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Organic nitrogen uptake by plants: reevaluation by position-specific labeling of amino acids

Reevaluation of organic N uptake by plants by position-specific labeling

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Abstract

Current studies suggest that many plants are able to take up not only inorganic nitrogen (N) but also organic N. We used the novel tool of position-specific isotope labeling to improve the quantification of intact amino acid uptake and to deepen our understanding of the processes occurring at the root-soil-microorganism interface. Position-specific 14C and 15N labeled alanine enabled us to trace the uptake of C from individual molecule positions by Zea mays, Lupinus albus and Cichorium intybus. Uniformly 14C labeled alanine and acetate and inorganic 15NH4 + and 15NO3 − were applied as controls. Equal uptake of uniformly 14C labeled alanine and acetate showed that plant uptake of low molecular weight organic substances (LMWOS) is independent of N in the molecule. 14C uptake from individual molecule positions of alanine strongly differed: this confirmed that soil microorganisms cleaved alanine within 6 h into transformation products, which were then taken up by the plants. Microbial utilization strongly outcompeted the plant uptake of LMWOS in agricultural soils. This study revealed that position-specific labeling is an innovative tool that enables separation of the intact uptake from the uptake of molecule fragments and improves the understanding of competing processes for LMWOS utilization in the rhizosphere.

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References

  • Adamczyk B, Smolander A, Kitunen V, Godlewski M (2012) Proteoid roots and exudation of proteases by plant roots. In: Vivanco J, Baluska F (eds) Secretions and exudates in biological systems. Springer, Berlin, pp 75–89

    Chapter  Google Scholar 

  • Amelung W, Bol R, Friedrich C (1999) Natural C-13 abundance: a tool to trace the incorporation of dung-derived carbon into soil particle-size fractions. Rapid Commun Mass Spectrom 13:1291–1294

    Article  Google Scholar 

  • Ameziane R, RichardMolard C, Deleens E, MorotGaudry JF, Limami AM (1997) Nitrate ((NO3)-N-15) limitation affects nitrogen partitioning between metabolic and storage sinks and nitrogen reserve accumulation in chicory (Cichorium intybus L.). Planta 202:303–312

    Article  Google Scholar 

  • Apostel C, Dippold M, Glaser B, Kuzyakov Y (2013) Biochemical pathways of amino acids in soil: assessment by position-specific labeling and 13C-PLFA analysis. Soil Biol Biochem 67:31–40

    Article  Google Scholar 

  • Bardgett RD, Streeter TC, Bol R (2003) Soil microbes compete effectively with plants for organic-nitrogen inputs to temperate grasslands. Ecology 84:1277–1287

    Article  Google Scholar 

  • Biernath C, Fischer H, Kuzyakov Y (2008) Root uptake of N-containing and N-free low molecular weight organic substances by maize: A (14)C/(15)N tracer study. Soil Biol Biochem 40:2237–2245

    Article  Google Scholar 

  • Blagodatskaya EV, Blagodatsky SA, Anderson TH, Kuzyakov Y (2009) Contrasting effects of glucose, living roots and maize straw on microbial growth kinetics and substrate availability in soil. Eur J Soil Sci 60:186–197

    Article  Google Scholar 

  • Chapin FSI, Moilanen L, Kielland K (1993) Preferential use of organic nitrogen for growth by a non-mycorrhizal arctic sedge. Nature (Lond.) 361:150–153

    Article  Google Scholar 

  • Day DA, Poole PS, Tyerman SD, Rosendahl L (2001) Ammonia and amino acid transport across symbiotic membranes in nitrogen-fixing legume nodules. Cell Mol Life Sci 58:61–71

    Article  Google Scholar 

  • Delgado-Baquerizo M, Covelo F, Gallardo A (2011) Dissolved organic nitrogen in Mediterranean ecosystems. Pedosphere 21:309–318

    Article  Google Scholar 

  • Demidchik V, Maathuis FJM (2007) Physiological roles of nonselective cation channels in plants: from salt stress to signalling and development. New Phytol 175:387–404

    Article  Google Scholar 

  • Dijkstra P, Blankinship JC, Selmants PC, Hart SC, Koch GW, Schwartz E, Hungate BA (2011a) Probing carbon flux patterns through soil microbial metabolic networks using parallel position-specific tracer labeling. Soil Biol Biochem 43:126–132

    Article  Google Scholar 

  • Dijkstra P, Dalder JJ, Selmants PC, Hart SC, Koch GW, Schwartz E, Hungate BA (2011b) Modeling soil metabolic processes using isotopologue pairs of position-specific C-13-labeled glucose and pyruvate. Soil Biol Biochem 43:1848–1857

    Article  Google Scholar 

  • Dippold M, Kuzyakov Y (2013) Biogeochemical transformations of amino acids in soil assessed by position-specific labelling. Plant Soil 373:385–401

    Article  Google Scholar 

  • Doerr N, Kaiser K, Sauheitl L, Lamersdorf N, Stange CF, Guggenberger G (2012) Fate of ammonium N-15 in a Norway spruce forest under long-term reduction in atmospheric N deposition. Biogeochemistry 107:409–422

    Article  Google Scholar 

  • Doubnerova V, Ryslava H (2011) What can enzymes of C-4 photosynthesis do for C-3 plants under stress? Plant Sci 180:575–583

    Article  Google Scholar 

  • FAO (2006) Guidelines for soil description. Food and Agriculture Organization of the United Nations (FAO), Rome

    Google Scholar 

  • Fischer H, Kuzyakov Y (2010) Sorption, microbial uptake and decomposition of acetate in soil: transformations revealed by position-specific C-14 labeling. Soil Biol Biochem 42:186–192

    Article  Google Scholar 

  • Fischer H, Meyer A, Fischer K, Kuzyakov Y (2007) Carbohydrate and amino acid composition of dissolved organic matter leached from soil. Soil Biol Biochem 39:2926–2935

    Article  Google Scholar 

  • Fischer H, Eckhardt K-U, Meyer A, Neumann G, Leinweber P, Fischer K, Kuzyakov Y (2010a) Rhizodeposition of maize: short-term carbon budget and composition. J Plant Nutr Soil Sci 173:67–79

    Article  Google Scholar 

  • Fischer H, Ingwersen J, Kuzyakov Y (2010b) Microbial uptake of low-molecular-weight organic substances out-competes sorption in soil. Eur J Soil Sci 61:504–513

    Article  Google Scholar 

  • Fry B (2006) Stable isotope ecology. Springer, New York

    Book  Google Scholar 

  • Gavrichkova O, Kuzyakov Y (2008) Ammonium versus nitrate nutrition of Zea mays and Lupinus albus: effect on root-derived CO2 efflux. Soil Biol Biochem 40:2835–2842

    Article  Google Scholar 

  • Gavrichkova O, Kuzyakov Y (2010) Respiration costs associated with nitrate reduction as estimated by (CO2)-C-14 pulse labeling of corn at various growth stages. Plant Soil 329:433–445

    Article  Google Scholar 

  • Ge T-D, Roberts P, Jones DL, Yang D-D, Song S-W, Lu B, Ming D, Huang D-F (2008) Influence of inorganic and organic nitrogen on enzymes of nitrogen assimilation and growth in tomato seedlings. J Hortic Sci Biotechnol 83:513–519

    Google Scholar 

  • Ge T, Song S, Roberts P, Jones DL, Huang D, Iwasaki K (2009) Amino acids as a nitrogen source for tomato seedlings: the use of dual-labeled (C-13, N-15) glycine to test for direct uptake by tomato seedlings. Environ Exp Bot 66:357–361

    Article  Google Scholar 

  • Glaser B (2005) Compound-specific stable-isotope (delta C-13) analysis in soil science. J Plant Nutr Soil Sci 168:633–648

    Article  Google Scholar 

  • Glass ADM, Britto DT, Kaiser BN, Kinghorn JR, Kronzucker HJ, Kumar A, Okamoto M, Rawat S, Siddiqi MY, Unkles SE, Vidmar JJ (2002) The regulation of nitrate and ammonium transport systems in plants. J Exp Bot 53:855–864

    Article  Google Scholar 

  • Godlewski M, Adamczyk B (2007) The ability of plants to secrete proteases by roots. Plant Physiol Biochem 45:657–664

    Article  Google Scholar 

  • Goupil P, Loncle D, Druart N, Bellettre A, Rambour S (1998) Influence of ABA on nitrate reductase activity and carbohydrate metabolism in chicory roots (Cichorium intybus L.). J Exp Bot 49:1855–1862

    Article  Google Scholar 

  • Hawkins HJ, Wolf G, Stock WD (2005) Cluster roots of Leucadendron laureolum (Proteaceae) and Lupinus albus (Fabaceae) take up glycine intact: an adaptive strategy to low mineral nitrogen in soils? Ann Bot 96:1275–1282

    Article  Google Scholar 

  • He HB, Li XB, Zhang W, Zhang XD (2011) Differentiating the dynamics of native and newly immobilized amino sugars in soil frequently amended with inorganic nitrogen and glucose. Eur J Soil Sci 62:144–151

    Article  Google Scholar 

  • Hermans C, Hammond JP, White PJ, Verbruggen N (2006) How do plants respond to nutrient shortage by biomass allocation? Trends Plant Sci 11:610–617

    Article  Google Scholar 

  • Herrmann AM, Coucheney E, Nunan N (2014) Isothermal microcalorimetry provides new insight into terrestrial carbon cycling. Environ Sci Technol 48:4344–4352

    Article  Google Scholar 

  • Hobbie JE, Hobbie EA (2012) Amino acid cycling in plankton and soil microbes studied with radioisotopes: measured amino acids in soil do not reflect bioavailability. Biogeochemistry 107:339–360

    Article  Google Scholar 

  • Hocking PJ, Jeffery S (2004) Cluster-root production and organic anion exudation in a group of old-world lupins and a new-world lupin. Plant Soil 258:135–150

    Article  Google Scholar 

  • Hodge A, Robinson D, Fitter A (2000) Are microorganisms more effective than plants at competing for nitrogen? Trends Plant Sci 5:304–308

    Article  Google Scholar 

  • Jamtgard S, Nasholm T, Huss-Danell K (2008) Characteristics of amino acid uptake in barley. Plant Soil 302:221–231

    Article  Google Scholar 

  • Jones DL (1999) Amino acid biodegradation and its potential effects on organic nitrogen capture by plants. Soil Biol Biochem 31:613–622

    Article  Google Scholar 

  • Jones DL, Willett VB (2006) Experimental evaluation of methods to quantify dissolved organic nitrogen (DON) and dissolved organic carbon (DOC) in soil. Soil Biol Biochem 38:991–999

    Article  Google Scholar 

  • Jones DL, Shannon D, Murphy DV, Farrar J (2004) Role of dissolved organic nitrogen (DON) in soil N cycling in grassland soils. Soil Biol Biochem 36:749–756

    Article  Google Scholar 

  • Jones DL, Healey JR, Willett VB, Farrar JF, Hodge A (2005a) Dissolved organic nitrogen uptake by plants—an important N uptake pathway? Soil Biol Biochem 37:413–423

    Article  Google Scholar 

  • Jones DL, Kemmitt SJ, Wright D, Cuttle SP, Bol R, Edwards AC (2005b) Rapid intrinsic rates of amino acid biodegradation in soils are unaffected by agricultural management strategy. Soil Biol Biochem 37:1267–1275

    Article  Google Scholar 

  • Jones DL, Shannon D, Junvee-Fortune T, Farrar JF (2005c) Plant capture of free amino acids is maximized under high soil amino acid concentrations. Soil Biol Biochem 37:179–181

    Article  Google Scholar 

  • Jones DL, Shannon D, Junvee-Fortune T, Farrarc JF (2005d) Plant capture of free amino acids is maximized under high soil amino acid concentrations. Soil Biol Biochem 37:179–181

    Article  Google Scholar 

  • Jones DL, Nguyen C, Finlay RD (2009) Carbon flow in the rhizosphere: carbon trading at the soil-root interface. Plant Soil 321:5–33

    Article  Google Scholar 

  • Kranabetter JM, Dawson CR, Dunn DE (2007) Indices of dissolved organic nitrogen, ammonium and nitrate across productivity gradients of boreal forests. Soil Biol Biochem 39:3147–3158

    Article  Google Scholar 

  • Kuzyakov YV (1996) Transformation of low-molecular nitrogen-containing compounds in soil. Eurasian Soil Sci 29:1333–1341

    Google Scholar 

  • Kuzyakov Y, Blagodatskaya E (2015) Microbial hotspots and hot moments in soil: concept & review. Soil Biol Biochem 83:184–199

    Article  Google Scholar 

  • Kuzyakov Y, Jones DL (2006) Glucose uptake by maize roots and its transformation in the rhizosphere. Soil Biol Biochem 38:851–860

    Article  Google Scholar 

  • Kuzyakov Y, Xu X (2013) Competition between roots and microorganisms for nitrogen: mechanisms and ecological relevance. New Phytol 198:656–669

    Article  Google Scholar 

  • Lipson DA, Raab TK, Schmidt SK, Monson RK (1999) Variation in competitive abilities of plants and microbes for specific amino acids. Biol Fertil Soils 29:257–261

    Article  Google Scholar 

  • Liu Y, Wu L, Baddeley JA, Watson CA (2011) Models of biological nitrogen fixation of legumes. Sustainable agriculture, vol 2. Springer, Berlin, pp 883–905

    Google Scholar 

  • Matson PA, Parton WJ, Power AG, Swift MJ (1997) Agricultural intensification and ecosystem properties. Science 277:504–509

    Article  Google Scholar 

  • Nasholm T, Persson J (2001) Plant acquisition of organic nitrogen in boreal forests. Physiol Plant 111:419–426

    Article  Google Scholar 

  • Nasholm T, Ekblad A, Nordin A, Giesler R, Hogberg M, Hogberg P (1998) Boreal forest plants take up organic nitrogen. Nature 392:914–916

    Article  Google Scholar 

  • Nasholm T, Huss-Danell K, Hogberg P (2001) Uptake of glycine by field grown wheat. New Phytol 150:59–63

    Article  Google Scholar 

  • Paungfoo-Lonhienne C, Visser J, Lonhienne TGA, Schmidt S (2012) Past, present and future of organic nutrients. Plant Soil 359:1–18

    Article  Google Scholar 

  • Persson J, Nasholm T (2001) A GC–MS method for determination of amino acid uptake by plants. Physiol Plant 113:352–358

    Article  Google Scholar 

  • Rasmussen J, Sauheitl L, Eriksen J, Kuzyakov Y (2010) Plant uptake of dual-labeled organic N biased by inorganic C uptake: results of a triple labeling study. Soil Biol Biochem 42:524–527

    Article  Google Scholar 

  • Roberts P, Jones DL (2012) Microbial and plant uptake of free amino sugars in grassland soils. Soil Biol Biochem 49:139–149

    Article  Google Scholar 

  • Roberts P, Bol R, Jones DL (2007) Free amino sugar reactions in soil in relation to soil carbon and nitrogen cycling. Soil Biol Biochem 39:3081–3092

    Article  Google Scholar 

  • Sauheitl L, Glaser B, Weigelt A (2009a) Advantages of compound-specific stable isotope measurements over bulk measurements in studies on plant uptake of intact amino acids. Rapid Commun Mass Spectrom 23:3333–3342

    Article  Google Scholar 

  • Sauheitl L, Glaser B, Weigelt A (2009b) Uptake of intact amino acids by plants depends on soil amino acid concentrations. Environ Exp Bot 66:145–152

    Article  Google Scholar 

  • Schimel JP, Chapin FS (1996) Tundra plant uptake of amino acid and NH4 + nitrogen in situ: Plants compete well for amino acid N. Ecology 77:2142–2147

    Article  Google Scholar 

  • Spohn M, Kuzyakov Y (2013) Phosphorus mineralization can be driven by microbial need for carbon. Soil Biol Biochem 61:69–75

    Article  Google Scholar 

  • Stahl VM, Beyschlag W, Werner C (2011) Dynamic niche sharing in dry acidic grasslands—a N-15-labeling experiment. Plant Soil 344:389–400

    Article  Google Scholar 

  • Streeter TC, Bol R, Bardgett RD (2000) Amino acids as a nitrogen source in temperate upland grasslands: the use of dual labelled (C-13, N-15) glycine to test for direct uptake by dominant grasses. Rapid Commun Mass Spectrom 14:1351–1355

    Article  Google Scholar 

  • Svennerstam H, Ganeteg U, Bellini C, Nasholm T (2007) Comprehensive screening of Arabidopsis mutants suggests the lysine histidine transporter 1 to be involved in plant uptake of amino acids. Plant Physiol 143:1853–1860

    Article  Google Scholar 

  • Szajdak L, Jezierski A, Cabrera ML (2003) Impact of conventional and no-tillage management on soil amino acids, stable and transient radicals and properties of humic and fulvic acids. Org Geochem 34:693–700

    Article  Google Scholar 

  • Thede B (2010) Use of isotopes in agrochemical research. J Labelled Compd Radiopharm 53:322–326

    Article  Google Scholar 

  • Tian J, Dippold M, Pausch J, Blagodatskaya E, Fan M, Li X, Kuzyakov Y (2013) Microbial response to rhizodeposition depending on water regimes in paddy soils. Soil Biol Biochem 65:195–203

    Article  Google Scholar 

  • Tischner R (2000) Nitrate uptake and reduction in higher and lower plants. Plant Cell Environ 23:1005–1024

    Article  Google Scholar 

  • van Hees PAW, Jones DL, Godbold DL (2002) Biodegradation of low molecular weight organic acids in coniferous forest podzolic soils. Soil Biol Biochem 34:1261–1272

    Article  Google Scholar 

  • van Hees PAW, Jones DL, Finlay R, Godbold DL, Lundstomd US (2005) The carbon we do not see—the impact of low molecular weight compounds on carbon dynamics and respiration in forest soils: a review. Soil Biol Biochem 37:1–13

    Article  Google Scholar 

  • Vitousek PM, Gosz JR, Grier CC, Melillo JM, Reiners WA, Todd RL (1979) Nitrate losses from disturbed ecosystems. Science 204:469–474

    Article  Google Scholar 

  • Vitousek PM, Aber JD, Howarth RW, Likens GE, Matson PA, Schindler DW, Schlesinger WH, Tilman D (1997) Human alteration of the global nitrogen cycle: sources and consequences. Ecol Appl 7:737–750

    Google Scholar 

  • Warren CR (2009) Does nitrogen concentration affect relative uptake rates of nitrate, ammonium, and glycine? J Plant Nutr Soil Sci 172:224–229

    Article  Google Scholar 

  • Warren CR (2012) Post-uptake metabolism affects quantification of amino acid uptake. New Phytol 193:522–531

    Article  Google Scholar 

  • Wegener F, Beyschlag W, Werner C (2010) The magnitude of diurnal variation in carbon isotopic composition of leaf dark respired CO2 correlates with the difference between delta C-13 of leaf and root material. Funct Plant Biol 37:849–858

    Article  Google Scholar 

  • Wegner LH (2014) Root pressure and beyond: energetically uphill water transport into xylem vessels? J Exp Bot 65:381–393

    Article  Google Scholar 

  • Weigelt A, King R, Bol R, Bardgett RD (2003) Inter-specific variability in organic nitrogen uptake of three temperate grassland species. J Plant Nutr Soil Sci 166:606–611

    Article  Google Scholar 

  • Weigelt A, Bol R, Bardgett RD (2005) Preferential uptake of soil nitrogen forms by grassland plant species. Oecologia 142:627–635

    Article  Google Scholar 

  • Werner C, Gessler A (2011) Diel variations in the carbon isotope composition of respired CO2 and associated carbon sources: a review of dynamics and mechanisms. Biogeosciences 8:2437–2459

    Article  Google Scholar 

  • Xu XL, Ouyang H, Cao GM, Richter A, Wanek W, Kuzyakov Y (2011) Dominant plant species shift their nitrogen uptake patterns in response to nutrient enrichment caused by a fungal fairy in an alpine meadow. Plant Soil 341:495–504

    Article  Google Scholar 

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Acknowledgments

This study was financed by Deutsche Forschungsgemeinschaft DFG (DFG KU 1184/19-1). We thank Ilse Thaufelder and Stefanie Bösel, technical staff at the University of Bayreuth and Martin-Luther University of Halle, respectively, and Cristina Cavedon for figure design.

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    Authors and Affiliations

    1. Department of Agroecosystem Research, BayCEER, University of Bayreuth, Bayreuth, Germany

      Daniel Moran-Zuloaga & Michaela Dippold

    2. Departamento de Mecánica y Ciencias de la Producción, Escuela Superior Politécnica del Litoral, Guayaquil, Ecuador

      Daniel Moran-Zuloaga

    3. Department of Agricultural Soil Science, Georg-August-University Goettingen, Göttingen, Germany

      Michaela Dippold & Yakov Kuzyakov

    4. Department of Soil Biogeochemistry, Institute of Agricultural and Nutritional Science, Martin-Luther University Halle-Wittenberg, Halle, Germany

      Bruno Glaser

    5. Department of Soil Science of Temperate Ecosystems, Georg-August University Goettingen, Göttingen, Germany

      Yakov Kuzyakov

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    1. Daniel Moran-Zuloaga
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    Correspondence to Daniel Moran-Zuloaga.

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    Daniel Moran-Zuloaga and Michaela Dippold have contributed equally.

    Responsible Editor: Melanie A. Vile

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    Supplementary Figure

    Percentage of 14C incorporation in soil and CO2 efflux after position-specific labeling with alanine. The alanine positions were C-1 (carboxyl group), C-2 (amino-bound group) and C-3 (methyl group). Letters indicate significant differences (p < 0.001) between alanine C positions. Supplementary material 1 (JPEG 1840 kb).

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    Moran-Zuloaga, D., Dippold, M., Glaser, B. et al. Organic nitrogen uptake by plants: reevaluation by position-specific labeling of amino acids. Biogeochemistry 125, 359–374 (2015). https://doi.org/10.1007/s10533-015-0130-3

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