Plant Growth Regulation

, Volume 36, Issue 1, pp 61–70 | Cite as

The use of the electrolyte leakage method for assessing cell membrane stability as a water stress tolerance test in durum wheat

  • Mohammed Bajji
  • Jean-Marie Kinet
  • Stanley LuttsEmail author


This work was carried out to adapt the electrolyte leakage technique todurum wheat and then to evaluate its relevance in the assessment of the cellmembrane stability as a mechanism of water stress tolerance in this species.Themethod currently used is based on in vitro desiccation ofleaf tissues by a solution of polyethylene glycol (PEG) and a subsequentmeasurement of electrolyte leakage into deionised water. It consists of threesuccessive steps: (1) a washing treatment to remove solutes from both leafsurfaces and cells damaged by cutting; (2) a stress period during which theleaftissues are plunged in a PEG-solution and (3) a rehydration period during whichafter-effects of the stress are evaluated. During the washing period, the majorpart of electrolytes was removed within 15 min. Varying the stressconditions influenced both the percent and the kinetics of electrolyte leakageduring rehydration. Electrolyte leakage exhibited a characteristic patternreflecting the condition of cellular membranes (repair and hardening). Inpractice, we recommend a 15-minute washing time, a10-hour stress period and 4 h of rehydration. Theextent of the cell membrane damage not only correlated well with the growthresponses of wheat seedlings belonging to various cultivars to withholdingwaterbut also with the recognised field performances of these cultivars. Therelativeproportion of endogenous ions lost in the effusate during the rehydration stepmay vary strongly according to the element analysed and the precise nutritionalstatus of the plant should therefore be considered. However, an increase ininorganic ion leakage does not fully explain the recorded PEG-induced increasein electrical conductivity (EC) during the subsequent rehydration step andorganic ions are probably also involved in such an increase.

Cell membrane injury Drought tolerance Electrolyte leakage Osmotic stress Polyethylene glycol Triticum durum 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Adam A.L., Gala A.A., Manninger K. and Barna B. 2000. Inhibition of the development of leaf rust (Puccinia recondita) by treatment of wheat with allopurinol and production of a hypersensitive-like reaction in a compatible host. Plant Pathology 49: 317–323.Google Scholar
  2. Agarie S., Hanaoka N., Kubota F., Agata W. and Kaufman B. 1995. Measurement of cell membrane stability evaluated by electrolyte leakage as a drought and heat tolerance test in rice (Oryza sativa L.). J. Fac. Agr. Kyushu Univ. 40: 233–240.Google Scholar
  3. Ali Dib T., Monneveux P., Acevedo E. and Nachit M.M. 1994. Evaluation of proline analysis and chlorophyll fluorescence quenching measurements as drought tolerance indicators in durum wheat (Triticum turgidum L. var. durum). Euphytica 79: 65–73.Google Scholar
  4. Bajji M. 1999. Etude des mécanismes de résistance au stress hydrique chez le blé dur (Triticum durum Desf.):. caractèrisation de cultivars différant par leurs niveaux de résistance á la sécheresse et de variants somaclonaux sélectionnés in vitro, PhD Dissertation, Université catholique de Louvain, Belgium.Google Scholar
  5. Bajji M., Lutts S. and Kinet J.M. 2000a. Physiological changes after exposure to and recovery from polyethylene glycol-induced water deficit in callus cultures issued from durum wheat (Triticum durum Desf.) cultivars differing in drought resistance. J. Plant Physiol. 156: 75–83.Google Scholar
  6. Bajji M., Lutts S. and Kinet J.M. 2000b. Physiological changes after exposure to and recovery from polyethylene glycol-induced water deficit in roots and leaves of durum wheat (Triticum durum Desf.) cultivars differing in drought resistance. J. Plant Physiol. 157: 100–108.Google Scholar
  7. Bajji M., Lutts S. and Kinet J.M. 2000c. Resistance to water stress in durum wheat: Comparison of cell and whole seedling behaviours. In: Royo C., Nachit M.M., Di Fonzo N. and Araus J.L. (eds), Durum wheat improvement in the Mediterranean region: New challenges. Options Méditerranéennes, pp. 22–34.Google Scholar
  8. Bandurska H. and Gniazdowska-Skoczek H. 1995. Cell membrane stability in two barley genotypes under water stress conditions. Acta Soc. Bot. Pol. 64: 29–32.Google Scholar
  9. Bandurska H., Stroinski A. and Zielezinska M. 1997. Effects of water deficit stress on membrane properties, lipid peroxidation and hydrogen peroxide metabolism in the leaves of barley genotypes. Acta Soc. Bot. Pol. 66: 177–183.Google Scholar
  10. Blum A. and Ebercon A. 1981. Cell membrane stability as a measure of drought and heat tolerance in wheat. Crop Sci. 21: 43–47.Google Scholar
  11. Borochov-Neori H. and Borochov A. 1991. Response of melon plants to salt. 1. Growth, morphology and root membrane, J. Plant Physiol. 139: 100–106.Google Scholar
  12. Chen Q., Zhang W.H. and Liu Y.L. 1999. Effect of NaCl, glutathione and ascorbic acid on function of tonoplast vesicles isolated from barley leaves. J. Plant Physiol. 155: 685–690.Google Scholar
  13. Coursolle C., Bigras F.J. and Margolis H.A. 2000. Assessment of root freezing damage of two-year-old white spruce, black spruce and jack pine seedlings. Scand. J. For. Res. 15: 343–353.Google Scholar
  14. De B. and Mukherjee A.K. 1996. Mercuric chloride induced membrane damage in tomato cultured cells. Biol. Plant. 38: 469–473.Google Scholar
  15. Flint H.L., Boyce B.R. and Beattie D.J. 1967. Index of injury-A useful expression of freezing injury to plant tissues as determined by the electrolytic method. Can. J. Plant Sci. 47: 229–230.Google Scholar
  16. Franca M.G.C., Thi A.T.P., Pimentel C., Rossiello R.O.P., Zuily Fodil Y. and Laffray D. 2000. Differences in growth and water relations among Phaseolus vulgaris cultivars in response to induced drought stress. Environm. Exp. Bot. 43: 227–237.Google Scholar
  17. Garty J., Weissman L., Tamir O., Beer S., Cohen Y., Karnieli A. et al. 2000. Comparison of five physiological parameters to assess the vitality of the lichen Ramalina lacera exposed to air pollution. Physiol. PLant. 109: 410–418.Google Scholar
  18. Ismail A.M. and Hall A.E. 1999. Reproductive-stage heat tolerance, leaf membrane thermostability and plant morphology in cowpea. Crop Sci. 39: 1762–1768.Google Scholar
  19. Knowles L., Trimble M.R. and Knowles N.R. 2001. Phosphorus status affects postharvest respiration, membrane permeability and lipid chemistry of European seedless cucumber fruits (Cucumis sativus L.). Postharvest Biol. and Technol. 21: 179–188.Google Scholar
  20. Lauriano J.A., Lidon F.C., Carvalho C.A., Campos P.S. and Matos M.D. 2000. Drought effects on membrane lipids and photosynthetic activity in different peanut cultivars. Photosynthetica 38: 7–12.Google Scholar
  21. Leopold A.C. and Vertucci C.W. 1986. Physiological attributes of desiccated seeds. In: Leopold A.C. (ed.), Membranes, Metabolism, and Dry Organisms. Comstoch Publishing Associates, Ithaca,/ New York, pp. 22–34.Google Scholar
  22. Leopold A.C., Musgrave M.E. and Williams K.M. 1981. Solute leakage resulting from leaf desiccation. Plant Physiol. 68: 1222–1225.Google Scholar
  23. Liu X.Z. and Huang B.R. 2000. Heat stress injury in relation to membrane lipid peroxidation in creeping bentgrass. Crop Sci. 40: 503–510.Google Scholar
  24. Maheswary M., Joshi D.K., Saha R., Nagarajan S. and Gambhir P.N. 1999. Transverse relaxation time of leaf water protons and membrane injury in wheat (Triticum aestivum L.) in response to high temperature. Ann. Bot. 84: 741–745.Google Scholar
  25. Martinoia E., Schramm M.J., Kaiser G., Kaiser W.M. and Heber U. 1986. Transport of anions in isolated vacuoles. 1. Permeability to anions and evidence for a Cl-uptake system, Plant Physiol. 80: 895–901.Google Scholar
  26. Nilsen E.T. and Orcutt D. 1996. The Physiology of Plants under Stress: Abiotic Factors. John Wiley & Sons, Inc., New York.Google Scholar
  27. Palliotti A. and Bongi G. 1996. Freezing injury in the olive leaf and effects of mefluidide treatment. J. Hort. Sci. 71: 57–63.Google Scholar
  28. Palta J.P., Levitt J. and Stadelmann E.J. 1977. Freezing injury in onion bulb cells. I. Evaluation of the conductivity methods and analysis of ion and sugar efflux from injured cells, Plant Physiol. 60: 393–397.Google Scholar
  29. Pràsil I. and Zàmecnìk J. 1998. The use of a conductivity measurement method for assessing freezing injury. I. Influence of leakage time, segment number, size and shape in a sample on evaluation of the degree of injury. Environ. Exp. Bot. 40: 1–10.Google Scholar
  30. Premachandra G.S. and Shimada T. 1987. The measurement of cell membrane stability using polyethylene glycol as a drought tolerance test in wheat. Jpn. J. Crop Sci. 56: 92–98.Google Scholar
  31. Premachandra G.S., Saneoka H. and Ogata S. 1989. Nutrio-physiological evaluation of polyethylene glycol test of cell membrane stability in maize. Crop Sci. 29: 1287–1292.Google Scholar
  32. Saelim S. and Zwiazek J.J. 2000. Preservation of thermal stability of cell membranes and gas exchange in high temperature-acclimated Xylia xylocarpa seedlings. J. Plant Physiol. 156: 380–385.Google Scholar
  33. Senaratna T., McKersie B.D. and Borochov A. 1987. Desiccation and free radical mediated changes in plant membranes. J. Exp. Bot. 38: 2005–2014.Google Scholar
  34. Shcherbakova A. and Kacperska A. 1983. Water stress injuries and tolerance as related to potassium efflux from winter rape hypocotyls. Physiol. Plant. 57: 296–300.Google Scholar
  35. Simane B., Struik P.C., Nachit M.M. and Peacock J.M. 1993. Ontogenic analysis of yield stability of durum wheat in water-limited environments. Euphytica 71: 211–219.Google Scholar
  36. Spencer D.F. and Ksander G.G. 1999. Influence of dilute acetic acid treatments on survival of monoecious Hydrilla tuber in the Oregon House Canal, California. J. Aqu. Plant Manag. 37: 67–71.Google Scholar
  37. Sreenivasulu N., Grimm B., Wobus U. and Weshke W. 2000. Differential response of antioxidant compounds to salinity stress in salt-tolerant and salt-sensitive seedlings of foxtail millet (Setaria italica). Physiol. Plant. 109: 435–442.Google Scholar
  38. Sriram S., Raguchander T., Babu S., Nandakumar R., Shanmugam V., Vidhyasekaran P. et al. 2000. Inactivation of phytotoxin produced by the rice sheath blight pathogen Rhizoctonia solani. Can. J. Microbiol. 46: 520–524.PubMedGoogle Scholar
  39. Stevanovic B., Sinzar J. and Glisic O. 1997. Electrolyte leakage differences between poikilohydrous and homoiohydrous species of Gesneriaceae. Biol. Plant. 40: 299–303.Google Scholar
  40. Sullivan C.Y. and Ross W.M. 1979. Selecting for drought and heat resistance in grain sorghum. In: Mussell H. and Staples R.C. (eds), Stress Physiology in Crop Plants. John Wiley and Sons, New York, pp. 263–281.Google Scholar
  41. Tamura A. 2000. Evaluation of freezing tolerance of whole plants in komatsuna (Brassica campestris L.) and spinach (Spinacia oleraceae L.). J. Jap. Soc. Hort. Sci. 69: 332–338.Google Scholar
  42. Vainola A. and Repo T. 2000. Impedance spectroscopy in frost hardiness evaluation of Rhododendron leaves. Ann. Bot. 86: 799–805.Google Scholar
  43. Vasquez-Tello A., Zuily-Fodil Y., Pham Thi A.T. and Viera Da Silva J.B. 1990. Electrolyte and Pi leakages and soluble sugar content as physiological tests for screening resistance to water stress in Phaseolus and Vigna species. J. Exp. Bot. 41: 827–832.Google Scholar
  44. Zwiazek J.J. and Blake T.J. 1990. Effects of preconditioning on electrolyte leakage and lipid composition in black spruce (Picea mariana) stressed with polyethylene glycol. Physiol. Plant. 79: 71–77.Google Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Mohammed Bajji
    • 1
  • Jean-Marie Kinet
    • 1
  • Stanley Lutts
    • 1
    Email author
  1. 1.Laboratoire de CytogénétiqueUniversité catholique de LouvainLouvain-la-NeuveBelgium

Personalised recommendations