Plant and Soil

, Volume 314, Issue 1–2, pp 197–210 | Cite as

Effects of elevated carbon dioxide and nitrogen fertilization on nitrate reductase activity in sweetgum and loblolly pine trees in two temperate forests

  • Susan M. Natali
  • Sergio A. Sañudo-Wilhelmy
  • Manuel T. Lerdau
Regular Article


Nitrogen (N) availability is a major factor limiting plant production in many terrestrial ecosystems and is a key regulator of plant response to elevated CO2. Plant N status is a function of both soil N availability and plant N uptake and assimilation capacity. As a rate-limiting step in nitrate assimilation, the reduction of nitrate is an important component of plant physiological response to elevated CO2 and terrestrial carbon sequestration. We examine the effects of elevated CO2 and N availability on the activity of nitrate reductase, the enzyme catalyzing the reduction of nitrate to nitrite, in two temperate forests—a closed canopy sweetgum (Liquidambar styraciflua) plantation in Tennessee (Oak Ridge National Laboratory (ORNL)) and a loblolly pine (Pinus taeda) stand in North Carolina (Duke). Both CO2 and N enrichment had species specific impacts on nitrate reductase activity (NaR). Elevated CO2 and N fertilization decreased foliar NaR in P. taeda, but there were no treatment effects on L. styraciflua NaR at ORNL or Duke. NaR in 1-year P. taeda needles was significantly greater than in 0-year old needles across treatments. P. taeda NaR was negatively correlated with bio-available molybdenum concentrations in soils, suggesting that CO2 and N-mediated changes in soil nutrient status may be altering soil-plant N-dynamics. The variation in response among species may reflect different strategies for acquiring N and suggests that elevated CO2 may alter plant N dynamics through changes in NaR.


Free-air carbon dioxide enrichment (FACE) Liquidambar styraciflua Micronutrients Molybdenum NO3 assimilation Pinus taeda 



nitrate reductase activity


free-air carbon dioxide enrichment


soil organic matter



We thank C. Iversen, R. Norby, R. Oren, and the staff at the FACE sites for field support, and R. Norby, S. Baines, and two anonymous reviewers for advice during manuscript preparation. This work was supported by the U. S. Department of Energy, Office of Science (BER), and fellowships from the National Science Foundation (S.M.N.) and Department of Energy (S.M.N.).


  1. Ainsworth EA, Long SP (2005) What have we learned from 15 years of free-air CO2 enrichment (FACE)? A meta-analytic review of the responses of photosynthesis, canopy properties and plant production to rising CO2. New Phytol 165:351–371PubMedCrossRefGoogle Scholar
  2. Andrews JA, Schlesinger WH (2001) Soil CO2 dynamics, acidification, and chemical weathering in a temperate forest with experimental CO2 enrichment. Global Biogeochem Cycles 15:149–162CrossRefGoogle Scholar
  3. BassiriRad H (2000) Kinetics of nutrient uptake by roots: responses to global change. New Phytol 147:155–169CrossRefGoogle Scholar
  4. BassiriRad H, Griffin KL, Strain BR, Reynolds JF (1996a) Effects of CO2 enrichment on growth and root (NH4 +)-N-15 uptake rate of loblolly pine and ponderosa pine seedlings. Tree Physiol 16:957–962PubMedGoogle Scholar
  5. BassiriRad H, Thomas RB, Reynolds JF, Strain BR (1996b) Differential responses of root uptake kinetics of NH4 + and NO3 - to enriched atmospheric CO2 concentration in field-grown loblolly pine. Plant Cell Environ 19:367–371CrossRefGoogle Scholar
  6. Bloom AJ, Caldwell RM, Finazzo J, Warner RL, Weissbart J (1989) Oxygen and carbon dioxide fluxes from barley shoots depend on nitrate assimilation. Plant Physiol 91:352–356PubMedGoogle Scholar
  7. Bloom AJ, Smart DR, Nguyen DT, Searles PS (2002) Nitrogen assimilation and growth of wheat under elevated carbon dioxide. Proc Natl Acad Sci USA 99:1730–1735PubMedCrossRefGoogle Scholar
  8. Campbell WH (1999) Nitrate reductase structure, function and regulation: Bridging the gap between biochemistry and physiology. Annu Rev Plant Physiol 50:277–303CrossRefGoogle Scholar
  9. Constable JVH, BassiriRad H, Lussenhop J, Zerihun A (2001) Influence of elevated CO2 and mycorrhizae on nitrogen acquisition: contrasting responses in Pinus taeda and Liquidambar styraciflua. Tree Physiol 21:83–91PubMedGoogle Scholar
  10. Cousins AB, Bloom AJ (2003) Influence of elevated CO2 and nitrogen nutrition on photosynthesis and nitrate photo-assimilation in maize (Zea mays L.). Plant Cell Environ 26:1525–1530CrossRefGoogle Scholar
  11. Crous KY, Ellsworth DS (2004) Canopy position affects photosynthetic adjustments to long-term elevated CO2 concentration (FACE) in aging needles in a mature Pinus taeda forest. Tree Physiol 24:961–970PubMedGoogle Scholar
  12. Ellsworth DS (1999) CO2 enrichment in a maturing pine forest: are CO2 exchange and water status in the canopy affected? Plant Cell Environ 22:461–472CrossRefGoogle Scholar
  13. Ellsworth DS, Reich PB, Naumburg ES, Koch GW, Kubiske ME, Smith SD (2004) Photosynthesis, carboxylation and leaf nitrogen responses of 16 species to elevated pCO2 across four free-air CO2 enrichment experiments in forest, grassland and desert. Glob Change Biol 10:2121–2138CrossRefGoogle Scholar
  14. Field CB, Behrenfeld MJ, Randerson JT, Falkowski P (1998) Primary production of the biosphere: Integrating terrestrial and oceanic components. Science 281:237–240PubMedCrossRefGoogle Scholar
  15. Filion M, Dutilleul P, Potvin C (2000) Optimum experimental design for Free-Air Carbon dioxide Enrichment (FACE) studies. Glob Change Biol 6:843–854CrossRefGoogle Scholar
  16. Finzi AC, Allen AS, DeLucia EH, Ellsworth DS, Schlesinger WH (2001) Forest litter production, chemistry, and decomposition following two years of free-air CO2 enrichment. Ecology 82:470–484Google Scholar
  17. Fontes RLF, Coelho HA (2005) Molybdenum determination in Mehlich-1 and Mehlich-3 soil test extracts and molybdenum adsorption in Brazilian soils. Commun Soil Sci Plann 36:2367–2381CrossRefGoogle Scholar
  18. Geiger M, Walch-Liu P, Engels C, Harnecker J, Schulze ED, Ludewig F et al (1998) Enhanced carbon dioxide leads to a modified diurnal rhythm of nitrate reductase activity in older plants, and a large stimulation of nitrate reductase activity and higher levels of amino acids in young tobacco plants. Plant Cell Environ 21:253–268CrossRefGoogle Scholar
  19. Geiger M, Haake V, Ludewig F, Sonnewald U, Stitt M (1999) The nitrate and ammonium nitrate supply have a major influence on the response of photosynthesis, carbon metabolism, nitrogen metabolism and growth to elevated carbon dioxide in tobacco. Plant Cell Environ 22:1177–1199CrossRefGoogle Scholar
  20. Gunderson CA, Sholtis JD, Wullschleger SD, Tissue DT, Hanson PJ, Norby RJ (2002) Environmental and stomatal control of photosynthetic enhancement in the canopy of a sweetgum (Liquidambar styraciflua L.) plantation during 3 years of CO2 enrichment. Plant Cell Environ 25:379–393CrossRefGoogle Scholar
  21. Guo SW, Zhou Y, Gao YX, Li Y, Shen QR (2007) New insights into the nitrogen form effect on photosynthesis and photorespiration. Pedosphere 17:601–610CrossRefGoogle Scholar
  22. Haynes RJ, Goh KM (1978) Ammonium and nitrate nutrition of plants. Biol Rev Camb Philos Soc 53:465–510CrossRefGoogle Scholar
  23. Hendrey GR, Ellsworth DS, Lewin KF, Nagy J (1999) A free-air enrichment system for exposing tall forest vegetation to elevated atmospheric CO2. Glob Change Biol 5:293–309CrossRefGoogle Scholar
  24. Hocking PJ, Meyer CP (1991) Effects of CO2 entichment and nitrogen stress on growth and partitioning of dry-matter and nitrogen in wheat and maize. Aust J Plant Physiol 18:339–356CrossRefGoogle Scholar
  25. Hungate BA, Stiling PD, Dijkstra P, Johnson DW, Ketterer ME, Hymus GJ et al (2004) CO2 elicits long-term decline in nitrogen fixation. Science 304:1291–1291PubMedCrossRefGoogle Scholar
  26. Huppe HC, Turpin DH (1994) Integration of carbon and nitrogen metabolism in plant and algal cells. Annu Rev Plant Physiol 45:577–607Google Scholar
  27. Jastrow JD, Miller RM, Matamala R, Norby RJ, Boutton TW, Rice CW et al (2005) Elevated atmospheric carbon dioxide increases soil carbon. Glob Change Biol 11:2057–2064CrossRefGoogle Scholar
  28. Johnson DW, Cheng W, Joslin JD, Norby RJ, Edwards NT, Todd DE (2004) Effects of elevated CO2 on nutrient cycling in a sweetgum plantation. Biogeochemistry 69:379–403CrossRefGoogle Scholar
  29. Kaiser WM, Kandlbinder A, Stoimenova M, Glaab J (2000) Discrepancy between nitrate reduction rates in intact leaves and nitrate reductase activity in leaf extracts: What limits nitrate reduction in situ? Planta 210:801–807PubMedCrossRefGoogle Scholar
  30. Kaiser BN, Gridley KL, Brady JN, Phillips T, Tyerman SD (2005) The role of molybdenum in agricultural plant production. Ann Bot (Lond) 96:745–754CrossRefGoogle Scholar
  31. Kruse J, Hetzger I, Mai C, Polle A, Rennenberg H (2003) Elevated pCO(2) affects N-metabolism of young poplar plants (Populus tremula x P. alba) differently at deficient and sufficient N-supply. New Phytol 157:65–81CrossRefGoogle Scholar
  32. Lang F, Kaupenjohann M (1999) Molybdenum fractions and mobilization kinetics in acid forest soils. J Plant Nutr Soil Sci 162:309–314CrossRefGoogle Scholar
  33. Lang F, Kaupenjohann M (2000) Molybdenum at German Norway spruce sites: contents and mobility. Can J For Res 30:1034–1040CrossRefGoogle Scholar
  34. Larigauderie A, Reynolds JF, Strain BR (1994) Root response to CO2 enrichment and nitrogen supply in loblolly pine. Plant Soil 165:21–32CrossRefGoogle Scholar
  35. Larios B, Aguera E, de la Haba P, Perez-Vicente R, Maldonado JM (2001) A short-term exposure of cucumber plants to rising atmospheric CO2 increases leaf carbohydrate content and enhances nitrate reductase expression and activity. Planta 212:305–312PubMedCrossRefGoogle Scholar
  36. Liu D, Clark JD, Crutchfield JD, Sims JL (1996) Effect of pH of ammonium oxalate extracting solutions on prediction of plant available molybdenum in soil. Commun Soil Sci Plann 27:2511–2541CrossRefGoogle Scholar
  37. Luo Y, Su B, Currie WS, Dukes JS, Finzi A, Hartwig U et al (2004) Progressive nitrogen limitation of ecosystem responses to rising atmospheric carbon dioxide. Bioscience 54:731–739CrossRefGoogle Scholar
  38. Marschner H (1995) Mineral Nutrition of Higher Plants. Academic Press, New York, p 889Google Scholar
  39. Matt P, Geiger M, Walch-Liu P, Engels C, Krapp A, Stitt M (2001) Elevated carbon dioxide increases nitrate uptake and nitrate reductase activity when tobacco is growing on nitrate, but increases ammonium uptake and inhibits nitrate reductase activity when tobacco is growing on ammonium nitrate. Plant Cell Environ 24:1119–1137CrossRefGoogle Scholar
  40. Mehlich A (1984) Mehlich-3 soil test extractant—a modification of Mehlich-2 extractant. Commun Soil Sci Plann 15:1409–1416CrossRefGoogle Scholar
  41. Melillo JM, McGuire AD, Kicklighter DW, Moore B, Vorosmarty CJ, Schloss AL (1993) Global climate change and terrestrial net primary production. Nature 363:234–240CrossRefGoogle Scholar
  42. Norby RJ, Iversen CM (2006) Nitrogen uptake, distribution, turnover, and efficiency of use in a CO2-enriched sweetgum forest. Ecology 87:5–14PubMedCrossRefGoogle Scholar
  43. Norby RJ, Todd DE, Fults J, Johnson DW (2001) Allometric determination of tree growth in a CO2-enriched sweetgum stand. New Phytol 150:477–487CrossRefGoogle Scholar
  44. Oh NH, Richter DD (2004) Soil acidification induced by elevated atmospheric CO2. Glob Change Biol 10:1936–1946CrossRefGoogle Scholar
  45. Oliva SR, Raitio H (2003) Review of cleaning techniques and their effects on the chemical composition of foliar samples. Boreal Environ Res 8:263–272Google Scholar
  46. Randall PJ (1969) Changes in nitrate and nitrate reductase levels on restoration of molybdenum to molybdenum-deficient plants. Aust J Agric Res 20:635–642CrossRefGoogle Scholar
  47. Satterthwaite FE (1946) An approximate distribution of estimates of variance components. Biom Bull 2:110–114CrossRefGoogle Scholar
  48. Schafer KVR, Oren R, Lai CT, Katul GG (2002) Hydrologic balance in an intact temperate forest ecosystem under ambient and elevated atmospheric CO2 concentration. Glob Change Biol 8:895–911CrossRefGoogle Scholar
  49. Searles PS, Bloom AJ (2003) Nitrate photo-assimilation in tomato leaves under short-term exposure to elevated carbon dioxide and low oxygen. Plant Cell Environ 26:1247–1255CrossRefGoogle Scholar
  50. Sechley KA, Yamaya T, Oaks A (1992) Compartmentation of nitrogen assimilation in higher plants. Int Rev Cytol 134:85–163CrossRefGoogle Scholar
  51. Sicher RC (2001) Responses of nitrogen metabolism in N-sufficient barley primary leaves to plant growth in elevated atmospheric carbon dioxide. Photosynth Res 68:193–201PubMedCrossRefGoogle Scholar
  52. Sims JL, Evarsi F (1997) Testing for molybdenum availability in soils. In: Gupta UC (ed) Molybdenum in Agriculture. Cambridge University Press, Cambridge, pp 111–130Google Scholar
  53. Sinsabaugh RL, Saiya-Corka K, Long T, Osgood MP, Neher DA, Zak DR et al (2003) Soil microbial activity in a Liquidambar plantation unresponsive to CO2-driven increases in primary production. Appl Soil Ecol 24:263–271CrossRefGoogle Scholar
  54. Smart DR, Ritchie K, Bloom AJ, Bugbee BB (1998) Nitrogen balance for wheat canopies (Triticum aestivum cv. Veery 10) grown under elevated and ambient CO2 concentrations. Plant Cell Environ 21:753–763PubMedCrossRefGoogle Scholar
  55. Springer CJ, DeLucia EH, Thomas RB (2005) Relationships between net photosynthesis and foliar nitrogen concentrations in a loblolly pine forest ecosystem grown in elevated atmospheric carbon dioxide. Tree Physiol 25:385–394PubMedGoogle Scholar
  56. Stark JM, Hart SC (1997) High rates of nitrification and nitrate turnover in undisturbed coniferous forests. Nature 385:61–64CrossRefGoogle Scholar
  57. Stiefel EL (2002) The biogeochemistry of molybdenum and tungsten. In: Sigel H (ed) Molybdenum and Tungsten: their roles in biological processes. Marcel Dekker, New York, pp 1–29Google Scholar
  58. Stitt M, Krapp A (1999) The interaction between elevated carbon dioxide and nitrogen nutrition: the physiological and molecular background. Plant Cell Environ 22:583–621CrossRefGoogle Scholar
  59. Stout PR, Meagher WR (1948) Studies of the molybdenum nutrition of plants with radioactive molybdenum. Science 108:471–473PubMedCrossRefGoogle Scholar
  60. Templer PH, Dawson TE (2004) Nitrogen uptake by four tree species of the Catskill Mountains, New York: Implications for forest N dynamics. Plant Soil 262:251–261CrossRefGoogle Scholar
  61. Vitousek PM, Howarth RW (1991) Nitrogen limitation on land and sea–how can it occur. Biogeochemistry 13:87–115CrossRefGoogle Scholar
  62. Yong ZH, Chen GY, Zhang DY, Chen Y, Chen J, Zhu JG et al (2007) Is photosynthetic acclimation to free-air CO2 enrichment (FACE) related to a strong competition for the assimilatory power between carbon assimilation and nitrogen assimilation in rice leaf? Photosynthetica 45:85–91CrossRefGoogle Scholar
  63. Zerihun A, McKenzie BA, Morton JD (1998) Photosynthate costs associated with the utilization of different nitrogen-forms: influence on the carbon balance of plants and shoot-root biomass partitioning. New Phytol 138:1–11CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Susan M. Natali
    • 1
  • Sergio A. Sañudo-Wilhelmy
    • 2
  • Manuel T. Lerdau
    • 3
  1. 1.Department of Ecology and Evolution, 650 Life SciencesState University of New York at Stony BrookStony BrookUSA
  2. 2.Marine and Environmental BiologyUniversity of Southern CaliforniaLos AngelesUSA
  3. 3.Blandy Experimental Farm, Department of Environmental SciencesUniversity of VirginiaCharlottesvilleUSA

Personalised recommendations