Photosynthesis Research

, Volume 139, Issue 1–3, pp 337–358 | Cite as

Distribution of Cd and other cations between the stroma and thylakoids: a quantitative approach to the search for Cd targets in chloroplasts

  • Eugene A. LysenkoEmail author
  • Alexander A. Klaus
  • Alexander V. Kartashov
  • Victor V. Kusnetsov
Original Article


Plant growth and photosynthetic activity are usually inhibited due to the overall action of Cd on a whole organism, though few cadmium cations can invade chloroplasts in vivo. We found that in vivo, the major portion of Cd in barley chloroplasts is located in the thylakoids (80%), and the minor portion is in the stroma (20%). Therefore, the electron-transport chain in the thylakoids would be the likely target for direct Cd action in vivo. In vitro, we found the distribution of Cd to be shifted to the stroma (40–60%). In barley chloroplasts, the major portions of Mg, Fe, Mn, and Cu were found to be located in the thylakoids, and most Ca, Zn, and K in the stroma. This finding was true for both control and Cu- or Fe-treated plants. Treatment with Cd affected the contents of all cations, and the largest portions of Ca and Zn were in the thylakoids. Alterations of the K and Mn contents were caused by Cd, Cu, or Fe treatment; the levels of other cations in chloroplasts were changed specifically by Cd treatment. The quantity of Cd in chloroplasts was small in comparison to that of Mg, Ca, and Fe. In thylakoids, the amount of Cd was similar to that of Cu and comparable to the levels of Zn and Mn. Accordingly, the possible targets for direct Cd action in thylakoids are the Mn cluster, plastocyanin, carbonic anhydrase, or FtsH protease. The quantity of Cd in thylakoids is sufficient to replace a cation nearly completely at one of these sites or partially (20–30%) at many of these sites.


Plant Cadmium Cations Chloroplast Thylakoid Stroma Cation distribution 



Ethylenediaminetetraacetic acid


Minimal Chl fluorescence in light


Maximal Chl fluorescence in light


Steady-state Chl fluorescence


Heavy metal




Nicotinamide adenine dinucleotide phosphate, oxidized form


Oxygen evolving complex of PSII




Maximal amplitude of P700 change in light


Polyphenol oxidase


Photosystem I


Photosystem II


Standard deviation


Standard error


Superoxide dismutase


Wild type (not mutant)



The work was supported by Grant No. 14-14-00584 from the Russian Science Foundation.

Supplementary material

11120_2018_528_MOESM1_ESM.pdf (75 kb)
Supplementary material 1 (PDF 75 KB)


  1. Aguirre G, Pilon M (2016) Copper delivery to chloroplast proteins and its regulation. Front Plant Sci. Google Scholar
  2. Andresen E, Küpper H (2013) Cadmium toxicity in plants. In: Sigel A, Sigel H, Sigel R (eds) Cadmium: from toxicity to essentiality. Metal ions in life sciences, vol 11. Springer, Dordrecht, pp 395–413. CrossRefGoogle Scholar
  3. Aravind P, Prasad MNV (2004) Carbonic anhydrase impairment in cadmium-treated Ceratophyllum demersum L. (free floating freshwater macrophyte): toxicity reversal by zinc. J Anal At Spectrom 19:52–57. CrossRefGoogle Scholar
  4. Baker AJM (1981) Accumulators and excluders—strategies in response of plants to heavy metals. J Plant Nutr 3:643–654. CrossRefGoogle Scholar
  5. Banci L, Bertini I, Cabelli DE, Hallewell RA, Tung JW, Viezzoli MS (1991) A characterization of copper/zinc superoxide dismutase mutants at position 124 Zinc-deficient proteins. FEBS J 196:123–128. Google Scholar
  6. Barcelo J, Vazquez MD, Poschenrieder Ch (1988) Structural and ultrastructural disorders in cadmium-treated bush bean plants (Phaseolus vulgaris L.). New Phytol 108:37–49. CrossRefGoogle Scholar
  7. Baryla A, Carrier P, Franck F, Coulomb C, Sahut C, Havaux M (2001) Leaf chlorosis in oilseed rape plants (Brassica napus) grown on cadmium-polluted soil: causes and consequences for photosynthesis and growth. Planta 212:696–709. CrossRefGoogle Scholar
  8. Baszynski T, Wajda L, Krol M, Wolinska D, Krupa Z, Tukendorf Z (1980) Photosynthetic activities of cadmium-treated tomato plants. Physiol Plant 48:365–370. CrossRefGoogle Scholar
  9. Bazzaz MB, Govindjee (1974) Effects of cadmium nitrate on spectral characteristics and light reactions of chloroplasts. Environ Lett 6:1–12CrossRefGoogle Scholar
  10. Bazzaz FA, Rolfe GL, Carlson RW (1974) Effect of Cd on photosynthesis and transpiration of excised leaves of corn and sunflower. Physiol Plant 32:373–376. CrossRefGoogle Scholar
  11. Blaby-Haas CE, Merchant SS (2013) Metal homeostasis: sparing and salvaging metals in chloroplasts. Encycl Inorg Bioinorg Chem. Google Scholar
  12. Burzynski M, Klobus G (2004) Changes of photosynthetic parameters in cucumber leaves under Cu, Cd, and Pb stress. Photosynthtica 42:505–510. CrossRefGoogle Scholar
  13. Chow WS, Fan D-Y, Oguchi R, Jia H, Losciale P, Park Y-I, He J, Öquist G, Shen Y-G, Anderson JM (2012) Quantifying and monitoring functional photosystem II and the stoichiometry of the two photosystems in leaf segments: approaches and approximations. Photosynth Res 113:63–74. CrossRefGoogle Scholar
  14. Church WB, Guss JM, Potter JJ, Freeman HC (1986) The crystal structure of mercury-substituted Poplar Plastocyanin at 1.9-Å Resolution. J Biol Chem 261:234–237.
  15. Ci D, Jiang D, Wollenweber B, Dai T, Jing Q, Cao W (2010) Cadmium stress in wheat seedlings: growth, cadmium accumulation and photosynthesis. Acta Physiol Plant 32:365–373. CrossRefGoogle Scholar
  16. Cullen JT, Maldonado MT (2013) Biogeochemistry of cadmium and its release to the environment. In: Sigel A, Sigel H, Sigel R (eds) Cadmium: from toxicity to essentiality. Metal ions in life sciences, vol 11. Springer, Dordrecht, pp 31–62. CrossRefGoogle Scholar
  17. Delperee C, Lutts S (2008) Growth inhibition occurs independently of cell mortality in tomato (Solanum lycopersicum) exposed to high cadmium concentrations. J Integr Plant Biol 50:300–310. CrossRefGoogle Scholar
  18. Demmig B, Gimmler H (1983) Properties of the isolated intact chloroplast at cytoplasmic K+ concentrations I. Light-induced cation uptake into intact chloroplasts is driven by an electrical potential difference. Plant Physiol 73:169–174. CrossRefGoogle Scholar
  19. Faller P, Kienzler K, Krieger-Liszkay A (2005) Mechanism of Cd2+ toxicity: Cd2+ inhibits photoactivation of photosystem II by competitive binding to the essential Ca2+ site. Biochim Biophys Acta 1706:158–164. CrossRefGoogle Scholar
  20. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875. CrossRefGoogle Scholar
  21. Gallego SM, Pena LB, Barcia RA, Azpilicueta CE, Iannone MF, Rosales EP, Zawoznik MS, Groppa MD, Benavides MP (2012) Unravelling cadmium toxicity and tolerance in plants: insight into regulatory mechanisms. Environ Exp Bot 83:33–46. CrossRefGoogle Scholar
  22. Gross EL (1993) Plastocyanin: structure and function. Photosynth Res 37:103–116. CrossRefGoogle Scholar
  23. Iglesias AA, Andreo CS (1984) Involvement of thiol groups in the activity of phosphoenolpyruvate carboxylase from maize leaves. Photosynth Res 5:215–226. CrossRefGoogle Scholar
  24. Jansson H, Hansson Ö (2008) Competitive inhibition of electron donation to photosystem 1 by metal-substituted plastocyanin. Biochim Biophys Acta 1777:1116–1121. CrossRefGoogle Scholar
  25. Jozefczak M, Bohler S, Schat H, Horemans N, Guisez Y, Remans T, Vangronsveld J, Cuypers A (2015) Both the concentration and redox state of glutathione and ascorbate influence the sensitivity of arabidopsis to cadmium. Ann Bot 116:601–612. CrossRefGoogle Scholar
  26. Klaus AA, Lysenko EA, Kholodova VP (2013) Maize plant growth and accumulation of photosynthetic pigments at short- and longterm exposure to cadmium. Russ J Plant Physiol 60:250–259. CrossRefGoogle Scholar
  27. Lee MJ, Ayaki H, Goji J, Kitamura K, Nishio H (2006) Cadmium restores in vitro splicing activity inhibited by zinc-depletion. Arch Toxicol 80:638. CrossRefGoogle Scholar
  28. Li EH, Miles CD (1975) Effects of cadmium on photoreaction II of chloroplasts. Plant Sci Lett 5:33–40. CrossRefGoogle Scholar
  29. Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382. CrossRefGoogle Scholar
  30. Lin Y-F, Aarts MGM (2012) The molecular mechanism of zinc and cadmium stress response in plants. Cell Mol Life Sci 69:3187–3206. CrossRefGoogle Scholar
  31. Lysenko EA, Klaus AA, Pshybytko NL, Kusnetsov VV (2015) Cadmium accumulation in chloroplasts and its impact on chloroplastic processes in barley and maize. Photosynth Res 125:291–303. CrossRefGoogle Scholar
  32. Maret W, Moulis J-M (2013) The bioinorganic chemistry of cadmium in the context of its toxicity. In: Sigel A, Sigel H, Sigel R (eds) Cadmium: from toxicity to essentiality. Metal ions in life sciences, vol 11. Springer, Dordrecht, pp 1–29. CrossRefGoogle Scholar
  33. Maury WJ, Huber SC, Moreland DE (1981) Effects of magnesium on intact chloroplasts II. Cation specificity and involvement of the envelope ATPase in (sodium) potassium/proton exchange across the envelope. Plant Physiol 68:1257–1263. CrossRefGoogle Scholar
  34. Nakatani HY, Barber J, Minski MJ (1979) The influence of the thylakoid membrane surface properties on the distribution of ions in chloroplasts. Biochem Biophys Acta 545:24–35. Google Scholar
  35. Nishimura K, Kato Y, Sakamoto W (2016) Chloroplast proteases: updates on proteolysis within and across suborganellar compartments. Plant Phys 171:2280–2293. Google Scholar
  36. Nobel PS (1969) Light-induced changes in the ionic content of chloroplasts in Pisum sativum. Biochem Biophys Acta 172:134–143. Google Scholar
  37. Pagliano C, Raviolo M, Dalla Vecchia F, Gabbrielli R, Gonnelli C, Rascio N, Barbato R, La Rocca N (2006) Evidence for PSII donor-side damage and photoinhibition induced by cadmium treatment on rice (Oryza sativa L.). J Photochem Photobiol B 84:70–78. CrossRefGoogle Scholar
  38. Pan J, Plant JA, Voulvoulis N, Oates CJ, Ihlenfeld C (2010) Cadmium levels in Europe: implications for human health. Environ Geochem Health 32:1–12. CrossRefGoogle Scholar
  39. Pätsikkä E, Kairavuo M, Šeršen F, Aro E-M, Tyystjärvi E (2002) Excess copper predisposes photosystem II to photoinhibition in vivo by outcompeting iron and causing decrease in leaf chlorophyll. Plant Physiol 129:1359–1367. CrossRefGoogle Scholar
  40. Pietrini F, Iannelli MA, Pasqualini S, Massacci A (2003) Interaction of cadmium with glutathione and photosynthesis in developing leaves and chloroplasts of Phragmites australis (Cav.) Trin. ex Steudel. Plant Physiol 133:829–837. CrossRefGoogle Scholar
  41. Pilon M, Ravet K, Tapken W (2011) The biogenesis and physiological function of chloroplast superoxide dismutases. Biochim Biophys Acta 1807:989–998. CrossRefGoogle Scholar
  42. Portis AR, Heldt HW (1976) Light-dependent changes of the Mg2+ concentration in the stroma in relation to the Mg2+ dependency of CO2 fixation. Biochim Biophys Acta 449:434–446. CrossRefGoogle Scholar
  43. Rascio N, Dalla Vecchia F, La Rocca N, Barbato R, Pagliano C, Raviolo M, Gonnelli C, Gabbrielli R (2008) Metal accumulation and damage in rice (cv. Vialone nano) seedlings exposed to cadmium. Environ Exp Bot 62:267–278. CrossRefGoogle Scholar
  44. Schneider A, Steinberger I, Herdean A, Gandini C, Eisenhut M, Kurz S, Morper A, Hoecker N, Rühle T, Labs M, Flügge U-I, Geimer S, Schmidt SB, Husted S, Weber APM, Spetea C, Leister D (2016) The evolutionarily conserved protein PHOTOSYNTHESIS AFFECTED MUTANT71 is required for efficient manganese uptake at the thylakoid membrane in Arabidopsis. Plant Cell 28:892–910. Google Scholar
  45. Seregin IV, Ivanov VB (2001) Physiological aspects of cadmium and lead toxic effects on higher plants. Russ J Plant Physiol 48:523–544. CrossRefGoogle Scholar
  46. Shi GR, Cai QS (2008) Photosynthetic and anatomic responses of peanut leaves to cadmium stress. Photosynthetica 46:627–630. CrossRefGoogle Scholar
  47. Shikanai T, Müller-Moulé P, Munekage Y, Niyogi KK, Pilon M (2003) PAA1, a P-Type ATPase of Arabidopsis, functions in copper transport in chloroplasts. Plant Cell 15:1333–1346. CrossRefGoogle Scholar
  48. Siedlecka A, Baszynski T (1993) Inhibition of electron flow around photosystem I in chloroplasts of Cd-treated maize plants is due to Cd-induced iron deficiency. Physiol Plant 87:199–202. CrossRefGoogle Scholar
  49. Sigfridsson KGV, Bernat G, Mamedov F, Styring S (2004) Molecular interference of Cd2+ with Photosystem II. Biochim Biophys Acta 1659:19–31. CrossRefGoogle Scholar
  50. Solti A, Kovács K, Müller B, Vázquez S, Hamar E, Pham HD, Tóth B, Abadia J, Fodor F (2016) Does a voltage-sensitive outer envelope transport mechanism contributes to the chloroplast iron uptake? Planta 244:1303–1313. CrossRefGoogle Scholar
  51. Tang L, Ying R-R, Jiang D, Zeng X-W, Morel J-L, Tang Y-T, Qiu R-L (2013) Impaired leaf CO2 diffusion mediates Cd-induced inhibition of photosynthesis in the Zn/Cd hyperaccumulator Picris divaricata. Plant Physiol Biochem 73:70–76. CrossRefGoogle Scholar
  52. Toth T, Zsiros O, Kis M, Garab G, Kovacs L (2012) Cadmium exerts its toxic effects on photosynthesis via a cascade mechanism in the cyanobacterium, Synechocystis PCC 6803. Plant Cell Environ 35:2075–2086. CrossRefGoogle Scholar
  53. Umena Y, Kawakami K, Shen JR, Kamiya N (2011) Crystal structure of oxygen-evolving photosystem II at a resolution of 1.9 Å. Nature 473:55–60. CrossRefGoogle Scholar
  54. Wang H, Zhao SC, Liu RL, Zhou W, Jin JY (2009) Changes of photosynthetic activities of maize (Zea mays L.) seedlings in response to cadmium stress. Photosynthetica 47:277–283. CrossRefGoogle Scholar
  55. Weigel HJ (1985) The effect of Cd2+ on photosynthetic reactions of mesophyll protoplasts. Physiol Plant 63:192–200. CrossRefGoogle Scholar
  56. Whatley FR, Ordin L, Arnon DI (1951) Distribution of micronutrient metals in leaves and chloroplast fragments. Plant Physiol 26:414–418. CrossRefGoogle Scholar
  57. Wu FB, Zhang GP, Yu JS (2003) Genotypic differences in effect of Cd on photosynthesis and chlorophyll fluorescence of barley (Hordeum vulgare L.). Bull Environ Contam Toxicol 71:1272–1281. Google Scholar
  58. Xu Y, Feng L, Jeffrey PD, Shi Y, Morel FM (2008) Structure and metal exchange in the cadmium carbonic anhydrase of marine diatoms. Nature 452:56–61. CrossRefGoogle Scholar
  59. Ying R-R, Qiu R-L, Tang Y-T, Hu P-J, Qiu H, Chen H-R, Shi T-H, Morel J-L (2010) Cadmium tolerance of carbon assimilation enzymes and chloroplast in Zn/Cd hyperaccumulator Picris divaricata. J Plant Phys 167:81–87. CrossRefGoogle Scholar
  60. Zhu R, Sheila M, Macfie SM, Ding Z (2005) Cadmium-induced plant stress investigated by scanning electrochemical microscopy. J Exp Bot 56:2831–2838. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Plant PhysiologyRussian Academy of SciencesMoscowRussia

Personalised recommendations