Ecological Research

, Volume 22, Issue 3, pp 475–484 | Cite as

Effect of elevated CO2 levels on leaf starch, nitrogen and photosynthesis of plants growing at three natural CO2 springs in Japan

  • Yusuke Onoda
  • Tadaki Hirose
  • Kouki Hikosaka
Original Article


Plant communities around natural CO2 springs have been exposed to elevated CO2 levels over many generations and give us a unique opportunity to investigate the effects of long-term elevated CO2 levels on wild plants. We searched for natural CO2 springs in cool temperate climate regions in Japan and found three springs that were suitable for studying long-term responses of plants to elevated levels of CO2: Ryuzin-numa, Yuno-kawa and Nyuu. At these CO2 springs, the surrounding air was at high CO2 concentration with no toxic gas emissions throughout the growth season, and there was natural vegetation around the springs. At each site, high-CO2 (HC) and low-CO2 (LC) plots were established, and three dominant species at the shrub layers were used for physiological analyses. Although the microenvironments were different among the springs, dicotyledonous species growing at the HC plots tended to have more starch and less nitrogen per unit dry mass in the leaves than those growing at the LC plots. In contrast, monocotyledonous species growing in the HC and LC plots had similar starch and nitrogen concentrations. Photosynthetic rates at the mean growth CO2 concentration were higher in HC plants than LC plants, but photosynthetic rates at a common CO2 concentration were lower in HC plants. Efficiency of water and nitrogen use of leaves at growth CO2 concentration was greatly increased in HC plants. These results suggest that natural plants growing in elevated CO2 levels under cool temperate climate conditions have down-regulated their photosynthetic capacity but that they increased photosynthetic rates and resource use efficiencies due to the direct effect of elevated CO2 concentration.


Down-regulation of photosynthesis Long-term response to elevated CO2 Photosynthetic nitrogen use efficiency Water use efficiency Wild plants 



The landowners of CO2 springs (Hakkoda-Onsen Co., Tashiro Bokuya-Chikusan Kumiai and Yudonosan shrine) kindly gave us a permission to use their land for this study. We thank Makoto Tsurumi (Hirosaki Univ.) and Atsuki Uchida (Nippon Geophysical Prospecting Co.) for geological information of Ryuzin-numa and Yuno-kawa, and Taku Noro (Ministry of Land, Infrastructure and Transport) for climate data at Hakkosawa meteorological station. We also thank Mark Lieffering (AgResearch, NZ) for his comments on the draft manuscript, Kouji Yonekura (Tohoku Univ.) for sorting plant species, and Yuko Yasumura, Hisashi Tsujisawa and Riichi Oguchi (Tohoku Univ.) for their technical assistance. Noriyuki Osada (Tohoku Univ.) kindly provided his data of soil nutrients and also gave us constructive comments on the draft manuscript. This study was partly supported by grants-in-aid from JSPS for Young Research Fellows (Y.O.), and from the Japan Ministry of Education, Culture, Sports, Science and Technology to K.H. and T.H.


  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 165:351–372PubMedCrossRefGoogle Scholar
  2. Ainsworth EA, Davey PA, Hymus GJ, Osborne CE, Rogers A, Blum H, Nosberger J, Long SP (2003) Is stimulation of leaf photosynthesis by elevated carbon dioxide concentration maintained in the long term? A test with Lolium perenne grown for ten years at two nitrogen fertilization levels under free air CO2 enrichment (FACE). Plant Cell Environ 26:705–714CrossRefGoogle Scholar
  3. Anonymous (1953) A plan to change land utilization in Ara-kawa and Komagome-gawa areas. Official document of Aomori City, Aomori (in Japanese)Google Scholar
  4. Arp WJ (1991) Effects of source-sink relations on photosynthetic acclimation to elevated carbon dioxide. Plant Cell Environ 14:869–876CrossRefGoogle Scholar
  5. Bernacchi CJ, Singsaas EL, Pimentel C, Portis AR Jr., Long SP (2001) Improved temperature response functions for models of Rubisco-limited photosynthesis. Plant Cell Environ 24:253–259CrossRefGoogle Scholar
  6. Blaschke L, Schulte M, Raschi A, Slee N, Rennenberg H, Polle A (2001) Photosynthesis, soluble and structural carbon compounds in two Mediterranean oak species (Quercus pubescens and Q. ilex) after lifetime growth at naturally elevated CO2 concentrations. Plant Biol 3:288–297CrossRefGoogle Scholar
  7. von Caemmerer S (2000) Biochemical models of leaf photosynthesis. CSIRO publishing, CanberraGoogle Scholar
  8. Chaves MM, Pereira JS, Cerasoli S, Clifton-Brown J, Miglietta F, Raschi A (1995) Leaf metabolism during summer drought in Quercus ilex trees with lifetime exposure to elevated CO2. J Biogeogr 22:255–259CrossRefGoogle Scholar
  9. Cook AC, Tissue DT, Roberts SW, Oechel WC (1998) Effects of long-term elevated [CO2] from natural CO2 springs on Nardus stricta: photosynthesis, biochemistry, growth and phenology. Plant Cell Environ 21:417–425CrossRefGoogle Scholar
  10. Cook AC, Vourlitis GL, Harazono Y (2000) Evaluating the potential for long-term elevated CO2 exposure studies using CO2 springs in Japan. J Agric Meteorol 56:31–40Google Scholar
  11. Cotrufo MF, Raschi A, Lanini M, Ineson P (1999) Decomposition and nutrient dynamics of Quercus pubescens leaf litter in a naturally enriched CO2 Mediterranean ecosystem. Func Ecol 13:343–351CrossRefGoogle Scholar
  12. Coviella CE, Trumble JT (1999) Effects of elevated atmospheric carbon dioxide on insect–plant Interactions. Conserv Biol 13:700–712CrossRefGoogle Scholar
  13. Drake BG, Gonzalez-Meler MA, Long SP (1997) More efficient plants—a consequence of rising atmospheric CO2. Ann Rev Plant Physiol Plant Mol Biol 48:609–639CrossRefGoogle Scholar
  14. Farquhar GD, von Caemmerer S, Berry JA (1980) A biochemical model of photosynthetic CO2 assimilation in leaves of C3 species. Planta 149:78–90CrossRefGoogle Scholar
  15. Fernandez MD, Pieters A, Donoso C, Tezara W, Azkue M, Herrera C, Rengifo E, Herrera A (1998) Effects of a natural source of very high CO2 concentration on the leaf gas exchange, xylem water potential and stomatal characteristics of plants of Spatiphylum cannifolium and Bauhinia multinervia. New Phytol 138:689–697CrossRefGoogle Scholar
  16. Gahrooee FR (1998) Impacts of elevated atmospheric CO2 on litter quality, litter decomposability and nitrogen turnover rate of two oak species in a Mediterranean forest ecosystem. Global Change Biol 4:667–677CrossRefGoogle Scholar
  17. Grill D, Muller M, Tausz M, Strnad B, Wonisch A, Raschi A (2004) Effects of sulphurous gases in two CO2 springs on total sulphur and thiols in acorns and oak seedlings. Atmos Environ 38:3775–3780CrossRefGoogle Scholar
  18. Hättenschwiler S, Miglietta F, Raschi A, Körner C (1997) Thirty years of in situ tree growth under elevated CO2—a model for future forest responses. Global Change Biol 3:463–471CrossRefGoogle Scholar
  19. Hikosaka K (2003) A model of dynamics of leaves and nitrogen in a plant canopy: an integration of canopy photosynthesis, leaf life span, and nitrogen use efficiency. Am Nat 162:149–164PubMedCrossRefGoogle Scholar
  20. Hikosaka K, Nagashima H, Harada Y, Hirose T (2001) A simple formulation of interaction between individuals competing for light in a monospecific stand. Func Ecol 15:642–646CrossRefGoogle Scholar
  21. Idso KE, Idso SB (1994) Plant responses to atmospheric CO2 enrichment in the face of environmental constraints: a review of the past 10 years’ research. Agric Forest Meteorol 69:153–203CrossRefGoogle Scholar
  22. Jones MB, Clifton Brown J, Raschi A, Miglietta F (1995) The effects on Arbutus unedo L. of long-term exposure to elevated CO2. Global Change Biol 1:295–302CrossRefGoogle Scholar
  23. Keeney DR, Nelson DW (1982) Nitrogen—Inorganic Forms. In: Page AL (Ed) Methods of soil analysis, part 2. American Society of Agronomy, Madison, pp 643–698Google Scholar
  24. Kimball BA, Pinter PJ, Garcia RL, Lamorte RL, Wall GW, Hunsaker DJ, Wechsung G, Wechsung F, Kartschall T (1995) Productivity and water use of wheat under free-air CO2 enrichment. Global Change Biol 1:429–442CrossRefGoogle Scholar
  25. Kimbara K (1992) Distribution map and catalogue of hot and mineral springs in Japan. Geological Survey of Japan, TsukubaGoogle Scholar
  26. Koch GW (1994) The use of natural situation of CO2 enrichment in studies of vegetation response to increasing atmospheric CO2. In: Schulze ED, Mooney HA (Eds) Design and execution of experiments on CO2 enrichment. Commission of the European Communities, Luxembourg, pp 381–392Google Scholar
  27. Kohut R (2003) The long-term effects of carbon dioxide on natural systems: issues and research needs. Environ Int 29:171–180PubMedCrossRefGoogle Scholar
  28. Körner C, Miglietta F (1994) Long term effects of naturally elevated CO2 on Mediterranean grassland and forest trees. Oecologia 99:343–351CrossRefGoogle Scholar
  29. Long SP (1991) Modification of the response of photosynthetic productivity to rising temperature by atmospheric carbon dioxide concentrations: has its importance been underestimated? Plant Cell Environ 14:729–740CrossRefGoogle Scholar
  30. Long SP, Ainsworth EA, Rogers A, Ort DR (2004) Rising atmospheric carbon dioxide: plants FACE the future. Annu Rev Plant Biol 55:591–628PubMedCrossRefGoogle Scholar
  31. Luo Y, Reynolds J (1999) Validity of extrapolating field CO2 experiments to predict carbon sequestration in natural ecosystems. Ecology 80:1568–1583Google Scholar
  32. Miglietta F, Raschi M (1993) Studying the effect of elevated CO2 in the open in a naturally enriched environment in Central Italy. Vegetation 104/105:391–400CrossRefGoogle Scholar
  33. Miglietta F, Raschi A, Bettarini I, Resti R, Selvi F (1993a) Natural CO2 springs in Italy: a resource for examining long-term response of vegetation to rising atmospheric CO2 concentrations. Plant Cell Environ 16:873–878CrossRefGoogle Scholar
  34. Miglietta F, Raschi A, Resti R, Resti R, Selvi F (1993b) Growth and onto-morphogenesis of soybean (Glycine max Merril) in an open, naturally CO2-enriched environment. Plant Cell Environ 16:909–918CrossRefGoogle Scholar
  35. Miglietta F, Raschi A, Bettarini I, Baldiani M, van Gardingen P (1994) Carbon dioxide springs and their use for experimentation. In: Schulze ED, Mooney HA (Eds) Design and execution of experiments on CO2 enrichment. Commission of the European communities, Luxembourg, pp 393–403Google Scholar
  36. Mott KA (1990) Sensing of atmospheric CO2 by plants. Plant Cell Environ 13:731–737CrossRefGoogle Scholar
  37. Nakano H, Makino A, Mae T (1998) The responses of Rubisco protein to long-term exposure to elevated CO2 in rice and bean leaves. Photosynth Mech Eff 5:3391–3394Google Scholar
  38. Newton PCD, Bell CC, Clark H (1996) Carbon dioxide emissions from mineral springs in Northland and the potential of these sites for studying the effects of elevated carbon dioxide on pastures. New Zeal J Agric Res 39:33–40Google Scholar
  39. Poorter H (1993) Interspecific variation in the growth response of plants to an elevated ambient CO2 concentration. Vegetation 104/105:77–97CrossRefGoogle Scholar
  40. Rapparini F, Baraldi R, Miglietta F, Loreto F (2004) Isoprenoid emission in trees of Quercus pubescens and Quercus ilex with lifetime exposure to naturally high CO2 environment. Plant Cell Environ 27:381–391CrossRefGoogle Scholar
  41. Rufty TW, Huber SC (1983) Changes in starch formation and activities of sucrose phosphate synthesis and cytoplasmic fructose-1–6-bisphosphatase in response to source-sink alterations. Plant Physiol 72:474–480PubMedCrossRefGoogle Scholar
  42. Scholefield PA, Doick KJ, Herbert BMJ, Hewitt CNS, Schnitzler JP, Pinelli P, Loreto F (2004) Impact of rising CO2 on emissions of volatile organic compounds: isoprene emission from Phragmites australis growing at elevated CO2 in a natural carbon dioxide spring. Plant Cell Environ 27:393–401CrossRefGoogle Scholar
  43. Sheen J. (1990) Metabolic repression of transcription in higher plants. Plant Cell 2:1027–1038PubMedCrossRefGoogle Scholar
  44. 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
  45. Stylinski CD, Oechel WC, Gamon JA, Tissue DT, Miglietta F, Raschi A (2000) Effects of lifelong [CO2] enrichment on carboxylation and light utilization of Quercus pubescens Willd. examined with gas exchange, biochemistry and optical techniques. Plant Cell Environ 23:1353–1362CrossRefGoogle Scholar
  46. Tognetti R, Giovannelli A, Longobucco A, Miglietta F, Raschi A. (1996) Water relations of oak species growing in the natural CO2 spring of Rapolano (central Italy). Ann Des Sci Forest 53:475–485Google Scholar
  47. Tognetti R, Longobucco A, Miglietta F, Raschi A (1998) Transpiration and stomatal behaviour of Quercus ilex plants during the summer in a Mediterranean carbon dioxide spring. Plant Cell Environ 21:613–622CrossRefGoogle Scholar
  48. Tognetti R, Longobucco A, Miglietta F, Raschi A (1999) Water relations, stomatal response and transpiration of Quercus pubescens trees during summer in a Mediterranean carbon dioxide spring. Tree Physiol 19:261–270PubMedGoogle Scholar
  49. Tognetti R, Cherubini P, Innes JL (2000) Comparative stem-growth rates of Mediterranean trees under background and naturally enhanced ambient CO2 concentrations. New Phytol 146:59–74CrossRefGoogle Scholar
  50. Tsurumi M, Hirabayashi J, Yoshimura K, Kondo H, Sasaki M, Ishida S (1999) Report on the volcanic gas in the Hakkoda Mountain. Official document of Aomori Prefecture, Aomori (in Japanese)Google Scholar
  51. Vodnik D, Pfanz H, Wittmann C, Macek I, Kastelec D, Turk B, Batic F (2002) Photosynthetic acclimation in plants growing near a carbon dioxide spring. Phyton 42:239–244Google Scholar
  52. Wand SJE, Midgley GF, Jones MH, Curtis PS (1999) Response of wild C4 and C3 grass (Poaceae) species to elevated atmospheric CO2 concentration: a meta-analytic test of current theories and perceptions. Global Change Biol 5:723–741CrossRefGoogle Scholar
  53. Wong SC (1990) Elevated atmospheric partial pressure of carbon dioxide and plant growth ii. non-structural carbohydrate content in cotton plants and its effect on growth parameters. Photosynth Res 23:171–180CrossRefGoogle Scholar
  54. Wong SC, Cowan IR, Farquhar GD (1979) Stomatal conductance correlates with photosynthetic capacity. Nature 282:424–426CrossRefGoogle Scholar

Copyright information

© The Ecological Society of Japan 2006

Authors and Affiliations

  1. 1.Graduate School of Life SciencesTohoku UniversityAoba, SendaiJapan
  2. 2.Section of Plant Ecology and Biodiversity, Institute of Environmental BiologyUtrecht UniversityUtrechtThe Netherlands
  3. 3.Department of International Agriculture DevelopmentTokyo University of AgricultureTokyoJapan

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