, Volume 19, Issue 1, pp 89–98 | Cite as

Nitrogen isotope discrimination in white spruce fed with low concentrations of ammonium and nitrate

Original Article


Differences in δ15N among ten white spruce [Picea glauca (Moench) Voss] families were examined in hydroponic experiments testing (1) three N sources [100 μM N as (i) NH4+, (ii) NO3 or (iii) NH4NO3] and (2) two supply regimes [200 μM NH4+ (i) maintained steadily or (ii) recurrently drawn-down]. In the N-source experiment, the NH4+ treatment resulted in superior growth and lower C/N ratios. Whole plant δ15N was higher on NH4+ and NH4NO3, reflecting higher NH4+ removal rates from the media. Families expressed differences in biomass, C/N, δ15N and δ13C. Family δ15N and δ13C were positively correlated in the NH4NO3 treatment and the steady-state regime. Supply regime did not affect total biomass, but higher root/shoot ratios implied N was more limiting under the draw-down regime. Family rank changed with supply regime, but not with N source. Analysis of media isotope enrichment during substrate depletion revealed relationships between net discrimination and external N concentration. Discrimination against 15NH4+ was about twice that of 15NO3. A simple model relating isotope discrimination to relative rates of ion efflux and influx predicted efflux/influx ratios consistent with published values for white spruce. We propose that genetic differences in discrimination are caused by different demands on assimilation and in uptake capacity which interact, influencing the balance between N influx and efflux.


Picea glauca Stable isotopes Fractionation Nitrogen assimilation Ion flux 



This research was supported by a Natural Sciences and Engineering Research Council (Canada) grant to R.D.G.


  1. Bergersen FJ, Peoples MB, Turner GL (1988) Isotopic discrimination during the accumulation of nitrogen by soybeans. Aust J Plant Physiol 15:407–420Google Scholar
  2. Brooks PD, Stark JM, McInteer BB, Preston T (1989) Diffusion method to prepare soil extracts for automated nitrogen-15 analysis. Soil Sci Soc Am J 53:1707–1711Google Scholar
  3. Cawse PA (1967) The determination of nitrate in soil solutions by ultraviolet spectrophotometry. Analyst 92:311–315CrossRefGoogle Scholar
  4. Comstock JP (2001) Steady-state isotopic fractionation in branched pathways using plant uptake of NO3 as an example. Planta 214:220–234PubMedGoogle Scholar
  5. Eghball B, Maranville JW (1993) Root development and nitrogen influx of corn genotypes grown under combined drought and nitrogen stresses. Agron J 85:147–152Google Scholar
  6. Evans RD (2001) Physiological mechanisms influencing plant nitrogen isotope composition. Trends Plant Sci 6:121–126CrossRefPubMedGoogle Scholar
  7. Evans RD, Bloom AJ, Sukrapanna SS, Ehleringer JR (1996) Nitrogen isotope composition of tomato (Lycopersicon esculentum Mill. cv.T-f) grown under ammonium or nitrate nutrition. Plant Cell Environ 19:1317–1323Google Scholar
  8. Farquhar GD, O’Leary MH, Berry JA (1982) On the relationship between carbon isotope discrimination and the intercellular carbon dioxide concentration in leaves. Aust J Plant Physiol 9:121–137Google Scholar
  9. Guy RD, Berry JA, Fogel ML, Hoering TC (1989) Differential fractionation of oxygen isotopes by cyanide-resistant and cyanide-sensitive respiration in plants. Planta 177:483–491Google Scholar
  10. Handley LL, Raven JA (1992) The use of natural abundance of nitrogen isotopes in plant physiology and ecology. Plant Cell Environ 15:965–985Google Scholar
  11. Handley LL, Robinson D, Forster BP, Ellis RP, Scrimgeour CM, Gordon DC, Nevo E, Raven JA (1997) Shoot δ15N correlates with genotype and salt stress in barley. Planta 201:100–102Google Scholar
  12. Henry BK, Atkin OK, Day DA, Millar AH, Menz RI, Farquhar GD (1999) Calculation of the oxygen isotope discrimination factor for studying plant respiration. Aust J Plant Physiol 26:773–780Google Scholar
  13. Hoch MP, Fogel ML, Kirchman DL (1992) Isotope fractionation associated with ammonium uptake by a marine bacterium. Limnol Oceanogr 37:1447–1459Google Scholar
  14. Högberg P (1997) Tansley Review No. 95. 15N natural abundance in soil-plant systems. New Phytol 137:179–203CrossRefGoogle Scholar
  15. Högberg P, Högberg MN, Quist ME, Ekblad A, Nasholm T (1999) Nitrogen isotope fractionation during nitrogen uptake by ectomycorrhizal and non-mycorrhizal Pinus sylvestris. New Phytol 142:569–576CrossRefGoogle Scholar
  16. Holmes RM, McClelland JW, Sigman DM, Fry B, Peterson BJ (1998) Measuring 15N–NH4+ in marine, estuarine and fresh waters: an adaptation of the ammonia diffusion method for samples with low ammonium concentrations. Mar Chem 60:235–243CrossRefGoogle Scholar
  17. Imsande J, Touraine B (1994) N demand and the regulation of nitrate uptake. Plant Physiol 105:3–7PubMedGoogle Scholar
  18. Johnson CM, Stout PR, Broyer TC, Carlton AB (1957) Comparative chlorine requirements of different plant species. Plant Soil 8:337–353Google Scholar
  19. Khan SN, Mulvaney RL, Mulvaney CS (1997) Accelerated diffusion methods for inorganic nitrogen analysis of soil extracts and water. Soil Sci Soc Am J 61:936–947Google Scholar
  20. King BJ, Siddiqi MY, Ruth TJ, Warner RL, Glass A (1993) Feedback regulation of nitrate influx in barley roots by nitrate, nitrite, and ammonium. Plant Physiol 102:1279–1286PubMedGoogle Scholar
  21. Kolb KJ, Evans RD (2003) Influence of nitrogen source and concentration on nitrogen isotopic discrimination in two barley genotypes (Hordeum vulgare L.). Plant Cell Environ 26:1431–1440CrossRefGoogle Scholar
  22. Kronzucker HJ, Siddiqi MY, Glass ADM (1995a) Compartmentation and flux characteristics of ammonium in spruce. Planta 196:691–698Google Scholar
  23. Kronzucker HJ, Siddiqi MY, Glass ADM (1995b) Compartmentation and flux characteristics of nitrate in spruce. Planta 196:674–682Google Scholar
  24. Kronzucker HJ, Siddiqi MY, Glass ADM (1997) Conifer root discrimination against soil nitrate and the ecology of forest succession. Nature 385:59–61CrossRefGoogle Scholar
  25. Ledgard SF, Woo KC, Bergersen FJ (1985) Isotopic fractionation during reduction of nitrate and nitrite by extracts of spinach leaves. Aust J Plant Physiol 12:631–640Google Scholar
  26. Li B, McKeand SE, Allen HL (1991) Genetic variation in nitrogen use efficiency of loblolly pine seedlings. For Sci 37:613–626Google Scholar
  27. Livingston NJ, Guy RD, Sun ZJ, Ethier GJ (1999) The effects of nitrogen stress on the stable carbon isotope composition, productivity and water use efficiency of white spruce (Picea glauca (Moench) Voss) seedlings. Plant Cell Environ 22:281–289CrossRefGoogle Scholar
  28. Mariotti A, Mariotti F, Champigny M-L, Amarger N, Moyse A (1982) Nitrogen isotope fractionation associated with nitrate reductase activity and uptake of NO3 by pearl millet. Plant Physiol 69:880–884Google Scholar
  29. Merbach M, Mirus E, Knof G, Remus R, Ruppel S, Russow R, Gransee A, Schulze J (1999) Release of carbon and nitrogen compounds by plant roots and their possible ecological importance. J Plant Nutr Soil Sci 162:373–383CrossRefGoogle Scholar
  30. Min X, Siddiqi MY, Guy RD, Glass ADM, Kronzucker HJ (1999) A comparative study of fluxes and compartmentation of nitrate and ammonium in early-successional tree species. Plant Cell Environ 22:821–830CrossRefGoogle Scholar
  31. Mulavaney RL, Khan SA (1999) Use of diffusion to determine inorganic N in a complex organic matrix. Soil Sci Soc Am J 63:240–246Google Scholar
  32. Needoba JA, Waser NA, Harrison PJ, Calvert SE (2003) Nitrogen isotope fractionation in 12 species of marine phytoplankton during growth on nitrate. Mar Ecol Prog Ser 255:81–91Google Scholar
  33. O’Leary MH (1993) Biochemical basis of carbon isotope fractionation. In: Ehleringer JR, Hall AE, Farquhar GD (eds) Stable isotopes and plant carbon-water relations. Academic, New York, pp 19–28Google Scholar
  34. Patterson TB, Guy RD, Dang QL (1997) Whole-plant nitrogen- and water-relations traits, and their associated trade-offs, in adjacent muskeg and upland boreal spruce species. Oecologia 110:160–168CrossRefGoogle Scholar
  35. Ricker WE (1984) Computation and uses of central trend lines. Can J Zool 62:1897–1905Google Scholar
  36. Robinson D (2001) δ15N as an integrator of the nitrogen cycle. Trends Ecol Evol 16:153–162CrossRefPubMedGoogle Scholar
  37. Robinson D, Handley LL, Scrimgeour CM, Gordon DC, Forster BP, Ellis RP (2000) Using stable isotope natural abundances (δ15N and δ13C) to integrate the stress responses of wild barley (Hordeum spontaneum C. Koch.) genotypes. J Exp Bot 51:41–50CrossRefPubMedGoogle Scholar
  38. Shearer G, Kohl DH (1989) Natural 15N abundance of NH4+, amide N, and total N in various fractions of nodules of peas, soybeans and lupins. Aust J Plant Physiol 16:305–313Google Scholar
  39. Solorzano L (1969) Determination of ammonia in natural waters by the phenolhypochlorite method. Limnol Oceanogr 14:799–801Google Scholar
  40. Sun ZJ, Livingston NJ, Guy RD, Ethier GJ (1996) Stable carbon isotopes as indicators of increased water use efficiency and productivity in white spruce (Picea glauca (Moench) Voss) seedlings. Plant Cell Environ 19:887–894Google Scholar
  41. Swiader JM, Chyan Y, Splittstoesser WE (1991) Genotypic differences in nitrogen uptake, dry matter production, and nitrogen distribution in pumpkins (Cucurbita moschata Poir.). J Plant Nutr 14:511–524Google Scholar
  42. Tirol-Padre A, Ladha JK, Singh U, Laureles E, Punzalan G, Akita S (1996) Grain yield performance of rice genotypes at suboptimal levels of soil N as affected by N uptake and utilization efficiency. Field Crops Res 46:127–143CrossRefGoogle Scholar
  43. Waser NA, Yu Z, Yin K, Nielsen B, Harrison PJ, Turpin DH, Calvert SE (1999) Nitrogen isotopic fractionation during a simulated diatom spring bloom: importance of N-starvation in controlling fractionation. Mar Ecol Prog Ser 179:291–296Google Scholar
  44. Yoneyama T, Kaneko A (1989) Variations in the natural abundance of 15N in nitrogenous fractions of the komatsuna plants supplied with nitrate. Plant Cell Physiol 30:957–962Google Scholar
  45. Yoneyama T, Omata T, Nakata S, Yazaki J (1991) Fractionation of nitrogen isotopes during the uptake and assimilation of ammonia by plants. Plant Cell Physiol 32:1211–1217Google Scholar
  46. Yoneyama T, Kamachi K, Yamaya T, Mae T (1993) Fractionation of nitrogen isotopes by glutamine synthetase isolated from spinach leaves. Plant Cell Physiol 34:489–491Google Scholar
  47. Yoneyama T, Matsumaru T, Usui K, Engelaar WMHG (2001) Discrimination of nitrogen isotopes during absorption of ammonium and nitrate at different nitrogen concentrations by rice (Oryza sativa L.) plants. Plant Cell Environ 24:133–139CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Department of Forest Sciences, Faculty of ForestryUniversity of British ColumbiaVancouverCanada

Personalised recommendations