Relationships among phosphorus, molybdenum and free-living nitrogen fixation in tropical rain forests: results from observational and experimental analyses

Abstract

Biological nitrogen (N) fixation is the primary source of “new” N to unmanaged ecosystems, and recent analyses suggest that terrestrial N inputs via free-living N fixation may be more important than previously assumed. This may be particularly true in some tropical rain forests, where free-living fixation could outpace symbiotic N fixation to represent the dominant source of new N inputs. However, our understanding of the controls over free-living N fixation in tropical rain forests remains poor, which directly constrains our ability to predict how N cycling will respond to changing environmental conditions. Although both phosphorus (P) and molybdenum (Mo) availability have been shown to limit free-living N fixation rates in the tropics, few studies have simultaneously explored P versus Mo limitation or the potential importance of P × Mo interactions. Here, an archived set of foliar, litter, and soil samples from a Costa Rican tropical rain forest provided an opportunity to simultaneously assess the relative strength of P versus Mo relationships with free-living N fixation rates. We also conducted a short-term, full-factorial (P × Mo) litter incubation experiment to directly assess nutrient limitation, allowing us to explore P and Mo controls over free-living N fixation rates using both observational and experimental approaches. We previously showed that N fixation rates were positively correlated with P concentrations in all substrates and, using the archived samples, we now show that Mo concentrations correlated with N fixation only in canopy leaves (where total Mo concentrations were extremely low). Likewise, fertilization with P alone (and not Mo) stimulated leaf litter N fixation rates. Thus, our results suggest that P availability dominantly controls free-living N fixation at this site, and when taken with data from other studies, our results suggest that attempts to identify “the nutrient” that limits N fixation in “the tropics” may be misguided. Rather, nutrient controls over free-living N fixation appear to be more nuanced—and the true nature of nutrient limitation to N fixation likely varies over a variety of scales across the vast tropical rain forest biome.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. Aber JD, Nadelhoffer KJ, Steudler P, Melillo JM (1989) Nitrogen saturation in northern forest ecosystems. Bioscience 39:378–386

    Article  Google Scholar 

  2. Barron AR, Wurzburger N, Bellenger JP, Wright SJ, Kraepiel AML, Hedin LO (2009) Molybdenum limitation of asymbiotic nitrogen fixation in tropical forest soils. Nat Geosci 2:42–45

    Article  Google Scholar 

  3. Barron AR, Purves DW, Hedin LO (2011) Facultative nitrogen fixation by canopy legumes in a lowland tropical forest. Oecologia 165:511–520

    Article  Google Scholar 

  4. Belnap J (1996) Soil surface disturbance in cold deserts: effects on nitrogenase activity in cyanobacterial-lichen soil crusts. Biol Fertil Soils 23:362–367

    Article  Google Scholar 

  5. Benner JW, Conroy S, Lunch CK, Toyoda N, Vitousek PM (2007) Phosphorus fertilization increases the abundance and nitrogenase activity of the cyanolichen Pseudocyphellaria corcata in Hawaiian montane forests. Biotropica 39:400–405

    Article  Google Scholar 

  6. Bern CR, Townsend AR, Farmer GL (2005) Unexpected dominance of parent material strontium in a tropical forest on highly weathered soils. Ecology 86:626–632

    Article  Google Scholar 

  7. Berrange JP, Thorpe RS (1988) The geology, geochemistry and emplacement of the cretaceous tertiary ophiolitic Nicoya complex of the Osa Peninsula, southern Costa Rica. Tectonophysics 147:193–199

    Article  Google Scholar 

  8. Bishop PE, Jarlenski DM, Hetherington DR (1982) Expression of an alternative nitrogen fixation system in Azotobacter vinelandii. J Bacteriol 150:1244–1251

    Google Scholar 

  9. Bowell RJ, Ansah RK (1993) Trace element budget in an African savannah ecosystem. Biogeochemistry 20:103–126

    Article  Google Scholar 

  10. Brookshire ENJ, Hedin L, Newbold JD, Sigman DM, Jackson JK (2012) Sustained losses of bioavailable nitrogen from montane tropical forests. Nat Geosci. doi:10.1038/ngeo1372

    Google Scholar 

  11. Cleveland CC, Townsend AR (2006) Nutrient additions to a tropical rain forest drive substantial carbon dioxide losses to the atmosphere. Proc Natl Acad Sci (USA) 104:10316–10321

    Article  Google Scholar 

  12. Cleveland CC, Townsend AR, Schimel DS, Fisher H, Howarth RW, Hedin LO, Perakis SS, Latty EF, von Fischer JC, Elseroad A, Wasson M (1999) Global patterns of terrestrial biological nitrogen (N2) fixation in natural ecosystems. Glob Biogeochem Cycles 13:623–645

    Article  Google Scholar 

  13. Cleveland CC, Townsend AR, Schmidt SK (2002) Phosphorus limitation of microbial processes in moist tropical forests: evidence from short-term laboratory incubations and field studies. Ecosystems 5:680–691

    Article  Google Scholar 

  14. Cleveland CC, Houlton BZ, Neill C, Reed SC, Townsend AR, Wang Y (2010) Using indirect methods to constrain symbiotic nitrogen fixation rates: a case study from an Amazonian rain forest. Biogeochemistry 99:1–13

    Article  Google Scholar 

  15. Cusack DF, Silver W, McDowell WH (2009) Biological nitrogen fixation in two tropical forests: ecosystem-level patterns and effects of nitrogen fertilization. Ecosystems 12:1299–1315

    Article  Google Scholar 

  16. Eisele KA, Schimel DS, Kapsutka LA, Parton WJ (1989) Effects of available phosphorus and nitrogen–phosphorus ratios on non-symbiotic dinitrogen fixation in tallgrass prairie soils. Oecologia 79:471–474

    Article  Google Scholar 

  17. Elser JJ, Bracken MES, Cleland EE, Gruner DS, Harpole WS, Hillebrand H, Ngai JT, Seabloom ET, Shurin JB, Smith JE (2007) Global analysis of nitrogen and phosphorus limitation of primary producers in freshwater, marine and terrestrial ecosystems. Ecol Lett 10:1135–1142

    Article  Google Scholar 

  18. Galloway JN et al (2004) Nitrogen cycles: past, present and future. Biogeochemistry 70:153–166

    Article  Google Scholar 

  19. Gutschick VP (1981) Evolved strategies in nitrogen acquisition by plants. Am Naturalist 118:607–637

    Article  Google Scholar 

  20. Hardy RFW, Holsten RD, Jackson EK, Burns RC (1968) The acetylene–ethylene assay for N2 fixation: laboratory and field evaluation. Plant Physiol 43:1185–1207

    Article  Google Scholar 

  21. Hedin LO, Vitousek PM, Matson PA (2003) Nutrient losses over four million years of tropical forests development. Ecology 84:2231–2255

    Article  Google Scholar 

  22. Hedin LO, Brookshire ENJ, Menge DNL, Barron AR (2009) The nitrogen paradox in tropical forest ecosystems. Annu Rev Ecol Evol Syst 40:613–635

    Article  Google Scholar 

  23. Hicks WT, Harmon ME, Griffiths RP (2003) Abiotic controls on nitrogen fixation and respiration in selected woody debris from the Pacific Northwest, USA. Ecoscience 10:66–73

    Google Scholar 

  24. Holdridge LR, Grenke WC, Hatheway WH, Lian T, Tosi JA (1971) Forest environments in tropical life zones: a pilot study. Oxford, Oxford

    Google Scholar 

  25. Houlton BZ, Sigman DM, Hedin LO (2006) Isotopic evidence for large gaseous nitrogen losses from tropical rainforests. Proc Natl Acad Sci (USA) 103:8745–8750

    Article  Google Scholar 

  26. Howarth RW, Marino R, Lane J, Cole JJ (1988) Nitrogen fixation in freshwater, estuarine, and marine ecosystems. 1. Rates and importance. Limnol Oceanogr 33:669–687

    Article  Google Scholar 

  27. Hungate BA, Dukes JS, Shaw MR, Luo Y, Field CB (2003) Nitrogen and climate change. Science 302:1512–1513

    Article  Google Scholar 

  28. Hungate BA et al (2004) CO2 elicits a long-term decline in nitrogen fixation. Science 304:1291

    Article  Google Scholar 

  29. John RJ, Dalling W, Harms KE, Yavitt JB, Stallard RF, Mirabello M, Hubbell SP, Valencia R, Navarrete H, Vallejo M, Foster RB (2007) Soil nutrients influence spatial distributions of tropical tree species. Proc Natl Acad Sci (USA) 104:864–869

    Article  Google Scholar 

  30. Kappelle M, Castro M, Acevedo H, Gonzalez L, Monge H (2002) Ecosystems of the Osa Conservation Area (ACOSA). Instituto Nacional Biodiversidad (INBio), Santo Domingo de Heredia

    Google Scholar 

  31. Kuo S (1996) Methods of soil analysis. Part 3: Chemical methods. In: Sparks DL (ed) Phosphorus. Soil Society of America, Madison, WI, pp 869–919

    Google Scholar 

  32. Lewis WM, Melack JM, McDowell WH, McClain M, Richey JE (1999) Nitrogen yields from undisturbed watersheds in the Americas. Biogeochemistry 46:149–162

    Google Scholar 

  33. Lilienfein J, Wilcke W, Ayarza MA, Vilela L, do Carmo Lima S, Zech W (2000) Chemical fractionation of phosphorus, sulphur, and molybdenum in Brazilian savannah Oxisols under different land use. Geoderma 96:31–46

    Article  Google Scholar 

  34. Madigan MT, Martinko JM, Parker J (eds) (2003) Brock biology of microorganisms, 10th edn. Pearson Education Inc., Upper Saddle River, NJ

    Google Scholar 

  35. McNamara NP, Black HIJ, Piearce TG, Reay DS, Ineson P (2008) The influence of afforestation and tree species on soil methane fluxes from shallow organic soils at the UK Gisburn Forest Experiment. Soil Use Manag 24:1–7

    Article  Google Scholar 

  36. Menge DNL, Pacala SW, Hedin LO (2009) Emergence and maintenance of nutrient limitation over multiple time scales in terrestrial ecosystems. Am Nat 174:465–477

    Article  Google Scholar 

  37. Menyailo OV, Hungate BA (2003) Interactive effects of tree species and soil moisture on methane consumption. Soil Biol Biochem 35:625–628

    Article  Google Scholar 

  38. Palm C, Sanchez P, Ahamed S, Awiti A (2007) Soils: a contemporary perspective. Annu Rev Environ Resour 32:99–129

    Article  Google Scholar 

  39. Pearson HL, Vitousek PM (2002) Soil phosphorus fractions and symbiotic nitrogen fixation across a substrate-age gradient in Hawaii. Ecosystems 5:587–596

    Article  Google Scholar 

  40. Porder S, Vitousek P, Chadwick O, Chamberlain C, Hilley G (2007) Uplift, erosion, and phosphorus limitation in terrestrial ecosystems. Ecosystems 10:158–170

    Article  Google Scholar 

  41. Raymond J, Siefert JL, Staples CR, Blankenship RE (2004) The natural history of nitrogen fixation. Mol Biol Evol 21:541–554

    Article  Google Scholar 

  42. Reed SC, Seastedt TR, Mann CM, Suding KN, Townsend AR, Cherwin KL (2007a) Phosphorus fertilization stimulates nitrogen fixation and increases inorganic nitrogen concentrations in a restored Prairie. Appl Soil Ecol 36:238–242

    Article  Google Scholar 

  43. Reed SC, Cleveland CC, Townsend AR (2007b) Controls over leaf litter and soil nitrogen fixation in two lowland tropical rain forests. Biotropica 39:585–592

    Article  Google Scholar 

  44. Reed SC, Cleveland CC, Townsend AR (2008) Tree species control rates of free-living nitrogen fixation in a tropical rain forest. Ecology 89:2924–2934

    Article  Google Scholar 

  45. Reed SC, Townsend AR, Cleveland CC, Nemergut DR (2010) Microbial community shifts influence patterns in tropical forest nitrogen fixation. Oecologia 164:521–531

    Article  Google Scholar 

  46. Reed SC, Cleveland CC, Townsend AR (2011) Functional ecology of free-living nitrogen fixation: a contemporary perspective. Annu Rev Ecol Evol Syst 42:489–512

    Article  Google Scholar 

  47. Sanchez PA, Bandy DE, Villachica JH, Nicholaides JJ (1982) Amazon basin soils: management for continuous crop production. Science 216:821–827

    Article  Google Scholar 

  48. Silvester WB (1989) Molybdenum limitation of asymbiotic nitrogen fixation in forests of Pacific Northwest America. Soil Biol Biochem 21:283–289

    Article  Google Scholar 

  49. Smith VH (1992) Effects of nitrogen:phosphorus supply ratios on nitrogen fixation in agricultural and pastoral ecosystems. Biogeochemistry 18:19–35

    Article  Google Scholar 

  50. Sprent JI, Sprent P (1990) Nitrogen fixing organisms. Chapman and Hall, London

    Google Scholar 

  51. Townsend AR, Cleveland CC, Asner GP, Bustamante MMC (2007) Controls over foliar N:P ratios in tropical rain forests. Ecology 88:107–118

    Article  Google Scholar 

  52. Townsend AR, Asner GP, Cleveland CC (2008) The biogeochemical heterogeneity of tropical forests. Trends Ecol Evol 23:424–431

    Article  Google Scholar 

  53. van Groenigen K-J, Six J, Hungate BA, de Graaff M-A, van Breemen N, van Kessel C (2006) Element interactions limit soil carbon storage. Proc Natl Acad Sci (USA) 103:6571–6574

    Article  Google Scholar 

  54. van Haren JLM, de Oliveira RC Jr, Restrepo-Coupe N, Hutyra L, de Camargo PB, Keller M, Saleska SR (2010) Do plant species influence soil CO2 and N2O fluxes in a diverse tropical forest? J Geophys Res 115:G03010

    Article  Google Scholar 

  55. Vitousek PM, Field CB (1999) Ecosystem constraints to symbiotic nitrogen fixers: a simple model and its implications. Biogeochemistry 46:179–202

    Google Scholar 

  56. Vitousek P, Hobbie S (2000) Heterotrophic nitrogen fixation in decomposing litter: patterns, mechanisms, and models. Ecology 75:418–429

    Article  Google Scholar 

  57. Vitousek PM, Howarth RW (1991) Nitrogen limitation on land and in the sea: how can it occur? Biogeochemistry 13:87–115

    Article  Google Scholar 

  58. Vitousek PM, Sanford RL Jr (1986) Nutrient cycling in moist tropical forest. Annu Rev Ecol Syst 17:137–167

    Article  Google Scholar 

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

    Google Scholar 

  60. Vitousek PM et al (2002) Towards an ecological understanding of biological nitrogen fixation. Biogeochemistry 57(58):1–45

    Article  Google Scholar 

  61. Wichard T, Mishra B, Myneni SCB, Bellenger J-P, Kraepiel AML (2009) Storage and bioavailability of molybdenum in soils increased by organic matter complexation. Nat Geosci 2:625–629

    Article  Google Scholar 

  62. Wurzburger N, Bellenger JP, Kraepiel AML, Hedin LO (2012) Molybdenum and phosphorus interact to constrain asymbiotic nitrogen fixation in tropical forests. PLos One 7:e33710

    Article  Google Scholar 

Download references

Acknowledgments

We thank A. Barron, L. Hedin, W. Bowman, J. Neff, T. Seastedt, D. Nemergut, and S. Schmidt for discussions that helped shape this research and to two anonymous reviewers whose suggestions significantly improved the paper. We thank W. Wieder and A. Vega for help with sample collection; J. Feis and N. Ascarrunz for laboratory assistance; P. Vitousek and B. Houlton for advice on a previous version of the manuscript; John Zobitz and Dan Liptzin for statistical advice; and H. and M. Michaud, F. Campos and the Organization for Tropical Studies (OTS) and Ministerio de Ambiente y Energia (MINAE) in Costa Rica for logistical support. We appreciate the help of J. Drexler, F. Luiszer and the CU Boulder Laboratory for Environmental and Geological Studies for their assistance with molybdenum extraction and analysis. Any use of trade names is for descriptive purposes only and does not imply endorsement by the U.S. Government. This work was supported by grants from the National Science Foundation and the Andrew W. Mellon Foundation to CC, AT, and SR.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Sasha C. Reed.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Reed, S.C., Cleveland, C.C. & Townsend, A.R. Relationships among phosphorus, molybdenum and free-living nitrogen fixation in tropical rain forests: results from observational and experimental analyses. Biogeochemistry 114, 135–147 (2013). https://doi.org/10.1007/s10533-013-9835-3

Download citation

Keywords

  • Costa Rica
  • Fertilization
  • Free-living nitrogen fixation
  • Micro-nutrient
  • Molybdenum
  • Nutrient limitation
  • Phosphorus
  • Rain forest