Photosynthesis Research

, Volume 138, Issue 2, pp 219–232 | Cite as

Different CO2 acclimation strategies in juvenile and mature leaves of Ottelia alismoides

  • Wen Min Huang
  • Hui Shao
  • Si Ning Zhou
  • Qin Zhou
  • Wen Long Fu
  • Ting Zhang
  • Hong Sheng Jiang
  • Wei Li
  • Brigitte GonteroEmail author
  • Stephen C. MaberlyEmail author
Original Article


The freshwater macrophyte, Ottelia alismoides, is a bicarbonate user performing C4 photosynthesis in the light, and crassulacean acid metabolism (CAM) when acclimated to low CO2. The regulation of the three mechanisms by CO2 concentration was studied in juvenile and mature leaves. For mature leaves, the ratios of phosphoenolpyruvate carboxylase (PEPC) to ribulose-bisphosphate carboxylase/oxygenase (Rubisco) are in the range of that of C4 plants regardless of CO2 concentration (1.5–2.5 at low CO2, 1.8–3.4 at high CO2). In contrast, results for juvenile leaves suggest that C4 is facultative and only present under low CO2. pH-drift experiments showed that both juvenile and mature leaves can use bicarbonate irrespective of CO2 concentration, but mature leaves have a significantly greater carbon-extracting ability than juvenile leaves at low CO2. At high CO2, neither juvenile nor mature leaves perform CAM as indicated by lack of diurnal acid fluctuation. However, CAM was present at low CO2, though the fluctuation of titratable acidity in juvenile leaves (15–17 µequiv g−1 FW) was slightly but significantly lower than in mature leaves (19–25 µequiv g−1 FW), implying that the capacity to perform CAM increases as leaves mature. The increased CAM activity is associated with elevated PEPC activity and large diel changes in starch content. These results show that in O. alismoides, carbon-dioxide concentrating mechanisms are more effective in mature compared to juvenile leaves, and C4 is facultative in juvenile leaves but constitutive in mature leaves.


Bicarbonate use C4 metabolism Carbon dioxide-concentrating mechanism (CCM) Crassulacean acid metabolism (CAM) Freshwater macrophyte Leaf maturity 





Crassulacean acid metabolism


Carbon dioxide-concentrating mechanism


Concentration of total inorganic carbon


Fresh weight


High CO2


Low CO2


NAD(P)-malic enzyme


Phosphoenol pyruvate


PEP carboxylase


Pyruvate phosphate dikinase


Ribulose 1,5-bisphosphate carboxylase-oxygenase


Standard deviation



This research was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB31000000), the Chinese Academy of Sciences President’s International Fellowship Initiative to SCM and BG (2015VBA023, 2016VBA006) and the National Key Research and Development Program of China (2016YFA0601000). We thank the two reviewers for their helpfull comments.

Supplementary material

11120_2018_568_MOESM1_ESM.docx (107 kb)
Supplementary material 1 (DOCX 107 KB)


  1. Adams P, Nelson DE, Yamada S, Chmara W, Jensen RG, Bohnert HJ, Griffiths H (1998) Growth and development of Mesembryanthemum crystallinum (Aizoaceae). New Phytol 138:171–190CrossRefGoogle Scholar
  2. Aulio K (1985) Differential expression of diel acid metabolism in two life forms of Littorella uniflora (L.) Aschers. New Phytol 100:533–536CrossRefGoogle Scholar
  3. Borland AM, Dodd AN (2002) Carbohydrate partitioning in crassulacean acid metabolism plants: reconciling potential conflicts of interest. Funct Plant Biol 29:707–716CrossRefGoogle Scholar
  4. Bowes G, Salvucci ME (1989) Plasticity in the photosynthetic carbon metabolism of submerged aquatic macrophytes. Aquat Bot 34:233–266CrossRefGoogle Scholar
  5. Bowes G, Rao SK, Estavillo GM, Reiskind JB (2002) C4 mechanisms in aquatic angiosperms: comparisons with terrestrial C4 systems. Funct Plant Biol 29:379–392CrossRefGoogle Scholar
  6. Brain RA, Solomon KR (2007) A protocol for conducting 7-day daily renewal tests with Lemna gibba. Nat Protoc 2:979–987CrossRefPubMedGoogle Scholar
  7. Casati P, Lara MV, Andreo CS (2000) Induction of a C4-like mechanism of CO2 fixation in Egeria densa, a submersed aquatic species. Plant Physiol 123:1611–1621CrossRefPubMedPubMedCentralGoogle Scholar
  8. Ceusters J, Borland AM, Taybi T, Frans M, Godts C, De Proft MP (2014) Light quality modulates metabolic synchronization over the diel phases of crassulacean acid metabolism. J Exp Bot 65:3705–3714CrossRefPubMedPubMedCentralGoogle Scholar
  9. Cook CDK, Urmi-König K (1984) A revision of the genus Ottelia (Hydrocharitaceae). 2. The species of Eurasia, Australasia and America. Aquat Bot 20:131–177CrossRefGoogle Scholar
  10. Guralnick LJ, Edwards GE, Ku MSB, Hockema B, Franceschi VR (2002) Photosynthetic and anatomical characteristics in the C4-crassulacean acid metabolism-cycling plant, Portulaca grandiflora. Funct Plant Biol 29:763–773CrossRefGoogle Scholar
  11. Hatch MD (1987) C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochim Biophys Acta 895:81–106CrossRefGoogle Scholar
  12. Holtum JAM, Hancock LP, Edwards EJ, Winter K (2017) Optional use of CAM photosynthesis in two C4 species, Portulaca cyclophylla and Portulaca digyna. J Plant Physiol 214:91–96CrossRefPubMedGoogle Scholar
  13. Hostrup O, Wiegleb G (1991) The influence of different CO2 concentrations in the light and the dark on diurnal malate rhythm and phosphoenolpyruvate carboxylase activities in leaves of Littorella uniflora (L.) Aschers. Aquat Bot 40:91–100CrossRefGoogle Scholar
  14. Huber SC (1983) Relation between photosynthetic starch formation and dry-weight partitioning between the shoot and root. Can J Bot 61:2709–2716CrossRefGoogle Scholar
  15. Jones MB (1975) The effect of leaf age on leaf resistance and CO2 exchange of the CAM plant Bryophyllum fedtschenkoi. Planta 123:91–96CrossRefPubMedGoogle Scholar
  16. Keeley JE (1981) Isoetes howelli—a submerged aquatic CAM plant. Am J Bot 68:420–424CrossRefGoogle Scholar
  17. Keeley JE, Rundel PW (2003) Evolution of CAM and C4 carbon concentrating mechanisms. Int J Plant Sci 164:S55–S77CrossRefGoogle Scholar
  18. Keeley JE, Walker CM, Mathews RP (1983) Crassulacean acid metabolism in Isoetes bolanderi in high elevation oligotrophic lakes. Oecologia 58:63–69CrossRefPubMedGoogle Scholar
  19. Kitajima M, Butler WL (1975) Quenching of chlorophyll fluorescence and primary photochemistry in chloroplasts by dibromothymoquinone. Biochim Biophys Acta 376:105–115CrossRefPubMedGoogle Scholar
  20. Klavsen SK, Maberly SC (2010) Effect of light and CO2 on inorganic carbon uptake in the invasive aquatic CAM plant Crassula helmsii. Funct Plant Biol 37:737–747CrossRefGoogle Scholar
  21. Klavsen SK, Madsen TV (2008) Effect of leaf age on CAM activity in Littorella uniflora. Aquat Bot 89:50–56CrossRefGoogle Scholar
  22. Klavsen SK, Madsen TV, Maberly SC (2011) Crassulacean acid metabolism in the context of other carbon-concentrating mechanisms in freshwater plants: a review. Photosynth Res 109:269–279CrossRefPubMedGoogle Scholar
  23. Koch K, Kennedy RA (1980) Characteristics of crassulacean acid metabolism in the succulent C4 dicot, Portulaca oleracea L. Plant Physiol 65:193–197CrossRefPubMedPubMedCentralGoogle Scholar
  24. Maberly SC (1996) Diel, episodic and seasonal changes in pH and concentrations of inorganic carbon in a productive lake. Freshw Biol 35:579–598CrossRefGoogle Scholar
  25. Maberly SC, Gontero B (2017) Ecological imperatives for aquatic CO2-concentrating mechanisms. J Exp Bot 68:3797–3814CrossRefPubMedGoogle Scholar
  26. Maberly SC, Madsen TV (2002) Use of bicarbonate ions as a source of carbon in photosynthesis by Callitriche hermaphroditica L. Aquat Bot 73:1–7CrossRefGoogle Scholar
  27. Maberly SC, Spence DHN (1983) Photosynthetic inorganic carbon use by freshwater plants. J Ecol 71:705–724CrossRefGoogle Scholar
  28. Madsen TV (1987) The effect of different growth conditions on dark and light carbon assimilation in Littorella uniflora. Physiol Plant 70:183–188CrossRefGoogle Scholar
  29. Madsen TV, Maberly SC, Bowes G (1996) Photosynthetic acclimation of submersed angiosperms to CO2 and HCO3 . Aquat Bot 53:15–30CrossRefGoogle Scholar
  30. Neuhaus HE, Schulte N (1996) Starch degradation in chloroplasts isolated from C3 or CAM (crassulacean acid metabolism)-induced Mesembryanthemum crystallinum L. Biochem J 318:945–953CrossRefPubMedPubMedCentralGoogle Scholar
  31. Nishida K (1978) Effect of leaf age on light and dark 14CO2 fixation in a CAM plant, Bryophyllum calycinum. Plant Cell Physiol 19:935–941CrossRefGoogle Scholar
  32. Nobel PS, Hartsok TL (1983) Relationships between photosynthetically active raditation, nocturnal acid accumulation, and CO2 uptake for a crassulacean acid metabolism plant, Opuntia ficus-indica. Plant Phys 71:71–75CrossRefGoogle Scholar
  33. Osmond CB (1978) Crassulacean acid metabolism: a curiosity in context. Ann Rev Plant Physiol 29:379–414CrossRefGoogle Scholar
  34. Paul MJ, Loos K, Stitt M, Ziegler P (1993) Starch-degrading enzymes during the induction of CAM in Mesembryanthemum crystallinum. Plant Cell Environ 16:531–538CrossRefGoogle Scholar
  35. Pedersen O, Rich SM, Pulido C, Cawthray GR, Colmer TD (2011) Crassulacean acid metabolism enhances underwater photosynthesis and diminishes photorespiration in the aquatic plant Isoetes australis. New Phytol 190:332–339CrossRefPubMedPubMedCentralGoogle Scholar
  36. Robe WE, Griffiths H (1990) Photosynthesis of Littorella uniflora grown under two PAR regimes: C3 and CAM gas exchange and the regulation of internal CO2 and O2 concentrations. Oecologia 85:128–136CrossRefPubMedPubMedCentralGoogle Scholar
  37. Sage RF (2016) A portrait of the C4 photosynthetic family on the 50th anniversary of its discovery: species number, evolutionary lineages, and Hall of Fame. J Exp Bot 67:4039–4056CrossRefPubMedPubMedCentralGoogle Scholar
  38. Salvucci ME, Bowes G (1983) Two photosynthetic mechanisms mediating the low photorespiratory state in submersed aquatic angiosperms. Plant Physiol 73:488–496CrossRefPubMedPubMedCentralGoogle Scholar
  39. Sand-Jensen K, Gordon DM (1986) Variable HCO3 affinity of Elodea canadensis Michaux in response to different HCO3 and CO2 concentrations during growth. Oecologia 70:426–432CrossRefGoogle Scholar
  40. Schulze W, Stitt M, Schulze ED, Neuhaus HE, Fichtner K (1990) A quantification of the significance of assimilatory starch for growth of Arabidopsis thaliana L. Heynh. Plant Physiol 95:890–895CrossRefGoogle Scholar
  41. Shao H, Gontero B, Maberly SC, Jiang HS, Cao Y, Li W, Huang WM (2017) Responses of Ottelia alismoides, an aquatic plant with three CCMs, to variable CO2 and light. J Exp Bot 68:3985–3995CrossRefPubMedPubMedCentralGoogle Scholar
  42. Silvera K, Neubig KM, Whitten WM, Williams NH, Winter K, Cushman JC (2010) Evolution along the crassulacean acid metabolism continuum. Funct Plant Biol 37:995–1010CrossRefGoogle Scholar
  43. Smith JAC, Bryce JH (1992) Metabolite compartmentation and transport in CAM plants. In: Tobin AK (ed) Plant organelles. Cambridge University Press, Cambridge, pp 141–167Google Scholar
  44. Smith AM, Zeeman SC (2006) Quantification of starch in plant tissues. Nat Protoc 1:1342–1345CrossRefPubMedGoogle Scholar
  45. Taybi T, Cushman JC, Borland AM (2002) Environmental, hormonal and circadian regulation of crassulacean acid metabolism expression. Funct Plant Biol 29:669–678CrossRefGoogle Scholar
  46. Ting IP, Hann J, Sipes D, Patel A, Walling LL (1993) Expression of P-enolpyruvate carboxylase and other aspects of CAM during the development of Peperomia camptotricha leaves. Bot Acta 106:313–319CrossRefGoogle Scholar
  47. Ting IP, Patel A, Kaur S, Hann J, Walling L (1996) Ontogenetic development of crassulacean acid metabolism as modified by water stress in Peperomia. In: Winter K, Smith JAC (eds) Crassulacean acid metabolism. Biochemistry, ecophysiology and evolution. Springer, Heidelberg, pp 204–215CrossRefGoogle Scholar
  48. Vadstrup M, Madsen TV (1995) Growth limitation of submerged aquatic macrophytes by inorganic carbon. Freshw Biol 34:411–419CrossRefGoogle Scholar
  49. von Willert DJ, Kirst GO, Treichel S, von Willert K (1976) The effect of leaf age and salt stress on malate accumulation and phosphoenolpyruvate carboxylase activity in Mesembryanthemum crystallinum. Plant Sci Lett 7:341–346CrossRefGoogle Scholar
  50. Wen H, Wagner J, Larcher W (1997) Growth and nocturnal acid accumulation during early ontogeny of Agave attenuate grown in nutrient solution and in vitro culture. Biol Plant 39:1–11CrossRefGoogle Scholar
  51. Winter K (1980) Day/night changes in the sensitivity of phosphoenolpyruvate carboxylase to malate during crassulacean acid metabolism. Plant Physiol 65:792–796CrossRefPubMedPubMedCentralGoogle Scholar
  52. Winter K, von Willert DJ (1972) NaCl-induzierter crassulaceen säurestoffwechsel bei Mesembryanthemum crystallinum. Z Pflanzenphysiol 67:166–170CrossRefGoogle Scholar
  53. Winter K, Garcia M, Holtum JAM (2008) On the nature of facultative and constitutive CAM: environmental and developmental control of CAM expression during early growth of Clusia, Kalanchoё, and Opuntia. J Exp Bot 59:1829–1840CrossRefPubMedGoogle Scholar
  54. Yin LY, Li W, Madsen TV, Maberly SC, Bowes G (2017) Photosynthetic inorganic carbon acquisition in 30 freshwater macrophytes. Aquat Bot 140:48–54CrossRefGoogle Scholar
  55. Yu LF, Yu D (2009) Responses of the threatened aquatic plant Ottelia alismoides to water level fluctuations. Fund Appl Limnol 175:295–300CrossRefGoogle Scholar
  56. Zhang YZ, Yin LY, Jiang HS, Li W, Gontero B, Maberly SC (2014) Biochemical and biophysical CO2 concentrating mechanisms in two species of freshwater macrophyte within the genus Ottelia (Hydrocharitaceae). Photosynth Res 121:285–297CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Key Laboratory of Aquatic Botany and Watershed Ecology, Wuhan Botanical GardenChinese Academy of SciencesWuhanChina
  2. 2.Hubei Key Laboratory of Wetland Evolution & Ecological Restoration, Wuhan Botanical GardenChinese Academy of SciencesWuhanChina
  3. 3.Aix Marseille Univ, CNRS, BIP, UMR 7281Marseille Cedex 09France
  4. 4.Sino-Danish CenterThe University of Chinese Academy of SciencesBeijingChina
  5. 5.School of Resources and Environmental ScienceHubei UniversityWuhanChina
  6. 6.School of Resource and Environmental EngineeringEast China University of Science and TechnologyShanghaiChina
  7. 7.Lake Ecosystems Group, Centre for Ecology & HydrologyLancaster Environment CentreBailriggUK

Personalised recommendations