, 233:177 | Cite as

The root microtubule cytoskeleton and cell cycle analysis through desiccation of Brassica napus seedlings

Original Article


Desiccation tolerance (DT) of orthodox seeds is reduced upon their germination. The main aim of this study was to estimate the range of rape seedling DT by examining the consequences of desiccation on the distribution, stability and orientation of microtubules in diverse cells. Using different parameters, such as relative water content (RWC), the tetrazolium viability test and electrolyte leakage, it has been demonstrated that a small percentage decrease in relative humidity can cause irreparable changes in membrane permeability, as well as in nuclear structure and microtubule cytoskeleton stability. Seedling root tips survived when exposed to low desiccation stress intensity, but small changes in microtubule behavior were observed. Cortical microtubules formed thick arrays, especially near the plasma membrane. Water loss also resulted in a reduction of the mitotic activity. More rapid desiccation caused microtubule depolymerization. Occasionally, abnormal tubulin aggregates were visible. Cell divisions were not detectable under these conditions. Due to the observable microtubule defects, the hypersensitivity of the microtubule cytoskeleton might be a useful and simple parameter for estimating environmental stress intensity.


Rape Canola Confocal microscopy Cytoskeleton Desiccation tolerance DNA integrity Microtubules TTC viability test 


  1. Alpert P, Oliver MJ (2002) Drying without dying. In: Black M, Pritchard HW (eds) Desiccation and survival in plants. CABI, UK, pp 3–43Google Scholar
  2. Bagniewska-Zadworna A, Zenkteler E, Czaczyk K, Osińska M (2007) The effect of dehydration with or without abscisic acid pretreatment on buds regeneration from Polypodium vulgare L. rhizomes. Acta Physiol Plant 29:47–56CrossRefGoogle Scholar
  3. Bartolo ME, Carter JV (1991) Microtubules in the mesophyll cells of nonacclimated and cold-acclimated spinach. Plant Physiol 97:175–181PubMedCrossRefGoogle Scholar
  4. Bewley JD (1979) Physiological aspects of desiccation-tolerance. Annu Rev Plant Physiol 30:195–238CrossRefGoogle Scholar
  5. Blancaflor EB, Hasenstein KH (1995) Growth and microtubule orientation of Zea mays roots subjected to osmotic stress. Int J Plant Sci 156:774–783PubMedCrossRefGoogle Scholar
  6. Buitink J, Vu BL, Satour P, Leprince O (2003) The re-establishment of desiccation tolerance in germinated radicles of Medicago truncatula Gaertn. seeds. Seed Sci Res 13:273–286CrossRefGoogle Scholar
  7. Clemensson-Lindell A (1994) Triphenyltetrazolium chloride as an indicator of fine-root vitality and environmental stress in coniferous forest stands: applications and limitations. Plant Soil 159:297–300CrossRefGoogle Scholar
  8. Comas LH, Eissenstat DM, Lakso AN (2000) Assessing root death and root system dynamics in a study of grape canopy pruning. New Phytol 147:171–178CrossRefGoogle Scholar
  9. Danon A, Delorme V, Mailhac N, Gallois P (2000) Plant programmed cell death: a common way to die. Plant Physiol Biochem 38:647–655CrossRefGoogle Scholar
  10. Dhonukshe P, Laxalt AM, Goedhart J, Gadella TW, Munnik T (2003) Phospholipase D activation correlates with microtubule reorganization in living plant cells. Plant Cell 15:2666–2679PubMedCrossRefGoogle Scholar
  11. Faria JMR, van Lammeren AAM, Hilhorst HWM (2004) Desiccation sensitivity and cell cycle aspects in seeds of Inga vera subsp. affinis. Seed Sci Res 14:165–178CrossRefGoogle Scholar
  12. Faria JMR, Buitink J, van Lammeren AAM, Hilhorst HWM (2005) Changes in DNA and microtubules during loss and re-establishment of desiccation tolerance in germinating Medicago truncatula seeds. J Exp Bot 56:2119–2130PubMedCrossRefGoogle Scholar
  13. Fischer R, Timberlake WE (1995) Aspergillus nidulans apsA (anucleate primary sterigmata) encodes a coiled-coil protein necessary for nuclear positioning and completion of asexual development. J Cell Biol 128:485–498PubMedCrossRefGoogle Scholar
  14. Gusta LV, Gao Y-P, Benning NT (2006) Freezing and desiccation tolerance of imbibed canola seeds. Physiol Plant 126:237–246CrossRefGoogle Scholar
  15. Huang JM (2001) Involvement of antioxidants and lipid peroxidation in the adaptation of two cool-season grasses to localized drought stress. Environ Exp Bot 45:105–114PubMedCrossRefGoogle Scholar
  16. Khokhlova LP, Olinevich OV, Raudaskokski M (2003) Reorganization of the microtubule and actin cytoskeleton in root cells of Triticum aestivum L. during low temperature and abscissic acid treatments. Cell Biol Int 27:211–212PubMedCrossRefGoogle Scholar
  17. Komis G, Apostolakos P, Galatis B (2001) Altered patterns of tubulin polymerization in dividing leaf cells of Chlorophyton comosum after a hyperosmotic treatment. New Phytol 149:193–207CrossRefGoogle Scholar
  18. Komis G, Apostolakos P, Galatis B (2002) Hyperosmotic stress induced formation of tubulin macrotubules in root-tip cells of Triticum turgidum: their probable involvement in protoplast volume control. Plant Cell Physiol 43:911–922PubMedCrossRefGoogle Scholar
  19. Lambert AM, Vantard M, Schmit AC, Stoeckel H (1991) Mitosis in plants. In: Lloyd CW (ed) The cytoskeletal basis of plant growth and form. Academic, San Diego, pp 199–226Google Scholar
  20. Lang-Pauluzzi I, Gunning BES (2000) A plasmolytic cycle: the fate of cytoskeletal elements. Protoplasma 212:174–185CrossRefGoogle Scholar
  21. Lü B, Gong Z, Zhang J, Liang J (2007) Microtubule dynamics in relation to osmotic stress-induced ABA accumulation in Zea mays roots. J Exp Bot 58:2565–2572PubMedCrossRefGoogle Scholar
  22. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–409PubMedCrossRefGoogle Scholar
  23. Müller J, Menzel D, Šamaj J (2007) Cell-type-specific disruption and recovery of the cytoskeleton in Arabidopsis thaliana epidermal root cells upon heat shock tress. Protoplasma 230:231–242PubMedCrossRefGoogle Scholar
  24. Panteris E, Galatis B (2005) The morphogenesis of lobed plant cells in the mesophyll and epidermis: organization and distinct roles of cortical microtubules and actin filaments. New Phytol 167:721–732PubMedCrossRefGoogle Scholar
  25. Pressel S, Ligrone R, Duckett JG (2006) Effects of de- and rehydration on food-conducting cells in the moss Polytrichum formosum: a cytological study. Ann Bot 98:67–76PubMedCrossRefGoogle Scholar
  26. Proctor MCF, Tuba Z (2002) Poikilohydry and homoihydry: antithesis or spectrum of possibilities? New Phytol. 156:327–349CrossRefGoogle Scholar
  27. Proctor MCF, Oliver MJ, Wood AJ, Alpert P, Stark LR, Cleavitt NL, Mischler BD (2007) Desiccation-tolerance in bryophytes: a review. Bryologist 110:595–621CrossRefGoogle Scholar
  28. Sargent JA, Mandi SS, Osborne DJ (1981) The loss of desiccation tolerance during germination: an ultrastructural and biochemical approach. Protoplasma 105:225–239CrossRefGoogle Scholar
  29. Senaratna T, McKersie BD (1983) Characterization of solute efflux from dehydration injured soybean (Glycine max L. Merr) seeds. Plant Physiol 72:911–914PubMedCrossRefGoogle Scholar
  30. Sieberer BJ, Ketelaar T, Esseling JJ, Emons AM (2005) Microtubules guide root hair tip growth. New Phytol 167:711–719PubMedCrossRefGoogle Scholar
  31. Śliwińska E (2003) Cell cycle and germination of fresh, dried and deteriorated sugarbeet seeds as indicators of optimal harvest time. Seed Sci Res 13:131–138CrossRefGoogle Scholar
  32. Smertenko A, Dráber P, Viklický V, Opatrný Z (1997) Heat stress affects the organization of microtubules and cell division in Nicotiana tabacum cells. Plant Cell Environ 20:1534–1542CrossRefGoogle Scholar
  33. Steponkus PL, Lanphear FO (1967) Refinement of the triphenyltetrazolium chloride method of determining cold injury. Plant Physiol 42:1423–1426PubMedCrossRefGoogle Scholar
  34. Sun WQ (2002) Methods for studying water relations under stress. In: Black M, Pritchard HW (eds) Desiccation and survival in plants. CABI, UKGoogle Scholar
  35. Sybenga J (1972) General cytogenetics. Elsevier, New York, USAGoogle Scholar
  36. Wang C, Li J, Yuan M (2007) Salt tolerance requires cortical microtubule reorganization in Arabidopsis. Plant Cell Physiol 48:1534–1547PubMedCrossRefGoogle Scholar
  37. Wasteneys GO (2003) Microtubules show their sensitive nature. Plant Cell Physiol 44:653–654PubMedCrossRefGoogle Scholar
  38. Wasteneys GO (2004) Progress in understanding the role of microtubules in plant cells. Curr Opin Cell Biol 7:651–660Google Scholar
  39. Wasteneys GO, Galway ME (2003) Remodeling the cytoskeleton for growth and form. Annu Rev Plant Biol 54:691–722PubMedCrossRefGoogle Scholar
  40. Zhang DH, Wadsworth P, Hepler PK (1990) Microtubule dynamics in living plant cells: confocal imaging of microinjected fluorescent brain tubulin. Proc Nat Acad Sci USA 87:8820–8824PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Department of General Botany, Institute of Experimental BiologyAdam Mickiewicz UniversityPoznańPoland

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