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Planta

, Volume 236, Issue 6, pp 1817–1829 | Cite as

Surface tip-to-base Ca2+ and H+ ionic fluxes are involved in apical growth and graviperception of the Phycomyces stage I sporangiophore

  • Branka D. ŽivanovićEmail author
Original Article

Abstract

Net fluxes of Ca2+ and H+ ions were measured non-invasively close to the surface of Phycomyces blakesleeanus sporangiophores stage I using ion-selective vibrating microelectrodes. The measurements were performed on a wild type (Wt) and a gravitropic mutant A909 kept in either vertical or tilted orientation. Microelectrodes were positioned 4 μm from the surface of sporangiophore, and ion fluxes were recorded from the apical (0–20 μm) and subapical (50–100 μm) regions. The magnitude and direction of ionic fluxes measured were dependent on the distance from the tip along the growing zone of sporangiophore. Vertically oriented sporangiophores displayed characteristic tip-to-base ion fluxes patterns. Ca2+ and H+ fluxes recorded from apical region of Wt sporangiophores were inward-directed, while ion fluxes from subapical locations occurred in both directions. In contrast to Wt, mutant A909 showed opposite (outward) direction of Ca2+ fluxes and reduced H+ influxes in the apical region. Following gravistimulation, the magnitude and direction of ionic fluxes were altered. Wt sporangiophore exhibited oppositely directed fluxes on the lower (influx) and the upper (efflux) sides of the cell, while mutant A909 did not show such patterns. A variable elongation growth in vertical position and reduced growth rate upon gravistimulation were observed in both strains. The data show that tip-growing sporangiophores exhibit a tip-to-base ion flux pattern which changes characteristically upon gravistimulation in Wt in contrast to the mutant A909 with a strongly reduced gravitropic response.

Keywords

Gravitropic bending Growth rate Ion-selective microelectrodes Ion fluxes Tip growth 

Abbreviations

APW

Artificial pond water

ASET

Automated Scanning Electrode Techniques

A909

Gravitropically defective mutant

Wt

Wild type

Notes

Acknowledgments

This study was performed in Botanisches Institutder Universität Karlsruhe (TH), Germany, through a DAAD scholarship and partial support by Grant 173040 from the Serbian Ministry of Education and Science. The author is grateful to Prof. Dr. M.H. Weisenseel for helpful discussions, Dr. G.Monshausen for help in setting up measuring line and processing data, and Ms. B. Schlicke and Mr. W. Müller for their excellent technical assistance. The author is also grateful to Prof. Dr. Ž. Vučinić and Prof. Dr. P. Galland for their critical reading and useful suggestions during preparation of the manuscript, and to M.Sc. D. Mutavdžić for help in statistical data analysis.

References

  1. Alcántara-Sánchez F, Reynaga-Peñal CG, Salcedo-Hernández R, Ruiz-Herrera J (2004) Possible role of ionic gradients in the apical growth of Neurospora crassa. Anton Leeuw 86:301–311CrossRefGoogle Scholar
  2. Arend M, Monshausen G, Wind C, Weisenseel MH, Fromm J (2004) Effect of potassium deficiency on the plasma membrane H+-ATPase of the wood ray parenchyma in poplar. Plant Cell Environ 27:1288–1296CrossRefGoogle Scholar
  3. Bartnicki-García S (2002) Hyphal tip growth: outstanding questions. In: Osiewacz HD (ed) Molecular biology of fungal development. Marcel Dekker, New York, pp 42–71Google Scholar
  4. Bergman K, Burke PV, Cerdá-Olmedo E, David CN, Delbrück M, Foster KW, Goodell EW, Heisenberg M, Meissner G, Zalokar M, Denninson DS, Shropshire WJR (1969) Phycomyces. Bacteriol Rev 33:99–157PubMedGoogle Scholar
  5. Bergman K, Eslava AP, Cerdá-Olmedo E (1973) Mutants of Phycomyces with abnormal phototropism. Mol Gen Genet 123:1–6PubMedCrossRefGoogle Scholar
  6. Braun M, Hemmersbach R (2008) Single-cell gravitropism and gravitaxis. In: Gilroy S, Masson PH (eds) Plant tropisms, 1st edn. Blackwell Publishing Professional, Iowa, pp 141–160Google Scholar
  7. Braun M, Limbach C (2006) Rhizoids and protonemata of characean algae: model cells for research on polarized growth and plant gravity sensing. Protoplasma 229:133–142PubMedCrossRefGoogle Scholar
  8. Campuzano V, Galland P, Senger H, Alvarez MI, Eslava AP (1994) Isolation and characterization of phototropism mutants of Phycomyces insensitive to ultraviolet light. Curr Genet 26:49–53PubMedCrossRefGoogle Scholar
  9. Campuzano V, Galland P, Eslava AP, Alvarez MI (1995) Genetic characterization of two phototropism mutants of Phycomyces with defects in the genes madI and mad. J Curr Genet 27:524–527CrossRefGoogle Scholar
  10. Campuzano V, Galland P, Alvarez MI, Eslava AP (1996) Blue-light receptor requirement for gravitropism, autochemotropism and ethylene response in Phycomyces. Photochem Photobiol 63:686–694PubMedCrossRefGoogle Scholar
  11. Castle ES (1940) Discontinuous growth of single plant cells measured at short intervals and the theory of intussusceptions. J Cell Comp Physiol 15:285–298CrossRefGoogle Scholar
  12. Edwards KL (1991) The gravitational response of Phycomyces with respect to gadolinium and factors affecting calcium function. 1989–1990 NASA Space/Gravitational Biology Accomplishments. NASA Tech Mem 4258:32–33Google Scholar
  13. Eibel P, Schimek C, Fries V, Grolig F, Schapat T, Schmidt W, Schneckenburger H, Ootaki T, Galland P (2000) Statoliths in Phycomyces: characterization of octahedral protein crystals. Fungal Genet Biol 29:211–220PubMedCrossRefGoogle Scholar
  14. Galland P, Fries V, Grolig F, Schmidt W (2007) Gravireception of the sporangiophore of Phycomyces blakesleeanus. Adv Space Res 39:1134–1139CrossRefGoogle Scholar
  15. Garrill A, Jackson SL, Lew RR, Heath IB (1993) Ion channel activity and tip growth: tip-localized stretch-activated channels generate an essential Ca2+ gradient in the oomycete Saprolegnia ferax. Eur J Cell Biol 60:358–365PubMedGoogle Scholar
  16. Geitmann A, Cresti M, Heath IB (eds) (2001) Cell biology of plant and fungal tip growth. IOS Press, NATO Science Series, AmsterdamGoogle Scholar
  17. Grolig F, Herkenrath H, Pumm T, Gross A, Galland P (2004) Gravity susception by buoyancy: floating lipid globules in sporangiophores of Phycomyces. Planta 218:658–667PubMedCrossRefGoogle Scholar
  18. Groves PM, Gamow RI (1975) Intracellular recordings from Phycomyces. Plant Physiol 55:946–947PubMedCrossRefGoogle Scholar
  19. Harold FM, Caldwell JH (1990) Tips and currents: electrobiology of apical growth. In: Heath LB (ed) Tip growth in plant and fungal cells. Academic, New York, pp 59–90Google Scholar
  20. Heath IB (1990) Tip growth in plant and fungal cells. Academic Press, San DiegoGoogle Scholar
  21. Heath IB (1995) Integration and regulation of hyphal tip growth. Can J Bot 73(Suppl 1):S131–S139CrossRefGoogle Scholar
  22. Holdaway-Clarke TL, Feijo JA, Hackett GA, Kunkel JG, Hepler PK (1997) Pollen tube growth and the intracellular-cytosolic calcium gradient oscillate in phase while extracellular calcium influx is delayed. Plant Cell 9:1999–2010PubMedGoogle Scholar
  23. Horie T, Schimek C, Ootaki T (1998) Gravitropic responses of Phycomyces and Pilobulus sporangiophores and possible mode of action. In: Abstract. 6th International Mycological Congress, Jerusalem, Israel, p 103Google Scholar
  24. Jackson SL, Heath IB (1993) Roles of calcium ions in hyphal tip growth. Microbiol Rev 57:367–382PubMedGoogle Scholar
  25. Lafay J-F, Matricon J (1985) Observation of growth rate fluctuations in Phycomyces blakesleeanus using a high resolution automated measurement system. Physiol Vég 23:929–934Google Scholar
  26. Leitz G, Schnepf E, Greulich KO (1995) Micromanipulation of statoliths in gravity-sensing Chara rhizoids by optical tweezers. Planta 197:278–288PubMedCrossRefGoogle Scholar
  27. Levina NN, Lew RR, Heath LB (1994) Cytoskeletal regulation of ion channel distribution in the tip-growing organism Saprolegnia ferax. J Cell Sci 107:127–134PubMedGoogle Scholar
  28. Levina NN, Lew RR, Hyde GJ, Heath IB (1995) The roles of Ca2+ and plasma membrane ion channels in hyphal tip growth of Neurospora crassa. J Cell Sci 108:3405–3417PubMedGoogle Scholar
  29. Lew RR (1999) Comparative analysis of Ca2+ and H+ flux magnitude and location along growing hyphae of Saprolegnia ferax and Neurospora crassa. Eur J Cell Biol 79:892–902CrossRefGoogle Scholar
  30. Limbach C, Hauslage J, Schäfer C, Braun M (2005) How to activate a plant gravireceptor. Early mechanisms of gravity sensing studied in Characean rhizoids during parabolic flights. Plant Physiol 139:1030–1040PubMedCrossRefGoogle Scholar
  31. López-Franco R, Bartnicki-Garciá S, Bracker CE (1994) Pulsed growth of fungal hyphal tips. Proc Natl Acad Sci USA 91:12228–12232PubMedCrossRefGoogle Scholar
  32. McGillivray AM, Gow NAR (1986) Applied electrical fields polarize the growth of mycelial fungi. J Gen Microbiol 131:751–756Google Scholar
  33. Messerli MA, Danuser G, Robinson KP (1999) Pulsatile influxes of H+, K+ and Ca2+ lag growth pulses of Lilium longiflorum pollen tubes. J Cell Sci 112:1497–1509PubMedGoogle Scholar
  34. Mitiku G, Edwards KL (1991) Ion channels of membrane vesicles from gravity-sensitive sporangiophores of Phycomyces. Am Soc Gravit Space Biol Bull 5:47Google Scholar
  35. Mogus MA, Wolken JJ (1974) Phycomyces: electrical response to light stimuli. Plant Physiol 53:512–513PubMedCrossRefGoogle Scholar
  36. Money NP (2001) The pulse of the machine—reevaluating tip-growth methodology. New Phytol 151:553–555CrossRefGoogle Scholar
  37. Monshausen GB, Messerli MA, Gilroy S (2008) Imaging of the yellow cameleon 3.6 indicator reveals that elevations in cytosolic Ca2+ follow oscillating increases in growth in root hairs of Arabidopsis. Plant Physiol 147:1690–1698PubMedCrossRefGoogle Scholar
  38. Pierson ES, Miller D, Callaham D, Van Aken J, Hackett G, Hepler PK (1996) Tip localized calcium entry fluctuates during pollen tube growth. Dev Biol 174:160–173PubMedCrossRefGoogle Scholar
  39. Reinhardt MO (1892) Das Wachstum der Pilz Hyphen. Jahrb Wissenschaft Bot 23:479–566Google Scholar
  40. Reiss H-D, Herth W (1979) Calcium gradients in tip growing plant cells visualized by chlortetracycline fluorescence. Planta 146:615–621CrossRefGoogle Scholar
  41. Ruiz-Herrera J, Martinez-Cardena G, Loarca F, Salcedo-Hernández R (2003) Analysis of phenomena involved in apical growth of Phycomyces blakesleeanus. Arch Microbiol 180:427–433PubMedCrossRefGoogle Scholar
  42. Sampson K, Lew RR, Heath B (2003) Time series analysis demonstrates the absence of pulsative hyphal growth. Microbiology 149:3111–3119PubMedCrossRefGoogle Scholar
  43. Schiefelbein JW, Shipley A, Rowse P (1992) Calcium influx at the tip of growing root-hair cells of Arabidopsis thaliana. Planta 187:455–459CrossRefGoogle Scholar
  44. Schimek C, Eibel P, Grolig F, Horie T, Ootaki T, Galland P (1999a) Gravitropism in Phycomyces: a role for sedimenting protein crystals and floating lipid globules. Planta 210:132–142PubMedCrossRefGoogle Scholar
  45. Schimek C, Eibel P, Horie T, Galland P, Ootaki T (1999b) Protein crystals in Phycomyces sporangiophores are involved in graviperception. Adv Space Biol 24:687–696CrossRefGoogle Scholar
  46. Schmidt W (2007) Advanced micro dual wavelength spectrophotometer (MDWS) extended to the near UV and IR range for measuring gravity-induced absorption changes (GIACs) during parabolic flights on the airbus A300-Zero-G. Micrograv Sci Technol 19:11–15CrossRefGoogle Scholar
  47. Schmidt W, Galland P (2000) Gravity-induced absorption changes in Phycomyces: a novel method for detecting primary responses of gravitropism. Planta 210:848–852PubMedCrossRefGoogle Scholar
  48. Schmidt W, Galland P (2004) Optospectroscopic detection of primary reactions associated with the graviperception of Phycomyces: effects of micro- and hypergravity. Plant Physiol 135:183–192PubMedCrossRefGoogle Scholar
  49. Silverman-Gavrila LB, Lew R (2001) Regulation of the tip-high [Ca2+] gradient in growing hyphae of the fungus Neurospora crassa. Eur Cell Biol 80:379–390CrossRefGoogle Scholar
  50. Stifler R (1961) Growth of Phycomyces immersed in water. Science 133:1022PubMedCrossRefGoogle Scholar
  51. Takeuchi Y, Schmid J, Caldwell JH, Harold FM (1988) Transcellular ion currents and extension of Neurospora crassa hyphae. J Membr Biol 101:33–41PubMedCrossRefGoogle Scholar
  52. Thornton RM (1968) The fine structure of Phycomyces II. Organization of the stage I sporangiophore apex. Protoplasma 66:269–285Google Scholar
  53. Weiss J, Weisenseel MH (1990) Blue light-induced changes in membrane potential and intracellular pH of Phycomyces hyphae. J Plant Physiol 136:78–85CrossRefGoogle Scholar
  54. Živanović B (2005) Ca2+ and H+ ion fluxes near the surface of gravitropically stimulated Phycomyces sporangiophore. Ann New York Acad Sci 1048:487–490CrossRefGoogle Scholar
  55. Živanović B, Köhler K, Galland P, Weisenseel MH (2001) Membrane potential and endogenous ion current of Phycomyces sporangiophores. Electro Magnetobiol 20:343–362Google Scholar
  56. Živić M, Popović M, Živanović B, Vučinić Ž (2005) A new model system for investigation of ionic channel in filamentous fungi: evidence for existence of two K+-permeable ionic channels in Phycomyces blakesleeanus. Ann New York Acad Sci 1048:491–495CrossRefGoogle Scholar
  57. Živić M, Popović M, Todorović N, Vučinić Ž (2009) Outwardly rectifying anionic channel from the plasma membrane of the fungus Phycomyces blakesleeanus. Eukaryot Cell 8:1439–1448PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Department for Life Sciences, Institute for Multidisciplinary ResearchUniversity of BelgradeBelgradeSerbia

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