Carbon partitioning in N2 fixing Medicago sativa plants exposed to different CO2 and temperature conditions
Iker Aranjuelo A B C , Juan J. Irigoyen B , Manuel Sánchez-Díaz B and Salvador Nogués AA Unitat de Fisologia Vegetal, Facultat de Biologia, Universitat de Barcelona, Diagonal 645, 08028, Barcelona, Spain.
B Departamento de Biología Vegetal, Sección Fisiología Vegetal (Assoicated Unit with the Spanish National Research Council, CSIC, EEAD, Zaragoza), Facultades de Ciencias y Farmacia, Universidad de Navarra, Irunlarrea s/n, 31008, Pamplona, Navarra, Spain.
C Corresponding author. Email: iker.aranjuelo@ub.edu
Functional Plant Biology 35(4) 306-317 https://doi.org/10.1071/FP07296
Submitted: 14 December 2007 Accepted: 22 April 2008 Published: 3 June 2008
Abstract
Many of the studies analysing the CO2 effect on plant development have been conducted in optimal growth conditions. Furthermore, although some of those studies suggest that legumes might show a steady productivity increase with rising CO2, the role of nodule activity on the plant responsiveness to predicted atmospheric CO2 enhancement is not well understood. In this study, C (metabolism and allocation) and N (nodule activity) interaction between the plant and the bacterial symbiont during the photosynthetic acclimation of N2-fixing alfalfa (Medicago sativa L. cv. Aragón) plants exposed to elevated CO2 and temperature conditions was analysed. The plants were grown in temperature gradient greenhouses (TGG) where, in the case of elevated CO2 treatments, the isotopic 13C/12C composition (δ13C) inside the TGG was modified. Compared with the corresponding temperature treatment, exposure to 700 μmol mol–1 CO2 enhanced dry mass (DM) of plants in elevated temperature treatments (26%), whereas no significant effect was detected in ambient temperature treatments. The δ13C data revealed that although all the carbon corresponding to leaf total organic matter (TOM) came from newly assimilated C, plants exposed to elevated CO2 did not develop strong sink activity (especially in ambient temperature conditions). Leaf carbohydrate build-up induced reduction in the Rubisco (E.C. 4.1.1.39) carboxylation capacity of plants. Despite this reduction in Rubisco content, plants exposed to elevated CO2 conditions maintained (at ambient temperature) or increased (at elevated temperature) photosynthetic rates (measured at growth conditions) by increasing N use efficiency. The larger C sink strength of nodules in plants grown at elevated CO2 and temperature conditions did not contribute towards overcoming photosynthetic acclimation. Further, the inhibitory effect of CO2 on nodule total activity was caused by a large depletion in total soluble protein (TSP) of nodules. Depletion of leaf N demand, together with the reduction in nodule carbohydrate availability (as reflected by the nodule starch concentration), negatively affected the nodule TSP content and enzymatic activity.
Additional keywords: acclimation, carbon and nitrogen isotopes, climate change, isotope discrimination, nitrogen fixation, nodule metabolism, 13C isotopic composition.
Acknowledgements
This work was supported in part by the Spanish Science and Education Ministry (BFU-2004–05096/BFI, AGL2004–00194/AGR and Juan de la Cierva research grant), the Fundación Universitaria de Navarra, Fundación Caja Navarra and by the European Project PERMED (INCO-CT-2004–509140). The temperature gradient greenhouses used in this study were funded by the Spanish Commission of Science and Technology (AMB96–0396). The authors wish to thank Dr R. Martínez-Carrasco and Dr P. Pérez for providing temperature gradient greenhouses facilities. The technical co-operation of A. Urdiain and A. Verdejo is also acknowledged.
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 Phytologist 165, 351–372.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Ainsworth EA, Rogers A
(2007) The response of photosynthesis and stomatal conductance to rising [CO2]: mechanisms and environmental interactions. Plant, Cell & Environment 30, 258–270.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Ainsworth EA,
Rogers A,
Nelson R, Long SP
(2004) Testing the ‘source–sink’ hypothesis of down-regulation of photosynthesis in elevated [CO2] in the field with single gene substitutions in Glycine max. Agricultural and Forest Meteorology 122, 85–94.
| Crossref | GoogleScholarGoogle Scholar |
Almeida JPF,
Hartwig UA,
Frehner M,
Nösberger J, Lüscher A
(2000) Evidence that P deficiency induces N feedback regulation of symbiotic N2 fixation in white clover (Trifolium repens L.). Journal of Experimental Botany 51, 1289–1297.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Aranjuelo I,
Pérez P,
Martínez-Carrasco R, Sánchez-Díaz M
(2005a) The use of temperature gradient tunnels for studying the effect of water availability, elevated CO2 and temperature on nodulated alfalfa plants growth and N2 fixation. Annals of Applied Biology 146, 51–60.
| Crossref | GoogleScholarGoogle Scholar |
Aranjuelo I,
Zita G,
Hernandez L,
Pérez P,
Martínez-Carrasco R, Sánchez-Díaz M
(2005b) Response of nodulated alfalfa to water supply, temperature and elevated CO2: photosynthetic down-regulation. Physiologia Plantarum 123, 348–358.
| Crossref | GoogleScholarGoogle Scholar |
Aranjuelo I,
Pérez P,
Martínez-Carrasco R, Sánchez-Díaz M
(2006) Response of nodulated alfalfa to water supply, temperature and elevated CO2: productivity and water relations. Environmental and Experimental Botany 55, 130–141.
| Crossref | GoogleScholarGoogle Scholar |
Arrese-Igor C,
González EM,
Gordon AJ,
Minchin FR,
Gálvez L,
Royuela M,
Cabrerizo PM, Aparicio-Tejo PM
(1999) Sucrose synthase and nodule nitrogen fixation under drought and other environmental stresses. Symbiosis 27, 1–24.
Avice JC,
Ourry A,
Lemaire G, Boucaud J
(1996) Nitrogen and carbon flows estimated by 15N and 13C pulse–chase labeling during regrowth of alfalfa. Plant Physiology 112, 281–290.
| PubMed |
Batts GR,
Morison JIL,
Ellis RH,
Hadley P, Wheeler TR
(1997) Effects of CO2 and temperature on growth and yield crops of winter wheat over four seasons. European Journal of Agronomy 7, 43–52.
| Crossref | GoogleScholarGoogle Scholar |
Bertrand A,
Prévost D,
Bigras FJ,
Lalande R,
Tremblay GF,
Tremblay GF,
Castonguay Y, Bélanger G
(2007) Alfalfa response to elevated atmospheric CO2 varies with the symbiotic rhizobial strain. Plant and Soil 301, 173–187.
| Crossref | GoogleScholarGoogle Scholar |
Bloom AJ,
Smart DR,
Nguyen DT, Searles PS
(2002) Nitrogen assimilation and growth of wheat under elevated carbon dioxide. Proceedings of the National Academy of Sciences USA 99, 1730–1735.
| Crossref | GoogleScholarGoogle Scholar |
Bradford MM
(1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein – dye binding. Analytical Biochemistry 72, 248–254.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
von Caemmerer S, Farquhar GD
(1981) Some relationships between the biochemistry of photosynthesis and the gas exchange of leaves. Planta 153, 376–387.
| Crossref | GoogleScholarGoogle Scholar |
Conroy J, Hocking P
(1993) Nitrogen nutrition of C3 plants at elevated atmospheric CO2 concentrations. Physiologia Plantarum 89, 570–576.
| Crossref | GoogleScholarGoogle Scholar |
Davey PA,
Olcer H,
Zakhleniuk O,
Bernacchi CJ,
Calfapietra C,
Long SP, Raines CA
(2006) Can fast-growing plantation trees escape biochemical down-regulation of photosynthesis when grown throughout their complete production cycle in the open air under elevated carbon dioxide? Plant, Cell & Environment 29, 1235–1244.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
De Luis I,
Irigoyen JJ, Sánchez-Díaz M
(1999) Elevated CO2 enhances plant growth in droughted N2-fixing alfalfa without improving water status. Physiologia Plantarum 107, 84–89.
| Crossref | GoogleScholarGoogle Scholar |
De Luis I,
Irigoyen JJ, Sánchez-Díaz M
(2002) Low vapour pressure deficit reduced beneficial effect of elevated CO2 on growth of N2-fixing alfalfa plants. Physiologia Plantarum 116, 497–502.
| Crossref | GoogleScholarGoogle Scholar |
De Vries GE,
In’tVeld P, Kijne JW
(1980) Production of organic acids in Pisum sativum, root nodules as a result of oxygen stress. Plant Scientific Letters 20, 115–123.
| Crossref | GoogleScholarGoogle Scholar |
Deroche ME,
Carrayol E, Jolivet E
(1983) Phosphoenolpyruvate carboxylase in legume nodules. Physiologie Vegetale 21, 1075–1081.
Drake BG,
González-Meler MA, Long SP
(1997) More efficient plants: a consequence of rising atmospheric CO2? Annual Review of Plant Physiology and Plant Molecular Biology 48, 609–639.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Du Cloux H,
Andre M,
Gerbaud A, Daguenet A
(1989) Wheat response to CO2 enrichment: effect on photosynthetic and photorespiratory characteristics. Photosynthetica 23, 145–153.
Erice G,
Irigoyen JJ,
Pérez P,
Martinez-Carrasco R, Sánchez-Díaz M
(2006) Effect of elevated CO2, temperature and drought on dry matter partitioning and photosynthesis before and after cutting of nodulated alfalfa. Plant Science 170, 1059–1067.
| Crossref | GoogleScholarGoogle Scholar |
Ethier GJ, Livingston NJ
(2004) On the need to incorporate sensitivity to CO2 transfer conductance into the Farquhar–von Caemmerer–Berry leaf photosynthesis model. Plant, Cell & Environment 27, 137–153.
| Crossref | GoogleScholarGoogle Scholar |
Farquhar GD, Sharkey TD
(1982) Stomatal conductance and photosynthesis. Annual Review of Plant Physiology 33, 317–345.
| Crossref | GoogleScholarGoogle Scholar |
Farquhar GD,
Hubick KT,
Condon AG, Richards RA
(1989) Carbon isotope fractionation and water-use efficiency. In ‘Stable isotopes in ecological research. Ecological Studies 68, 21–40.
Farrar JF, Williams ML
(1991) The effects of increased atmospheric carbon dioxide and temperature on carbon partitioning, source–sink relations and respiration. Plant, Cell & Environment 14, 819–830.
| Crossref | GoogleScholarGoogle Scholar |
Gebbing T,
Schnyder H, Kuhbauch W
(1998) Carbon mobilization in shoots parts and roots of wheat during grain filling: assessment by 13C/12C steady state labeling, growth analysis and balance sheets of reserves. Plant, Cell & Environment 21, 301–310.
| Crossref | GoogleScholarGoogle Scholar |
Geiger DR, Shieh W-J
(1988) Analysis partitioning of recently fixed and of reserve carbon in reproductive Phaseolus vulgaris L. plants. Plant, Cell & Environment 11, 777–783.
| Crossref | GoogleScholarGoogle Scholar |
Gessler A,
Keitel C,
Kodama N,
Weston C,
Winters AJ,
Keith H,
Grice K,
Leuning R, Farquhar GD
(2007) δ13C of organic matter transported from leaves to the roots in Eucaliptus delegatensis: short-term variations and relation to respired CO2. Functional Plant Biology 34, 692–706.
| Crossref | GoogleScholarGoogle Scholar |
Griffin KL,
Tissue DT,
Turnbull MH, Whitehead D
(2000) The onset of photosynthetic acclimation to elevated CO2 partial pressure in field-grown Pinus radiata D. Don. after 4 years. Plant, Cell & Environment 23, 1089–1098.
| Crossref | GoogleScholarGoogle Scholar |
Gunderson CA,
Norby RJ, Wullschleger SD
(1993) Foliar gas exchange of two deciduous hardwoods during three years of growth in elevated CO2: no loss of photosynthetic enhancement. Plant, Cell & Environment 16, 797–807.
| Crossref | GoogleScholarGoogle Scholar |
Hartwig UA
(1998) The regulation of symbiotic N2 fixation: a conceptual model of N feedback from ecosystem to the gene expression level. Perspectives in Plant Ecology, Evolution and Systematics 1, 92–120.
| Crossref | GoogleScholarGoogle Scholar |
Hartwig UA,
Heim I,
Lüscher A, Nösberger J
(1994) The nitrogen-sink is involved in the regulation of nitrogenase activity in white clover after defoliation. Physiologia Plantarum 92, 375–382.
| Crossref | GoogleScholarGoogle Scholar |
Idso SB, Kimball BA
(1992) Effects of atmospheric CO2 enrichment on photosynthesis, respiration and growth of four orange trees. Plant Physiology 99, 341–343.
| PubMed |
Irigoyen JJ,
Emerich DW, Sánchez-Díaz M
(1992) Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Physiologia Plantarum 84, 55–60.
| Crossref | GoogleScholarGoogle Scholar |
Jarvis CE, Walker JRL
(1993) Simultaneous, rapid, spectrophotometric determination of total starch, amylose and amylopectin. Journal of the Science of Food and Agriculture 63, 53–57.
| Crossref | GoogleScholarGoogle Scholar |
Jifon JL, Wolfe DW
(2002) Photosynthetic acclimation to elevated CO2 in Phaseolus vulgaris L. is altered by growth response to nitrogen supply. Global Change Biology 8, 1018–1027.
| Crossref | GoogleScholarGoogle Scholar |
King BJ,
Layzell DB, Canvin DT
(1986) The role of dark carbon dioxide fixation in root nodules of soybean. Plant Physiology 81, 200–205.
| PubMed |
Klumpp K,
Schäufele R,
Lötscher M,
Lattanzi FA,
Feneis W, Schnyder H
(2005) C-isotope composition of CO2 respired by shoots and roots: fractionation during dark respiration? Plant, Cell & Environment 28, 241–250.
| Crossref | GoogleScholarGoogle Scholar |
Laemmli UK
(1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Lawlor DW, Mitchell RAC
(1991) The effect of increasing CO2 on crop photosynthesis and productivity: a review of field studies. Plant, Cell & Environment 14, 807–818.
| Crossref | GoogleScholarGoogle Scholar |
Lee TD,
Reich PB, Tjoelker MG
(2003) Legume presence increases photosynthesis and N concentrations of co-occurring non-fixers but does not modulate their responsiveness to carbon dioxide enrichment. Oecologia 137, 22–31.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Long SP
(1991) Modification of the response of photosynthetic productivity to rising temperature by atmospheric CO2 concentration: has its importance been underestimated? Plant, Cell & Environment 14, 729–739.
| Crossref | GoogleScholarGoogle Scholar |
Long SP,
Ainsworth EA,
Rogers A, Ort DR
(2004) Rising atmospheric carbon dioxide: plants FACE the future. Annual Review of Plant Biology 55, 591–628.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Luo Y,
Sue B,
Currie WS,
Duke JS, Finzi A , et al.
(2004) Progressive nitrogen limitation of ecosystem response to rising atmospheric carbon dioxide. Bioscience 54, 731–739.
| Crossref | GoogleScholarGoogle Scholar |
Lüscher A,
Hartwig UA,
Suter D, Nösberger J
(2000) Direct evidence that symbiotic N2 fixation in fertile grassland is an evidence trait for a strong response of plants to elevated atmospheric CO2. Global Change Biology 6, 655–662.
| Crossref | GoogleScholarGoogle Scholar |
Martín del Molino IM,
Martínez-Carrasco R,
Pérez P,
Hernández L,
Morcuende R, Sánchez de la Puente L
(1995) Influence of nitrogen supply and sink strength on changes in leaf nitrogen compounds during senescence in two wheat cultivars. Physiologia Plantarum 95, 51–58.
| Crossref | GoogleScholarGoogle Scholar |
Matt P,
Krapp A,
Haake V,
Mock HP, Stitt M
(2002) Decreased Rubisco activity leads to dramatic changes of nitrate metabolism, amino acid metabolism and the levels of phenylpropanoids and nicotine in tobacco antisense RBCS transformants. The Plant Journal 30, 663–677.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Moore BD,
Cheng SH,
Sims D, Seeman JR
(1999) The biochemical and molecular basis for photosynthetic acclimation to elevated atmospheric CO2. Plant, Cell & Environment 22, 567–582.
| Crossref | GoogleScholarGoogle Scholar |
Morison JIL, Lawlor DW
(1999) Interactions between increasing CO2 concentration and temperature on plant growth. Plant, Cell & Environment 22, 659–682.
| Crossref | GoogleScholarGoogle Scholar |
Murphy PM
(1986) Effect of light and atmospheric carbon dioxide concentration on nitrogen fixation by herbage legumes. Plant and Soil 95, 399–409.
| Crossref | GoogleScholarGoogle Scholar |
Nogués S,
Tcherkez G,
Cornic G, Ghashghaie J
(2004) Respiratory carbon metabolism following illumination in intact French bean leaves using 13C/12C isotope labelling. Plant Physiology 136, 3245–3254.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Nogués S,
Damesin C,
Tcherkez G,
Maunoury F,
Cornic G, Ghashghaie J
(2006) 13C/12C isotope labelling to study leaf carbon respiration and allocation in twigs of field-grown beech trees. Rapid Communications in Mass Spectrometry 20, 219–226.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Nogués S,
Aranjuelo I,
Pardo A, Azcon-Bieto J
(2008) Assessing the stable-carbon isotopic composition of intercellular CO2 in a CAM plant using gas chromatography-combustion-isotope ratio mass spectrometry. Rapid Communications in Mass Spectrometry 22, 1017–1022.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Norby RJ,
Wullschleger SD,
Gunderson CA,
Johnson DW, Ceulemans R
(1999) Tree responses to rising CO2 in field experiments: implications for the future forest. Plant, Cell & Environment 22, 683–714.
| Crossref | GoogleScholarGoogle Scholar |
Rawson HM
(1995) Yield responses of two genotypes to carbon dioxide and temperature in field studies using temperature gradient tunnels. Australian Journal of Plant Physiology 22, 23–32.
Reich PB,
Hungate BA, Luo Y
(2006) Carbon-nitrogen interactions in terrestrial ecosystems in response to rising atmospheric carbon dioxide. Annual Review of Ecology, Evolution and Systematics 37, 611–636.
| Crossref | GoogleScholarGoogle Scholar |
Rogers A,
Fischer BC,
Bryant J,
Frehmer M,
Blum H,
Raines CA, Long SP
(1998) Acclimation of photosynthesis to elevated CO2 under low-nitrogen nutrition is affected by the capacity for assimilate utilization. Perennial ryegrass under free air CO2 enrichment. Plant Physiology 118, 683–689.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Rogers A,
Gibon Y,
Stitt M,
Morgan PB,
Bernacchi CJ,
Ort DR, Long SP
(2006) Increased C availability at elevated carbon dioxide concentration improves N assimilation in legume. Plant, Cell & Environment 29, 1651–1658.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Schubert S,
Serraj R,
Plies-Balzer E, Mengel K
(1995) Effect of drought stress on growth, sugar concentrations and amino acid accumulation in N2-fixing alfalfa. Journal of Plant Physiology 146, 541–546.
Serraj R,
Sinclair TR, Allen LH
(1998) Soybean nodulation and N2 fixation response to drought under carbon dioxide enrichment. Plant, Cell & Environment 21, 491–500.
| Crossref | GoogleScholarGoogle Scholar |
Serraj R,
Sinclair TR, Purcell LC
(1999) Symbiotic N2 fixation response to drought. Journal of Experimental Botany 50, 143–155.
| Crossref | GoogleScholarGoogle Scholar |
Sgherri CLM,
Quartacci MF,
Menconi M,
Raschi A, Navari-izzo F
(1998) Interactions between drought and elevated CO2 on alfalfa plants. Journal of Plant Physiology 152, 118–124.
Soussana JF, Hartwig UA
(1996) The effects of elevated CO2 on symbiotic N2 fixation; a link between the carbon and nitrogen cycles in grassland ecosystems. Plant and Soil 187, 312–332.
Stitt M, Krapp A
(1999) The interaction between elevated carbon dioxide and nitrogen nutrition: the physiological and molecular background. Plant, Cell & Environment 22, 583–621.
| Crossref | GoogleScholarGoogle Scholar |
Stock WD, Evans JR
(2006) Effects of water availbility, nitrogen supply and atmospheric CO2 concentraions on plant nitrogen natural abundance values. Functional Plant Biology 33, 219–227.
| Crossref | GoogleScholarGoogle Scholar |
Sun J,
Gibson KM,
Kiirats O,
Okita TW, Edwards GE
(2002) Interactions of nitrate and CO2 enrichment on growth, carbohydrates, and Rubisco in Arabidopsis starch mutants. Significance of starch and hexose. Plant Physiology 130, 1573–1583.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Tcherkez G, Farquhar GD
(2005) Carbon isotope predictions for enzymes involved in the primary carbon metabolism of plant leaves. Functional Plant Biology 32, 277–291.
| Crossref | GoogleScholarGoogle Scholar |
Udvardi MK, Day DA
(1997) Metabolic transport across symbiotic membranes of legume nodules. Annual Review of Plant Physiology and Plant Molecular Biology 48, 493–523.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Vance CP, Heichel GH
(1991) Carbon in N2 fixation: limitation or exquisite adaptation. Annual Review of Plant Physiology and Plant Molecular Biology 42, 373–392.
| Crossref | GoogleScholarGoogle Scholar |
Vance CP, Stade S
(1984) Alfalfa root nodule carbon dioxide fixation. II. Partial purification and characterization of root nodule phosphoenolpyruvate carboxylase. Plant Physiology 75, 261–264.
| PubMed |
Voisin AS,
Salon C,
Jeudy C, Warembourg FR
(2003) Symbiotic N2 fixation activity in relation to C economy of Pisum sativum L. as a function of plant phenology. Journal of Experimental Botany 54, 2733–2744.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Walsh KB
(1995) Physiology of the legume nodule and its response to stress. Soil Biology & Biochemistry 27, 637–655.
| Crossref | GoogleScholarGoogle Scholar |
West JB,
Lambers JHR,
Lee TD,
Hobbie SE, Reich PB
(2005) Legume species identity and soil nitrogen supply determined symbiotic nitrogen-fixation responses to elevated atmospheric [CO2]. New Phytologist 167, 523–530.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Wolfe DW,
Gifford RM,
Hilbert D, Luo Y
(1998) Integration of photosynthetic acclimation to CO2 at the whole plant level. Global Change Biology 4, 879–893.
| Crossref | GoogleScholarGoogle Scholar |
Woodrow IW
(1994) Optimal acclimation of the C3 photosynthetic system under enhanced CO2. Photosynthesis Research 39, 401–412.
| Crossref | GoogleScholarGoogle Scholar |
Zanetti S,
Hartwig UA, Nösberger J
(1998) Elevated atmospheric CO2 does not affect per se the preference for symbiotic nitrogen as opposed to mineral nitrogen of Trifolium repens L. Plant, Cell & Environment 21, 623–630.
| Crossref | GoogleScholarGoogle Scholar |