Studies on stomatal function, epicuticular wax and stem-root transition region of polyethylene glycol-treated and nontreatedin vitro grape plantlets

  • Massoma Ali-Ahmad
  • Harrison G. Hughes
  • Farida Safadi
Cell Biology


Scanning electron microscopy, light microscopy, and gravimetric analysis was used to evaluate stomatal function, epicuticular wax, and the stem-root transition region of grape (Vitis sp. ‘Valiant’) plantlets grownin vitro, polyethylene glycoltreatedin vitro, and greenhouse-grown plants. Scanning electron microscopic studies of leaf surfaces ofin vitro-grown plants showed widely open stomata as compared to leaf stomata of polyethylene glycol-treatedin vitro-cultured and greenhouse-grown plants. Ultrastructurally, leaf epicuticular wax ofin vitro plants was less dense than in their polyethylene-treated and greenhouse counterparts. Quantitatively,in vitro-grown plants had reduced epicuticular was as compared to polyethylene glycol-treated and greenhouse-grown plants. Light microscopic studies showed no obvious differences in the vascular connections in the stem-root transition region ofin vitro-cultured, polyethylene glycol-treatedin vitro-cultured, and greenhouse-grown plants. It is therefore likely that the rapid wilting and desiccation observed after transplantingin vitro grape plantlets is due to their defective stomatal function and reduced epicuticular wax and may not be due to poor water transport associated with vascular connection.

Key words

acclimatization grape Vitis sp. ‘Valiant’ in vitro micropropagation epicuticular wax 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ali-Ahmad, M.; Hughes, H. G. Stomatal function of in vitro grape plantlets treated with and without polyethylene glycol. HortScience 29:547(Abstr.); 1994a.Google Scholar
  2. Ali-Ahmad, M.; Hughes, H. G. Epicuticular wax structure of in vitro grape plantlets treated with and without polyethylene glycol. HortScience 29:547(Abstr.); 1994b.Google Scholar
  3. Baker, E. A. The influence on leaf wax development inBrassica oleracea var. Gemmifera. New Phytol. 73:955–966; 1974.CrossRefGoogle Scholar
  4. Brainerd, K. E.; Fuchigami, L. H. Acclimatization of aseptically cultured apple plants to low relative humidity. J. Hort. Sci. 106:516–518; 1981.Google Scholar
  5. Capellades, M.; Fontarnau, R.; Carulla, C.; Debergh, P. Environment influences anatomy of stomata and epidermal cells in tissue-culturedRosa multiflora. J. Hort. Sci. 115:141–145; 1990.Google Scholar
  6. Cheng, T. Y. Clonal propagation of woody plant species through tissue culture technique. Proc. Int. Plant Prop. Soc. 28:139–155; 1978.Google Scholar
  7. Dami, I.; Hughes, H. G. PEG effects on growth and water loss of micropropagated grape. HortScience 26:725(Abstr.); 1991.Google Scholar
  8. Donnelly, D. J.; Skelton, F. E.; Daubeny, H. A. External leaf features of tissue cultured ‘Silvan’ blackberry. HortScience 21:306–308; 1986.Google Scholar
  9. Donnelly, D. J.; Skelton, F. E.; Nelles, J. E. Hydathode anatomy and adaxial water loss in micropropagated ‘Silvan’ blackberry. J. Hort. Sci. 112:566–569; 1987.Google Scholar
  10. Fuchigami, L. H.; Cheng, T. Y.; Soeldner, A. Abaxial transpiration and water loss in aseptically cultured plum. J. Hort. Sci. 106:519–522; 1981.Google Scholar
  11. Grout, B. W. W. Effects of light and temperature on the composition of epicuticular wax of barley leaves. Phytochemistry 14:921–929; 1975.CrossRefGoogle Scholar
  12. Grout, B. W. W.; Aston, M. J. Transplanting of cauliflower plants regenerated from meristem culture. 1. Water loss and water transfer related to changes in leaf wax and to xylem regeneration. Hort. Res. 17:1–7; 1977.Google Scholar
  13. Hasegawa, P. M.; Bressan, A.; Handa, S., et al. Cellular mechanism of tolerance to water stress. HortScience 19:371–377; 1984.Google Scholar
  14. Heyser, J. W.; Nabors, M. W. Growth, water content, and solute accumulation of two tobacco cell lines cultured on sodium chloride, dextran, and polyethylene glycol. Plant Physiol. 68:1454–1459; 1981.PubMedCrossRefGoogle Scholar
  15. Jensen, W. A. Botanical histochemistry. Principles and practice. San Francisco: Freeman and Co.; 1962:407.Google Scholar
  16. Johansen, D. A. Botanical microtechnique. New York: McGraw-Hill; 1940:523.Google Scholar
  17. Lee, N.; Wetzstein, H. Y.; Sommer, H. E. Quantum flux density effect on the anatomy and surface morphology of in vitro and in vivo development of sweetgum leaves. J. Hort. Sci. 113:167–171; 1988.Google Scholar
  18. Marin, J. A.; Gella, R. Is desiccation the cause of poor survival rate in the acclimatization of micropropagatedPrunus cerasus L.? Acta Hort. 230:105–112; 1988.Google Scholar
  19. Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 155:473–497; 1962.CrossRefGoogle Scholar
  20. Pierik, P. L. M. Handicaps for the scale commercial application of micropropagation. Acta Hort. 230:63–71; 1988.Google Scholar
  21. Safadi F. Physiological studies in acclimatization of in vitro tobacco plantlets. Ph.D. Dissertation. Colorado State University, Fort Collins. 1992:1–147.Google Scholar
  22. Safadi, F.; Hughes, H. G. Comparison of diffusive resistance of polyethylene glycol treated and non treated tobacco plantlets. HortScience 25:293 (Abstr.); 1991.Google Scholar
  23. Shackle, K. A.; Novello, V.; Sutter, E. G. Stomatal function and cuticular conductance in whole tissue-cultured apple shoots. J. Hort. Sci. 115:468–472; 1990.Google Scholar
  24. Short, K. C.; Warburton, J.; Roberts, A. V. In vitro hardening of cultured cauliflower and chrysanthemum plantlets to humidity. Acta Hort. 212:329–334; 1987.Google Scholar
  25. Smith, E. F.; Palta, J. P.; McCown, B. H. Comparative anatomy and physiology of microculture, seedlings, and greenhouse-grown Asian white birch. J. Hort. Sci. 111:437–442; 1986.Google Scholar
  26. Sutter, E. Stomatal and cuticular water loss from apple, cherry and sweetgum plants after removal from in vitro culture. J. Hort. Sci. 113:234–238; 1988.Google Scholar
  27. Sutter, E.; Langhans, R. W. Epicuticular wax formation on carnation plantlets regenerated from shoot tip culture. J. Hort. Sci. 104:493–496; 1979.Google Scholar
  28. Sutter, E.; Langhans, R. W. Formation of epicuticular wax and its effects on water loss in cabbage plants regenerated from shoot tips culture. Can. J. Bot. 60:2896–2908; 1982.CrossRefGoogle Scholar
  29. Sutter, E.; Nevello, V.; Shackel, K. Physiological and anatomical aspects of water stress of cultured plants. Acta Hort. 230:113–119; 1988.Google Scholar
  30. Wardle, K.; Dobbs, E. B.; Short, K. C. In vitro acclimatization of aseptically cultured plantlets to humidity. J. Hort. Sci. 108:386–389; 1983.Google Scholar
  31. Whitecross, M. I.; Armstrong, D. J. Environment effects on epicuticular wax ofBrassica napus L. Aust. J. Bot. 20:87–95; 1972.CrossRefGoogle Scholar
  32. Zaid, A.; Hughes, H. G. Water loss and polyethylene glycol-mediated acclimatization of in vitro-grown seedlings of five cultivars of date palm (Phoenix dactylifera L.) plantlets. Plant Cell Rep. 14:385–388; 1995.CrossRefGoogle Scholar
  33. Ziv, M.; Meir, G.; Halvey, A. H. Factors influencing the production of hardened glaucous carnation plants in vitro. Plant Cell Tissue Organ Cult. 2:55–65; 1983.CrossRefGoogle Scholar

Copyright information

© Society for In Vitro Biology 1998

Authors and Affiliations

  • Massoma Ali-Ahmad
    • 1
  • Harrison G. Hughes
    • 2
  • Farida Safadi
    • 3
  1. 1.Plant Biotechnology Laboratory, Division of Agricultural SciencesFlorida A & M UniversityTallahassee
  2. 2.Department of Horticulture and Landscape ArchitectureColorado State UniversityFort Collins
  3. 3.Department of BiologyColorado State UniversityFort Collins

Personalised recommendations