Membrane Excitation and Cytoplasmic Streaming as Modulators of Photosynthesis and Proton Flows in Characean Cells

  • A. A. Bulychev


Internodal cells of Chara corallina represent a unique model system to study interactions between photosynthesis, membrane excitation, and cytoplasmic streaming, as well as the role of these processes in generation and regulation of functional patterns in green cells and tissues. It is established that the inflow of cytoplasm from darkened cell parts promotes photosynthetic activity of chloroplasts residing at intermediate irradiance, whereas the arrival of cytoplasm from illuminated regions suppresses this activity and enhances nonphotochemical quenching. The vectorial movement of the “irradiated” cytoplasm induces functional asymmetry around the light spot (pattern formation) both in the chloroplast layer and in the plasma membrane. The messenger transported between illuminated and shaded cell parts was found to move at the velocity of cytoplasmic streaming. The effects of membrane excitation (action potential) on photosynthesis and membrane H+ transport are area specific; they are mediated by different mechanisms under physiological conditions and in the presence of some redox-cycling compounds. The influence of action potential on chlorophyll fluorescence under spot illumination appears to involve the activation of Ca2+-mediated pathways and the suppression of metabolite exchange between darkened and illuminated cell parts due to the stoppage of cyclosis. The cytoplasmic flow from darkened to illuminated cell parts seems to enhance interactions between respiratory and light-dependent metabolism, which promotes photosynthesis and protects chloroplasts from photooxidative damage under excess light.


Methyl Viologen Action Potential Generation Cytoplasmic Streaming Alkaline Zone Characean Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



area of inspection where external pH and chlorophyll fluorescence are measured in Chara internodal cell;


action potential;


cytochalasin B;




methyl viologen;


nonphotochemical quenching;


cytosolic pH;


pH near the cell surface in the outer medium;


plasma membrane;


photon flux density;


photosystems I and II;


reactive oxygen species;


effective quantum yield of electron transport in PSII



This work was supported by the Russian Foundation of Basic Research


  1. Asada K (1999) The water–water cycle in chloroplasts: scavenging of active oxygen and dissipation of excess photons. Ann Rev Plant Physiol Plant Mol Biol 50:601–639CrossRefGoogle Scholar
  2. Baker NR (2008) Chlorophyll fluorescence: a probe of photosynthesis in vivo. Annu Rev Plant Biol 59:89–113PubMedCrossRefGoogle Scholar
  3. Beilby MJ (2007) Action potential in charophytes. Int Rev Cytol 257:43–82. doi: 10.1016/S0074-7696(07)57002-6 PubMedCrossRefGoogle Scholar
  4. Beilby MJ, Mimura T, Shimmen T (1993) The proton pump, high pH channels, and excitation: voltage clamp studies of intact and perfused cells of Nitellopsis obtusa. Protoplasma 175:144–152CrossRefGoogle Scholar
  5. Berestovsky GN, Kataev AA (2005) Voltage-gated calcium and Ca2+-activated chloride channels and Ca2+ transients: voltage-clamp studies of perfused and intact cells of Chara. Eur Biophys J 34:973–986. doi: 10.1007/s00249-005-0477-9 PubMedCrossRefGoogle Scholar
  6. Bisson MA, Walker NA (1980) The Chara plasmalemma at high pH. Electrical measurements show rapid specific passive uniport of H+ or OH. J Membr Biol 56:1–7CrossRefGoogle Scholar
  7. Borowitzka MA (1987) Calcification in algae: mechanisms and the role of metabolism. Crit Rev Plant Sci 6:1–45CrossRefGoogle Scholar
  8. Bradley MO (1973) Microfilaments and cytoplasmic streaming: inhibition of streaming with cytochalasin. J Cell Sci 12:327–343PubMedGoogle Scholar
  9. Braun M, Foissner I, Lühring H, Schubert H, Thiel G (2007) Characean algae: still a valid model system to examine fundamental principles in plants. Progr Bot 68:193–220CrossRefGoogle Scholar
  10. Bulychev AA, Dodonova SO (2011) Effects of cyclosis on chloroplast–cytoplasm interactions revealed with localized lighting in characean cells at rest and after electrical excitation. Biochim Biophys Acta 1807:1221–1230. doi: 10.1016/j.bbabio.2011.06.009 PubMedCrossRefGoogle Scholar
  11. Bulychev AA, Kamzolkina NA (2006) Differential effects of plasma membrane electric excitation on H+ fluxes and photosynthesis in characean cells. Bioelectrochemistry 69:209–215. doi: 10.1016/j.bioelechem.2006.03.001 PubMedCrossRefGoogle Scholar
  12. Bulychev AA, Krupenina NA (2008a) Action potential opens access for the charged cofactor to the chloroplasts of Chara corallina cells. Russ J Plant Physiol 55:175–184. doi: 10.1134/S1021443708020039 CrossRefGoogle Scholar
  13. Bulychev AA, Krupenina NA (2008b) Facilitated permeation of methyl viologen into chloroplasts in situ during electric pulse generation in excitable plant cell membranes. Biochem (Moscow), Suppl Series A: Membr Cell Biol 2:387–394. doi: 10.1134/S1990747808040132 CrossRefGoogle Scholar
  14. Bulychev AA, Krupenina NA (2008c) Effects of plasma membrane excitation on spatially distributed H+ fluxes, photosynthetic electron transport and non-photochemical quenching in the plant cell. In: Bernstein EM (ed) Bioelectrochemistry research developments. Nova Science Publishers, New YorkGoogle Scholar
  15. Bulychev AA, Krupenina NA (2009) Transient removal of alkaline zones after excitation of Chara cells is associated with inactivation of high conductance in the plasmalemma. Plant Signal Behav 4:727–734PubMedCrossRefGoogle Scholar
  16. Bulychev AA, Krupenina NA (2010) Physiological implications of action potential in characean cell: effects on pH bands and spatial pattern of photosynthesis. In: DuBois ML (ed) Action potential: biophysical and cellular context, initiation, phases and propagation. Nova Science Publishers, New YorkGoogle Scholar
  17. Bulychev AA, Vredenberg WJ (2003) Spatio-temporal patterns of photosystem II activity and plasma-membrane proton flows in Chara corallina cells exposed to overall and local illumination. Planta 218:143–151. doi: 10.1007/s00425-003-1084-6 PubMedCrossRefGoogle Scholar
  18. Bulychev AA, Cherkashin AA, Rubin AB, Vredenberg WJ, Zykov VS, Müller SC (2001a) Comparative study on photosynthetic activity of chloroplasts in acid and alkaline zones of Chara corallina. Bioelectrochemistry 53:225–232PubMedCrossRefGoogle Scholar
  19. Bulychev AA, Polezhaev AA, Zykov SV, Pljusnina TY, Riznichenko GY, Rubin AB, Jantoss W, Zykov VS, Müller SC (2001b) Light-triggered pH banding profile in Chara cells revealed with a scanning pH microprobe and its relation to self-organization phenomena. J Theor Biol 212:275–294. doi: 10.1006/jtbi.2001.2375 PubMedCrossRefGoogle Scholar
  20. Bulychev AA, Zykov SV, Rubin AB, Müller SC (2003) Transitions from alkaline spots to regular bands during pH pattern formation at the plasmalemma of Chara cells. Eur Biophys J 32:144–153. doi: 10.1007/s00249-003-0280-4 PubMedGoogle Scholar
  21. Bulychev AA, Kamzolkina NA, Luengviriya J, Rubin AB, Müller SC (2004) Effect of a single excitation stimulus on photosynthetic activity and light-dependent pH banding in Chara cells. J Membr Biol 202:11–19. doi: 10.1007/s00232-004-0716-5 PubMedCrossRefGoogle Scholar
  22. Bulychev AA, Van den Wijngaard PWJ, De Boer AH (2005) Spatial coordination of chloroplast and plasma membrane activities in Chara cells and its disruption through inactivation of 14-3-3 proteins. Biochemistry (Moscow) 70:55–61Google Scholar
  23. Coelho SMB, Brownlee C, Bothwell JHF (2008) A tip-high, Ca2+-interdependent, reactive oxygen species gradient is associated with polarized growth in Fucus serratus zygotes. Planta 227:1037–1046. doi: 10.1007/s00425-007-0678-9 PubMedCrossRefGoogle Scholar
  24. Davies E (2006) Electrical signals in plants: facts and hypotheses. In: Volkov A (ed) Plant electrophysiology theory and methods. Springer, BerlinGoogle Scholar
  25. Dodge A (1989) Herbicides interacting with photosystem I. In: Dodge A (ed) Herbicides and plant metabolism. Cambridge University Press, CambridgeGoogle Scholar
  26. Dodonova SO, Bulychev AA (2011) Cyclosis-related asymmetry of chloroplast–plasma membrane interactions at the margins of illuminated area in Chara corallina cells. Protoplasma 248(4):737–749. doi: 10.1007/s00709-010-0241-6 PubMedCrossRefGoogle Scholar
  27. Dodonova SO, Krupenina NA, Bulychev AA (2010) Suppression of the plasma membrane H+-conductance on the background of high H+-pump activity in dithiothreitol-treated Chara cells. Biochem (Moscow), Suppl Series A Membr Cell Biol 4:389–396. doi: 10.1134/S1990747810040094 CrossRefGoogle Scholar
  28. Dorn A, Weisenseel MH (1984) Growth and the current pattern around internodal cells of Nitella flexilis L. J Exp Bot 35:373–383CrossRefGoogle Scholar
  29. Eremin A, Bulychev A, Krupenina NA, Mair T, Hauser MJB, Stannarius R, Müller S, Rubin AB (2007) Excitation-induced dynamics of external pH pattern in Chara corallina cells and its dependence on external calcium concentration. Photochem Photobiol Sci 6:103–109. doi: 10.1039/b607602e PubMedCrossRefGoogle Scholar
  30. Feijo JA, Sainhas J, Hackett GR, Kunkel JG, Hepler PK (1999) Growing pollen tubes possess a constitutive alkaline band in the clear zone and a growth-dependent acidic tip. J Cell Biol 144:483–496PubMedCrossRefGoogle Scholar
  31. Felle HH (1998) The apoplastic pH of the Zea mays root cortex as measured with pH-sensitive microelectrodes: aspects of regulation. J Exp Bot 49:987–995Google Scholar
  32. Finazzi G, Johnson GN, Dallosto L, Joliot P, Wollman F-A, Bassi R (2004) A zeaxanthin-independent nonphotochemical quenching mechanism localized in the photosystem II core complex. Proc Natl Acad Sci U S A 101:12375–12380. doi: 10.1073/pnas.0404798101 PubMedCrossRefGoogle Scholar
  33. Foissner I (2004) Microfilaments and microtubules control the shape, motility, and subcellular distribution of cortical mitochondria in characean internodal cells. Protoplasma 224:145–157. doi: 10.1007/s00709-004-0075-1 PubMedCrossRefGoogle Scholar
  34. Foissner I, Wasteneys GO (2007) Wide-ranging effects of eight cytochalasins and latrunculin A and B on intracellular motility and actin filament reorganization in characean internodal cells. Plant Cell Physiol 48:585–597. doi: 10.1093/pcp/pcm030 PubMedCrossRefGoogle Scholar
  35. Fromm J (2006) Long-distance electrical signaling and physiological functions in higher plants. In: Volkov A (ed) Plant electrophysiology theory and methods. Springer, BerlinGoogle Scholar
  36. Fromm J, Lautner S (2007) Electrical signals and their physiological significance in plants. Plant, Cell Environ 30:249–257. doi: 10.1111/j.1365-3040.2006.01614.x CrossRefGoogle Scholar
  37. Goldstein RE, Tuval I, Van de Meent J-W (2008) Microfluidics of cytoplasmic streaming and its implications for intracellular transport. Proc Natl Acad Sci U S A 105:3663–3667. doi: 10.1073/pnas.0707223105 PubMedCrossRefGoogle Scholar
  38. Gow NAR, Kropf DL, Harold FM (1984) Growing hyphae of Achlya bisexualis generate a longtitudinal pH gradient in the surrounding medium. J Gen Microbiol 130:2967–2974PubMedGoogle Scholar
  39. Grams TEE, Lautner S, Felle HH, Matyssek R, Fromm J (2009) Heat-induced electrical signals affect cytoplasmic and apoplastic pH as well as photosynthesis during propagation through the maize leaf. Plant, Cell Environ 32:319–326. doi: 10.1111/j.1365-3040.2008.01922.x CrossRefGoogle Scholar
  40. Hager A (2003) Role of the plasma membrane H+-ATPase in auxin-induced elongation growth: historical and new aspects. J Plant Res 116:483–505. doi: 10.1007/s10265-003-0110-x PubMedCrossRefGoogle Scholar
  41. Hansen U-P, Moldaenke C, Tabrizi H, Ramm D (1993) The effect of transthylakoid proton uptake on cytosolic pH and the imbalance of ATP and NADPH/H+ production as measured by CO2− and light-induced depolarization of the plasmalemma. Plant Cell Physiol 34:681–695Google Scholar
  42. Harada A, Shimazaki K (2009) Measurement of changes in cytosolic Ca2+ in Arabidopsis guard cells and mesophyll cells in response to blue light. Plant Cell Physiol 50:360–373. doi: 10.1093/pcp/pcn203 PubMedCrossRefGoogle Scholar
  43. Jansson C, Northen T (2010) Calcifying cyanobacteria—the potential of biomineralization for carbon capture and storage. Curr Opin Biotechnol 21:1–7. doi: 10.1016/j.copbio.2010.03.017 CrossRefGoogle Scholar
  44. Johnson CH, Shingles R, Ettinger WF (2006) Regulation and role of calcium fluxes in the chloroplast. In: Wise RR, Hoober JK (eds) The structure and function of plastids. Springer, DordrechtGoogle Scholar
  45. Kamiya N (1959) Protoplasmic streaming. Springer, WienCrossRefGoogle Scholar
  46. Koziolek C, Grams TEE, Schreiber U, Matyssek R, Fromm J (2003) Transient knockout of photosynthesis mediated by electrical signals. New Phytol 161:715–722. doi: 10.1046/j.1469-8137.2003.00985.x CrossRefGoogle Scholar
  47. Krol E, Dziubinska H, Trebacz K (2010) What do plants need action potentials for? In: DuBois ML (ed) Action potential: biophysical and cellular context, initiation, phases and propagation. Nova Science Publisher, New YorkGoogle Scholar
  48. Krupenina NA, Bulychev AA (2007) Action potential in a plant cell lowers the light requirement for non-photochemical energy-dependent quenching of chlorophyll fluorescence. Biochim Biophys Acta 1767:781–788. doi: 10.1016/j.bbabio.2007.01.004 PubMedCrossRefGoogle Scholar
  49. Krupenina NA, Bulychev AA, Roelfsema MRG, Schreiber U (2008) Action potential in Chara cells intensifies spatial patterns of photosynthetic electron flow and non-photochemical quenching in parallel with inhibition of pH banding. Photochem Photobiol Sci 7:681–688. doi: 10.1039/b802243g PubMedCrossRefGoogle Scholar
  50. Krupenina NA, Bulychev AA, Schreiber U (2011) Chlorophyll fluorescence images demonstrate variable pathways in the effects of plasma membrane excitation on electron flow in chloroplasts of Chara cells. Protoplasma 248:513–522. doi: 10.1007/s00709-010-0198-5 PubMedCrossRefGoogle Scholar
  51. Lucas WJ (1975a) The influence of light intensity on the activation and operation of the hydroxyl efflux system of Chara corallina. J Exp Bot 26:347–360CrossRefGoogle Scholar
  52. Lucas WJ (1975b) Photosynthetic fixation of 14carbon by internodal cells of Chara corallina. J Exp Bot 26:331–346CrossRefGoogle Scholar
  53. Lucas WJ, Dainty J (1977) Spatial distribution of functional OH carriers along a characean internodal cell: determined by the effect of cytochalasin B on H14CO3 assimilation. J Membr Biol 32:75–92PubMedCrossRefGoogle Scholar
  54. Lucas WJ, Nuccitelli R (1980) HCO3 and OH transport across the plasmalemma of Chara: spatial resolution obtained using extracellular vibrating probe. Planta 150:120–131CrossRefGoogle Scholar
  55. Lunevsky VS, Zherelova OM, Vostrikov IY, Berestovsky GN (1983) Excitation of characeae cell membranes as a result of activation of calcium and chloride channels. J Membr Biol 72:43–58CrossRefGoogle Scholar
  56. Marten I, Deeken R, Hedrich R, Roelfsema MRG (2010) Light-induced modification of plant plasma membrane ion transport. Plant Biol 12:64–79. doi: 10.1111/j.1438-8677.2010.00384.x PubMedCrossRefGoogle Scholar
  57. McConnaughey T (1991) Calcification in Chara corallina: CO2 hydroxylation generates protons for bicarbonate assimilation. Limnol Oceanogr 36:619–628CrossRefGoogle Scholar
  58. Metraux JP, Richmond PA, Taiz L (1980) Control of cell elongation in Nitella by endogeneous cell wall pH gradients. Multiaxial extensibility and growth studies. Plant Physiol 65:204–210PubMedCrossRefGoogle Scholar
  59. Muto S, Izawa S, Miyachi S (1982) Light-induced Ca2+ uptake by intact chloroplasts. FEBS Lett 139:250–254CrossRefGoogle Scholar
  60. Nobel PS (2005) Physicochemical and environmental plant physiology. Academic, LondonGoogle Scholar
  61. Ogata K, Toko K, Fujiyoshi T, Yamafuji K (1987) Electric inhomogeneity in membrane of characean internode influenced by light-dark transition, O2, N2, CO2-free air and extracellular pH. Biophys Chem 26:71–81PubMedCrossRefGoogle Scholar
  62. Palmgren MG (1998) Protein gradients and plant growth: role of the plasma membrane H+-ATPase. Adv Bot Res 28:1–70CrossRefGoogle Scholar
  63. Pavlovic A, Slovakova L, Pandolfi C, Mancuso S (2011) On the mechanism underlying photosynthetic limitation upon trigger hair irritation in the carnivorous plant Venus flytrap (Dionaea muscipula Ellis). J Exp Bot 62:1991–2000. doi: 10.1093/jxb/erq404 PubMedCrossRefGoogle Scholar
  64. Pickard WF (2003) The role of cytoplasmic streaming in symplastic transport. Plant, Cell Environ 26:1–15CrossRefGoogle Scholar
  65. Plieth C, Tabrizi H, Hansen U-P (1994) Relationship between banding and photosynthetic activity in Chara corallina as studied by the spatially different induction curves of chlorophyll fluorescence observed by an image analysis system. Physiol Plant 91:205–211CrossRefGoogle Scholar
  66. Plyusnina TY, Lavrova AI, Riznichenko GY, Rubin AB (2005) Modeling the pH and the transmembrane potential banding along the cell membrane of alga Chara corallina. Biophysics 50:434–440Google Scholar
  67. Prins HBA, Snel JFH, Zanstra PE, Helder RJ (1982) The mechanism of bicarbonate assimilation by the polar leaves of Potamogeton and Elodea. CO2 concentrations at the leaf surface. Plant, Cell Environ 5:207–214Google Scholar
  68. Rayle DL, Cleland DL (1992) The acid growth theory of auxin-induced cell elongation is alive and well. Plant Physiol 99:1271–1274PubMedCrossRefGoogle Scholar
  69. Schmölzer PM, Höftberger M, Foissner I (2011) Plasma membrane domains participate in pH banding of Chara internodal cells. Plant Cell Physiol 52:1274–1288. doi: 10.1093/pcp/pcr074 PubMedCrossRefGoogle Scholar
  70. Schreiber U (2004) Pulse-amplitude (PAM) fluorometry and saturation pulse method. In: Papageorgiou G, Govindjee (eds) Chlorophyll a fluorescence: a signature of photosynthesis. Kluwer Academic Publishers, DordrechtGoogle Scholar
  71. Schurr U, Walter A, Rascher U (2006) Functional dynamics of plant growth and photosynthesis—from steady-state to dynamics—from homogeneity to heterogeneity. Plant, Cell Environ 29:340–352. doi: 10.1111/j.1365-3040.2005.01490.x CrossRefGoogle Scholar
  72. Sehnke PC, DeLille JM, Ferl RJ (2002) Consummating signal transduction: the role of 14-3-3 proteins in the completion of signal-induced transitions in protein activity. Plant Cell 14:S339–S354. doi: 10.1105/tpc.010430 PubMedGoogle Scholar
  73. Shepherd VA, Beilby MJ, Khazaaly SAS, Shimmen T (2008) Mechano-perception in Chara cells: the influence of salinity and calcium on touch-activated receptor potentials, action potentials and ion transport. Plant, Cell Environ 31:1575–1591CrossRefGoogle Scholar
  74. Shimmen T, Wakabayashi A (2008) Involvement of membrane potential in alkaline band formation by internodal cells of Chara corallina. Plant Cell Physiol 49:1614–1620. doi: 10.1093/pcp/pcn136 PubMedCrossRefGoogle Scholar
  75. Shimmen T, Yamamoto A (2002) Induction of a new alkaline band at a target position in internodal cells of Chara corallina. Plant Cell Physiol 43:980–983PubMedCrossRefGoogle Scholar
  76. Shimmen T, Yokota E (2004) Cytoplasmic streaming in plants. Curr Opin Cell Biol 16:68–72. doi: 10.1016/ PubMedCrossRefGoogle Scholar
  77. Siebke K, Weis E (1995) Assimilation images of leaves of Glechoma hederacea: analysis of non-synchronous stomata related oscillations. Planta 196:155–165CrossRefGoogle Scholar
  78. Smith JR, Walker NA (1985) Effects of pH and light on the membrane conductance measured in the acid and basic zones of Chara. J Membr Biol 83:193–205CrossRefGoogle Scholar
  79. Spear DG, Barr JK, Barr CE (1969) Localization of hydrogen ion and chloride ion fluxes in Nitella. J Gen Physiol 54:397–414PubMedCrossRefGoogle Scholar
  80. Stahlberg R, Cosgrove DJ (1997) The propagation of slow wave potentials in pea epicotyls. Plant Physiol 113:209–217PubMedGoogle Scholar
  81. Takakura T, Fang W (2002) Climate under cover. Kluwer, DordrechtCrossRefGoogle Scholar
  82. Tazawa M (2003) Cell physiological aspects of the plasma membrane electrogenic H+ pump. J Plant Res 116:419–442. doi: 10.1007/s10265-003-0109-3 PubMedCrossRefGoogle Scholar
  83. Thiel G, Wacke M, Foissner I (2002) Ca2+ mobilization from internal stores in electrical membrane excitation in Chara. Progr Bot 64:217–233CrossRefGoogle Scholar
  84. Van Sambeek JW, Pickard BG (1976) Mediation of rapid electrical, metabolic, transpirational, and photosynthetic changes by factors released from wounds. III. Measurements of CO2 and H2O flux. Can J Bot 54:2662–2671CrossRefGoogle Scholar
  85. Verchot-Lubicz J, Goldstein RE (2010) Cytoplasmic streaming enables the distribution of molecules and vesicles in large plant cells. Protoplasma 240:99–107. doi: 10.1007/s00709-009-0088-x PubMedCrossRefGoogle Scholar
  86. Walker NA, Smith FA, Cathers IR (1980) Bicarbonate assimilation by fresh-water charophytes and higher plants. I. Membrane transport of bicarbonate ions is not proven. J Membr Biol 57:51–58CrossRefGoogle Scholar
  87. Williamson RE, Ashley CC (1982) Free Ca2+ and cytoplasmic streaming in the alga Chara. Nature 296:647–650PubMedCrossRefGoogle Scholar

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© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  1. 1.Department of Biophysics, BiologyMoscow State UniversityMoscowRussia

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