberi and Hylocomi...

  17. Combined inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases phosphorus use efficiency for symbiotic nitrogen fixation in common bean (Phaseolus vulgaris L.)

    PubMed Central

    Tajini, Fatma; Trabelsi, Mustapha; Drevon, Jean-Jacques

    2011-01-01

    This study compared the response of common bean (Phaseolus vulgaris L.) to arbuscular mycorrhizal fungi (AMF) and rhizobia strain inoculation. Two common bean genotypes i.e. CocoT and Flamingo varying in their effectiveness for nitrogen fixation were inoculated with Glomus intraradices and Rhizobium tropici CIAT899, and grown for 50 days in soil–sand substrate in glasshouse conditions. Inoculation of common bean plants with the AM fungi resulted in a significant increase in nodulation compared to plants without inoculation. The combined inoculation of AM fungi and rhizobia significantly increased various plant growth parameters compared to simple inoculated plants. In addition, the combined inoculation of AM fungi and rhizobia resulted in significantly higher nitrogen and phosphorus accumulation in the shoots of common bean plants and improved phosphorus use efficiency compared with their controls, which were not dually inoculated. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve the efficiency in phosphorus use for symbiotic nitrogen fixation especially under phosphorus deficiency. PMID:23961175

  18. Combined inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases phosphorus use efficiency for symbiotic nitrogen fixation in common bean (Phaseolus vulgaris L.).

    PubMed

    Tajini, Fatma; Trabelsi, Mustapha; Drevon, Jean-Jacques

    2012-04-01

    This study compared the response of common bean (Phaseolus vulgaris L.) to arbuscular mycorrhizal fungi (AMF) and rhizobia strain inoculation. Two common bean genotypes i.e. CocoT and Flamingo varying in their effectiveness for nitrogen fixation were inoculated with Glomus intraradices and Rhizobium tropici CIAT899, and grown for 50 days in soil-sand substrate in glasshouse conditions. Inoculation of common bean plants with the AM fungi resulted in a significant increase in nodulation compared to plants without inoculation. The combined inoculation of AM fungi and rhizobia significantly increased various plant growth parameters compared to simple inoculated plants. In addition, the combined inoculation of AM fungi and rhizobia resulted in significantly higher nitrogen and phosphorus accumulation in the shoots of common bean plants and improved phosphorus use efficiency compared with their controls, which were not dually inoculated. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve the efficiency in phosphorus use for symbiotic nitrogen fixation especially under phosphorus deficiency. PMID:23961175

  19. Dinitrogen fixation and dissolved organic nitrogen fueled primary production and particulate export during the VAHINE mesocosms experiment (New Caledonia lagoon)

    NASA Astrophysics Data System (ADS)

    Berthelot, H.; Moutin, T.; L'Helguen, S.; Leblanc, K.; Hélias, S.; Grosso, O.; Leblond, N.; Charrière, B.; Bonnet, S.

    2015-03-01

    In the oligotrophic ocean characterized by nitrate (NO3-) depletion in surface waters, dinitrogen (N2) fixation and dissolved organic nitrogen (DON) can represent significant nitrogen (N) sources for the ecosystem. Here we deployed in New Caledonia large in situ mesocosms in order to investigate (1) the contribution of N2 fixation and DON use to primary production (PP) and particle export and (2) the fate of the freshly produced particulate organic N (PON) i.e. whether it is preferentially accumulated and recycled in the water column or exported out of the system. The mesocosms were fertilized with phosphate (P) in order to prevent P-limitation and promote N2 fixation. The diazotrophic community was dominated by diatoms-diazotrophs associations (DDAs) during the first part of the experiment for 10 days (P1) followed by the unicellular N2-fixing cyanobacteria UCYN-C the 9 last days (P2) of the experiment. N2 fixation rates averaged 9.8 ± 4.0 and 27.7 ± 8.6 nM d-1 during P1 and P2, respectively. NO3- concentrations (< 40 nM) in the mesocosms were a negligible source of N indicating that N2 fixation was the main driver of new production all along the experiment. The contribution of v fixation to PP was not significantly different (p > 0.05) during P1 (9.0 ± 3.3%) and P2 (12.6 ± 6.1%). However, the e ratio that quantifies the efficiency of a system to export particulate organic carbon (POCexport) compared to PP (e ratio = POCexport/PP) was significantly higher (p < 0.05) during P2 (39.7 ± 24.9%) than during P1 (23.9 ± 20.2%) indicating that the production sustained by UCYN-C was more efficient at promoting C export than the production sustained by DDAs. During P1, PON was stable and the total amount of N provided by N2 fixation (0.10 ± 0.02 μM) was not significantly different (p > 0.05) from the total amount of PON exported (0.10 ± 0.04 μM), suggesting a rapid and probably direct export of the recently fixed N2 by the DDAs. During P2, both PON concentrations

  20. Inhibition of nitrogen fixation in symbiotic Medicago truncatula upon Cd exposure is a local process involving leghaemoglobin

    PubMed Central

    Marino, Daniel

    2013-01-01

    Leguminous biological nitrogen fixation (BNF) is very sensitive to environmental fluctuations. It is still contentious how BNF is regulated under stress conditions. The local or systemic control of BNF and the role played by reactive oxygen species (ROS) in such regulation have still not been elucidated completely. Cadmium, which belongs to the so-called heavy metals, is one of the most toxic substances released into the environment. The mechanisms involved in Cd toxicity are still not completely understood but the overproduction of ROS is one of its characteristic symptoms. In this work, we used a split-root system approach to study nodule BNF and the antioxidant machinery’s response to the application of a mild Cd treatment on one side of a nodulated Medicago truncatula root system. Cd induced the majority of nodule antioxidants without generating any oxidative damage. Cd treatment also provoked BNF inhibition exclusively in nodules directly exposed to Cd, without provoking any effect on plant shoot biomass or chlorophyll content. The overall data suggest that the decline in BNF was not due to a generalized breakdown of the plant but to control exerted through leghaemoglobin/oxygen availability, affecting nitrogenase function. PMID:24151304

  1. A JAZ Protein in Astragalus sinicus Interacts with a Leghemoglobin through the TIFY Domain and Is Involved in Nodule Development and Nitrogen Fixation.

    PubMed

    Li, Yixing; Xu, Meng; Wang, Ning; Li, Youguo

    2015-01-01

    Leghemoglobins (Lbs) play an important role in legumes-rhizobia symbiosis. Lbs bind O2 and protect nitrogenase activity from damage by O2 in nodules, therefore, they are regarded as a marker of active nitrogen fixation in nodules. Additionally, Lbs are involved in the nitric oxide (NO) signaling pathway, acting as a NO scavenger during nodule development and nitrogen fixation. However, regulators responsible for Lb expression and modulation of Lb activity have not been characterized. In our previous work, a Jasmonate-Zim-domain (JAZ) protein interacting with a Lb (AsB2510) in Astragalus sinicus was identified and designated AsJAZ1. In this study, the interaction between AsJAZ1 and AsB2510 was verified using a yeast two-hybrid system and in vitro Glutathione S-transferase (GST) pull-down assays, resulting in identification of the interaction domain as a TIFY (previously known as zinc-finger protein expressed in inflorescence meristem, ZIM) domain. TIFY domain is named after the most conserved amino acids within the domain. Bimolecular fluorescence complementation (BiFC) was used to confirm the interaction between AsJAZ1 and AsB2510 in tobacco cells, demonstrating that AsJAZ1-AsB2510 interaction was localized to the cell membrane and cytoplasm. Furthermore, the expression patterns and the symbiotic phenotypes of AsJAZ1 were investigated. Knockdown of AsJAZ1 expression via RNA interference led to decreased number of nodules, abnormal development of bacteroids, accumulation of poly-x-hydroxybutyrate (PHB) and loss of nitrogenase activity. Taken together, our results suggest that AsJAZ1 interacts with AsB2510 and participates in nodule development and nitrogen fixation. Our results provide novel insights into the functions of Lbs or JAZ proteins during legume-rhizobia symbiosis.

  2. A JAZ Protein in Astragalus sinicus Interacts with a Leghemoglobin through the TIFY Domain and Is Involved in Nodule Development and Nitrogen Fixation.

    PubMed

    Li, Yixing; Xu, Meng; Wang, Ning; Li, Youguo

    2015-01-01

    Leghemoglobins (Lbs) play an important role in legumes-rhizobia symbiosis. Lbs bind O2 and protect nitrogenase activity from damage by O2 in nodules, therefore, they are regarded as a marker of active nitrogen fixation in nodules. Additionally, Lbs are involved in the nitric oxide (NO) signaling pathway, acting as a NO scavenger during nodule development and nitrogen fixation. However, regulators responsible for Lb expression and modulation of Lb activity have not been characterized. In our previous work, a Jasmonate-Zim-domain (JAZ) protein interacting with a Lb (AsB2510) in Astragalus sinicus was identified and designated AsJAZ1. In this study, the interaction between AsJAZ1 and AsB2510 was verified using a yeast two-hybrid system and in vitro Glutathione S-transferase (GST) pull-down assays, resulting in identification of the interaction domain as a TIFY (previously known as zinc-finger protein expressed in inflorescence meristem, ZIM) domain. TIFY domain is named after the most conserved amino acids within the domain. Bimolecular fluorescence complementation (BiFC) was used to confirm the interaction between AsJAZ1 and AsB2510 in tobacco cells, demonstrating that AsJAZ1-AsB2510 interaction was localized to the cell membrane and cytoplasm. Furthermore, the expression patterns and the symbiotic phenotypes of AsJAZ1 were investigated. Knockdown of AsJAZ1 expression via RNA interference led to decreased number of nodules, abnormal development of bacteroids, accumulation of poly-x-hydroxybutyrate (PHB) and loss of nitrogenase activity. Taken together, our results suggest that AsJAZ1 interacts with AsB2510 and participates in nodule development and nitrogen fixation. Our results provide novel insights into the functions of Lbs or JAZ proteins during legume-rhizobia symbiosis. PMID:26460857

  3. Plant Growth-Promoting Rhizobacteria Inoculation to Enhance Vegetative Growth, Nitrogen Fixation and Nitrogen Remobilisation of Maize under Greenhouse Conditions.

    PubMed

    Kuan, Khing Boon; Othman, Radziah; Abdul Rahim, Khairuddin; Shamsuddin, Zulkifli H

    2016-01-01

    Plant growth-promoting rhizobacteria (PGPR) may provide a biological alternative to fix atmospheric N2 and delay N remobilisation in maize plant to increase crop yield, based on an understanding that plant-N remobilisation is directly correlated to its plant senescence. Thus, four PGPR strains were selected from a series of bacterial strains isolated from maize roots at two locations in Malaysia. The PGPR strains were screened in vitro for their biochemical plant growth-promoting (PGP) abilities and plant growth promotion assays. These strains were identified as Klebsiella sp. Br1, Klebsiella pneumoniae Fr1, Bacillus pumilus S1r1 and Acinetobacter sp. S3r2 and a reference strain used was Bacillus subtilis UPMB10. All the PGPR strains were tested positive for N2 fixation, phosphate solubilisation and auxin production by in vitro tests. In a greenhouse experiment with reduced fertiliser-N input (a third of recommended fertiliser-N rate), the N2 fixation abilities of PGPR in association with maize were determined by 15N isotope dilution technique at two harvests, namely, prior to anthesis (D50) and ear harvest (D65). The results indicated that dry biomass of top, root and ear, total N content and bacterial colonisations in non-rhizosphere, rhizosphere and endosphere of maize roots were influenced by PGPR inoculation. In particular, the plants inoculated with B. pumilus S1r1 generally outperformed those with the other treatments. They produced the highest N2 fixing capacity of 30.5% (262 mg N2 fixed plant-1) and 25.5% (304 mg N2 fixed plant-1) of the total N requirement of maize top at D50 and D65, respectively. N remobilisation and plant senescence in maize were delayed by PGPR inoculation, which is an indicative of greater grain production. This is indicated by significant interactions between PGPR strains and time of harvests for parameters on N uptake and at. % 15Ne of tassel. The phenomenon is also supported by the lower N content in tassels of maize treated with

  4. Plant Growth-Promoting Rhizobacteria Inoculation to Enhance Vegetative Growth, Nitrogen Fixation and Nitrogen Remobilisation of Maize under Greenhouse Conditions.

    PubMed

    Kuan, Khing Boon; Othman, Radziah; Abdul Rahim, Khairuddin; Shamsuddin, Zulkifli H

    2016-01-01

    Plant growth-promoting rhizobacteria (PGPR) may provide a biological alternative to fix atmospheric N2 and delay N remobilisation in maize plant to increase crop yield, based on an understanding that plant-N remobilisation is directly correlated to its plant senescence. Thus, four PGPR strains were selected from a series of bacterial strains isolated from maize roots at two locations in Malaysia. The PGPR strains were screened in vitro for their biochemical plant growth-promoting (PGP) abilities and plant growth promotion assays. These strains were identified as Klebsiella sp. Br1, Klebsiella pneumoniae Fr1, Bacillus pumilus S1r1 and Acinetobacter sp. S3r2 and a reference strain used was Bacillus subtilis UPMB10. All the PGPR strains were tested positive for N2 fixation, phosphate solubilisation and auxin production by in vitro tests. In a greenhouse experiment with reduced fertiliser-N input (a third of recommended fertiliser-N rate), the N2 fixation abilities of PGPR in association with maize were determined by 15N isotope dilution technique at two harvests, namely, prior to anthesis (D50) and ear harvest (D65). The results indicated that dry biomass of top, root and ear, total N content and bacterial colonisations in non-rhizosphere, rhizosphere and endosphere of maize roots were influenced by PGPR inoculation. In particular, the plants inoculated with B. pumilus S1r1 generally outperformed those with the other treatments. They produced the highest N2 fixing capacity of 30.5% (262 mg N2 fixed plant-1) and 25.5% (304 mg N2 fixed plant-1) of the total N requirement of maize top at D50 and D65, respectively. N remobilisation and plant senescence in maize were delayed by PGPR inoculation, which is an indicative of greater grain production. This is indicated by significant interactions between PGPR strains and time of harvests for parameters on N uptake and at. % 15Ne of tassel. The phenomenon is also supported by the lower N content in tassels of maize treated with

  5. Plant Growth-Promoting Rhizobacteria Inoculation to Enhance Vegetative Growth, Nitrogen Fixation and Nitrogen Remobilisation of Maize under Greenhouse Conditions

    PubMed Central

    Kuan, Khing Boon; Othman, Radziah; Abdul Rahim, Khairuddin; Shamsuddin, Zulkifli H.

    2016-01-01

    Plant growth-promoting rhizobacteria (PGPR) may provide a biological alternative to fix atmospheric N2 and delay N remobilisation in maize plant to increase crop yield, based on an understanding that plant-N remobilisation is directly correlated to its plant senescence. Thus, four PGPR strains were selected from a series of bacterial strains isolated from maize roots at two locations in Malaysia. The PGPR strains were screened in vitro for their biochemical plant growth-promoting (PGP) abilities and plant growth promotion assays. These strains were identified as Klebsiella sp. Br1, Klebsiella pneumoniae Fr1, Bacillus pumilus S1r1 and Acinetobacter sp. S3r2 and a reference strain used was Bacillus subtilis UPMB10. All the PGPR strains were tested positive for N2 fixation, phosphate solubilisation and auxin production by in vitro tests. In a greenhouse experiment with reduced fertiliser-N input (a third of recommended fertiliser-N rate), the N2 fixation abilities of PGPR in association with maize were determined by 15N isotope dilution technique at two harvests, namely, prior to anthesis (D50) and ear harvest (D65). The results indicated that dry biomass of top, root and ear, total N content and bacterial colonisations in non-rhizosphere, rhizosphere and endosphere of maize roots were influenced by PGPR inoculation. In particular, the plants inoculated with B. pumilus S1r1 generally outperformed those with the other treatments. They produced the highest N2 fixing capacity of 30.5% (262 mg N2 fixed plant−1) and 25.5% (304 mg N2 fixed plant−1) of the total N requirement of maize top at D50 and D65, respectively. N remobilisation and plant senescence in maize were delayed by PGPR inoculation, which is an indicative of greater grain production. This is indicated by significant interactions between PGPR strains and time of harvests for parameters on N uptake and at. % 15Ne of tassel. The phenomenon is also supported by the lower N content in tassels of maize treated

  6. Contrasted reactivity to oxygen tensions in Frankia sp. strain CcI3 throughout nitrogen fixation and assimilation.

    PubMed

    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

  7. 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

  8. Rates of fixation by lightning of carbon and nitrogen in possible primitive atmospheres

    NASA Technical Reports Server (NTRS)

    Chameides, W. L.; Walker, J. C. G.

    1981-01-01

    A thermochemical-hydrodynamic model of the production of trace species by electrical discharges has been used to estimate the rates of fixation of C and N by lightning in the primitive atmosphere. Calculations for various possible mixtures of CH4, CO2, CO, N2, H2, and H2O reveal that the prime species produced were probably HCN and NO and that the key parameter determining the rates of fixation was the ratio of C atoms to O atoms in the atmosphere. Atmospheres with C more abundant than O have large HCN fixation rates, in excess of 10 to the 17th molecules/J, but small NO yields. However, when O is more abundant than C, the NO fixation rate approaches 10 to the 17th molecules/J while the HCN yield is small. The implications for the evolution of life are discussed.

  9. 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

  10. 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

    2015-12-01

    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.

  11. Site-specific recombination of nitrogen-fixation genes in cyanobacteria by XisF-XisH-XisI complex: structures and models

    PubMed Central

    Golden, James W.; Pascual, Jaime; Xu, Dong; Cheltsov, Anton

    2014-01-01

    Nitrogen fixation is an important process that converts atmospheric gaseous nitrogen, a form plants cannot utilize, into ammonia that can be easily assimilated. Large serine recombinase XisF (fdxN element site-specific recombinase), together with controlling factors XisH and XisI, plays a critical role in the expression of nitrogen fixation genes of certain Anabaena and Nostoc species of cyanobacteria. All three proteins are required to excise the fdxN DNA element from the chromosome in differentiating heterocysts for the expression of nitrogen fixation related genes. We report the first crystal structures of XisH and XisI proteins, both adopting novel protein folds. Based on the analysis of their sequences and structures, we propose that XisH and XisI proteins function as endonucleases and recombination directionality factors (RDFs), respectively. PMID:25179344

  12. Response of nitrogen fixation in relation to nodule carbohydrate metabolism in Medicago ciliaris lines subjected to salt stress.

    PubMed

    Ben Salah, Imène; Albacete, Alfonso; Martínez Andújar, Cristina; Haouala, Rabiaa; Labidi, Nehla; Zribi, Fethia; Martinez, Vicente; Pérez-Alfocea, Francisco; Abdelly, Chedly

    2009-03-15

    The effect of salt stress on nitrogen fixation, in relation to sucrose transport towards nodules and other sink organs and the potential of sucrose breakdown by nodules, was investigated in two lines of Medicago ciliaris. Under salt stress conditions, the two lines showed a decrease of total biomass production, but TNC 1.8 was less affected by salt than TNC 11.9. The chlorophyll content was not changed in TNC 1.8, in contrast to TNC 11.9. Shoot, root, and nodule biomass were also affected in the two lines, but TNC 1.8 exhibited the higher potentialities of biomass production of these organs. Nitrogen fixation also decreased in the two lines, and was more sensitive to salt than growth parameters. TNC 1.8 consistently exhibited the higher values of nitrogen fixation. Unlike nodules, leaves of both lines were well supplied in nutrients with some exceptions. Specifically, the calcium content decreased in the sensitive line leaves, and the nodule magnesium content was not changed in either line. The tolerant line accumulated more sodium in its leaves. The two lines did not show any differences in the nodule sodium content. Sucrose allocation towards nodules was affected by salt in the two lines, but this constraint did not seem to affect the repartition of sucrose between sink organs. Salt stress induced perturbations in nodule sucrolytic activities in the two lines. It inhibited sucrose synthase, but the inhibition was more marked in TNC 11.9; alkaline/neutral activity was not altered in TNC 1.8, whereas it decreased more than half in TNC 11.9. Thus, the relative tolerance of TNC 1.8 to salt stress could be attributed to a better use of these photoassimilates by nodules and a better supply of bacteroids in malate. The hypothesis of a competition for sucrose between nodules and other sink organs under salt stress could not be verified. PMID:18804311

  13. Nitrogen fixation at passivated Fe nanoclusters supported by an oxide surface: Identification of viable reaction routes using density functional calculations

    NASA Astrophysics Data System (ADS)

    Šljivančanin, Željko; Brune, Harald; Pasquarello, Alfredo

    2009-08-01

    Using density-functional calculations, we investigate the possibility of ammonia synthesis at supported Fe nanoclusters along catalytic routes closely resembling those in biological nitrogen fixation. To achieve similar catalytic conditions as at the active site of the enzyme nitrogenase, the clusters are passivated with either S or N atoms. From calculated potential-energy profiles for the N2 hydrogenation, we find that low-temperature synthesis of ammonia is viable at the clusters passivated by N atoms due to the strong binding energy of the N2 molecule in the initial adsorption step.

  14. Rhythm of carbon and nitrogen fixation in unicellular cyanobacteria under turbulent and highly aerobic conditions.

    PubMed

    Krishnakumar, S; Gaudana, Sandeep B; Viswanathan, Ganesh A; Pakrasi, Himadri B; Wangikar, Pramod P

    2013-09-01

    Nitrogen fixing cyanobacteria are being increasingly explored for nitrogenase-dependent hydrogen production. Commercial success however will depend on the ability to grow these cultures at high cell densities. Photo-limitation at high cell densities leads to hindered photoautotrophic growth while turbulent conditions, which simulate flashing light effect, can lead to oxygen toxicity to the nitrogenase enzyme. Cyanothece sp. strain ATCC 51142, a known hydrogen producer, is reported to grow and fix nitrogen under moderately oxic conditions in shake flasks. In this study, we explore the growth and nitrogen fixing potential of this organism under turbulent conditions with volumetric oxygen mass transfer coefficient (KL a) values that are up to 20-times greater than in shake flasks. In a stirred vessel, the organism grows well in turbulent regime possibly due to a simulated flashing light effect with optimal growth at Reynolds number of approximately 35,000. A respiratory burst lasting for about 4 h creates anoxic conditions intracellularly with near saturating levels of dissolved oxygen in the extracellular medium. This is concomitant with complete exhaustion of intracellular glycogen storage and upregulation of nifH and nifX, the genes encoding proteins of the nitrogenase complex. Further, the rhythmic oscillations in exhaust gas CO2 and O2 profiles synchronize faithfully with those in biochemical parameters and gene expression thereby serving as an effective online monitoring tool. These results will have important implications in potential commercial success of nitrogenase-dependent hydrogen production by cyanobacteria.

  15. Simultaneous quantification of active carbon- and nitrogen-fixing communities and estimation of fixation rates using fluorescence in situ hybridization and flow cytometry.

    PubMed

    McInnes, Allison S; Shepard, Alicia K; Raes, Eric J; Waite, Anya M; Quigg, Antonietta

    2014-11-01

    Understanding the interconnectivity of oceanic carbon and nitrogen cycles, specifically carbon and nitrogen fixation, is essential in elucidating the fate and distribution of carbon in the ocean. Traditional techniques measure either organism abundance or biochemical rates. As such, measurements are performed on separate samples and on different time scales. Here, we developed a method to simultaneously quantify organisms while estimating rates of fixation across time and space for both carbon and nitrogen. Tyramide signal amplification fluorescence in situ hybridization (TSA-FISH) of mRNA for functionally specific oligonucleotide probes for rbcL (ribulose-1,5-bisphosphate carboxylase/oxygenase; carbon fixation) and nifH (nitrogenase; nitrogen fixation) was combined with flow cytometry to measure abundance and estimate activity. Cultured samples representing a diversity of phytoplankton (cyanobacteria, coccolithophores, chlorophytes, diatoms, and dinoflagellates), as well as environmental samples from the open ocean (Gulf of Mexico, USA, and southeastern Indian Ocean, Australia) and an estuary (Galveston Bay, Texas, USA), were successfully hybridized. Strong correlations between positively tagged community abundance and (14)C/(15)N measurements are presented. We propose that these methods can be used to estimate carbon and nitrogen fixation in environmental communities. The utilization of mRNA TSA-FISH to detect multiple active microbial functions within the same sample will offer increased understanding of important biogeochemical cycles in the ocean.

  16. Comparative genomic analysis of N2-fixing and non-N2-fixing Paenibacillus spp.: organization, evolution and expression of the nitrogen fixation genes.

    PubMed

    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

    2014-03-01

    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 σ(70)-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.

  17. Comparative genomic analysis of N2-fixing and non-N2-fixing Paenibacillus spp.: organization, evolution and expression of the nitrogen fixation genes.

    PubMed

    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

    2014-03-01

    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 σ(70)-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

  18. 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

  19. sup 15 NO sub 3 assimilation and its inhibitory effect on symbiotic nitrogen fixation in peanut

    SciTech Connect

    Stanfill, S.B.; Wells, R.; Israel, D.W.; Rufty, T.W. )

    1990-05-01

    To assess the inhibitory effect of nitrate on the contribution of symbiotic N fixation to total plant N, cultivars of different nodulation capacity were monitored in a growth chamber study. Plants inoculated with Bradyrhizobium sp. (Arachis) strain NC 70.1 were grown in a nutrient solution containing 0, 2.5, 5 or 10 mM NO{sub 3} enriched with 2.5 atom % {sup 15}N. Plant harvests at 30 and 60 DAP provided tissue for measurement of growth, total N, NO{sub 3} and {sup 15}N partitioning. Nitrogenase activity was estimated via C{sub 2}H{sub 2} reduction. Data indicates that plant growth was associated to NO{sub 3} concentration. Average nodule weight and N plant{sup {minus}1} decreased in excess of 2.5mM NO{sub 3}. Specific nitrogenase activity diminished markedly with application of NO{sub 3} with a decline from 40.2 to 25.0 {mu}moles C{sub 2}H{sub 2} g hr{sup {minus}1} at 0 and 2.5mM NO{sub 3}, respectively. Nitrate and fixed N assimilation patterns will be elucidated by {sup 15}N analysis.

  20. 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

  1. Effect on Heterocyst Differentiation of Nitrogen Fixation in Vegetative Cells of the Cyanobacterium Anabaena variabilis ATCC 29413

    PubMed Central

    Thiel, Teresa; Pratte, Brenda

    2001-01-01

    Heterocysts are terminally differentiated cells of some filamentous cyanobacteria that fix nitrogen for the entire filament under oxic growth conditions. Anabaena variabilis ATCC 29413 is unusual in that it has two Mo-dependent nitrogenases; one, called Nif1, functions in heterocysts, while the second, Nif2, functions under anoxic conditions in vegetative cells. Both nitrogenases depended on expression of the global regulatory protein NtcA. It has long been thought that a product of nitrogen fixation in heterocysts plays a role in maintenance of the spaced pattern of heterocyst differentiation. This model assumes that each cell in a filament senses its own environment in terms of nitrogen sufficiency and responds accordingly in terms of differentiation. Expression of the Nif2 nitrogenase under anoxic conditions in vegetative cells was sufficient to support long-term growth of a nif1 mutant; however, that expression did not prevent differentiation of heterocysts and expression of the nif1 nitrogenase in either the nif1 mutant or the wild-type strain. This suggested that the nitrogen sufficiency of individual cells in the filament did not affect the signal that induces heterocyst differentiation. Perhaps there is a global mechanism by which the filament senses nitrogen sufficiency or insufficiency based on the external availability of fixed nitrogen. The filament would then respond by producing heterocyst differentiation signals that affect the entire filament. This does not preclude cell-to-cell signaling in the maintenance of heterocyst pattern but suggests that overall control of the process is not controlled by nitrogen insufficiency of individual cells. PMID:11114927

  2. Cyanobacterial symbionts diverged in the late Cretaceous towards lineage-specific nitrogen fixation factories in single-celled phytoplankton

    PubMed Central

    Cornejo-Castillo, Francisco M.; Cabello, Ana M.; Salazar, Guillem; Sánchez-Baracaldo, Patricia; Lima-Mendez, Gipsi; Hingamp, Pascal; Alberti, Adriana; Sunagawa, Shinichi; Bork, Peer; de Vargas, Colomban; Raes, Jeroen; Bowler, Chris; Wincker, Patrick; Zehr, Jonathan P.; Gasol, Josep M.; Massana, Ramon; Acinas, Silvia G.

    2016-01-01

    The unicellular cyanobacterium UCYN-A, one of the major contributors to nitrogen fixation in the open ocean, lives in symbiosis with single-celled phytoplankton. UCYN-A includes several closely related lineages whose partner fidelity, genome-wide expression and time of evolutionary divergence remain to be resolved. Here we detect and distinguish UCYN-A1 and UCYN-A2 lineages in symbiosis with two distinct prymnesiophyte partners in the South Atlantic Ocean. Both symbiotic systems are lineage specific and differ in the number of UCYN-A cells involved. Our analyses infer a streamlined genome expression towards nitrogen fixation in both UCYN-A lineages. Comparative genomics reveal a strong purifying selection in UCYN-A1 and UCYN-A2 with a diversification process ∼91 Myr ago, in the late Cretaceous, after the low-nutrient regime period occurred during the Jurassic. These findings suggest that UCYN-A diversified in a co-evolutionary process, wherein their prymnesiophyte partners acted as a barrier driving an allopatric speciation of extant UCYN-A lineages. PMID:27002549

  3. Cyanobacterial symbionts diverged in the late Cretaceous towards lineage-specific nitrogen fixation factories in single-celled phytoplankton.

    PubMed

    Cornejo-Castillo, Francisco M; Cabello, Ana M; Salazar, Guillem; Sánchez-Baracaldo, Patricia; Lima-Mendez, Gipsi; Hingamp, Pascal; Alberti, Adriana; Sunagawa, Shinichi; Bork, Peer; de Vargas, Colomban; Raes, Jeroen; Bowler, Chris; Wincker, Patrick; Zehr, Jonathan P; Gasol, Josep M; Massana, Ramon; Acinas, Silvia G

    2016-03-22

    The unicellular cyanobacterium UCYN-A, one of the major contributors to nitrogen fixation in the open ocean, lives in symbiosis with single-celled phytoplankton. UCYN-A includes several closely related lineages whose partner fidelity, genome-wide expression and time of evolutionary divergence remain to be resolved. Here we detect and distinguish UCYN-A1 and UCYN-A2 lineages in symbiosis with two distinct prymnesiophyte partners in the South Atlantic Ocean. Both symbiotic systems are lineage specific and differ in the number of UCYN-A cells involved. Our analyses infer a streamlined genome expression towards nitrogen fixation in both UCYN-A lineages. Comparative genomics reveal a strong purifying selection in UCYN-A1 and UCYN-A2 with a diversification process ∼91 Myr ago, in the late Cretaceous, after the low-nutrient regime period occurred during the Jurassic. These findings suggest that UCYN-A diversified in a co-evolutionary process, wherein their prymnesiophyte partners acted as a barrier driving an allopatric speciation of extant UCYN-A lineages.

  4. 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.

  5. Cyanobacterial symbionts diverged in the late Cretaceous towards lineage-specific nitrogen fixation factories in single-celled phytoplankton.

    PubMed

    Cornejo-Castillo, Francisco M; Cabello, Ana M; Salazar, Guillem; Sánchez-Baracaldo, Patricia; Lima-Mendez, Gipsi; Hingamp, Pascal; Alberti, Adriana; Sunagawa, Shinichi; Bork, Peer; de Vargas, Colomban; Raes, Jeroen; Bowler, Chris; Wincker, Patrick; Zehr, Jonathan P; Gasol, Josep M; Massana, Ramon; Acinas, Silvia G

    2016-01-01

    The unicellular cyanobacterium UCYN-A, one of the major contributors to nitrogen fixation in the open ocean, lives in symbiosis with single-celled phytoplankton. UCYN-A includes several closely related lineages whose partner fidelity, genome-wide expression and time of evolutionary divergence remain to be resolved. Here we detect and distinguish UCYN-A1 and UCYN-A2 lineages in symbiosis with two distinct prymnesiophyte partners in the South Atlantic Ocean. Both symbiotic systems are lineage specific and differ in the number of UCYN-A cells involved. Our analyses infer a streamlined genome expression towards nitrogen fixation in both UCYN-A lineages. Comparative genomics reveal a strong purifying selection in UCYN-A1 and UCYN-A2 with a diversification process ∼91 Myr ago, in the late Cretaceous, after the low-nutrient regime period occurred during the Jurassic. These findings suggest that UCYN-A diversified in a co-evolutionary process, wherein their prymnesiophyte partners acted as a barrier driving an allopatric speciation of extant UCYN-A lineages. PMID:27002549

  6. A Novel Fixation System for Acetabular Quadrilateral Plate Fracture: A Comparative Biomechanical Study

    PubMed Central

    Zha, Guo-Chun; Sun, Jun-Ying; Dong, Sheng-Jie; Zhang, Wen; Luo, Zong-Ping

    2015-01-01

    This study aims to assess the biomechanical properties of a novel fixation system (named AFRIF) and to compare it with other five different fixation techniques for quadrilateral plate fractures. This in vitro biomechanical experiment has shown that the multidirectional titanium fixation (MTF) and pelvic brim long screws fixation (PBSF) provided the strongest fixation for quadrilateral plate fracture; the better biomechanical performance of the AFRIF compared with the T-shaped plate fixation (TPF), L-shaped plate fixation (LPF), and H-shaped plate fixation (HPF); AFRIF gives reasonable stability of treatment for quadrilateral plate fracture and may offer a better solution for comminuted quadrilateral plate fractures or free floating medial wall fracture and be reliable in preventing protrusion of femoral head. PMID:25802849

  7. Organization and regulation of the genes for nitrogen fixation in Rhodopseudomonas capsulata: Progress report, June 5, 1987-June 4, 1988

    SciTech Connect

    Haselkorn, R.

    1988-02-01

    We have cloned a number of fragments of DNA containing genes necessary for nitrogen fixation from the photosynthetic bacterium Rhodobacter capsulatus. The nif genes are locally clustered but the clusters are on non-neighboring DNA restriction fragments. We sought to determine the physical linkage of these fragments, to determine their relationship with the corresponding nif genes of Klebsiella, and to determine the nucleotide sequence of some of the fragments. So far we have identified six or seven regulatory genes among these, using a nifH::lac fusion. Four of the regulatory genes are required for expression of nifH. Two of these, nifR1 and nifR2, have sequences homologous to ntrC and ntrB of enteric bacteria. A third, nifR4, has sequence homology, in the C-terminal region, to the ntrA genes of Rhizobium and Klebsiella. Constitutive expression of nifR4 in R. capsulatus, from a plasmid clone, complemented a nifR4 chromosomal mutant but not a nifR1 mutant. Moreover, both oxygen and ammonia regulation of nitrogenase were maintained under these conditions. These results are consistent with a model requiring both nifR1 and nifR4 for nitrogenase gene expression; they rule out our earlier suggestion that nifR1 is needed only to turn on nifR4. Current efforts are focused on the purification of RNA polymerase and the products of nifR1, nifR2, and nifR4 to study nif gene transcription in vitro, particularly with the goal of determining the role of DNA supercoiling in transcription.

  8. Association of nitrogen fixation to water use efficiency and yield traits of peanut

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Improvement of N2 fixation might be an effective strategy in peanut breeding for high yield under drought stress conditions. However, under water limited conditions peanut varieties having high water-use efficiency (WUE) are favorable. A pot experiment was conducted under greenhouse conditions at ...

  9. Effect of nitrogen fixation, nitrogen fertilization and viral infection on yield, tannin and protein contents and in vitro protein digestibility of faba bean.

    PubMed

    Babiker, E E; el Sheikh, E A; Osman, A J; el Tinay, A H

    1995-04-01

    A field investigation of two faba bean cultivars (cv.), Agabat and Silaim, showed that bean yellow mosaic virus (BYMV) infection reduced (p < or = 0.001) yield (Kg/ha), protein content and in vitro protein digestibility (IVPD) but increased (p < or = 0.05) tannin content (mg/100 ml). Nitrogen fertilization with viral infection significantly reduced yield and IVPD for cv. Silaim and increased (p < or = 0.05) protein and tannin contents. Nitrogen fertilization alone was found to increase (p < or = 0.05) yield, protein and tannin contents but slightly reduced IVPD. Rhizobium inoculation with viral infection significantly decreased yield per unit area, protein content and IVPD, but increased (p < or = 0.05) tannin content. Rhizobium inoculation alone significantly increased (p < or = 0.001) yield and tannin content and slightly increased protein content but decreased IVPD. The results indicated that nitrogen fertilization or nitrogen fixation increased yield, protein and tannin contents and decreased IVPD. Viral infection had an adverse effect on yield, protein content and IVPD but had no effect on tannin content.

  10. Anoxygenic Photosynthesis and Nitrogen Fixation by a Microbial Mat Community in a Bahamian Hypersaline Lagoon

    PubMed Central

    Pinckney, J. L.; Paerl, H. W.

    1997-01-01

    Simultaneous measurements of photosynthesis (both oxygenic and anoxygenic) and N(inf2) fixation were conducted to discern the relationships between photosynthesis, N(inf2) fixation, and environmental factors potentially regulating these processes in microbial mats in a tropical hypersaline lagoon (Salt Pond, San Salvador Island, Bahamas). Major photoautotrophs included cyanobacteria, purple phototrophic bacteria, and diatoms. Chemosystematic photopigments were used as indicators of the relative abundance of mat phototrophs. Experimental manipulations consisted of light and dark incubations of intact mat samples exposed to the photosystem II inhibitor DCMU [3-(3,4-dichlorophenyl)-1,1-dimethylurea], a dissolved organic carbon source (D-glucose), and normal seawater (37(permil)). Photosynthetic rates were measured by both O(inf2) and (sup14)C methods, and nitrogenase activity (NA) was estimated by the acetylene reduction assay. Moderate reductions in salinity (from 74 to 37(permil)) had no measurable effect on photosynthesis, O(inf2) consumption, or NA. CO(inf2) fixation in DCMU-amended samples was (symbl)25% of that in the control (nonamended) samples and demonstrated photosynthetic activity by anoxygenic phototrophs. NA in DCMU-amended samples, which was consistently higher (by a factor of 2 to 3) than the other (light and dark) treatments, was also attributed to purple phototrophic bacteria. The ecological implication is that N(inf2) fixation by anoxygenic phototrophs (purple phototrophic bacteria and possibly cyanobacteria) may be regulated by the activity of oxygenic phototrophs (cyanobacteria and diatoms). Consortial interactions that enhance the physiological plasticity of the mat community may be a key for optimizing production, N(inf2) fixation, and persistence in these extreme environments. PMID:16535506

  11. Utilization of net photosynthate for nitrogen fixation and protein production in an annual legume.

    PubMed

    Herridge, D F; Pate, J S

    1977-11-01

    The economy of C and N in nodulated cowpea (Vigna unguiculata [L.] Walp.) was described in terms of fixation of CO(2) and N(2), respiratory losses of C, and the production of dry matter and protein.Net daytime gain of C by the shoot (net photosynthesis) rose to a maximum at flowering and then declined sharply due to abscission of leaves. Maximum N fixation occurred 10 days prior to maximum net photosynthesis. Shedding of nodules reduced fixation to zero by midfruiting. Fifty per cent of the plant's N and 37% of its net photosynthate were assimilated before flowering; 39% of plant N was incorporated into seed dry matter.Respiration of nodules and roots utilized 24% of the C from net photosynthate assimilated over the growth cycle; night respiration of shoots, 20%; dry matter production in seeds, 17%; and dry matter production in other plant parts, 39%. The proportion of net photosynthate translocated to the nodulated root decreased from 41 to 14% during growth. Developing fruits were major competitors for translocate. Nodules consumed 9% of the C from the plant's total net photosynthate, 43% of which was respired, 6% made into dry matter, and 51% returned to the shoot with N fixation products.For every 1 g N fixed, net photosynthate equivalent to 6.8 g carbohydrate was consumed by nodules, 25.7 g carbohydrate by the nodulated root. Translocate was used most efficiently for N fixation in late vegetative growth when nodules were most active and their carbohydrate supply still adequate.During vegetative growth and early flowering (0 to 78 days after sowing) cowpea consumed 17.2 g net photosynthate (as carbohydrate) for every gram of protein synthesized in its shoot. The comparable conversion in seed production was 32.5 g net photosynthate/g seed protein or 6.6 g/g seed dry matter.

  12. Utilization of Net Photosynthate for Nitrogen Fixation and Protein Production in an Annual Legume 1

    PubMed Central

    Herridge, David F.; Pate, John S.

    1977-01-01

    The economy of C and N in nodulated cowpea (Vigna unguiculata [L.] Walp.) was described in terms of fixation of CO2 and N2, respiratory losses of C, and the production of dry matter and protein. Net daytime gain of C by the shoot (net photosynthesis) rose to a maximum at flowering and then declined sharply due to abscission of leaves. Maximum N fixation occurred 10 days prior to maximum net photosynthesis. Shedding of nodules reduced fixation to zero by midfruiting. Fifty per cent of the plant's N and 37% of its net photosynthate were assimilated before flowering; 39% of plant N was incorporated into seed dry matter. Respiration of nodules and roots utilized 24% of the C from net photosynthate assimilated over the growth cycle; night respiration of shoots, 20%; dry matter production in seeds, 17%; and dry matter production in other plant parts, 39%. The proportion of net photosynthate translocated to the nodulated root decreased from 41 to 14% during growth. Developing fruits were major competitors for translocate. Nodules consumed 9% of the C from the plant's total net photosynthate, 43% of which was respired, 6% made into dry matter, and 51% returned to the shoot with N fixation products. For every 1 g N fixed, net photosynthate equivalent to 6.8 g carbohydrate was consumed by nodules, 25.7 g carbohydrate by the nodulated root. Translocate was used most efficiently for N fixation in late vegetative growth when nodules were most active and their carbohydrate supply still adequate. During vegetative growth and early flowering (0 to 78 days after sowing) cowpea consumed 17.2 g net photosynthate (as carbohydrate) for every gram of protein synthesized in its shoot. The comparable conversion in seed production was 32.5 g net photosynthate/g seed protein or 6.6 g/g seed dry matter. PMID:16660179

  13. Nitrogen Fixation by Thermophilic Blue-Green Algae (Cyanobacteria): Temperature Characteristics and Potential Use in Biophotolysis

    PubMed Central

    Miyamoto, Kazuhisa; Hallenbeck, Patrick C.; Benemann, John R.

    1979-01-01

    Thermophilic, nitrogen-fixing, blue-green algae (cyanobacteria) were investigated for use in biophotolysis. Three strains of Mastigocladus laminosus were tested and were found to be equally effective in biophotolysis as judged by nitrogenase activity. The alga, M. laminosus NZ-86-m, which was chosen for further study, grew well in the temperature range from 35 to 50°C, with optimum growth at 45°C, at which temperature acetylene reduction activity was also greatest. The maximum tolerable temperature was 55°C. Acetylene reduction activity was saturated at a light intensity of 1 × 104 ergs cm−2 s−1. Atmospheric oxygen tension was found to be slightly inhibitory to acetylene reduction of both slowly growing and exponentially growing cultures. Nonsterile continuous cultures, which were conducted to test problems of culture maintenance, could be operated for 2 months without any significant decrease in nitrogenase activity or contamination by other algae. Nitrogen-starved cultures of M. laminosus NZ-86-m produced hydrogen at comparable rates to Anabaena cylindrica. The conversion efficiency of light to hydrogen energy at maximum rates of hydrogen production was 2.7%. PMID:16345353

  14. 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

  15. Novel labeling technique illustrates transfer of 15N2 from Sphagnum moss to vascular plants via diazotrophic nitrogen fixation

    NASA Astrophysics Data System (ADS)

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

    2013-12-01

    We used 15N2 gas to trace nitrogen (N) from biological N2-fixation to vascular plant uptake in an Alberta bog in order to determine if neighboring bog plants acquire recently fixed N from diazotrophs associating with Sphagnum mosses. Recent evidence indicates high rates of N2-fixation in Sphagnum mosses of Alberta bogs (Vile et al. 2013). Our previous work has shown that mosses can assimilate fixed N from associated diazotrophs as evidenced by the high N content of mosses despite minimal inputs from atmospheric deposition, retranslocation, and N mineralization. Therefore, the potential exists for vascular plants to obtain N from ';leaky' tissues of live mosses, however, this phenomenon has not been tested previously. Here we document the potential for relatively rapid transfer to vascular plants of N fixed by Sphagnum moss-associated diazotrophs. We utilized the novel approach of incubating mosses in 15N2 to allow the process of diazotrophic N2-fixation to mechanistically provide the 15N label, which is subsequently transferred to Sphagnum mosses. The potential for vascular bog natives to tap this N was assessed by planting the vascular plants in the labeled moss. Sphagnum mosses (upper 3 cm of live plants) were incubated in the presence of 98 atom % 15N2 gas for 48 hours. Two vascular plants common to Alberta bogs; Picea mariana and Vaccinium oxycoccus were then placed in the labeled mosses, where the mosses served as the substrate. Tissue samples from these plants were collected at three time points during the incubation; prior to 15N2 exposure (to determine natural abundance 15N), and at one and two months after 15N2 exposure. Roots and leaves were separated and run separately on a mass spectrometer to determine 15N concentrations. Sphagnum moss capitula obtained N from N2-fixation (δ15N of -2.43 × 0.40, 122.76 × 23.78, 224.92 × 68.37, 143.74 × 54.38 prior to, immediately after, and at 1 and 2 months after exposure to 15N2, respectively). Nitrogen was

  16. Identification of the Sources of Energy for Nitrogen Fixation and Physiological Characterization of Nitrogen-Fixing Members of a Marine Microbial Mat Community

    PubMed Central

    Bebout, Brad M.; Fitzpatrick, Matthew W.; Paerl, Hans W.

    1993-01-01

    Experimental manipulations of a microbial mat community were performed to determine sources of energy and reductant used for nitrogen fixation and to physiologically characterize the responsible diazotrophs. The dominant photolithotrophic members of this community were nonheterocystous cyanobacteria, but other potential nitrogen-fixing microorganisms were also present. Pronounced diel variability in rates of acetylene reduction was observed, with nighttime rates a factor of three to four higher than daytime rates. Acetylene reduction measured at night was dependent upon the occurrence of oxygenic photosynthesis the preceding day; mats incubated in the dark during the daytime reduced acetylene at rates comparable to those of light-incubated mats but were not able to reduce acetylene at the normally high rates the following night. The addition of various exogenous carbon compounds to these dark-incubated mats did not elicit nighttime acetylene reduction. Nighttime acetylene reduction apparently proceeds under anoxic conditions in these mats; the highest rates of acetylene reduction occur late at night. Additions of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (an inhibitor of oxygenic photosynthesis) to mats resulted in a pronounced stimulation of acetylene reduction during the day, but acetylene reduction the next night proceeded at greatly reduced rates (relative to untreated mats). This daytime stimulation, under the 3-(3,4-dichlorophenyl)-1,1-dimethylurea-induced anoxic conditions in the experimentally treated mats, was light dependent. These results suggest that nitrogen fixation in these mats may be attributed to the activities of nonheterocystous cyanobacteria utilizing storage products of oxygenic photosynthesis under anoxic conditions at night. PMID:16348935

  17. Preferential remineralization of dissolved organic phosphorus and non-Redfield DOM dynamics in the global ocean: Impacts on marine productivity, nitrogen fixation, and carbon export

    NASA Astrophysics Data System (ADS)

    Letscher, Robert T.; Moore, J. Keith

    2015-03-01

    Selective removal of nitrogen (N) and phosphorus (P) from the marine dissolved organic matter (DOM) pool has been reported in several regional studies. Because DOM is an important advective/mixing pathway of carbon (C) export from the ocean surface layer and its non-Redfieldian stoichiometry would affect estimates of marine export production per unit N and P, we investigated the stoichiometry of marine DOM and its remineralization globally using a compiled DOM data set. Marine DOM is enriched in C and N compared to Redfield stoichiometry, averaging 317:39:1 and 810:48:1 for C:N:P within the degradable and total bulk pools, respectively. Dissolved organic phosphorus (DOP) is found to be preferentially remineralized about twice as rapidly with respect to the enriched C:N stoichiometry of marine DOM. Biogeochemical simulations with the Biogeochemical Elemental Cycling model using Redfield and variable DOM stoichiometry corroborate the need for non-Redfield dynamics to match the observed DOM stoichiometry. From our model simulations, preferential DOP remineralization is found to increase the strength of the biological pump by ~9% versus the case of Redfield DOM cycling. Global net primary productivity increases ~10% including an increase in marine nitrogen fixation of ~26% when preferential DOP remineralization and direct utilization of DOP by phytoplankton are included. The largest increases in marine nitrogen fixation, net primary productivity, and carbon export are observed within the western subtropical gyres, suggesting the lateral transfer of P in the form of DOP from the productive eastern and poleward gyre margins may be important for sustaining these processes downstream in the subtropical gyres.

  18. Phenotypic plasticity and its genetic regulation for yield, nitrogen fixation and δ13C in chickpea crops under varying water regimes.

    PubMed

    Sadras, Victor O; Lake, Lachlan; Li, Yongle; Farquharson, Elizabeth A; Sutton, Tim

    2016-07-01

    We measured yield components, nitrogen fixation, soil nitrogen uptake and carbon isotope composition (δ(13)C) in a collection of chickpea genotypes grown in environments where water availability was the main source of yield variation. We aimed to quantify the phenotypic plasticity of these traits using variance ratios, and to explore their genetic basis using FST genome scan. Fifty-five genes in three genomic regions were found to be under selection for plasticity of yield; 54 genes in four genomic regions for the plasticity of seeds per m(2); 48 genes in four genomic regions for the plasticity of δ(13)C; 54 genes in two genomic regions for plasticity of flowering time; 48 genes in five genomic regions for plasticity of nitrogen fixation and 49 genes in three genomic regions for plasticity of nitrogen uptake from soil. Plasticity of yield was related to plasticity of nitrogen uptake from soil, and unrelated to plasticity of nitrogen fixation, highlighting the need for closer attention to nitrogen uptake in legumes. Whereas the theoretical link between δ(13)C and transpiration efficiency is strong, the actual link with yield is erratic due to trade-offs and scaling issues. Genes associated with plasticity of δ(13)C were identified that may help to untangle the δ(13)C-yield relationship. Combining a plasticity perspective to deal with complex G×E interactions with FST genome scan may help understand and improve both crop adaptation to stress and yield potential. PMID:27296246

  19. Phenotypic plasticity and its genetic regulation for yield, nitrogen fixation and δ13C in chickpea crops under varying water regimes.

    PubMed

    Sadras, Victor O; Lake, Lachlan; Li, Yongle; Farquharson, Elizabeth A; Sutton, Tim

    2016-07-01

    We measured yield components, nitrogen fixation, soil nitrogen uptake and carbon isotope composition (δ(13)C) in a collection of chickpea genotypes grown in environments where water availability was the main source of yield variation. We aimed to quantify the phenotypic plasticity of these traits using variance ratios, and to explore their genetic basis using FST genome scan. Fifty-five genes in three genomic regions were found to be under selection for plasticity of yield; 54 genes in four genomic regions for the plasticity of seeds per m(2); 48 genes in four genomic regions for the plasticity of δ(13)C; 54 genes in two genomic regions for plasticity of flowering time; 48 genes in five genomic regions for plasticity of nitrogen fixation and 49 genes in three genomic regions for plasticity of nitrogen uptake from soil. Plasticity of yield was related to plasticity of nitrogen uptake from soil, and unrelated to plasticity of nitrogen fixation, highlighting the need for closer attention to nitrogen uptake in legumes. Whereas the theoretical link between δ(13)C and transpiration efficiency is strong, the actual link with yield is erratic due to trade-offs and scaling issues. Genes associated with plasticity of δ(13)C were identified that may help to untangle the δ(13)C-yield relationship. Combining a plasticity perspective to deal with complex G×E interactions with FST genome scan may help understand and improve both crop adaptation to stress and yield potential.

  20. 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.

  1. Nitrogen and Oxygen Isotopic Studies of the Marine Nitrogen Cycle.

    PubMed

    Casciotti, Karen L

    2016-01-01

    The marine nitrogen cycle is a complex web of microbially mediated reactions that control the inventory, distribution, and speciation of nitrogen in the marine environment. Because nitrogen is a major nutrient that is required by all life, its availability can control biological productivity and ecosystem structure in both surface and deep-ocean communities. Stable isotopes of nitrogen and oxygen in nitrate and nitrite have provided new insights into the rates and distributions of marine nitrogen cycle processes, especially when analyzed in combination with numerical simulations of ocean circulation and biogeochemistry. This review highlights the insights gained from dual-isotope studies applied at regional to global scales and their incorporation into oceanic biogeochemical models. These studies represent significant new advances in the use of isotopic measurements to understand the modern nitrogen cycle, with implications for the study of past ocean productivity, oxygenation, and nutrient status.

  2. Nitrogen and Oxygen Isotopic Studies of the Marine Nitrogen Cycle

    NASA Astrophysics Data System (ADS)

    Casciotti, Karen L.

    2016-01-01

    The marine nitrogen cycle is a complex web of microbially mediated reactions that control the inventory, distribution, and speciation of nitrogen in the marine environment. Because nitrogen is a major nutrient that is required by all life, its availability can control biological productivity and ecosystem structure in both surface and deep-ocean communities. Stable isotopes of nitrogen and oxygen in nitrate and nitrite have provided new insights into the rates and distributions of marine nitrogen cycle processes, especially when analyzed in combination with numerical simulations of ocean circulation and biogeochemistry. This review highlights the insights gained from dual-isotope studies applied at regional to global scales and their incorporation into oceanic biogeochemical models. These studies represent significant new advances in the use of isotopic measurements to understand the modern nitrogen cycle, with implications for the study of past ocean productivity, oxygenation, and nutrient status.

  3. Nitrogen and Oxygen Isotopic Studies of the Marine Nitrogen Cycle.

    PubMed

    Casciotti, Karen L

    2016-01-01

    The marine nitrogen cycle is a complex web of microbially mediated reactions that control the inventory, distribution, and speciation of nitrogen in the marine environment. Because nitrogen is a major nutrient that is required by all life, its availability can control biological productivity and ecosystem structure in both surface and deep-ocean communities. Stable isotopes of nitrogen and oxygen in nitrate and nitrite have provided new insights into the rates and distributions of marine nitrogen cycle processes, especially when analyzed in combination with numerical simulations of ocean circulation and biogeochemistry. This review highlights the insights gained from dual-isotope studies applied at regional to global scales and their incorporation into oceanic biogeochemical models. These studies represent significant new advances in the use of isotopic measurements to understand the modern nitrogen cycle, with implications for the study of past ocean productivity, oxygenation, and nutrient status. PMID:26747521

  4. Experimental Determination of the Respiration Associated with Soybean/Rhizobium Nitrogenase Function, Nodule Maintenance, and Total Nodule Nitrogen Fixation 1

    PubMed Central

    Rainbird, Ross M.; Hitz, William D.; Hardy, Ralph W. F.

    1984-01-01

    The total metabolic cost of soybean (Glycine max L. Mer Clark) nodule nitrogen fixation was empirically separated into respiration associated with electron flow through nitrogenase and respiration associated with maintenance of nodule function. Rates of CO2 evolution and H2 evolution from intact, nodulated root systems under Ar:O2 atmospheres decreased in parallel when plants were maintained in an extended dark period. While H2 evolution approached zero after 36 hours of darkness at 22°C, CO2 evolution rate remained at 38° of the rate measured in light. Of the remaining CO2 evolution, 62% was estimated to originate from the nodules and represents a measure of nodule maintenance respiration. The nodule maintenance requirement was temperature dependent and was estimated at 79 and 137 micromoles CO2 (per gram dry weight nodule) per hour at 22°C and 30°C, respectively. The cost of N2 fixation in terms of CO2 evolved per electron pair utilized by nitrogenase was estimated from the slope of H2 evolution rate versus CO2 evolution rate. The cost was 2 moles CO2 evolved per mole H2 evolved and was independent of temperature. In this symbiosis, nodule maintenance consumed 22% of total respiratory energy while the functioning of nitrogenase consumed a further 52%. The remaining respiratory energy was calculated to be associated with ammonia assimilation, transport of reduced N, and H2 evolution. PMID:16663599

  5. Measurement of symbiotic nitrogen-fixation in leguminous host-plants grown in heavy metal-contaminated soils amended with sewage sludge.

    PubMed

    Obbard, J P; Jones, K C

    2001-01-01

    Rates of nitrogen fixation by Rhizobium in symbiosis with leguminous host-plants including white clover, broad bean and peas have been established in soils that have been amended experimentally with heavy metal-contaminated sewage sludges. Results from 15N-dilution experiments for the measurement of N2 fixation have shown that adverse heavy metal effects are apparent on symbiotic N2 fixation rates for white clover grown in inter-specific competition with ryegrass under mixed sward conditions, compared to white clover grown in pure sward. Further experiments on broad bean and pea indicated a significant, but minor-inhibitory metal-related effect on the rate of N2 fixation compared to untreated soils and soils amended with a relatively uncontaminated sludge. The implications of the results with respect to sludge utilisation in agriculture are discussed.

  6. Crystal structure of cce_0566 from Cyanothece 51142, a protein associated with nitrogen fixation in the DUF269 family

    SciTech Connect

    Buchko G. W.; Robinson H.

    2012-02-01

    The crystal structure for cce{_}0566 (171 aa, 19.4 kDa), a DUF269 annotated protein from the diazotrophic cyanobacterium Cyanothece sp. ATCC 51142, was determined to 1.60 {angstrom} resolution. Cce{_}0566 is a homodimer with each molecule composed of eight {alpha}-helices folded on one side of a three strand anti-parallel {beta}-sheet. Hydrophobic interactions between the side chains of largely conserved residues on the surface of each {beta}-sheet hold the dimer together. The fold observed for cce{_}0566 may be unique to proteins in the DUF269 family, hence, the protein may also have a function unique to nitrogen fixation. A solvent accessible cleft containing conserved charged residues near the dimer interface could represent the active site or ligand-binding surface for the protein's biological function.

  7. Cellulose decomposition and associated nitrogen fixation by mixed cultures of Cellulomonas gelida and Azospirillum species or Bacillus macerans

    SciTech Connect

    Halsall, D.M.; Gibson, A.H.

    1985-10-01

    Mixed cultures of Cellulomonas gelida plus Azospirillum lipoferum or Azospirillum brasilense and C. gelida plus Bacillus macerans were shown to degrade cellulose and straw and to utilize the energy-yielding products to fix atmospheric nitrogen. This cooperative process was followed over 30 days in sand-based cultures in which the breakdown of 20% of the cellulose and 28 to 30% of the straw resulted in the fixation of 12 to 14.6 mg of N per g of cellulose and 17 to 19 mg of N per g of straw consumed. Cellulomonas species have certain advantages over aerobic cellulose-degrading fungi in being able to degrade cellulose at oxygen concentrations as low as 1% O/sub 2/ (vol/vol) which would allow a close association between cellulose-degrading and microaerobic diazotrophic microorganisms. Cultures inoculated with initially different proportions of A. brasilense and C. gelida all reached a stable ratio of approximately 1 Azospirillum/3 Cellulomonas cells.

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

    DOE PAGES

    Moyes, Andrew B.; Kueppers, Lara M.; Pett-Ridge, Jennifer; Carper, Dana L.; Vandehey, Nick; O'Neil, James; Frank, A. Carolin

    2016-02-01

    Coniferous forest nitrogen (N) budgets indicate unknown sources of N. A consistent association between limber pine (Pinus flexilis) and potential N2-fixing acetic acid bacteria (AAB) indicates that native foliar endophytes may supply subalpine forests with N.

  9. Nitrogen fixation in an anticyclonic eddy in the oligotrophic North Pacific Ocean.

    PubMed

    Fong, Allison A; Karl, David M; Lukas, Roger; Letelier, Ricardo M; Zehr, Jonathan P; Church, Matthew J

    2008-06-01

    Mesoscale physical processes (for example eddies, frontal meanders and planetary waves) can play important roles in controlling ocean biogeochemistry. We examined spatial variations in upper ocean (0-100 m) nutrient inventories, N(2) fixing microorganism diversity and abundance, and rates of N(2) fixation in an anticyclonic eddy near Station ALOHA (22 degrees 45' N, 158 degrees 00' W) in the North Pacific Subtropical Gyre (NPSG). In July 2005, satellite-based sea surface altimetry and ocean color observation revealed an anticyclonic eddy with enhanced chlorophyll in the upper ocean in the vicinity of Station ALOHA. Within the eddy, near-surface ocean chlorophyll concentrations were approximately 5-fold greater than in the surrounding waters. Inventories of nitrate and phosphate in the eddy were similar to the concentrations historically observed at Station ALOHA, while silicic acid inventories were significantly depleted (one-way analysis of variance, P<0.01). Quantitative PCR determinations of nifH gene copies revealed relatively high abundances of several N(2) fixing cyanobacteria, including Trichodesmium spp., Crocosphaera watsonii and Richelia intracellularis. Reverse transcriptase PCR (RT-PCR) amplified nitrogenase (nifH) gene transcripts were cloned and sequenced to examine the diversity of active N(2) fixing microorganisms; these clone libraries were dominated by sequence-types 97%-99% identical to the filamentous cyanobacteria Trichodesmium spp. Near-surface ocean rates of N(2) fixation were 2-18 times greater (averaging 8.6+/-5.6 nmol N per l per day) than previously reported measurements at Station ALOHA. These results suggest that mesoscale physical variability can play an important role in modifying the abundances of N(2) fixing microorganisms and associated rates of N(2) fixation in open ocean ecosystems.

  10. Expanding the regulatory network that controls nitrogen fixation in Sinorhizobium meliloti: elucidating the role of the two-component system hFixL-FxkR.

    PubMed

    Reyes-González, Alma; Talbi, Chouhra; Rodríguez, Susana; Rivera, Patricia; Zamorano-Sánchez, David; Girard, Lourdes

    2016-06-01

    In Sinorhizobium meliloti, nitrogen fixation is regulated in response to oxygen concentration through the FixL-FixJ two-component system (TCS). Besides this conserved TCS, the field isolate SM11 also encodes the hFixL-FxkR TCS, which is responsible for the microoxic response in Rhizobium etli. Through genetic and physiological assays, we evaluated the role of the hFixL-FxkR TCS in S. meliloti SM11. Our results revealed that this regulatory system activates the expression of a fixKf orthologue (fixKa), in response to low oxygen concentration. Null mutations in either hFixL or FxkR promote upregulation of fixK1, a direct target of FixJ. Furthermore, the absence of this TCS translates into higher nitrogen fixation values as well as higher expression of fixN1 in nodules. Individual mutations in each of the fixK-like regulators encoded in the S. meliloti SM11 genome do not completely restrict fixN1 or fixN2 expression, pointing towards redundancy among these regulators. Both copies of fixN are necessary to achieve optimal levels of nitrogen fixation. This work provides evidence that the hFixL-FxkR TCS is activated in response to low oxygen concentration in S. meliloti SM11 and that it negatively regulates the expression of fixK1, fixN1 and nitrogen fixation. PMID:27010660

  11. MucR Is Required for Transcriptional Activation of Conserved Ion Transporters to Support Nitrogen Fixation of Sinorhizobium fredii in Soybean Nodules.

    PubMed

    Jiao, Jian; Wu, Li Juan; Zhang, Biliang; Hu, Yue; Li, Yan; Zhang, Xing Xing; Guo, Hui Juan; Liu, Li Xue; Chen, Wen Xin; Zhang, Ziding; Tian, Chang Fu

    2016-05-01

    To achieve effective symbiosis with legume, rhizobia should fine-tune their background regulation network in addition to activating key genes involved in nodulation (nod) and nitrogen fixation (nif). Here, we report that an ancestral zinc finger regulator, MucR1, other than its paralog, MucR2, carrying a frameshift mutation, is essential for supporting nitrogen fixation of Sinorhizobium fredii CCBAU45436 within soybean nodules. In contrast to the chromosomal mucR1, mucR2 is located on symbiosis plasmid, indicating its horizontal transfer potential. A MucR2 homolog lacking the frameshift mutation, such as the one from S. fredii NGR234, can complement phenotypic defects of the mucR1 mutant of CCBAU45436. RNA-seq analysis revealed that the MucR1 regulon of CCBAU45436 within nodules exhibits significant difference compared with that of free-living cells. MucR1 is required for active expression of transporters for phosphate, zinc, and elements essential for nitrogenase activity (iron, molybdenum, and sulfur) in nodules but is dispensable for transcription of key genes (nif/fix) involved in nitrogen fixation. Further reverse genetics suggests that S. fredii uses high-affinity transporters to meet the demand for zinc and phosphate within nodules. These findings, together with the horizontal transfer potential of the mucR homolog, imply an intriguing evolutionary role of this ancestral regulator in supporting nitrogen fixation.

  12. Expanding the regulatory network that controls nitrogen fixation in Sinorhizobium meliloti: elucidating the role of the two-component system hFixL-FxkR.

    PubMed

    Reyes-González, Alma; Talbi, Chouhra; Rodríguez, Susana; Rivera, Patricia; Zamorano-Sánchez, David; Girard, Lourdes

    2016-06-01

    In Sinorhizobium meliloti, nitrogen fixation is regulated in response to oxygen concentration through the FixL-FixJ two-component system (TCS). Besides this conserved TCS, the field isolate SM11 also encodes the hFixL-FxkR TCS, which is responsible for the microoxic response in Rhizobium etli. Through genetic and physiological assays, we evaluated the role of the hFixL-FxkR TCS in S. meliloti SM11. Our results revealed that this regulatory system activates the expression of a fixKf orthologue (fixKa), in response to low oxygen concentration. Null mutations in either hFixL or FxkR promote upregulation of fixK1, a direct target of FixJ. Furthermore, the absence of this TCS translates into higher nitrogen fixation values as well as higher expression of fixN1 in nodules. Individual mutations in each of the fixK-like regulators encoded in the S. meliloti SM11 genome do not completely restrict fixN1 or fixN2 expression, pointing towards redundancy among these regulators. Both copies of fixN are necessary to achieve optimal levels of nitrogen fixation. This work provides evidence that the hFixL-FxkR TCS is activated in response to low oxygen concentration in S. meliloti SM11 and that it negatively regulates the expression of fixK1, fixN1 and nitrogen fixation.

  13. Effects of decomposing rice straw on growth of and nitrogen fixation by Rhizobium

    SciTech Connect

    Rice, E.L.; Huang, C.Y.; Lin, C.Y.

    1981-03-01

    Five phenolic compounds produced in decomposing rice straw and sterile extracts of decomposing rice straw in soil were very inhibitory to growth of three strains of Rhizobium. The effects were additive and in several instances synergistic. The phenolic compounds also reduced nodule numbers and hemoglobin content of the nodules in two bean (Phaseohus vulgaris) varieties. Extracts of decomposing rice straw in soil (same concentration as in the soil) significantly reduced N/sub 2/ fixation (acetylene reduction) in Bush Black Seeded beans. This may explain in part the great reduction in soybean yields in Taiwan following rice crops when the rice stubble is left in the field.

  14. Interactions between nitrogen fixation and osmoregulation in the methanogenic archaeon Methanosarcina barkeri 227

    SciTech Connect

    Brabban, A.D.; Orcutt, E.N.; Zinder, S.H.

    1999-03-01

    The nitrogenase enzyme complex of Methanosarcina barkeri 227 was found to be more sensitive to NaCl than previously studied molybdenum nitrogenases are, with total inhibition of activity occurring at 190 mM NaCl, compared with >600 mM NaCl for Azotobacter vinelandii and Clostridium pasteurianum nitrogenases. Na{sup +} and K{sup +} had equivalent effects, whereas Mg{sup 2+} was more inhibitory than either monovalent cation, even on a per-charge basis. The anion Cl{sup {minus}} was more inhibitory than acetate was. Because M. barkeri 227 is a facultative halophile, the authors examined the effects of external salt on growth and diazotrophy and found that inhibition of growth was not greater with N{sub 2} than with NH{sub 4}{sup +}. Cells grown with N{sub 2} and cells grown with NH{sub 4}{sup +} produced equal concentrations of {alpha}-glutamate at low salt concentrations and equal concentrations of N{sup {var_epsilon}}-acetyl-{beta}-lysine at NaCl concentrations greater than 500 mM. Despite the high energetic cost of fixing nitrogen for these osmolytes, the authors obtained no evidence that there is a shift towards nonnitrogenous osmolytes during diazotrophic growth. In vitro nitrogenase enzyme assays showed that at a low concentration potassium glutamate enhanced activity but at higher concentrations this compound inhibited activity; 50% inhibition occurred at a potassium glutamate concentration of approximately 400 mM.

  15. Easterly denitrification signal and nitrogen fixation feedback documented in the western Pacific sediments

    NASA Astrophysics Data System (ADS)

    Jia, Guodong; Li, Zhiyang

    2011-12-01

    A sedimentary δ15N record in the equatorial western Pacific (WP) shows glacial-interglacial variability from 6.2 to 11.2‰ during the last two climatic cycles, similar to the denitrification record in the eastern tropical Pacific (ETP). Contrastively, a record in the South China Sea (SCS) exhibits less changes from 4.4 to 6.4‰ and is quite alike previously published results in marginal seas in the WP. By ruling out several possible causes for the δ15N variability, the δ15N record in the equatorial WP is interpreted as the source nitrate δ15N signals advected from the ETP. Comparison of several δ15N records for the last 25 ka distributed in the WP brings out a pattern of northward decrease in δ15N values and variability from the equator to off Mindano and then to marginal seas, supposed to be caused by the northward increase of local N2 fixation. Therefore, the less glacial-interglacial changes in some δ15N records in the WP could imply that the glacial decrease in subsurface δ15N due to less denitrification in source waters from the ETP would have been isotopically compensated by a synchronous decrease in local N2 fixation.

  16. A new Open Top Chamber designed to test in situ effects of climatic and atmospheric changes on nitrogen fixation in boreal forest floor mosses

    NASA Astrophysics Data System (ADS)

    Bringuier, Charline; Bradley, Robert; Bellenger, Jean-Philippe; Morin, Hubert; Lindo, Zoë

    2014-05-01

    Biological nitrogen fixation (BNF) by cyanobacteria dwelling in forest floor moss layers is an important determinant of boreal black spruce forest productivity. Recent studies have suggested that these BNF rates may increase with increasing atmospheric CO2 and increasing temperature, as predicted by current weather models. This potential increase in BNF may be offset, however, by increasing atmospheric deposition of nitrogen, or by increasing demands for phosphorus (i.e. driving nodular ATP content) and for micronutrients such as Mo, Va and Fe (i.e. co-factors of nitrogenase enzyme). In order to study the relative and interactive effects of these factors controlling in situ BNF rates in boreal forest floor moss layers, a new Open Top Chamber (OTC) was developed in summer of 2013. The chambers measure 1.60 cm dia. × 60 cm height, and are equipped with an automated CO2 delivery system designed to maintain atmospheric daytime CO2 concentrations at 800 ppm, as well as buried heating coils that increase soil temperature by 4 ° C for 3 weeks in springtime. These 2 experimental factors are crossed in a full factorial (2 × 2) design that is replicated in 4 complete blocks. Each of the 16 OTCs is divided into 4 compartments, each of which are assigned 1 of 4 sub-plot factors. These include chronic additions of either atmospheric nitrogen, phosphorus, micronutrients or a non-amended control. Staring in summer 2014, a series of measurements will be made to assess the effects of treatments on BNF rates, cyanobacterial colonization and soil nitrogen cycling. Our poster will describe in detail the design and operation of the OTCs, as well as their construction and maintenance costs.

  17. Influence of heterogeneous ammonium availability on bacterial community structure and the expression of nitrogen fixation and ammonium transporter genes during in situ bioremediation of uranium-contaminated groundwater.

    PubMed

    Mouser, Paula J; N'Guessan, A Lucie; Elifantz, Hila; Holmes, Dawn E; Williams, Kenneth H; Wilkins, Michael J; Long, Philip E; Lovley, Derek R

    2009-06-15

    The influence of ammonium availability on bacterial community structure and the physiological status of Geobacter species during in situ bioremediation of uranium-contaminated groundwater was evaluated. Ammonium concentrations varied by 2 orders of magnitude (< 4 to 400 microM) across th study site. Analysis of 16S rRNA sequences suggested that ammonium may have been one factor influencing the community composition prior to acetate amendment with Rhodoferax species predominating over Geobacter species with higher ammonium and Dechloromonas species dominating at the site with lowest ammonium. However, once acetate was added and dissimilatory metal reduction was stimulated, Geobacter species became the predominant organisms at all locations. Rates of U(VI) reduction appeared to be more related to acetate concentrations rather than ammonium levels. In situ mRNA transcript abundance of the nitrogen fixation gene, nifD, and the ammonium transporter gene, amtB, in Geobacter species indicated that ammonium was the primary source of nitrogen during uranium reduction. The abundance of amtB was inversely correlated to ammonium levels, whereas nifD transcript levels were similar across all sites examined. These results suggest that nifD and amtB expression are closely regulated in response to ammonium availability to ensure an adequate supply of nitrogen while conserving cell resources. Thus, quantifying nifD and amtB transcript expression appears to be a useful approach for monitoring the nitrogen-related physiological status of subsurface Geobacter species. This study also emphasizes the need for more detailed analysis of geochemical and physiological interactions at the field scale in order to adequately model subsurface microbial processes during bioremediation.

  18. Influence of heterogeneous ammonium availability on bacterial community structure and the expression of nitrogen fixation and ammonium transporter genes during in situ bioremediation of uranium-contaminated groundwater

    SciTech Connect

    Mouser, P.J.; N'Guessan, A.L.; Elifantz, H.; Holmes, D.E.; Williams, K.H.; Wilkins, M.J.; Long, P.E.; Lovley, D.R.

    2009-04-01

    The impact of ammonium availability on microbial community structure and the physiological status and activity of Geobacter species during in situ bioremediation of uranium-contaminated groundwater was evaluated. Ammonium concentrations varied by as much as two orders of magnitude (<4 to 400 {micro}M) across the study site. Analysis of 16S rRNA gene sequences suggested that ammonium influenced the composition of the microbial community prior to acetate addition with Rhodoferax species predominating over Geobacter species at the site with the highest ammonium, and Dechloromonas species dominating at sites with lowest ammonium. However, once acetate was added, and dissimilatory metal reduction was stimulated, Geobacter species became the predominant organisms at all locations. Rates of U(VI) reduction appeared to be more related to the concentration of acetate that was delivered to each location rather than the amount of ammonium available in the groundwater. In situ mRNA transcript abundance of the nitrogen fixation gene, nifD, and the ammonium importer gene, amtB, in Geobacter species indicated that ammonium was the primary source of nitrogen during in situ uranium reduction, and that the abundance of amtB transcripts was inversely correlated to ammonium levels across all sites examined. These results suggest that nifD and amtB expression by subsurface Geobacter species are closely regulated in response to ammonium availability to ensure an adequate supply of nitrogen while conserving cell resources. Thus, quantifying nifD and amtB expression appears to be a useful approach for monitoring the nitrogen-related physiological status of Geobacter species in subsurface environments during bioremediation. This study also emphasizes the need for more detailed analysis of geochemical/physiological interactions at the field scale, in order to adequately model subsurface microbial processes.

  19. Influence of heterogeneous ammonium availability on bacterial community structure and the expression of nitrogen fixation and ammonium transporter genes during in situ bioremediation of uranium-contaminated groundwater

    SciTech Connect

    Mouser, Paula; N'guessan, Lucie A.; Elifantz, H.; Holmes, Dawn; Williams, Kenneth H.; Wilkins, Michael J.; Long, Philip E.; Lovley, Derek R.

    2009-06-15

    The influence of ammonium availability on bacterial community structure and the physiological status of Geobacter species during in situ bioremediation of uranium-contaminated groundwater was evaluated. Ammonium concentrations varied by two orders of magnitude (<4 to 400 μM) across the study site. Analysis of 16S rRNA sequences suggested that ammonium may have been one factor influencing the community composition prior to acetate amendment with Rhodoferax species predominating over Geobacter species with higher ammonium and Dechloromonas species dominating at the site with lowest ammonium. However, once acetate was added, and dissimilatory metal reduction was stimulated, Geobacter species became the predominant organisms at all locations. Rates of U(VI)-reduction appeared to be more related to acetate concentrations rather than ammonium levels. In situ mRNA transcript abundance of the nitrogen fixation gene, nifD, and the ammonium transporter gene, amtB, in Geobacter species indicated that ammonium was the primary source of nitrogen during uranium reduction. The abundance of amtB was inversely correlated to ammonium levels whereas nifD transcript levels were similar across all sites examined. These results suggest that nifD and amtB expression are closely regulated in response to ammonium availability to ensure an adequate supply of nitrogen while conserving cell resources. Thus, quantifying nifD and amtB transcript expression appears to be a useful approach for monitoring the nitrogen-related physiological status of subsurface Geobacter species and. This study also emphasizes the need for more detailed analysis of geochemical/physiological interactions at the field scale, in order to adequately model subsurface microbial processes during bioremediation.

  20. 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.

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

    PubMed

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

    2014-06-01

    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.

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

    PubMed

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

    2014-06-01

    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

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

    PubMed

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

    2015-08-18

    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.

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

    PubMed

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

    2015-08-18

    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

  5. Effect of Rhizobium sp. BARIRGm901 inoculation on nodulation, nitrogen fixation and yield of soybean (Glycine max) genotypes in gray terrace soil.

    PubMed

    Alam, Faridul; Bhuiyan, M A H; Alam, Sadia Sabrina; Waghmode, Tatoba R; Kim, Pil Joo; Lee, Yong Bok

    2015-01-01

    Soybean plants require high amounts of nitrogen, which are mainly obtained from biological nitrogen fixation. A field experiment was conducted by soybean (Glycine max) genotypes, growing two varieties (Shohag and BARI Soybean6) and two advanced lines (MTD10 and BGM02026) of soybean with or without Rhizobium sp. BARIRGm901 inoculation. Soybean plants of all genotypes inoculated with Rhizobium sp. BARIRGm901 produced greater nodule numbers, nodule weight, shoot and root biomass, and plant height than non-inoculated plants. Similarly, inoculated plants showed enhanced activity of nitrogenase (NA) enzyme, contributing to higher nitrogen fixation and assimilation, compared to non-inoculated soybean plants in both years. Plants inoculated with Rhizobium sp. BARIRGm901 also showed higher pod, stover, and seed yield than non-inoculated plants. Therefore, Rhizobium sp. BARIRGm901 established an effective symbiotic relationship with a range of soybean genotypes and thus increased the nodulation, growth, and yield of soybean grown in gray terrace soils in Bangladesh.

  6. 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

  7. Multi-Site Evidence for Marine Nitrogen Fixation in Mid-Cretaceous Black Shales

    NASA Astrophysics Data System (ADS)

    Yum, J.; Meyers, P. A.; Bernasconi, S.

    2004-12-01

    High concentrations of organic carbon in Cretaceous black shales imply levels of sustained export production of organic matter that are unknown in the modern ocean where marine productivity is usually limited by availability of dissolved nitrate. However, if a mid-water anoxic zone expands upward into the photic zone, then nitrogen-fixing cyanobacteria can flourish. These organisms produce organic matter having an isotopic composition close to atmospheric nitrogen (0 per mil). We have compared the carbon and nitrogen isotopic and total organic carbon compositions of Albian to Santonian black shale sequences from the Demerara Rise in the equatorial Atlantic, the Kerguelan Plateau in the southern Indian Ocean, the Hatteras Rise in the western North Atlantic Ocean, the Angola Basin in the eastern South Atlantic Ocean, and the Cape Verde Rise in the eastern North Atlantic Ocean . Nitrogen isotope compositions that become lighter as organic carbon concentrations increase indicate that organic matter production was enhanced by a consortium of primary producers that included nitrogen-fixers. Expansion of an intensified oxygen minimum zone into the photic zone probably permitted coexistence of algae and of cyanobacteria, the latter functioning best under low-oxygen conditions and not being limited by nitrate availability. Improved preservation of the exported organic matter in an intensified near-surface oxygen minimum zone is implied by C/N ratios that increase to 40 as organic carbon concentrations increase. Periods of wetter climate evidently created periods of increased surface stratification of Cretaceous oceans that led to enhanced cyanobacterial primary productivity, magnified organic matter export, and deposition of the organic-carbon-rich black shales. Our multi-site comparison suggests that climate-related gradients in the degree of surface stratification led to associated gradients in export production of organic matter.

  8. Occurrence of benthic microbial nitrogen fixation coupled to sulfate reduction in the seasonally hypoxic Eckernförde Bay, Baltic Sea

    NASA Astrophysics Data System (ADS)

    Bertics, V. J.; Löscher, C. R.; Salonen, I.; Dale, A. W.; Schmitz, R. A.; Treude, T.

    2012-06-01

    Despite the worldwide occurrence of marine hypoxic regions, benthic nitrogen (N) cycling within these areas is poorly understood and it is generally assumed that these areas represent zones of intense fixed N loss from the marine system. Sulfate reduction can be an important process for organic matter degradation in sediments beneath hypoxic waters and many sulfate-reducing bacteria (SRB) have the genetic potential to fix molecular N (N2). Therefore, SRB may supply fixed N to these systems, countering some of the N lost via microbial processes such as denitrification and anaerobic ammonium oxidation. The objective of this study was to evaluate if N2-fixation, possibly by SRB, plays a role in N cycling within the seasonally hypoxic sediments from Eckernförde Bay, Baltic Sea. Monthly samplings were performed over the course of one year to measure N2-fixation and sulfate reduction rates, to determine the seasonal variations in bioturbation (bioirrigation) activity and important benthic geochemical profiles, such as sulfur and N compounds, and to monitor changes in water column temperature and oxygen concentrations. Additionally, at several time points, rates of benthic denitrification were also measured and the active N-fixing community was examined via molecular tools. Integrated rates of N2-fixation and sulfate reduction showed a similar seasonality pattern, with highest rates occurring in August (approx. 22 and 880 nmol cm-3 d-1 of N and SO42-, respectively) and October (approx. 22 and 1300 nmol cm-3 d-1 of N and SO42-, respectively), and lowest rates occurring in February (approx. 8 and 32 nmol cm-3 d-1 of N and SO42-, respectively). These rate changes were positively correlated with bottom water temperatures and previous reported plankton bloom activities, and negatively correlated with bottom water oxygen concentrations. Other variables that also appeared to play a role in rate determination were bioturbation, bubble irrigation and winter storm events

  9. Comparative studies on nitrogen budgets of closed shrimp polyculture systems

    NASA Astrophysics Data System (ADS)

    Qi, Zhen-Xiong; Li, De-Shang; Zhang, Man-Ping; Dong, Shuang-Lin

    2001-09-01

    April to October, 1997 comparative studies on the nitrogen budgets of closed shrimp polyculture systems showed that, in all the studied polyculture systems, nitrogen from feeds and fertilizers were the main input items, which comprised 70.7% 83.9% of the total input nitrogen 3.2% 7.4% of which was provided by nitrogen fixation. It was in monoculture enclosures (Y-4, Y-11 and Y-12) that the percentage reached the maximum value. The output nitrogen in harvested products comprised 10.8% 24.6% of total input nitrogen, and the highest percentage, 24.6%, was found in shrimp-fish-tagelus polyculture systems. In shrimp monoculture and shrimp-fish polyculture systems, they were 19.1% and 21.9% respectively. The nitrogen utilization efficiency was different and varied from 12.2% was found in shrimp-tagelus polyculture systems. The lowest, 12.2%, was found in shrimp monoculture systems. All the nitrogen utilization efficiencies in shrimp-fish systems or shrimp-scallop systems seemed to be higher than that of the monoculture system, but they showed little statistical difference. The main outputs of nitrogen were found in sediment mud, and comprised 48.2% 60.8% of the total, input, the lowest percentage was found in shrimp-fish-tagelus polyculture systems, and the highest percentage in shrimp-scallop systems. During the experiment, nitrogen lost through denitrification and ammonia volatilization comprised 1.9% 6. 2%, averaged 2.8%, of the total input, and the loss through seepage comprised 5.9% 8.9% of the total. The estimated nitrogen attached to the enclosure wall comprised 3.7% 13.3% of the total, and was highest in shrimp monoculture systems. Compared with the classic shrimp farming industry, the closed shrimp polyculture systems may improve the nitrogen utilization efficiency, and hence reduce the environmental impacts on coastal waters. The nitrogen discharging rates for all the studied polyculture systems ranged from 3.0% to 6. 0% of total input nitrogen.

  10. Diversity of Ammonia Oxidation (amoA) and Nitrogen Fixation (nifH) Genes in Lava Caves of Terceira, Azores, Portugal

    PubMed Central

    Hathaway, Jennifer J. Marshall; Sinsabaugh, Robert L.; Dapkevicius, Maria De Lurdes N. E.; Northup, Diana E.

    2015-01-01

    Lava caves are an understudied ecosystem in the subterranean world, particularly in regard to nitrogen cycling. The diversity of ammonia oxidation (amoA) and nitrogen fixation (nifH) genes in bacterial mats collected from lava cave walls on the island of Terceira (Azores, Portugal) was investigated using denaturing gradient gel electrophoresis (DGGE). A total of 55 samples were collected from 11 lava caves that were selected with regard to surface land use. Land use types above the lava caves were categorized into pasture, forested, and sea/urban, and used to determine if land use influenced the ammonia oxidizing and nitrogen fixing bacterial communities within the lava caves. The soil and water samples from each lava cave were analyzed for total organic carbon, inorganic carbon, total nitrogen, ammonium, nitrate, phosphate and sulfate, to determine if land use influences either the nutrient content entering the lava cave or the nitrogen cycling bacteria present within the cave. Nitrosospira-like sequences dominated the ammonia-oxidizing bacteria (AOB) community, and the majority of the diversity was found in lava caves under forested land. The nitrogen fixation community was dominated by Klebsiella pneumoniae-like sequences, and diversity was evenly distributed between pasture and forested land, but very little overlap in diversity was observed. The results suggest that land use is impacting both the AOB and the nitrogen fixing bacterial communities. PMID:26778867

  11. Rhizobium-Legume Symbiosis and Nitrogen Fixation under Severe Conditions and in an Arid Climate

    PubMed Central

    Zahran, Hamdi Hussein

    1999-01-01

    Biological N2 fixation represents the major source of N input in agricultural soils including those in arid regions. The major N2-fixing systems are the symbiotic systems, which can play a significant role in improving the fertility and productivity of low-N soils. The Rhizobium-legume symbioses have received most attention and have been examined extensively. The behavior of some N2-fixing systems under severe environmental conditions such as salt stress, drought stress, acidity, alkalinity, nutrient deficiency, fertilizers, heavy metals, and pesticides is reviewed. These major stress factors suppress the growth and symbiotic characteristics of most rhizobia; however, several strains, distributed among various species of rhizobia, are tolerant to stress effects. Some strains of rhizobia form effective (N2-fixing) symbioses with their host legumes under salt, heat, and acid stresses, and can sometimes do so under the effect of heavy metals. Reclamation and improvement of the fertility of arid lands by application of organic (manure and sewage sludge) and inorganic (synthetic) fertilizers are expensive and can be a source of pollution. The Rhizobium-legume (herb or tree) symbiosis is suggested to be the ideal solution to the improvement of soil fertility and the rehabilitation of arid lands and is an important direction for future research. PMID:10585971

  12. Bradyrhizobium japonicum cytochrome c550 is required for nitrate respiration but not for symbiotic nitrogen fixation.

    PubMed Central

    Bott, M; Thöny-Meyer, L; Loferer, H; Rossbach, S; Tully, R E; Keister, D; Appleby, C A; Hennecke, H

    1995-01-01

    Bradyrhizobium japonicum possesses three soluble c-type cytochromes, c550, c552, and c555. The genes for cytochromes c552 (cycB) and c555 (cycC) were characterized previously. Here we report the cloning, sequencing, and mutational analysis of the cytochrome c550 gene (cycA). A B. japonicum mutant with an insertion in cycA failed to synthesize a 12-kDa c-type cytochrome. This protein was detectable in the cycA mutant complemented with cloned cycA, which proves that it is the cycA gene product. The cycA mutant, a cycB-cycC double mutant, and a cycA-cycB-cycC triple mutant elicited N2-fixing root nodules on soybean (Nod+ Fix+ phenotype); hence, none of these three cytochromes c is essential for respiration supporting symbiotic N2 fixation. However, cytochrome c550, in contrast to cytochromes c552 and c555, was shown to be essential for anaerobic growth of B. japonicum, using nitrate as the terminal electron acceptor. PMID:7721713

  13. A phosphate transport system is required for symbiotic nitrogen fixation by Rhizobium meliloti.

    PubMed Central

    Bardin, S; Dan, S; Osteras, M; Finan, T M

    1996-01-01

    The bacterium Rhizobium meliloti forms N2-fixing root nodules on alfalfa plants. The ndvF locus, located on the 1,700-kb pEXO megaplasmid of R. meliloti, is required for nodule invasion and N2 fixation. Here we report that ndvF contains four genes, phoCDET, which encode an ABC-type transport system for the uptake of Pi into the bacteria. The PhoC and PhoD proteins are homologous to the Escherichia coli phosphonate transport proteins PhnC and PhnD. The PhoT and PhoE proteins are homologous to each other and to the E. coli phosphonate transport protein PhnE. We show that the R. meliloti phoD and phoE genes are induced in response to phosphate starvation and that the phoC promoter contains two elements which are similar in sequence to the PHO boxes present in E. coli phosphate-regulated promoters. The R. meliloti ndvF mutants grow poorly at a phosphate concentration of 2 mM, and we hypothesize that their symbiotic phenotype results from their failure to grow during the nodule infection process. Presumably, the PhoCDET transport system is employed by the bacteria in the soil environment, where the concentration of available phosphate is normally 0.1 to 1 microM. PMID:8755882

  14. Multispecies methods of testing for toxicity: Use of the Rhizobium-legume symbiosis in nitrogen fixation and correlations between responses by algae and terrestrial plants

    SciTech Connect

    Garten, C.T. Jr.

    1989-01-01

    Responses of the Rhizobium-legume symbiotic relationship to long-term (5- to 7-week) and short-term (2-week) exposures of copper sulfate, 2,4-dichlorophenoxyacetic acid, and streptomycin sulfate were examined in bush beans and clover. Toxic effects were evaluated by comparing plant biomass (yield), nodulation success, nitrogen fixation rate as indicated by acetylene reduction, and plant nitrogen content in controls and in plants exposed to various concentrations of the chemicals. Plants with long-term continuous root exposure were affected more than plants with short-term exposure, as indicated by reductions in nitrogen fixation rates and plant growth. Although rates of acetylene reduction (nitrogen fixation) were depressed, plants biomass and the numbers of root nodules produced were simpler, less expensive indices of exposure. In a second test, the responses of algae (Selenastrum capricornutum and Chlorella vulgaris) and terrestrial plants (radishes, barley, bush beans, and soybeans) to 21 different herbicides were compared to evaluate the use of a short-term (96-h) algal growth inhibition test for identifying chemicals potentially toxic to terrestrial plants.

  15. Biomechanical Study of Acetabular Tridimensional Memoryalloy Fixation System

    NASA Astrophysics Data System (ADS)

    Liu, Xin-Wei; Xu, Shuo-Gui; Zhang, Yun-Tong; Zhang, Chun-Cai

    2011-07-01

    We developed the acetabular tridimensional memoryalloy fixation system (ATMFS), which is made of NiTi shape memory alloy, according to the specific mechanical properties of biological memory material, NiTi shape memory alloy and measured distribution of contact area and pressure between the acetabulum and the femoral head of cadaveric pelvis. Seven formalin-preserved cadaveric pelves were used for this investigation. Pressure-sensitive film was used to measure contact area and pressure within the anterior, superior, and posterior regions of the acetabulum. The pelves were loaded under the following four conditions: (1) intact; (2) following a creation posterior wall fracture defect; (3) following reduction and standard internal fixation with reconstruction plate; and (4) following reduction and internal fixation with a new shape memory alloy device named ATMFS. A posterior wall fracture was created along an arc of 40° to 90° about the acetabulur rim. Creation of a posterior wall defect resulted in increased load in the superior acetabulum (1485 N) as compared to the intact condition (748 N, P = 0.009). Following reduction and internal fixation, the load distributed to the superior acetabulum (1545 N) was not statistically different from the defect condition. Following the fixation with ATMFS, the load seen at the superior region of the actabulum (964 N) was familiar with fixation with reconstruction plate and was not different from intact state ( P = 0.45). These data indicate that the use of ATMFS as a fracture internal fixation device resulted a partial restoration of joint loading parameters toward the intact state. ATMFS fixation may result in a clinical benefit.

  16. Interactions between Nitrogen Fixation and Methane Cycling in Northern Minnesota Peat Bogs

    NASA Astrophysics Data System (ADS)

    Warren, M. J.; Gaby, J. C.; Lin, X.; Morton, P. L.; Kostka, J. E.; Glass, J. B.

    2014-12-01

    Peatlands cover only 3% of the Earth's surface, yet store a third of soil carbon. Increasing global temperatures have the potential to change peatlands from a net sink to a net source of atmospheric carbon. N is a limiting nutrient in oligotrophic Sphagnum-dominated peatlands and biological N2 fixation likely supplies a significant but unknown fraction of N inputs. Moreover, environmental controls on diazotrophic community composition in N-limited peatlands are poorly constrained. Thus, improved understanding of feedbacks between the CH4 and N cycles is critical for predicting future changes to CH4 flux from peat bogs. We coupled measurements of N2 fixation activity measured by the acetylene (C2H2) reduction assay (ARA) with molecular analyses of expression and diversity of nifH genes encoding the molybdenum (Mo)-containing nitrogenase from two peat bogs in the Marcell Experimental Forest, Minnesota, USA. The top 10 cm of peat was sampled from the high CH4 flux S1 bog and the low CH4 flux Zim bog in April and June 2014. Despite similar N concentrations in the top 10 cm of both bogs (0.5-1.0 μM NO2-+NO3- and 2-3 μM NH4+), the S1 bog displayed variable ARA activity (1-100 nmol C2H4 h-1 g-1) whereas the Zim bog had consistently low ARA activity (<1 nmol C2H4 h-1 g-1). Highest ARA activity was measured in June from S1 bog hollows with higher moisture content incubated without O2 in the light (20-100 nmol C2H4 h-1 g-1). Dissolved Fe (1-25 μM) was higher in hollow vs. hummock samples, and at S1 vs. Zim bog, while dissolved V (4-14 nM) was consistently higher than Mo (1-4 nM), suggesting that alternative V or Fe-containing nitrogenases might be present in these bogs. In contrast, Cu, an essential micronutrient for aerobic methanotrophs, was higher in hummocks (25-48 nM) than hollows (6-17 nM). The facultative methanotroph Methylocella was the dominant diazotroph in the S1 bog based on high throughput next generation sequencing of nifH cDNA amplicons. Given previous

  17. Nitrogen-Dependent Carbon Fixation by Picoplankton In Culture and in the Mississippi River

    SciTech Connect

    Aubrey Smith; Marguerite W. Coomes; Thomas E. Smith

    2005-04-30

    The pepc gene, which encodes phosphoenolpyruvate carboxylase (PEPC), of the marine cyanobacterium Synechococcus PCC 7002, was isolated and sequenced. PEPC is an anaplerotic enzyme, but it may also contribute to overall CO2 fixation through β-carboxylation reactions. A consensus sequence generated by aligning the pepc genes of Anabaena variabilis, Anacystis nidulans and Synechocystis PCC 6803 was used to design two sets of primers that were used to amplify segments of Synechococcus PCC 7002 pepc. In order to isolate the gene, the sequence of the PCR product was used to search for the pepc nucleotide sequence from the publicly available genome of Synechococcus PCC 7002. At the time, the genome for this organism had not been completed although sequences of a significant number of its fragments are available in public databases. Thus, the major challenge was to find the pepc gene among those fragments and to complete gaps as necessary. Even though the search did not yield the complete gene, PCR primers were designed to amplify a DNA fragment using a high fidelity thermostable DNA polymerase. An open reading frame (ORF) consisting of 2988 base pairs coding for 995 amino acids was found in the 3066 bp PCR product. The pepc gene had a GC content of 52% and the deduced protein had a calculated molecular mass of 114,049 Da. The amino acid sequence was closely related to that of PEPC from other cyanobacteria, exhibiting 59-61% identity. The sequence differed significantly from plant and E. coli PEPC with only 30% homology. However, comparing the Synechococcus PCC 7002 sequence to the recently resolved E. coli PEPC revealed that most of the essential domains and amino acids involved in PEPC activity were shared by both proteins. The recombinant Synechococcus PCC 7002 PEPC was expressed in E. coli.

  18. Enhancement of the nitrogen fixation efficiency of genetically-engineered Rhizobium with high catalase activity.

    PubMed

    Orikasa, Yoshitake; Nodasaka, Yoshinobu; Ohyama, Takuji; Okuyama, Hidetoshi; Ichise, Nobutoshi; Yumoto, Isao; Morita, Naoki; Wei, Min; Ohwada, Takuji

    2010-10-01

    The vktA catalase gene, which had been cloned from Vibrio rumoiensis S-1T having extraordinarily high catalase activity, was introduced into the root nodule bacterium, Rhizobium leguminosarum bv. phaseoli USDA 2676. The catalase activity of the vktA-transformed R. leguminosarum cells (free-living) was three orders in magnitude higher than that of the parent cells and this transformant could grow in a higher concentration of exogenous hydrogen peroxide (H2O2). The vktA-transformant was inoculated to the host plant (Phaseolus vulgaris L.) and the nodulation efficiency was evaluated. The results showed that the nitrogen-fixing activity of nodules was increased 1.7 to 2.3 times as compared to the parent. The levels of H2O2 in nodules formed by the vktA-transformant were decreased by around 73%, while those of leghemoglobins (Lba and Lbb) were increased by 1.2 (Lba) and 2.1 (Lbb) times compared with the parent. These results indicated that the increase of catalase activity in rhizobia could be useful to improve the nitrogen-fixing efficiency of nodules by the reduction of H2O2 content concomitantly with the enhancement of leghemoglobins contents.

  19. Global changes in the transcript and metabolic profiles during symbiotic nitrogen fixation in phosphorus-stressed common bean plants.

    PubMed

    Hernández, Georgina; Valdés-López, Oswaldo; Ramírez, Mario; Goffard, Nicolas; Weiller, Georg; Aparicio-Fabre, Rosaura; Fuentes, Sara Isabel; Erban, Alexander; Kopka, Joachim; Udvardi, Michael K; Vance, Carroll P

    2009-11-01

    Phosphorus (P) deficiency is widespread in regions where the common bean (Phaseolus vulgaris), the most important legume for human consumption, is produced, and it is perhaps the factor that most limits nitrogen fixation. Global gene expression and metabolome approaches were used to investigate the responses of nodules from common bean plants inoculated with Rhizobium tropici CIAT899 grown under P-deficient and P-sufficient conditions. P-deficient inoculated plants showed drastic reduction in nodulation and nitrogenase activity as determined by acetylene reduction assay. Nodule transcript profiling was performed through hybridization of nylon filter arrays spotted with cDNAs, approximately 4,000 unigene set, from the nodule and P-deficient root library. A total of 459 genes, representing different biological processes according to updated annotation using the UniProt Knowledgebase database, showed significant differential expression in response to P: 59% of these were induced in P-deficient nodules. The expression platform for transcription factor genes based in quantitative reverse transcriptase-polymerase chain reaction revealed that 37 transcription factor genes were differentially expressed in P-deficient nodules and only one gene was repressed. Data from nontargeted metabolic profiles indicated that amino acids and other nitrogen metabolites were decreased, while organic and polyhydroxy acids were accumulated, in P-deficient nodules. Bioinformatics analyses using MapMan and PathExpress software tools, customized to common bean, were utilized for the analysis of global changes in gene expression that affected overall metabolism. Glycolysis and glycerolipid metabolism, and starch and Suc metabolism, were identified among the pathways significantly induced or repressed in P-deficient nodules, respectively.

  20. Variability of projected terrestrial biosphere responses to elevated levels of atmospheric CO2 due to uncertainty in biological nitrogen fixation

    NASA Astrophysics Data System (ADS)

    Meyerholt, J.; Zaehle, S.; Smith, M. J.

    2015-12-01

    Including a terrestrial nitrogen (N) cycle in Earth system models has led to substantial attenuation of predicted biosphere-climate feedbacks. However, the magnitude of this attenuation remains uncertain. A particularly important, but highly uncertain process is biological nitrogen fixation (BNF), which is the largest natural input of N to land ecosystems globally. In order to quantify this uncertainty, and estimate likely effects on terrestrial biosphere dynamics, we applied six alternative formulations of BNF spanning the range of process formulations in current state-of-the-art biosphere models within a common framework, the O-CN model: a global map of static BNF rates, two empirical relationships between BNF and other ecosystem variables (net primary productivity (NPP) and evapotranspiration), two process-oriented formulations based on plant N status, and an optimality-based approach. We examined the resulting differences in model predictions under ambient and elevated atmospheric [CO2] and found that the predicted global BNF rates and their spatial distribution for contemporary conditions were broadly comparable, ranging from 95 to 134 Tg N yr-1 (median 119 Tg N yr-1), despite distinct regional patterns associated with the assumptions of each approach. Notwithstanding, model responses in BNF rates to elevated levels of atmospheric [CO2] (+200 ppm) ranged between -4 Tg N yr-1 (-3 %) and 56 Tg N yr-1 (+42 %) (median 7 Tg N yr-1 (+8 %)). As a consequence, future projections of global ecosystem carbon storage (+281 to +353 Pg C, or +13 to +16 %), as well as N2O emission (-1.6 to +0.5 Tg N yr-1, or -19 to +7 %) differed significantly across the different model formulations. Our results emphasize the importance of better understanding the nature and magnitude of BNF responses to change-induced perturbations, particularly through new empirical perturbation experiments and improved model representation.

  1. Variability of projected terrestrial biosphere responses to elevated levels of atmospheric CO2 due to uncertainty in biological nitrogen fixation

    NASA Astrophysics Data System (ADS)

    Meyerholt, Johannes; Zaehle, Sönke; Smith, Matthew J.

    2016-03-01

    Including a terrestrial nitrogen (N) cycle in Earth system models has led to substantial attenuation of predicted biosphere-climate feedbacks. However, the magnitude of this attenuation remains uncertain. A particularly important but highly uncertain process is biological nitrogen fixation (BNF), which is the largest natural input of N to land ecosystems globally. In order to quantify this uncertainty and estimate likely effects on terrestrial biosphere dynamics, we applied six alternative formulations of BNF spanning the range of process formulations in current state-of-the-art biosphere models within a common framework, the O-CN model: a global map of static BNF rates, two empirical relationships between BNF and other ecosystem variables (net primary productivity and evapotranspiration), two process-oriented formulations based on plant N status, and an optimality-based approach. We examined the resulting differences in model predictions under ambient and elevated atmospheric [CO2] and found that the predicted global BNF rates and their spatial distribution for contemporary conditions were broadly comparable, ranging from 108 to 148 Tg N yr-1 (median: 128 Tg N yr-1), despite distinct regional patterns associated with the assumptions of each approach. Notwithstanding, model responses in BNF rates to elevated levels of atmospheric [CO2] (+200 ppm) ranged between -4 Tg N yr-1 (-3 %) and 56 Tg N yr-1 (+42 %) (median: 7 Tg N yr-1 (+8 %)). As a consequence, future projections of global ecosystem carbon (C) storage (+281 to +353 Pg C, or +13 to +16 %) as well as N2O emission (-1.6 to +0.5 Tg N yr-1, or -19 to +7 %) differed significantly across the different model formulations. Our results emphasize the importance of better understanding the nature and magnitude of BNF responses to change-induced perturbations, particularly through new empirical perturbation experiments and improved model representation.

  2. 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.

  3. Protein turnover and plant RNA and phosphorus requirements in relation to nitrogen fixation.

    PubMed

    Raven, John A

    2012-06-01

    Phosphorus (P) is the proximate (immediate) limiting element for primary productivity in some habitats, and is generally the ultimate limiting element for primary productivity. Although RNA can account for over half of the non-storage P in photosynthetic organisms, some primary producers have more ribosomes than the minimum needed for the observed rate of net protein synthesis; some of this RNA may be needed for protein turnover. Two cases of protein turnover which can occur at a much faster rate than the bulk protein turnover are those of photodamaged photosystem II and O(2)-damaged nitrogenase. While RNA involved in photosystem II repair accounts for less than 1% of the non-storage P in photosynthetic organisms, a maximum, of 12% of non-storage P could occur in RNA associated with replacement of damaged nitrogenase and/or O(2) damage avoidance mechanism in diazotrophic (N(2) fixing) organisms. There is a general trend in published data towards lower P use efficiency (g dry matter gain per day per mol P in the organism) for photosynthetic diazotrophic organisms growing under P limitation with N(2) as their nitrogen source, rather than with NH(4)(+), urea or NO(3)(-). Additional work is needed to examine the generality of a statistically verified decrease in P use efficiency for diazotrophic growth relative to growth on other nitrogen sources and, if this is confirmed, further investigation of the mechanism is needed. The outcome of such work would be important for relating the global distribution of diazotrophy to P availability. There are no known P acquisition mechanisms specific to diazotrophs. Phosphorus (P) is the proximate (immediate) limiting element for primary productivity in some habitats, and is generally the ultimate limiting element for primary productivity. Although RNA can account for over half of the non-storage P in photosynthetic organisms, some primary producers have more ribosomes than the minimum needed for the observed rate of net protein

  4. Trichodesmium – a widespread marine cyanobacterium with unusual nitrogen fixation properties

    PubMed Central

    Bergman, Birgitta; Sandh, Gustaf; Lin, Senjie; Larsson, John; Carpenter, Edward J

    2013-01-01

    The last several decades have witnessed dramatic advances in unfolding the diversity and commonality of oceanic diazotrophs and their N2-fixing potential. More recently, substantial progress in diazotrophic cell biology has provided a wealth of information on processes and mechanisms involved. The substantial contribution by the diazotrophic cyanobacterial genus Trichodesmium to the nitrogen influx of the global marine ecosystem is by now undisputable and of paramount ecological importance, while the underlying cellular and molecular regulatory physiology has only recently started to unfold. Here, we explore and summarize current knowledge, related to the optimization of its diazotrophic capacity, from genomics to ecophysiological processes, via, for example, cellular differentiation (diazocytes) and temporal regulations, and suggest cellular research avenues that now ought to be explored. PMID:22928644

  5. Predictable and efficient carbon sequestration in the North Pacific Ocean supported by symbiotic nitrogen fixation

    PubMed Central

    Karl, David M.; Church, Matthew J.; Dore, John E.; Letelier, Ricardo M.; Mahaffey, Claire

    2012-01-01

    The atmospheric and deep sea reservoirs of carbon dioxide are linked via physical, chemical, and biological processes. The last of these include photosynthesis, particle settling, and organic matter remineralization, and are collectively termed the “biological carbon pump.” Herein, we present results from a 13-y (1992–2004) sediment trap experiment conducted in the permanently oligotrophic North Pacific Subtropical Gyre that document a large, rapid, and predictable summertime (July 15–August 15) pulse in particulate matter export to the deep sea (4,000 m). Peak daily fluxes of particulate matter during the summer export pulse (SEP) average 408, 283, 24.1, 1.1, and 67.5 μmol·m−2·d−1 for total carbon, organic carbon, nitrogen, phosphorus (PP), and biogenic silica, respectively. The SEP is approximately threefold greater than mean wintertime particle fluxes and fuels more efficient carbon sequestration because of low remineralization during downward transit that leads to elevated total carbon/PP and organic carbon/PP particle stoichiometry (371:1 and 250:1, respectively). Our long-term observations suggest that seasonal changes in the microbial assemblage, namely, summertime increases in the biomass and productivity of symbiotic nitrogen-fixing cyanobacteria in association with diatoms, are the main cause of the prominent SEP. The recurrent SEP is enigmatic because it is focused in time despite the absence of any obvious predictable stimulus or habitat condition. We hypothesize that changes in day length (photoperiodism) may be an important environmental cue to initiate aggregation and subsequent export of organic matter to the deep sea. PMID:22308450

  6. Tight coupling of root-associated nitrogen fixation and plant photosynthesis in the salt marsh Spartina alterniflora and carbon dioxide enhancement of Nitrogenase activity

    SciTech Connect

    Whiting, G.J.; Gandy, E.L.; Yoch, D.C.

    1986-07-01

    The coupling of root-associated nitrogen fixation and plant photosynthesis was examined in the salt marsh grass Spartina alterniflora. In both field experiments and hydroponic assay chambers, nitrogen fixation associated with the roots was rapidly enhanced by stimulating plant photosynthesis. A kinetic analysis of acetylene reduction activity (ARA) showed that a five-to-sixfold stimulation occurred within 10 to 60 min after the plant leaves were exposed to light or increase CO/sub 2/ concentrations (with the light held constant). In field experiments, CO/sub 2/ enrichment increased plant-associated ARA by 27%. Further evidence of the dependence of ARA on plant photosynthate was obtained when activity in excised roots was shown to decrease after young greenhouse plants were placed in the dark. Seasonal variation in the ARA of excised plant roots from field cores appears to be related to the annual cycle of net photosynthesis in S. alterniflora.

  7. The Azospirillum brasilense rpoN gene is involved in nitrogen fixation, nitrate assimilation, ammonium uptake, and flagellar biosynthesis.

    PubMed

    Milcamps, A; Van Dommelen, A; Stigter, J; Vanderleyden, J; de Bruijn, F J

    1996-05-01

    The rpoN (ntrA) gene (encoding sigma 54) of Azospirillum brasilense Sp7 was isolated by using conserved rpoN primers and the polymerase chain reaction, and its nucleotide sequence was determined. The deduced amino acid sequence of the RpoN protein was found to share a high degree of homology with other members of the sigma 54 family. Two additional open reading frames were found in the Azospirillum brasilense rpoN region, with significant similarity to equivalent regions surrounding the rpoN locus in other bacteria. An rpoN mutant of Azospirillum brasilense Sp7 was constructed by gene replacement and found to be defective in nitrogen fixation, nitrate assimilation, and ammonium uptake. Lack of ammonium uptake was also found in previously isolated Azospirillum brasilense ntrB and ntrC mutants, further supporting the role of the ntr system in this process. In addition, the rpoN mutant was found to be nonmotile, suggesting a role of RpoN in Azospirillum brasilense flagellar biosynthesis.

  8. Culture-Independent Characterization of a Gene Responsible for Nitrogen Fixation in the Symbiotic Microbial Community in the Gut of the Termite Neotermes koshunensis

    PubMed Central

    Noda, Satoko; Ohkuma, Moriya; Usami, Ron; Horikoshi, Koki; Kudo, Toshiaki

    1999-01-01

    Expression of the nitrogen fixation gene, nifH, in the gut of the termite Neotermes koshunensis was characterized without cultivation. nifH cDNA was directly amplified from mRNA of the mixed microbial population in the gut by reverse transcription (RT)-PCR. Analyses of the RT-PCR products revealed that, among the diverse nifH sequences, only a few corresponding to an alternative nitrogenase (encoded by the anf gene) were preferentially transcribed in the termite gut. Expression of the anf gene was further investigated quantitatively under several termite feeding conditions by competitive PCR. The levels of expression of the anf gene were largely congruent with the nitrogen fixation activity displayed by the termite. The amounts of the genomic anf gene in the population showed no significant change, indicating that the level of expression was critical for nitrogen fixation activity. Interestingly, no significant decrease in the expression level was observed when the diet contained molybdenum (Mo), which represses ordinary anf genes. A 3.6-kb DNA region downstream of the anf gene was isolated and found to contain reading frames homologous to anfH, anfD, and anfG of the Bacteria domain which encode subunits of an alternative nitrogenase having no Mo as a cofactor. This DNA region also contained reading frames encoding glnB-like proteins, which is a common feature of the nitrogenase genes of the Archaea domain. These results indicate that the anf group of nitrogenase genes is the most important group of genes responsible for nitrogen fixation in N. koshunensis and that the anf gene possesses novel features with respect to the regulation of its expression and its gene organization. PMID:10543806

  9. Culture-independent characterization of a gene responsible for nitrogen fixation in the symbiotic microbial community in the gut of the termite Neotermes koshunensis.

    PubMed

    Noda, S; Ohkuma, M; Usami, R; Horikoshi, K; Kudo, T

    1999-11-01

    Expression of the nitrogen fixation gene, nifH, in the gut of the termite Neotermes koshunensis was characterized without cultivation. nifH cDNA was directly amplified from mRNA of the mixed microbial population in the gut by reverse transcription (RT)-PCR. Analyses of the RT-PCR products revealed that, among the diverse nifH sequences, only a few corresponding to an alternative nitrogenase (encoded by the anf gene) were preferentially transcribed in the termite gut. Expression of the anf gene was further investigated quantitatively under several termite feeding conditions by competitive PCR. The levels of expression of the anf gene were largely congruent with the nitrogen fixation activity displayed by the termite. The amounts of the genomic anf gene in the population showed no significant change, indicating that the level of expression was critical for nitrogen fixation activity. Interestingly, no significant decrease in the expression level was observed when the diet contained molybdenum (Mo), which represses ordinary anf genes. A 3.6-kb DNA region downstream of the anf gene was isolated and found to contain reading frames homologous to anfH, anfD, and anfG of the Bacteria domain which encode subunits of an alternative nitrogenase having no Mo as a cofactor. This DNA region also contained reading frames encoding glnB-like proteins, which is a common feature of the nitrogenase genes of the Archaea domain. These results indicate that the anf group of nitrogenase genes is the most important group of genes responsible for nitrogen fixation in N. koshunensis and that the anf gene possesses novel features with respect to the regulation of its expression and its gene organization. PMID:10543806

  10. Measurement of nitrogen fixation by soybean in the field using the ureide and natural sup 15 N abundance methods. [Glycine max (L. )

    SciTech Connect

    Herridge, D.F. ); Bergersen, F.J.; Peoples, M.B. )

    1990-06-01

    Nitrogen fixation by field-grown soybean (Glycine max (L.) Merrill) was assessed by the natural {sup 15}N abundance and ureide methods. The field sites (five) and genotypes (six, plus two levels of inoculation on Bragg) were chosen to provide a range of proportions of plant N derived from nitrogen fixation (P). Genotypes K466, K468, nts 1007, and nts 1116 and Davis were included on the basis of their reported tolerance of the suppressive effects of nitrate on nodulation and nitrogen fixation. Bragg was included as a nitrate-sensitive genotype. Seeds of all genotypes were inoculated at sowing with Bradyrhizobium japonicum CB1809 (USDA 136). Amounts of nitrate in the soil profile (0-1.2 meter depth) at sowing ranged from 70 (site 3) to 278 kilograms per hectare (site 5), resulting in large effects on plant nodulation, on the {delta}{sup 15}N values of nodulated plants, on the relative abundance of ureide-N in vacuum-extracted sap (VES) and stem extracts, and finally on the estimates of P. There was no relationship between amount of soil nitrate at sowing and the {delta}{sup 15}N of the plant-available soil N. Correlation matrices of the measured and calculated parameters indicated generally weak correlations between crop growth (dry matter and N) and the parameters of symbiotic activity (nodule weight, {delta}{sup 15}N, relative ureide-N); correlations were strong and highly significant between nodulation and the measures of nitrogen fixation. Estimates of P ranged between 0 and 68% ({delta}{sup 15}N) and between 6 and 56% (ureide) and were highly correlated.

  11. Nitrogen fixation associated with development and localization of mixed populations of Cellulomonas species and Azospirillium brasilense grown on cellulose or wheat straw

    SciTech Connect

    Halsall, D.M.; Goodchild, D.J.

    1986-04-01

    Mixed cultures of Cellulomonas sp. and Azospirillum brasilense were grown with straw or cellulose as the carbon source under conditions favoring the fixation of atmospheric nitrogen. Rapid increases in cell numbers, up to 10/sup 9/ cells per g of substrate, were evident after 4 and 5 days of incubation at 30 degrees C for cellulose and straw, respectively. Nitrogen fixation (detected by acetylene reduction measured on parallel cultures) commenced after 2 and 4 days of incubation for straw and cellulose, respectively, and continued for the duration of the experiment. Pure cultures of Cellulomonas sp. showed an increase in cell numbers, but CO/sub 2/ production was low, and acetylene reduction was not detected on either cellulose or straw. Pure cultures of A. brasilense on cellulose showed an inital increase in cell numbers (10/sup 7/ cells per g of substrate) over 4 days, followed by a decline presumably caused by the exhaustion of available carbon substrate. On straw, A. brasilense increased to 10/sup 9/ cells per g of substrate over 5 days and then declined slowly; this growth was accompanied by acetylene reduction. Scanning electron micrographs of straw incubated with a mixture under the above conditions for 8 days showed cells of both species in close proximity to each other. Evidence was furnished that the close spatial relatioship of cells from the two species facilitated the mutally beneficial association between them and thus increased the efficiency with which the products of straw breakdown were used for nitrogen fixation. 17 references.

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

    PubMed Central

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

    2016-01-01

    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. PMID:27162150

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

    PubMed

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

    2016-01-01

    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. PMID:27162150

  14. 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.

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

    PubMed

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

    2016-05-10

    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.

  16. 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

  17. Prebiotic Nitrogen Fixation by FeS Reduction of Nitrite Under Acidic Conditions

    NASA Technical Reports Server (NTRS)

    Summers, David P.; Mead, Susan C. (Technical Monitor)

    1999-01-01

    Theories for the origin of life require the availability of reduced nitrogen for the formation of such species as amino acid and nucleic acids. In a strongly reducing atmosphere, compounds essential to the chemical evolution of life, such as amino acids, can form by reactions between HCN, NH3, and carbonyl compounds produced in spark discharges. However, under non-reducing atmospheres, electric discharges produced NO rather than HCN or NH3. This raises the questions of; how ammonia can be formed under a neutral atmosphere, and what conditions are needed such formation to occur? On possibility is the conversion of NO into nitric and nitrous acids (through HNO) and rained into the oceans. The reduction of nitrite by aqueous Fe(II) (6 Fe(+2) + 7 H(+) + NO2(-) yields 6 Fe(III) + 2 H2O + NH3) such as was present on the early Earth could then have produced ammonia. However, this reaction does not proceed at pHs less than 7.3. An alternative is reduction by other forms of Fe(II), such as FeS. We will present results that show that FeS can reduce nitrite to ammonia at pHs as low as pH 5 under a variety of conditions.

  18. Diversity of Heterotrophic Nitrogen Fixation Genes in a Marine Cyanobacterial Mat

    PubMed Central

    Zehr, J. P.; Mellon, M.; Braun, S.; Litaker, W.; Steppe, T.; Paerl, H. W.

    1995-01-01

    The diversity of nitrogenase genes in a marine cyanobacterial mat was investigated through amplification of a fragment of nifH, which encodes the Fe protein of the nitrogenase complex. The amplified nifH products were characterized by DNA sequencing and were compared with the sequences of nitrogenase genes from cultivated organisms. Phylogenetic analysis showed that similar organisms clustered together, with the exception that anaerobic bacteria clustered together, even though they represented firmicutes, (delta)-proteobacteria, and (gamma)-proteobacteria. Mat nifH sequences were most closely related to those of the anaerobes, with a few being most closely related to the cluster of (gamma)-proteobacteria containing Klebsiella and Azotobacter species. No cyanobacterial nifH sequences were found from the mat collected in November when Microcoleus chthonoplastes was the dominant cyanobacterium, but sequences closely related to the cyanobacterium Lyngbya lagerheimeii were found during summer, when a Lyngbya strain was dominant. The results indicate that there is a high diversity of heterotrophic nitrogen-fixing organisms in marine cyanobacterial mats. PMID:16535068

  19. Will elevated carbon dioxide concentration amplify the benefits of nitrogen fixation in legumes?

    SciTech Connect

    Rogers, A.; Ainsworth, E. A.; Leakey, A. D. B.

    2009-11-01

    Growth at elevated [CO{sub 2}] stimulates photosynthesis and increases carbon (C) supply in all C3 species. A sustained and maximal stimulation in productivity at elevated [CO{sub 2}] requires an enhanced nutrient supply to match the increase in C acquisition. The ability of legumes to exchange C for nitrogen (N) with their N{sub 2}-fixing symbionts has led to the hypothesis that legumes will have a competitive advantage over nonleguminous species when grown at elevated [CO{sub 2}]. On balance, evidence suggests that in managed systems, legumes are more responsive to elevated [CO{sub 2}] than other plants (e.g. Ainsworth and Long, 2005); however, in natural ecosystems, nutrient availability can limit the response of legumes to elevated [CO{sub 2}] (Hungate et al., 2004; van Groenigen et al., 2006). Here, we consider these observations, outline the mechanisms that underlie them, and examine recent work that advances our understanding of how legumes respond to growth at elevated [CO{sub 2}]. First we highlight the global importance of legumes and provide a brief overview of the symbiotic relationship.

  20. Nitrogen fixation in forested soils by non-leguminous nitrogen-fixing plants and by non-symbiotic soil organisms. Final report, October 1, 1977-September 30, 1982

    SciTech Connect

    Lane, C.L.

    1982-01-01

    Studies using mixtures of nitrogen-fixing plants mixed with loblolly pine showed no growth increase on a good site but a growth increase on a poor site. Soil nutrient studies showed that soils beneath alder, eleagnus, wax myrtle and red cedar had a higher nutrient concentration than soils not influenced by the crown. Monthly nutrient concentrations were quite variable.

  1. Fungicide Enhancement of Nitrogen Fixation and Colonization of Phaseolus vulgaris by Rhizobium phaseoli.

    PubMed

    Lennox, L B; Alexander, M

    1981-02-01

    The number and weight of pods and the weight and nitrogen content of the tops of beans (Phaseolus vulgaris) derived from seeds inoculated with a thiram-resistant strain of Rhizobium phaseoli were increased if the seeds were treated with thiram before sowing in soil. A greater percentage of the nodules on 21-day-old plants were derived from the resistant strain, more nodules were formed, and these nodules were more effective in the presence of the fungicide than in its absence. These differences in nodule numbers were no longer present in 56-day-old plants, and only a small percentage of the nodules contained the resistant strain. The abundance of the fungicide-tolerant R. phaseoli increased rapidly soon after planting the seed and subsequently fell markedly, but the rate of decline was less if the seeds had been treated with the chemical. Protozoa also proliferated if thiram had not been applied to the seed, but their numbers were deleteriously influenced by thiram. Bdellovibrio, bacteriophages, and lytic micro-organisms acting on R. phaseoli were rare under these conditions. Ciliates and flagellated protozoa were initially suppressed by planting thiram-coated bean seeds in nonsterile soil, but the former were inhibited longer than the latter and the ciliate numbers never fully recovered if the seeds were treated with the fungicide. The resistant strain grew well in sterile soil also inoculated with a protozoa-free mixture of soil microorganisms whether thiram was added or not, but after an initial rise in numbers, its abundance fell if the mixture contained protozoa; the rate of this fall was delayed by the fungicide. The numbers of R. phaseoli were consistently less in sterile soil inoculated with the rhizobium plus a mixture of soil microorganisms containing ciliates and other protozoa than if the inoculum contained other protozoa but no ciliates. These results suggest that a suppression of protozoa, and possibly especially the ciliates, accounts for the

  2. Fungicide Enhancement of Nitrogen Fixation and Colonization of Phaseolus vulgaris by Rhizobium phaseoli

    PubMed Central

    Lennox, Linda B.; Alexander, Martin

    1981-01-01

    The number and weight of pods and the weight and nitrogen content of the tops of beans (Phaseolus vulgaris) derived from seeds inoculated with a thiram-resistant strain of Rhizobium phaseoli were increased if the seeds were treated with thiram before sowing in soil. A greater percentage of the nodules on 21-day-old plants were derived from the resistant strain, more nodules were formed, and these nodules were more effective in the presence of the fungicide than in its absence. These differences in nodule numbers were no longer present in 56-day-old plants, and only a small percentage of the nodules contained the resistant strain. The abundance of the fungicide-tolerant R. phaseoli increased rapidly soon after planting the seed and subsequently fell markedly, but the rate of decline was less if the seeds had been treated with the chemical. Protozoa also proliferated if thiram had not been applied to the seed, but their numbers were deleteriously influenced by thiram. Bdellovibrio, bacteriophages, and lytic micro-organisms acting on R. phaseoli were rare under these conditions. Ciliates and flagellated protozoa were initially suppressed by planting thiram-coated bean seeds in nonsterile soil, but the former were inhibited longer than the latter and the ciliate numbers never fully recovered if the seeds were treated with the fungicide. The resistant strain grew well in sterile soil also inoculated with a protozoa-free mixture of soil microorganisms whether thiram was added or not, but after an initial rise in numbers, its abundance fell if the mixture contained protozoa; the rate of this fall was delayed by the fungicide. The numbers of R. phaseoli were consistently less in sterile soil inoculated with the rhizobium plus a mixture of soil microorganisms containing ciliates and other protozoa than if the inoculum contained other protozoa but no ciliates. These results suggest that a suppression of protozoa, and possibly especially the ciliates, accounts for the

  3. Isolation and characterization of symbiotic mutants of bradyrhizobium sp. (Arachis) strain NC92: mutants with host-specific defects in nodulation and nitrogen fixation.

    PubMed Central

    Wilson, K J; Anjaiah, V; Nambiar, P T; Ausubel, F M

    1987-01-01

    Random transposon Tn5 mutagenesis of Bradyrhizobium sp. (Arachis) strain NC92, a member of the cowpea cross-inoculation group, was carried out, and kanamycin-resistant transconjugants were tested for their symbiotic phenotype on three host plants: groundnut, siratro, and pigeonpea. Two nodulation (Nod- phenotype) mutants were isolated. One is unable to nodulate all three hosts and appears to contain an insertion in one of the common nodulation genes (nodABCD); the other is a host-specific nodulation mutant that fails to nodulate pigeonpea, elicits uninvaded nodules on siratro, and elicits normal, nitrogen-fixing nodules on groundnut. In addition, nine mutants defective in nitrogen fixation (Fix- phenotype) were isolated. Three fail to supply symbiotically fixed nitrogen to all three host plants. Surprisingly, nodules elicited by one of these mutants exhibit high levels of acetylene reduction activity, demonstrating the presence of the enzyme nitrogenase. Three more mutants have partially effective phenotypes (Fix +/-) in symbiosis with all three host plants. The remaining three mutants fail to supply fixed nitrogen to one of the host plants tested while remaining partially or fully effective on the other two hosts; two of these mutants are Fix- in pigeonpea and Fix +/- on groundnut and on siratro, whereas the other one is Fix- on groundnut but Fix+ on siratro and on pigeonpea. These latter mutants also retain significant nodule acetylene reduction activity, even in the ineffective symbioses. Such bacterial host-specific fixation (Hsf) mutants have not previously been reported. Images PMID:3032910

  4. A comparative study of fixation techniques for type II fractures of the odontoid process.

    PubMed

    Graziano, G; Jaggers, C; Lee, M; Lynch, W

    1993-12-01

    Primary screw fixation of a Type II odontoid fracture or non-union is an attractive alternative to posterior atlanto-axial arthrodesis in that normal cervical motion can be maintained. Eight cervical cadaver spines, ranging in age from 17-90 years, were used for study. Type II fractures of the dens were created using an osteotome. Simulated fractures were fixed using one or two 3.5-mm bone screws. After testing each screw fixation technique, the screws were removed and a posterior C1-C2 brooks sublaminar wiring was performed using four 18-gauge wires with wooden blocks to simulate bone graft. No significant differences were found between bending and torsional stiffnesses for the one-screw and two-screw specimens. No significant differences were found between one- and two-screw fixation when compared with primary C1-C2 wiring in torsion. One- or two-screw fixation was as stiff as primary C1-C2 wiring in bending. One or two screws offers similar stability for fixation for a dens fracture. One- and two-screw fixation is at least as stiff as primary C1-C2 wiring in torsion and one- or two-screw fixation is stiffer than primary C1-C2 wiring in bending. PMID:8303437

  5. Multispecies methods of testing for toxicity: use of the Rhizobium-legume symbiosis in nitrogen fixation and correlations between responses by algae and terrestrial plants

    SciTech Connect

    Garten Jr, Charles T

    1991-12-01

    Responses of the Rhizobium-legume symbiotic relationship to long-term (5- to 7-week) and short-term (2-week) exposures of copper sulfate, 2,4-dichlorophenoxyacetic acid, and streptomycin sulfate were examined in bush beans and clover. Toxic effects were evaluated by comparing plant biomass (yield), nodulation success, nitrogen fixation rate as indicated by acetylene reduction, and plant nitrogen content in controls and in plants exposed to various concentrations of the chemicals. Plants with long-term continuous root exposure were affected more than plants with short-term exposure, as indicated by reductions in nitrogen fixation rates and plant growth. Although rates of acetylene reduction (nitrogen fixation) were depressed, plant biomass and the numbers of root nodules produced were simpler, less expensive indices of exposure. In a second test, the responses of algae (Selenastrum capricornutum and Chlorella vulgaris) and terrestrial plants (radishes, barley, bush beans, and soybeans) to 21 different herbicides were compared to evaluate the use of a short-term (96-h) algal growth inhibition test for identifying chemicals potentially toxic to terrestrial plants. Two test end points were evaluated: (1) the highest concentration resulting in no statistically significant reduction in the growth rate of algae or terrestrial plants, and (2) the concentration at which algal growth or terrestrial plant biomass was reduced by 50%. Test results were correlated for the two algae and for all pairs of vascular plants. However, results of the algal tests did not correlate with those of the terrestrial plants. Furthermore, according to tests with Selenastrum capricornutum, there was only a 50% chance of successfully identifying herbicide levels that reduced terrestrial plant biomass. For the herbicides and species examined, short-term toxicity tests with algae were not good indicators of toxicity to terrestrial plants.

  6. Enhanced nitrogen fixation in a Rhizobium etli ntrC mutant that overproduces the Bradyrhizobium japonicum symbiotic terminal oxidase cbb{sub 3}

    SciTech Connect

    Soberon, M.; Lopez, O.; Morera, C.; Girard, M.L.; Tabche, M.L.; Miranda, J.

    1999-05-01

    The ntrC gene codes for a transcriptional activator protein that modulates gene expression in response to nitrogen. The cytochrome production pattern of a Rhizobium etli ntrC mutant (CFN2012) was studied. CO difference spectral analysis of membranes showed that CFN2012 produced a terminal oxidase similar to the symbiotic terminal oxidase of bacteroids in free-living cells under aerobic conditions, with a characteristic trough at 553 nm. CFN2012 produced two c-type cytochromes with molecular masses of 27 and 32 kDa in contrast with the wild-type strain, which produced only a 32-kDa c-tye cytochrome. The expression levels of the R. etli fix/NOQP operon, which codes for terminal oxidase cbb{sub 3}, were not affected by the ntrC mutation. However, the production levels of the two c-type cytochromes (27 and 32 kDa) were enhanced at least eightfold when the Bradyrhizobium japonicum fixNOQP operon was expressed in CFN2012 from the nptII promoter (pMSfix{sup c}), suggesting that these proteins are subunits FixO (27 kDa) and FixP (32 kDa) of cbb{sub 3} and that CFN2012/pMSfix{sup c} overproduced this terminal oxidase. CFN2012/pMSfix{sup c} showed a significant increase in its symbiotic performance as judged by the determination of nitrogenase activities of plants inoculated with this strain, suggesting that the overproduction of cbb{sub 3} terminal oxidase correlates with an enhancement in symbiotic nitrogen fixation.

  7. A Functional myo-Inositol Dehydrogenase Gene Is Required for Efficient Nitrogen Fixation and Competitiveness of Sinorhizobium fredii USDA191 To Nodulate Soybean (Glycine max [L.] Merr.)

    PubMed Central

    Jiang, Guoqiao; Krishnan, Ammulu Hari; Kim, Yong-Woong; Wacek, Thomas J.; Krishnan, Hari B.

    2001-01-01

    Inositol derivative compounds provide a nutrient source for soil bacteria that possess the ability to degrade such compounds. Rhizobium strains that are capable of utilizing certain inositol derivatives are better colonizers of their host plants. We have cloned and determined the nucleotide sequence of the myo-inositol dehydrogenase gene (idhA) of Sinorhizobium fredii USDA191, the first enzyme responsible for inositol catabolism. The deduced IdhA protein has a molecular mass of 34,648 Da and shows significant sequence similarity with protein sequences of Sinorhizobium meliloti IdhA and MocA; Bacillus subtilis IolG, YrbE, and YucG; and Streptomyces griseus StrI. S. fredii USDA191 idhA mutants revealed no detectable myo-inositol dehydrogenase activity and failed to grow on myo-inositol as a sole carbon source. Northern blot analysis and idhA-lacZ fusion expression studies indicate that idhA is inducible by myo-inositol. S. fredii USDA191 idhA mutant was drastically affected in its ability to reduce nitrogen and revealed deteriorating bacteroids inside the nodules. The number of bacteria recovered from such nodules was about threefold lower than the number of bacteria isolated from nodules initiated by S. fredii USDA191. In addition, the idhA mutant was also severely affected in its ability to compete with the wild-type strain in nodulating soybean. Under competitive conditions, nodules induced on soybean roots were predominantly occupied by the parent strain, even when the idhA mutant was applied at a 10-fold numerical advantage. Thus, we conclude that a functional idhA gene is required for efficient nitrogen fixation and for competitive nodulation of soybeans by S. fredii USDA191. PMID:11274120

  8. Sequences of nifX, nifW, nifZ, nifB and two ORF in the Frankia nitrogen fixation gene cluster.

    PubMed

    Harriott, O T; Hosted, T J; Benson, D R

    1995-08-01

    The actinomycete Frankia alni fixes N2 in root nodules of several non-leguminous plants. It is one of the few known N2-fixing members of the high-GC Gram+ lineage of prokaryotes. Thus, we have undertaken a study of its nitrogen fixation gene (nif) organization to compare with that of the more extensively characterized proteobacteria. A cosmid (pFN1) containing the nif region of Fa CpI1 was isolated from a cosmid library using the nifHDK genes of Fa CpI1 as a probe. A 4.5-kb BamHI fragment that mapped downstream from the previously characterized nifHDK genes was cloned and sequenced. Based on nt and aa sequence similarities to nif from other N2-fixing bacteria, eight ORF were identified and designated nifX, orf3, orf1, nifW, nifZ, nifB, orf2 and nifU. A region that hybridized to Rhizobium meliloti and Klebsiella pneumoniae nifA did not appear to contain a nifA-like gene. We have revised the map of the Fa nif region to reflect current information. PMID:7642138

  9. Nitrogenase phylogeny and the molybdenum dependence of nitrogen fixation in Methanococcus maripaludis.

    PubMed Central

    Kessler, P S; McLarnan, J; Leigh, J A

    1997-01-01

    We studied the effects of molybdenum, vanadium, and tungsten on the diazotrophic growth of Methanococcus maripaludis. Mo stimulated growth, with a maximal response at 4.0 microM, while V had no effect at any concentration tested. W specifically inhibited diazotrophic growth in the presence of Mo. Coupling the results of our analysis and other known metal requirements with phylogenies derived from nifD and nifK genes revealed distinct clusters for Mo-, V-, and Fe-dinitrogenases and suggested that most methanogens also have molybdenum-type nitrogenases. PMID:8990309

  10. Comparison of Hallux Interphalangeal Joint Arthrodesis Fixation Techniques: A Retrospective Multicenter Study.

    PubMed

    Thorud, Jakob C; Jolley, Tyler; Shibuya, Naohiro; Lew, Eric; Britt, Matthew; Butterfield, Ted; Boike, Alan; Hardy, Mark; Brancheau, Steven P; Motley, Travis; Jupiter, Daniel C

    2016-01-01

    Few studies have investigated the complications that occur after hallux interphalangeal joint arthrodesis. The present study evaluated complications in 152 patients aged 18 to 80 years from 2005 to 2012 from 4 different academic institutions after hallux interphalangeal joint arthrodesis. Overall, 65.8% of the patients had ≥1 complication. Infections occurred in 16.5%, dehiscence in 12.5%, and reoperations in 27.0%. The clinical nonunion rate was ≥17.8%, and the radiographic nonunion rate was ≥13.8%. After logistic regression analysis, only the study site and peripheral neuropathy were associated with having ≥1 complication (p < .01 and p < .05, respectively). Single screw fixation compared with other fixation did not have a statistically significant influence on the postoperative complications. However, when fixation was expanded to 4 categories, single screw fixation had lower infection and reoperation rates than either crossed Kirschner wires or other fixation category but not compared with crossed screws on multivariate logistic regression analysis. Although additional studies are warranted, the findings from the present study might aid in both the prognosis of complications and the support of the use of a single screw over crossed Kirchner wire fixation in hallux interphalangeal joint arthrodesis.

  11. Inhibition of nitrogen fixation in alfalfa by arsenate, heavy metals, fluoride, and simulated Acid rain.

    PubMed

    Porter, J R; Sheridan, R P

    1981-07-01

    The acute effects of aqueous solutions of As, Cd, Cu, Pb, F, and Zn ions at concentrations from 0.01 to 100 micrograms per milliliter and solutions adjusted to pH 2 to 6 with nitric or sulfuric acid were studied with respect to acetylene reduction, net photosynthesis, respiration rate, and chlorophyll content in Vernal alfalfa (Medicago sativa L. cv. Vernal). The effects of the various treatments on acetylene reduction varied from no demonstrable effect by any concentration of F(-) and 42% inhibition by 100 micrograms Pb(2+) per milliliter, to 100% inhibition by 10 micrograms Cd(2+) per milliliter and 100 micrograms per milliliter As, Cu(2+), and Zn(2+) ions. Zn(2+) showed statistically significant inhibition of activity at 0.1 micrograms per milliliter. Acid treatments were not inhibitory above pH 2, at which pH nitric acid inhibited acetylene reduction activity more than did sulfuric acid. The inhibition of acetylene reduction by these ions was Zn(2+) > Cd(2+) > Cu(2+) > AsO(3) (-) > Pb(2+) > F(-). The sensitivity of acetylene reduction to the ions was roughly equal to the sensitivity of photosynthesis, respiration, and chlorophyll content when Pb(2+) was applied, but was 1,000 times more sensitive to Zn(2+). The relationship of the data to field conditions and industrial pollution is discussed.

  12. Inhibition of Nitrogen Fixation in Alfalfa by Arsenate, Heavy Metals, Fluoride, and Simulated Acid Rain

    PubMed Central

    Porter, John R.; Sheridan, Richard P.

    1981-01-01

    The acute effects of aqueous solutions of As, Cd, Cu, Pb, F, and Zn ions at concentrations from 0.01 to 100 micrograms per milliliter and solutions adjusted to pH 2 to 6 with nitric or sulfuric acid were studied with respect to acetylene reduction, net photosynthesis, respiration rate, and chlorophyll content in Vernal alfalfa (Medicago sativa L. cv. Vernal). The effects of the various treatments on acetylene reduction varied from no demonstrable effect by any concentration of F− and 42% inhibition by 100 micrograms Pb2+ per milliliter, to 100% inhibition by 10 micrograms Cd2+ per milliliter and 100 micrograms per milliliter As, Cu2+, and Zn2+ ions. Zn2+ showed statistically significant inhibition of activity at 0.1 micrograms per milliliter. Acid treatments were not inhibitory above pH 2, at which pH nitric acid inhibited acetylene reduction activity more than did sulfuric acid. The inhibition of acetylene reduction by these ions was Zn2+ > Cd2+ > Cu2+ > AsO3− > Pb2+ > F−. The sensitivity of acetylene reduction to the ions was roughly equal to the sensitivity of photosynthesis, respiration, and chlorophyll content when Pb2+ was applied, but was 1,000 times more sensitive to Zn2+. The relationship of the data to field conditions and industrial pollution is discussed. PMID:16661858

  13. 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

  14. 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.

  15. One member of a gro-ESL-like chaperonin multigene family in Bradyrhizobium japonicum is co-regulated with symbiotic nitrogen fixation genes.

    PubMed Central

    Fischer, H M; Babst, M; Kaspar, T; Acuña, G; Arigoni, F; Hennecke, H

    1993-01-01

    This report is concerned with the structural characterization and genetic regulation of new bacterial groES and groEL chaperonin genes, and presents two novelties. The first is the discovery that the nitrogen fixing soybean root nodule bacterium, Bradyrhizobium japonicum, unlike all other prokaryotes investigated so far, possesses a multigene family consisting of five very similar, though not identical, groESL-like genes. The second novelty relates to the finding that these five homologues are expressed to different degrees and, in particular, that one family member (namely groESL3) is induced by a mechanism that does not involve the well-known heat shock response. By contrast, the groESL3 genes are co-regulated together with symbiotic nitrogen fixation genes, in that they are activated by the nitrogen fixation regulatory protein NifA at low oxygen conditions and transcribed from a -24/-12 promoter by the sigma 54 RNA polymerase. Two other members of the groESL gene family are apparently expressed constitutively at different levels, and yet another one is strongly induced by high temperature. As an attractive hypothesis it follows that B. japonicum may modulate its cellular contents of GroES- and GroEL-like chaperonins in response to specific environmental conditions and physiological needs. Images PMID:8101485

  16. Fixation time does not affect expression of HER2/neu: a pilot study.

    PubMed

    Ibarra, Julio A; Rogers, Lowell W

    2010-10-01

    It is said that HER2/neu expression by immunohistochemical analysis varies with the time of fixation. The purpose of this pilot study was to determine the impact of the length of fixation in 10% buffered formalin on the expression of HER2/neu by immunohistochemical analysis. We studied tissue samples from 10 invasive breast cancer cases after fixation for 3, 48, 72, 96, and 120 hours. The tissue was processed immediately after fixation, resembling routine practice. The 50 resulting blocks were then batch stained with PATHWAY HER2/neu clone 4B5 rabbit monoclonal antibody using the Ventana Ultraview DAB detection kit in a Ventana BenchMark XT processor (Ventana, Tucson, AZ). The stained slides were reviewed and scored. We found no significant difference in the intensity of the stain or the percentage of cells stained regardless of the time in fixation. Fixation times between 3 and 120 hours in 10% buffered formalin do not appear to have an impact on the expression of HER2/neu by immunohistochemical analysis.

  17. 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

  18. Occurrence of benthic microbial nitrogen fixation coupled to sulfate reduction in the seasonally hypoxic Eckernförde Bay, Baltic Sea

    NASA Astrophysics Data System (ADS)

    Bertics, V. J.; Löscher, C. R.; Salonen, I.; Dale, A. W.; Gier, J.; Schmitz, R. A.; Treude, T.

    2013-03-01

    Despite the worldwide occurrence of marine hypoxic regions, benthic nitrogen (N) cycling within these areas is poorly understood and it is generally assumed that these areas represent zones of intense fixed N loss from the marine system. Sulfate reduction can be an important process for organic matter degradation in sediments beneath hypoxic waters and many sulfate-reducing bacteria (SRB) have the genetic potential to fix molecular N (N2). Therefore, SRB may supply fixed N to these systems, countering some of the N lost via microbial processes, such as denitrification and anaerobic ammonium oxidation. The objective of this study was to evaluate if N2 fixation, possibly by SRB, plays a role in N cycling within the seasonally hypoxic sediments from the Eckernförde Bay, Baltic Sea. Monthly samplings were performed over the course of one year to measure nitrogenase activity (NA) and sulfate reduction rates, to determine the seasonal variations in bioturbation (bioirrigation) activity and important benthic geochemical profiles, such as sulfur and N compounds, and to monitor changes in water column temperature and oxygen concentrations. Additionally, at several time points, the active N-fixing community was examined via molecular tools. Integrated rates of N2 fixation (approximated from NA) and sulfate reduction showed a similar seasonality pattern, with highest rates occurring in August (approx. 22 and 880 nmol cm-3 d-1 of N and SO42-, respectively) and October (approx. 22 and 1300 nmol cm-3 d-1 of N and SO42- respectively), and lowest rates occurring in February (approx. 8 and 32 nmol cm-3 d-1 of N and SO42-, respectively). These rate changes were positively correlated with bottom water temperatures and previous reported plankton bloom activities, and negatively correlated with bottom water oxygen concentrations. Other variables that also appeared to play a role in rate determination were bioturbation, bubble irrigation and winter storm events. Molecular analysis

  19. Evaluation of Tibial Condyle Fractures Treated with Ilizarov Fixation, A Prospective Study

    PubMed Central

    Reddy R, Sandeep; Shah, Harshad M; Golla, Dinesh Kumar; Ganesh D J, Niranthara; Kumar P, Ashok

    2014-01-01

    Background: Tibial plateau fractures are associated with significant soft tissue injuries which increases the risks of complications and must be considered when managing tibial plateau fractures. Various modalities of treatment are available for treatment of these fractures but Ilizarov fixation has a special advantage over others. Review of literature shows many studies of Ilizarov fixation in the treatment of tibial plateau fractures with variable results. Aim of our study was to evaluate tibial condyle fractures treated by Ilizarov fixation. Materials and Methods: Study included 43 patients with Schatzker type II and above tibial plateau fractures treated by ilizarov fixation. Standard trauma evaluation, a meticulous musculoskeletal and neurologic examination was carried out. All patients underwent Ilizarov fixation by same team of surgeons. Clinicoradilogical assessment of the patients carried out at regular intervals. Results: Our study included 43 cases of tibial plateau of various types except type I. Mean time for radiological union was 24.51 wk (range 15 to 32 wk). Mean fixator period was 26.6 wk( 16-34 wk). The functional results were measured by Lyshom’s and Hohl and Luck score. The mean Lyshom’s score was at the end of one year was 82.16. At end of one year by Hohl and Luck grading 11 patients had fair, 23 had good and 9 had excellent results. Conclusion: High energy tibial plateau fractures can be definitively treated with Ilizarov external fixation. Treatment with this method gives good union rates and less risk of infection. Closed reduction, minimal soft tissue damage and early mobilization are the key to low complications. PMID:25584250

  20. Growth and nitrogen fixation of legumes at increased salinity under field conditions: implications for the use of green manures in saline environments

    PubMed Central

    Bruning, Bas; van Logtestijn, Richard; Broekman, Rob; de Vos, Arjen; González, Andrés Parra; Rozema, Jelte

    2015-01-01

    The use of legumes as green manure can potentially increase crop productivity in saline environments and thus contribute to the sustainability of agricultural systems. Here, we present results from a field experiment conducted in the Netherlands that addressed the efficiency of nitrogen (N) fixation by a legume at varying salinities. We grew Melilotus officinalis in an agricultural field using drip irrigation with water salinity varying in electrical conductivity between 1.7 and 20 dS m−1. In the experiment, nearly 100 % of total plant N in M. officinalis was derived from symbiotic fixation at all but the highest salinity level (20 dS m−1). Our results indicated that this species derived substantial amounts of N via symbiotic fixation, the N becoming available in the soil (and thus available to crops) when cultivated legumes senesce and decompose. Based on the growth performance of M. officinalis and its ability to fix N at moderate soil salinity in our field experiments, we identified this species as a promising source for green manure in saline agriculture in temperate regions. PMID:25661201

  1. Efficient Visible Light Nitrogen Fixation with BiOBr Nanosheets of Oxygen Vacancies on the Exposed {001} Facets.

    PubMed

    Li, Hao; Shang, Jian; Ai, Zhihui; Zhang, Lizhi

    2015-05-20

    Even though the well-established Haber-Bosch process has been the major artificial way to "fertilize" the earth, its energy-intensive nature has been motivating people to learn from nitrogenase, which can fix atmospheric N2 to NH3 in vivo under mild conditions with its precisely arranged proteins. Here we demonstrate that efficient fixation of N2 to NH3 can proceed under room temperature and atmospheric pressure in water using visible light illuminated BiOBr nanosheets of oxygen vacancies in the absence of any organic scavengers and precious-metal cocatalysts. The designed catalytic oxygen vacancies of BiOBr nanosheets on the exposed {001} facets, with the availability of localized electrons for π-back-donation, have the ability to activate the adsorbed N2, which can thus be efficiently reduced to NH3 by the interfacial electrons transferred from the excited BiOBr nanosheets. This study might open up a new vista to fix atmospheric N2 to NH3 through the less energy-demanding photochemical process.

  2. Efficient Visible Light Nitrogen Fixation with BiOBr Nanosheets of Oxygen Vacancies on the Exposed {001} Facets.

    PubMed

    Li, Hao; Shang, Jian; Ai, Zhihui; Zhang, Lizhi

    2015-05-20

    Even though the well-established Haber-Bosch process has been the major artificial way to "fertilize" the earth, its energy-intensive nature has been motivating people to learn from nitrogenase, which can fix atmospheric N2 to NH3 in vivo under mild conditions with its precisely arranged proteins. Here we demonstrate that efficient fixation of N2 to NH3 can proceed under room temperature and atmospheric pressure in water using visible light illuminated BiOBr nanosheets of oxygen vacancies in the absence of any organic scavengers and precious-metal cocatalysts. The designed catalytic oxygen vacancies of BiOBr nanosheets on the exposed {001} facets, with the availability of localized electrons for π-back-donation, have the ability to activate the adsorbed N2, which can thus be efficiently reduced to NH3 by the interfacial electrons transferred from the excited BiOBr nanosheets. This study might open up a new vista to fix atmospheric N2 to NH3 through the less energy-demanding photochemical process. PMID:25874655

  3. Three-dimensional stabilization provided by the external spinal fixator compared to two internal fixation devices: a biomechanical in vitro flexibility study.

    PubMed

    Lund, Teija; Nydegger, Thomas; Rathonyi, Gabor; Nolte, Lutz-Peter; Schlenzka, Dietrich; Oxland, Thomas R

    2003-10-01

    We performed an in vitro study to investigate the stabilization (i.e. motion reduction) provided by the external spinal fixator (ESF), and to compare the three configurations of the ESF with two internal fixation techniques. Six human cadaveric lumbar spine specimens (L3-S1) were subjected to multidirectional flexibility testing in six configurations: (1) intact, (2) ESF in neutral, (3) ESF in distraction, (4) ESF in compression, (5) translaminar facet screw fixation, and (6) internal transpedicular fixation. Both the ESF and the internal fixation systems stabilized the specimens from L4 to S1. In each testing configuration, pure bending moments of flexion-extension, bilateral axial rotation, and bilateral lateral bending were applied to the uppermost vertebra stepwise to a maximum of 10 Nm. The rigid body motion between the vertebrae was measured using an optoelectronic camera system, and custom software was used to calculate the intervertebral rotations. For each applied motion in all testing configurations, the total range of motion (ROM) of L4-S1 is reported. All three ESF configurations stabilized the spine significantly when compared to the intact specimen. The ESF in compression provided significantly more stabilization in flexion-extension than the two other ESF configurations, but no other significant differences were found between the three ESF modes. In flexion-extension the ESF stabilized the spine significantly when compared with the two internal fixation devices. Only in bilateral lateral bending was the ESF inferior to internal transpedicular fixation in providing stabilization. The results of the present study suggest that the ESF provides a high degree of stabilization for preoperative assessment of selected low back pain patients. Whether other non-mechanical factors affect the pain relief experienced by the patients remains unknown.

  4. 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

  5. Backbone 1H, 13C, and 15N NMR assignments for the Cyanothece 51142 protein cce_0567: a protein associated with nitrogen fixation in the DUF683 family

    SciTech Connect

    Buchko, Garry W.; Sofia, Heidi J.

    2008-06-01

    The recently sequenced genome of the diurnal cyanobacterium Cyanothece sp. PCC 51142 (contig 83.1_1_243_746) contains the sequence for an hypothetical protein that falls into the DUF683 family. As observed for the other 54 DUF683 proteins currently listed in the GenBank database, this 78-residue (9.0 kDa) protein in Cyanothece is also found in a nitrogen fixation gene cluster suggesting that it is involved in the process. To date no structural information exists for any of the proteins in the DUF683 family. In an effort to elucidate the biochemical role DUF683 may play in nitrogen fixation and to obtain structural information for a member of the DUF683 protein family, a construct containing DUF683 from Cyanothece 51142 was generated, expressed, purified, and the solution properties characterized. A total rotational correlation time (tc) of 17.1 ns was estimated by nuclear magnetic resonance (NMR) spectroscopy suggesting a molecular weight of ~ 40 kDa, an observation dictating that DUF683 is a tetramer in solution. Using triple-labeled (2H, 13C, 15N) and residue-specific 15N-labeled amino acids (L, K, V, and E/Q) samples, most of the backbone and side chain resonances for DUF683 were assigned. The 13C alpha chemical shifts and NOESY NMR data indicate that the protein is helical from K18-E75.

  6. The novel regulatory ncRNA, NfiS, optimizes nitrogen fixation via base pairing with the nitrogenase gene nifK mRNA in Pseudomonas stutzeri A1501.

    PubMed

    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

    2016-07-26

    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.

  7. A simple method for the formalin fixation of lungs in toxicological pathology studies.

    PubMed

    Qu, Wen-Sheng; Yin, Ji-Ye; Wang, He-Mei; Dong, Yan-Sheng; Ding, Ri-Gao

    2015-10-01

    Optimized lung preparation for detailed structural evaluation is required to improve consistency in preclinical safety evaluation, differences of opinion exist among regulatory agency personnel regarding the optimal methods for routine formalin fixation of lungs from rodent toxicology studies. The simple tracheal ligation fixation method emphasizes tracheal ligation before opening the thorax instead of attempting to re-inflate after lung collapse when opening the thorax. Photomicrographs of this method demonstrated an unprecedented ability to maintain the natural lung architecture, in contrast to the unavoidable changes in the alveolar environment by the intratracheal instillation and vascular perfusion methods. In addition, a comparison of fixation methods on lung morphology in a rodent model of LPS-induced acute lung injury demonstrated that the tracheal ligation fixation method may provide a standard approach for morphometry. Additionally, a TUNEL assay was used to determine the degree of autolysis, which revealed that the autolysis was insignificant in the central areas of each lobe of the lung compared to the lung periphery by tracheal ligation fixation. In conclusion, our novel modified method, which avoids the disadvantages of generating artifacts, fulfills the requirement of preserving the clear, natural morphology of the lung making it suitable and worthy of recommendation for toxicological studies in a good laboratory practice (GLP) lab.

  8. A simple method for the formalin fixation of lungs in toxicological pathology studies.

    PubMed

    Qu, Wen-Sheng; Yin, Ji-Ye; Wang, He-Mei; Dong, Yan-Sheng; Ding, Ri-Gao

    2015-10-01

    Optimized lung preparation for detailed structural evaluation is required to improve consistency in preclinical safety evaluation, differences of opinion exist among regulatory agency personnel regarding the optimal methods for routine formalin fixation of lungs from rodent toxicology studies. The simple tracheal ligation fixation method emphasizes tracheal ligation before opening the thorax instead of attempting to re-inflate after lung collapse when opening the thorax. Photomicrographs of this method demonstrated an unprecedented ability to maintain the natural lung architecture, in contrast to the unavoidable changes in the alveolar environment by the intratracheal instillation and vascular perfusion methods. In addition, a comparison of fixation methods on lung morphology in a rodent model of LPS-induced acute lung injury demonstrated that the tracheal ligation fixation method may provide a standard approach for morphometry. Additionally, a TUNEL assay was used to determine the degree of autolysis, which revealed that the autolysis was insignificant in the central areas of each lobe of the lung compared to the lung periphery by tracheal ligation fixation. In conclusion, our novel modified method, which avoids the disadvantages of generating artifacts, fulfills the requirement of preserving the clear, natural morphology of the lung making it suitable and worthy of recommendation for toxicological studies in a good laboratory practice (GLP) lab. PMID:26388042

  9. RANDOMIZED PROSPECTIVE STUDY COMPARING TRANSVERSE AND EXTRACORTICAL FIXATION IN ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION

    PubMed Central

    da Silva Guarilha, Eduardo; de Andrade Fígaro Caldeira, Paulo Roberto; de Almeida Lira Neto, Ozorio; Navarro, Marcelo Schmidt; Milani, Antonio; Filho, Mario Carneiro

    2015-01-01

    Objective: This study had the objective of prospectively comparing transverse fixation (Cross-Pin™) with extracortical fixation (EZLoc™) for the femur, in surgical reconstruction of the anterior cruciate ligament, from a clinical, biomechanical and functional point of view. Methods: Between April 2007 and November 2009, 50 patients with acute or chronic anterior cruciate ligament injuries underwent arthroscopic reconstruction using the homologous flexor tendons (gracilis and semitendinosus). Randomization of the femoral fixation method was done by means of a draw at the time of the procedure. Patients were excluded if they presented multiple ligament lesions, fractures, previous surgery, autoimmune disease and impairment of the contralateral knee. The Lysholm scale, SF36 quality-of-life questionnaire and KT1000™ arthrometer were used. Results: After a mean follow-up of 18.1 months, there were no statistically significant differences between the groups regarding the Lysholm scale and KT1000™ measurements. However, the SF36 questionnaire showed a statistical difference such that transverse fixation was superior regarding pain and vitality. Conclusion: Both techniques were shown to be efficient for transfemoral fixation, but with almost no statistically significant difference between them. We believe that new studies will be necessary for better understanding of these differences. PMID:27042646

  10. Fixation light hue bias revisited: implications for using adaptive optics to study color vision.

    PubMed

    Hofer, H J; Blaschke, J; Patolia, J; Koenig, D E

    2012-03-01

    Current vision science adaptive optics systems use near infrared wavefront sensor 'beacons' that appear as red spots in the visual field. Colored fixation targets are known to influence the perceived color of macroscopic visual stimuli (Jameson, D., & Hurvich, L. M. (1967). Fixation-light bias: An unwanted by-product of fixation control. Vision Research, 7, 805-809.), suggesting that the wavefront sensor beacon may also influence perceived color for stimuli displayed with adaptive optics. Despite its importance for proper interpretation of adaptive optics experiments on the fine scale interaction of the retinal mosaic and spatial and color vision, this potential bias has not yet been quantified or addressed. Here we measure the impact of the wavefront sensor beacon on color appearance for dim, monochromatic point sources in five subjects. The presence of the beacon altered color reports both when used as a fixation target as well as when displaced in the visual field with a chromatically neutral fixation target. This influence must be taken into account when interpreting previous experiments and new methods of adaptive correction should be used in future experiments using adaptive optics to study color.

  11. Incorporating the Role of Nitrogen in the Noah-MP Land Surface Model for Climate and Environmental Studies

    NASA Astrophysics Data System (ADS)

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

    2014-12-01

    Noah-MP (Niu et al., 2011; Yang et al., 2011) is the next generation land surface model for the Weather Research and Forecasting (WRF) meteorological model and the Climate Forecast Systems in the National Centers for Environmental Prediction. While Noah-MP does not currently contain a dynamic nitrogen cycle, this can readily be updated with the interactive vegetation canopy option. In this study, Noah-MP is coupled with the Fixation & Uptake of Nitrogen (FUN) model (Fisher et al., 2010) for the above ground processes and the soil nitrogen model from the Soil and Water Assessment Tool (SWAT) for the below ground processes. This combines FUN's state-of-the-art concept of the carbon cost theory and SWAT's strength in representing the anthropogenic effects on the nitrogen cycle. The processes employed from FUN are the nitrogen uptake and fixation of plants, both of which are directly linked to the plant productivity. If passive nitrogen uptake cannot meet the nitrogen demand, plants have to spend part of the photosynthesized carbon production on nitrogen acquisition. The processes employed from SWAT are nitrogen mineralization, nitrification, immobilization, volatilization, atmospheric deposition, and leaching. In addition, the modified universal soil loss equation is used to more accurately account for the nitrogen removal in sediment caused by surface runoff. Because human input of nitrogen greatly changes the nitrogen cycle, a simple nitrogen fertilization approach is also applied to crops. Preliminary results show that Noah-MP is capable of simulating the dynamics of the major nitrogen pools. Further comprehensive evaluation of the new model will be conducted at one or more experimental sites.

  12. 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.

  13. 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