Advertisement

Plant and Soil

, Volume 399, Issue 1–2, pp 415–426 | Cite as

Do techniques based on 15N enrichment and 15N natural abundance give consistent estimates of the symbiotic dependence of N2-fixing plants?

  • Phillip M. ChalkEmail author
  • Caio T. Inácio
  • Fabiano C. Balieiro
  • Janaina R. C. Rouws
Review Article

Abstract

Aims

The primary aim of this review is to determine if methods based on 15N enrichment (E) and 15N natural abundance (NA) give consistent estimates of the proportional dependence of N2-fixing species on biological N2 fixation (P atm), and secondly to attempt to explain any inconsistencies that may be found.

Methods

Published estimates of the symbiotic dependence of N2-fixing plants based on E and NA techniques applied in the same experiment were compared across scales from glasshouse pots to field plots to landscapes in agricultural and forest ecosystems, which included grain legumes, pasture and forage legumes, and woody perennials. A meta-analysis of the published data was based on correlation coefficients, box-plots and confidence intervals of means.

Results

In some studies, estimates were reference plant dependent for both E and NA techniques, indicating temporal and/or spatial variations in the natural and artificial distribution of 15N, which can sometimes result in erroneous negative estimates of symbiotic dependence. While significant correlations were obtained between E and NA estimates of P atm for each of the three groups of N2-fixing species, the probability that the methods provided estimates of P atm within −5 to +5 % of each other was 0.29 or was 0.54 within −10 to +10 % of each other.

Conclusions

We have identified a number of interacting factors that may contribute to the inconsistent agreement between estimates of P atm by E and NA techniques, which underlines the need for a re-examination of the fundamental assumptions on which each method is based, and whether those assumptions are valid in any given situation.

Keywords

Symbiotic dependence 15N enrichment 15N natural abundance δ15Legumes Alder 

Notes

Acknowledgments

The senior author thanks the Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) for a visiting scientist fellowship (Pesquisador Visitante No. 101.466/2014) and EMBRAPA-Solos as the host Institution.

References

  1. Amarger N, Mariotti A, Mariotti F, Durr JC, Bourguignon C, Lagacherie B (1979) Estimate of symbiotically fixed nitrogen in field grown soybeans using variations in 15N natural abundance. Plant Soil 52:269–280CrossRefGoogle Scholar
  2. Androsoff GL, van Kessel C, Pennock DJ (1995) Landscape-scale estimates of dinitrogen fixation by Pisum sativum by nitrogen-15 natural abundance and enriched isotope dilution. Biol Fertil Soils 20:33–40CrossRefGoogle Scholar
  3. Bergersen FJ, Turner GL (1983) An evaluation of 15N methods for estimating nitrogen fixation in a subterranean clover-perennial ryegrass sward. Crop Past Sci 34:391–401CrossRefGoogle Scholar
  4. Boddey RM, Peoples MB, Palmer B, Dart PJ (2000) Use of the 15N natural abundance technique to quantify biological nitrogen fixation by woody perennials. Nutr Cycl Agroecosyst 57:235–270CrossRefGoogle Scholar
  5. Bouillet JP, Laclau JP, Gonçalves JLM, Moreira MZ, Trivelin PCO, Jourdan C, Silva EV, Piccolo MC, Tsai SM, Galiana A (2008) Mixed-species plantations of Acacia mangium and Eucalyptus grandis in Brazil. 2: nitrogen accumulation in the stands and biological N2 fixation. For Ecol Manag 255:3918–3930CrossRefGoogle Scholar
  6. Bremer E, van Kessel C (1990) Appraisal of the nitrogen-15 natural abundance method for quantifying dinitrogen fixation. Soil Sci Soc Am J 54:404–411CrossRefGoogle Scholar
  7. Brendel O, Wheeler C, Handley L (1997) A statistical comparison of the two-source δ15N and 15N isotope dilution methods for estimating plant N2-fixation using Trifolium pratense and Lolium perenne. Funct Plant Biol 24:631–636Google Scholar
  8. Burchill W, James EK, Li D, Lanigan GJ, Williams M, Iannetta PPM, Humphreys J (2014) Comparisons of biological nitrogen fixation in association with white clover (Trifolium repens L.) under four fertiliser nitrogen inputs as measured using two 15N techniques. Plant Soil 385:287–302CrossRefGoogle Scholar
  9. Cadisch G, Hairiah K, Giller KE (2000) Applicability of the natural 15N abundance technique to measure N2 fixation in Arachis hypogaea grown on an Ultisol. NJAS - Wageningen J Life Sci 48:31–45CrossRefGoogle Scholar
  10. Carranca C, de Varennes A, Rolston DE (1999) Biological nitrogen fixation estimated by 15N dilution, natural 15N abundance, and N difference techniques in a subterranean clover-grass sward under Mediterranean conditions. Eur J Agron 10:81–89CrossRefGoogle Scholar
  11. Chalk PM (1985) Estimation of N2 fixation by isotope dilution: an appraisal of techniques involving 15N enrichment and their application. Soil Biol Biochem 17:389–410CrossRefGoogle Scholar
  12. Chalk PM (1996) Estimation of N2 fixation by 15N isotope dilution – The A-value approach. Soil Biol Biochem 28:1123–1130CrossRefGoogle Scholar
  13. Chalk PM, Ladha JK (1999) Estimation of legume symbiotic dependence: an evaluation of techniques based on 15N dilution. Soil Biol Biochem 31:1901–1917CrossRefGoogle Scholar
  14. Chalk PM, Smith CJ, Hopmans P, Hamilton SD (1996) A yield-independent, 15N-isotope dilution method to estimate legume symbiotic dependence without a non-fixing reference plant. Biol Fertil Soils 23:196–199CrossRefGoogle Scholar
  15. Chalk PM, Inácio CT, Craswell ET, Chen D (2015) On the usage of absolute (x) and relative (δ) values of 15N abundance. Soil Biol Biochem 85:51–53CrossRefGoogle Scholar
  16. Domenach AM, Kurdali F, Bardin R (1989) Estimation of symbiotic dinitrogen fixation in alder forest by the method based on natural 15N abundance. Plant Soil 118:51–59CrossRefGoogle Scholar
  17. Doughton JA, Vallis I, Saffigna PG (1992) An indirect method for estimating 15N isotope fractionation during nitrogen fixation by a legume under field conditions. Plant Soil 144:23–29CrossRefGoogle Scholar
  18. Doughton JA, Saffigna PG, Vallis I, Mayer RJ (1995) Nitrogen fixation in chickpea. II. Comparison of 15N enrichment and 15N natural abundance methods for estimating nitrogen fixation. Crop Past Sci 46:225–236CrossRefGoogle Scholar
  19. Eriksen J, Høgh-Jensen H (1998) Variations in the natural abundance of 15N in ryegrass/white clover shoot material as influenced by cattle grazing. Plant Soil 205:67–76CrossRefGoogle Scholar
  20. Evans J, O’Connor GE, Turner GL, Bergersen FJ (1987) Influence of mineral nitrogen on nitrogen fixation by lupin (Lupinus angustifolius) as assessed by 15N isotope dilution methods. Field Crop Res 17:109–120CrossRefGoogle Scholar
  21. Feigin A, Kohl DH, Shearer G, Commoner B (1974) Variation in the natural nitrogen-15 abundance in nitrate mineralized during incubation of several Illinois soils. Soil Sci Soc Am J 38:90–95CrossRefGoogle Scholar
  22. Hairiah K, Van Noordwijk M, Cadisch G (2000) Quantification of biological N2 fixation by hedgerow trees in Northern Lampung. NJAS – Wageningen J Life Sci 48:47–59CrossRefGoogle Scholar
  23. Hansen JP, Vinther FP (2001) Spatial variability of symbiotic N2 fixation in grass-white clover pastures estimated by the 15N isotope dilution method and the natural 15N abundance method. Plant Soil 230:257–266CrossRefGoogle Scholar
  24. Høgh-Jensen H, Schjoerring JK (1994) Measurement of biological dinitrogen fixation in grassland: comparison of the enriched 15N dilution and the natural 15N abundance methods at different nitrogen application rates and defoliation frequencies. Plant Soil 166:153–163CrossRefGoogle Scholar
  25. Hossain SA, Waring SA, Strong WM, Dalal RC, Weston EJ (1995) Estimates of nitrogen fixation by legumes in alternate cropping systems at Warra, Queensland, using enriched-15N dilution and natural 15N abundance techniques. Crop Past Sci 46:493–505CrossRefGoogle Scholar
  26. Huss-Danell K, Chaia E (2005) Use of different plant parts to study N2 fixation with 15N techniques in field-grown red clover (Trifolium pratense). Physiol Plant 125:21–31CrossRefGoogle Scholar
  27. Huss-Danell K, Chaia E, Carlsson G (2007) N2 fixation and nitrogen allocation to above and below ground plant parts in red clover-grasslands. Plant Soil 299:215–226CrossRefGoogle Scholar
  28. Issah G, Kimaro AA, Kort J, Knight JD (2014) Quantifying biological nitrogen of agroforestry shrub species using 15N dilution techniques under greenhouse conditions. Agrofor Syst 88:607–617CrossRefGoogle Scholar
  29. Jacot KA, Lüscher A, Nösberger J, Hartwig UA (2000) Symbiotic N2 fixation of various legume species along an altitudinal gradient in the Swiss Alps. Soil Biol Biochem 32:1043–1052CrossRefGoogle Scholar
  30. Kohl DH, Shearer G, Harper JE (1980) Estimates of N2-fixation based on differences in the natural abundance of I5N in nodulating and non-nodulating isolines of soybeans. Plant Physiol 66:61–65CrossRefPubMedPubMedCentralGoogle Scholar
  31. Kurdali F, Domenach AM, Bardin R (1990) Alder-poplar associations: determination of plant nitrogen sources by isotope techniques. Biol Fertil Soils 9:321–329CrossRefGoogle Scholar
  32. Ledgard SF, Steele KW (1992) Biological nitrogen fixation in mixed legume/grass pastures. Plant Soil 141:137–153CrossRefGoogle Scholar
  33. Ledgard SF, Simpson JR, Freney JR, Bergersen FJ (1985a) a) Field evaluation of 15N techniques for estimating nitrogen fixation in legume-grass associations. Crop Past Sci 36:247–258CrossRefGoogle Scholar
  34. Ledgard SF, Simpson JR, Freney JR, Bergersen FJ (1985b) b) Effect of reference plant on estimation of nitrogen fixation by subterranean clover using 15N methods. Crop Past Sci 36:663–676CrossRefGoogle Scholar
  35. McAuliffe C, Chamblee DS, Uribe-Arango H, Woodhouse WW Jr (1958) Influence of inorganic nitrogen on nitrogen fixation by legumes as revealed by N15. Agron J 50:334–337CrossRefGoogle Scholar
  36. Oberson A, Nanzer S, Bosshard C, Dubois D, Mäder P, Frossard E (2007) Symbiotic N2 fixation by soybean in organic and conventional cropping systems estimated by 15N dilution and 15N natural abundance. Plant Soil 290:69–83CrossRefGoogle Scholar
  37. Ofori F, Pate JS, Stern WR (1987) Evaluation of N2-fixation and nitrogen economy of a maize/cowpea intercrop system using15N dilution methods. Plant Soil 102:149–160CrossRefGoogle Scholar
  38. Okito A, Alves BRJ, Urquiaga S, Boddey RM (2004) Isotopic fractionation during N2 fixation by four tropical legumes. Soil Biol Biochem 36:1179–1190CrossRefGoogle Scholar
  39. Pate JS, Unkovich MJ, Armstrong EL, Stanford P (1994) Selection of reference plants for 15N natural abundance assessment of N2 fixation by crop and pasture legumes in south-west Australia. Crop Past Sci 45:133–147CrossRefGoogle Scholar
  40. Pauferro N, Guimarães AP, Jantalia CP, Urquiaga S, Alves BJR, Boddey RM (2010) 15N natural abundance of biologically fixed N2 in soybean is controlled more by the Bradyrhizobium strain than by the variety of the host plant. Soil Biol Biochem 42:1694–1700CrossRefGoogle Scholar
  41. Peoples MB, Palmer B, Lilley DM, Duc LM, Herridge DF (1996) Application of 15N and xylem ureide methods for assessing N2 fixation of three shrub legumes periodically pruned for forage. Plant Soil 182:125–137CrossRefGoogle Scholar
  42. Riffkin PA, Quigley PE, Kearney GA, Cameron FJ, Gault RR, Peoples MB, Theis JE (1999) Factors associated with biological nitrogen fixation in dairy pastures in south-western Victoria. Crop Past Sci 50:261–272CrossRefGoogle Scholar
  43. Shearer G, Kohl DH (1986) N2 fixation in field settings: estimates based on natural 15N abundance. Funct Plant Biol 13:699–756Google Scholar
  44. Somado EA, Kuehne RF (2006) Appraisal of the 15N-isotope dilution and 15N natural abundance methods for quantifying nitrogen fixation by flood-tolerant green manure legumes. Afr J Biotechnol 5:1210–1214Google Scholar
  45. Stevenson FC, van Kessel C, Knight JD (1995) Dinitrogen fixation in pea: controls at the landscape- and micro-scale. Soil Sci Soc Am J 59:1603–1611CrossRefGoogle Scholar
  46. Tobita S, Ito O, Matsunaga R, Rao TP, Rego TJ, Johansen C, Yoneyama T (1994) Field evaluation of nitrogen fixation and use of nitrogen fertilizer by sorghum/pigeonpea intercropping on an Alfisol in the Indian semi-arid tropics. Biol Fertil Soils 17:241–248CrossRefGoogle Scholar
  47. Unkovich MJ, Pate JS (2000) An appraisal of recent field measurements of symbiotic N2 fixation by annual legumes. Field Crop Res 211:211–228CrossRefGoogle Scholar
  48. Unkovich MJ, Pate JS, Sanford P, Armstrong EL (1994) Potential precision of the δ15N natural abundance method in field estimates of nitrogen fixation by crop and pasture legumes in S.W. Australia. Crop Past Sci 45:119–132CrossRefGoogle Scholar
  49. Unkovich M, Herridge D, Peoples M, Cadisch G, Boddey R., Giller K, Alves B, Chalk, P (2008) Measuring Plant-Associated Nitrogen Fixation in Agricultural Systems. Australian Centre for International Agricultural Research, Canberra. 258 pp. http://aciar.gov.au/files/node/10169/mn136_measuring_plant_associated_nitrogen_fixation_19979.pdf
  50. Wanek W, Arndt SK (2002) Difference in δ15N signatures between nodulated roots and shoots of soybean is indicative of the contribution of symbiotic N2 fixation to plant N. J Exp Bot 371:1109–1118CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Phillip M. Chalk
    • 1
    Email author
  • Caio T. Inácio
    • 2
  • Fabiano C. Balieiro
    • 1
  • Janaina R. C. Rouws
    • 3
  1. 1.Embrapa-SolosRio de JaneiroBrazil
  2. 2.Departamento de SolosUniversidade Federal Rural do Rio de JaneiroSeropédicaBrazil
  3. 3.Embrapa-AgrobiologiaSeropédicaBrazil

Personalised recommendations