Acta Biologica Hungarica

, Volume 66, Issue 4, pp 419–435 | Cite as

Morphological and Physiological Changes in Esterase and Lipid Peroxidation of Two Bean Cultivars Pre-Soaked With Potassium Nitrate Under Salt Stress

  • Mohamed A. K. Shaddad
  • Gaber K. Abd El-BakiEmail author
  • Mostafa Doaa
  • Rafat Al-Shimaa


Two broad bean cultivars (Vicia faba CV Nobaria3 and Vicia faba CV Sakha3) were obtained from Mallwi Agriculture Research Center, El Minia Governorate, Egypt. The seeds were divided into two groups, the first group soaked with distilled water, while the second group were soaked with 3 niM KN03, respectively, for 4 hours. Seeds were sown and left to grow for 3 weeks then treated with different concentrations of NaCl (0.0, 40, 80, 120 and 160 mM) by top irrigation, then they left to grow further for 65 days from sowing. Plant samples were collected for some measurements: leaf area, plant height, root length, fresh and dry weight, photo synthetic pigments, carotenoids, soluble sugars, soluble proteins, total free amino acids, esterase enzyme, as well as MDA (malondialdehyde) content. Salinity reduced both fresh and dry weight in two broad bean cultivars, this reduction were more pronounced in Sakha3 than Nobaria3. Seed pre-soaking with KN03 resulted in enhancement of fresh and dry weight production in both cultivars especially at 40 mM NaCl. Photosynthetic pigments were substantially affected by salt treatment while the carotenoids were increased, seed pre-soaking with KN03 improved these components. The soluble sugars, amino acids as well as soluble proteins showed various responses with increasing salinity in the cultivars, seed pre-soaking with KN03 has improved these parameters to some extent. The shoots of two cultivars exhibited significant accumulation of MDA, compared to roots exposed to the highest salinity levels. Pre-soaking seeds with KN03 did not improve MDA in shoots but enhanced it in roots, however, in most cases still lower than the absolute control. The assessment of the esterase isozyme profiles on 7.5% native polyacrylamide gel revealed the presence of 13 isoforms in two faba bean plants in response to KNO3 pre-soaking and treatments with different concentrations of NaCl.


Amino acids chlorophylls esterase enzyme lipid peroxidation salinity Vicia faba 









(ethylene diaminetertaacetic acid)




(leaf area)




(trichloroacetic acid)


(water content)


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Argerich, C. A., Bradford, K. J. (1989) The effects of priming and aging on seed vigour in tomato. J. Exp. Bot. 40, 599–607.CrossRefGoogle Scholar
  2. 2.
    Ashraf M., Foolad, M. R. (2005) Pre-sowing seed treatment - A shotgun approach to improve germination, plant growth, and crop yield under saline and non-saline conditions. Adv. Agron. 88, 223–265.CrossRefGoogle Scholar
  3. 3.
    Azooz, M. (2009) Salt stress mitigation by seed priming with salicylic acid in two faba bean genotypes differing in salt tolerance. Int. J. Agric. and Biol. Engin. 11, 343–350.Google Scholar
  4. 4.
    Badour, S. S. A. (1959) Analitisch-chemische Untersuchung des Kaliummangels bei Chlorella in Vergleich mit anderen Mangel-Zustanden. Ph.D. Dissertation, Gottingen.Google Scholar
  5. 5.
    Bajehbaj, A. A. (2010) The effects of NaCl priming on salt tolerance in sunflower germination and seedling grown under salinity conditions. Afr J. Biotech. 9, 1764–1770.CrossRefGoogle Scholar
  6. 6.
    Bandehagh, A., Salekdeh, G. H., Toorchi, M. (2011) Comparative proteomic analysis of canola leaves under salinity stress’. Proteomics 11, 1965–1975.PubMedCrossRefPubMedCentralGoogle Scholar
  7. 7.
    Boursiac, Y., Chen, S., Luu, D. T., Sorieul, M., Dries, N., Maurel, C. (2005) Early effects of salinity on water transport in Arabidopsis roots: molecular and cellular features of aquaporin expression. Plant Physiol. 139, 790–805.PubMedPubMedCentralCrossRefGoogle Scholar
  8. 8.
    Bradford, K. J. (1986) Priming to improve germination under stress conditions. Hort. Sci. 21, 1105–1112.Google Scholar
  9. 9.
    Chandler, P. M., Robertson, M. (1994) Gene expression regulated by abscisic acid and its relation to stress tolerance’. Annu. Rev. Plant Physiol. Plant mol. Biol. 45, 113–141.CrossRefGoogle Scholar
  10. 10.
    Coppens, L., Dewitte, D. (1990) Esterase and peroxidase zymograms from barley (Hordeum vulgare L.) callus as a biochemical marker system of embryogenesis and organogenesis. Plant Science 67, 97–105.CrossRefGoogle Scholar
  11. 11.
    Cosgrove, D. J. (2001) Wall structure and wall loosening. A look backwards and forwards. Plant Physiol. 125, 131–134.PubMedPubMedCentralCrossRefGoogle Scholar
  12. 12.
    Cummins, I., Burnet, M., Edwards, R. (2001) Biochemical characterization of esterases active in hydrolysing xenobiotics in wheat and competing weeds. Physiol. Plant. 113, 477–485.CrossRefGoogle Scholar
  13. 13.
    De Lacerda, C. F., Cambraia, J., Oliva, M. A., Ruiz, H. A. (2003) Osmotic adjustment in roots and leaves of two sorghum genotypes under NaCl stress. Braz. J. Plant Physiol. 15, 113–118.CrossRefGoogle Scholar
  14. 14.
    Demir Kaya, M., Okcu, G., Atak, M., Cikili, Y., Kolsarici, O. (2006) Seed treatment to overcome salt and drought stress during germination in sunflower (Helianthus. annuus L.). J. Eur. Agron. 24, 291–295.CrossRefGoogle Scholar
  15. 15.
    Ebrahim, M. K. (2005) Amelioration of sucrose-metabolism and yield changes, in storage roots of NaCl-stressed sugar beet, by ascorbic acid. Agrochimica, XLIX (3-4), 93–103.Google Scholar
  16. 16.
    Fales, F. W. (1951) The assimilation and degradation of carbohydrates by yeast cells. J. Biol. Chem. 193, 113–124.PubMedGoogle Scholar
  17. 17.
    FAO (2008) FAO Land and Plant Nutrition Management Service, Scholar
  18. 18.
    Gadallah, M. A. (1999) Effects of proline and glycinebetaine on Viciafaba in response to salt stress. Biol. Plant 42, 249–257.CrossRefGoogle Scholar
  19. 19.
    Gao, Y. P., Young, L., Bonham-Smith, P., Gusta, L. V. (1999) Characterization and expression of plasma and tonoplast membrane aquaporins in primed seed of Brassica napus during germination under stress conditions. Plant Mol. Biol. 40, 635–644.PubMedCrossRefGoogle Scholar
  20. 20.
    Gigova, L., Gacheva, G., Ivanova, N., Pilarski, P. ( 2012) Effects of temperature on synechocystis sp. r10 (cyanoprokaryota) at two irradiance levels, i. effect on growth, biochemical composition and defense enzyme activities. Gen. Plant Physiol. V2, 24–37.Google Scholar
  21. 21.
    Guan, Y. J., Hu, J., Wang, X. J., Shao, C. X. (2009) Seed priming with chitosan improves maize germination and seedling growth in relation to physiological changes under low temperature stress. J. Zhejiang Univ-Sci. B 10, 427–433.PubMedPubMedCentralCrossRefGoogle Scholar
  22. 22.
    Hamada, A. M., El-Enany, A. E. (1994) Effect of NaCl salinity on growth, pigment and mineral element contents, and gas exchange of broad bean and pea plants. Biol. Plant. 36, 75–81.CrossRefGoogle Scholar
  23. 23.
    Harris, D., Rashid, A., Miraj, G., Arif M., Shah, H. (2007) On-farm’ seed priming with zinc sulphate solution -A cost-effective way to increase the maize yields of resource poor farmers. Field Crops Res. 102, 119–127.CrossRefGoogle Scholar
  24. 24.
    Heath, R. L., Packer, L. (1968) Photoperoxidation in isolated chloroplast. 1. Kinetics and stiochiom-etry of fatty acid peroxidation. Arch. Bioch. Biophys. 125, 189–198.CrossRefGoogle Scholar
  25. 25.
    Hus, J. L., Sung, J. M. (1997) Antioxidant role of glutatnione associated with accelerated agina and hydration of triploid Watermelon seeds. Physiol Plant. 100, 967–974.CrossRefGoogle Scholar
  26. 26.
    Hussein, M. M., Abd El-Rheem, K. M., Khaled, S. M., Youssef, R. A. (2011) Growth and nutrients status of wheat as affected by ascorbic acid and water salinity. Nature and Science 9, 64–69.Google Scholar
  27. 27.
    Jyotsna, V., Srivastava, A. K. (1998) Physiological basis of salt stress resistance in pigeon pea (Cajanuscajan L.)-II. Pre-sowing seed soaking treatment in regulating early seedling metabolism during seed germination. Plant Physiol. Biochem. 25, 89–94.Google Scholar
  28. 28.
    Khan, M. A., Ahmed, M. Z., Hameed, A. (2006) Effect of sea salt and L-ascorbic acid on the seed germination of halophytes. J. Arid Environ. 67, 535–540.CrossRefGoogle Scholar
  29. 29.
    Khosravinejad, H. F. R., Farboondia, T. (2008) Effect of salinity on photosynthetic pigments, respiration and water content in barley varieties. Pah. J. Biol. Sci. 11, 2438–2442.CrossRefGoogle Scholar
  30. 30.
    Lima, A. L. S., DaMatta, F. M., Pinheiro, H. A., Totola, M. R., Loureiro, M. E. (2002) Photochemical responses and oxidative stress in two clones of Coffea canephora under water deficit conditions. Environ. Exp. Bot. 47, 239–247.CrossRefGoogle Scholar
  31. 31.
    Lowery, O. H., Rosebrough, N. H., Farr, A. L., Randall, R. J. (1951) Protein measurements with the folin phenol reagent. J. Biol. Chem. 193, 291–297.Google Scholar
  32. 32.
    McDonald, M. B. (1999) Seed deterioration: physiology, repair and assessment. Seed Sci. Technol. 27, 177–237.Google Scholar
  33. 33.
    Metzner, H., Rau, H., Senger, H. (1965) Untersuchungen zur synchronisierbarkareit einzelener-pig-ment. Mangel Mutanten von Chlorella. Planta 65, 186–194.CrossRefGoogle Scholar
  34. 34.
    Moeinrad, H. (2008) The relationship between some physiological traits and salt tolerance in pistachio genotypes. Desert. 13, 129–136.Google Scholar
  35. 35.
    Mohammadi, G. R., Dezfuli, M. P. M., Sharifzadeh, F. (2008) Seed invigoration techniques to improve germination and early growth of inbred line of maize under salinity and drought stress. Gen. Appl. Plant Physiol. 34, 215–226.Google Scholar
  36. 36.
    Moore, S., Stein, W. (1948) Partition chromatography of amino acids on starch. Annual. N.Y. Acad Sci. 49, 265–278.CrossRefGoogle Scholar
  37. 37.
    Mukherjee, S., Bhattacharyya, P., Duttagupta, A. K. (2004) Heavy metal levels and esterase variations between metal-exposed and unexposed duckweed Lemna minor: field and laboratory studies. Environ Interactions 30, 811–814.Google Scholar
  38. 38.
    Munns, R., Brady, C. J., Barlow, E. W. (1979) Solute accumulation in the apex and leaves of wheat during water stress. Aust. Plant Physiol. 6, 379–389.Google Scholar
  39. 39.
    OlfaBaatour, R., Kaddour, W., Aidi Wannes, M., Lachaal Marzouk, B. (2009) Salt effects on the growth, mineral nutrition, essential oil yield and composition of marjoram (Origanum majorana). Acta Physiol. Plant. 10, 0374-4.Google Scholar
  40. 40.
    Qadir, M., Tubeileh, A., Akhtar, J., Larbi, A., Minhas, P. S., Khan, M. A. (2008) Productivity enhancement of salt-affected environments through crop diversification. Land Degradation Develop. 19, 429–453.CrossRefGoogle Scholar
  41. 41.
    Roy, N. K., Srivastava, A. K. (2000) Adverse effect of salt stress conditions on chlorophyll content in wheat (Triticum aestivum L.) leaves and its amelioration through pre-soaking treatments. Indian J. Agric. Sci. 70, 777–778.Google Scholar
  42. 42.
    Sallam, H. A. (1999) Effect of some seed-soaking treatments on growth and chemical components of faba bean plants under saline conditions. Ann. Agric. Sci. (Cairo). 44, 159–171.Google Scholar
  43. 43.
    Sarkar, R. K., Malik, G. C. (2001) Effect of foliar spray of potassium nitrate and calcium nitrate on grass pea (Lathyrus sativus L.) grown in rice fallows. LathyrusLathyrithm Newsletter 2, 47–48.Google Scholar
  44. 44.
    Schlegel, H. G. (1956) The recovery of organic acid by Chlorella in the light. Planta 47, 510–526.CrossRefGoogle Scholar
  45. 45.
    Takhti, S., Shekafandeh, A. (2012) Effect of different seed priming on germination rate and seedling growth of Ziziphus Spina-Christi. Adv. Environ. Biol. 6, 159–164.Google Scholar
  46. 46.
    Tanksley, S. D., Orton, T. J. (eds) (1983) Isoenzymes in plant genetics and breeding. Part A, Elsevier Amsterdam, New York.Google Scholar
  47. 47.
    Wang, Z. Q, Yuan, Y. Z., Ou, J. Q., Lin, Q. H., Zhang, C. F. (2007) Glutamine synthetase and gluta-mate dehydrogenase contribute differentially to proline accumulation in leaves of wheat (Triticum aestivum) seedlings exposed to different salinity. Original Research Article. J. of Plant Physiol. 164, 695–701.CrossRefGoogle Scholar
  48. 48.
    Wiersma, T. V., Bailey, T. B. (1975) Estimation of leaflet, trifoliate and total leaf area of soybean. Agron. J. 176, 26–30.CrossRefGoogle Scholar
  49. 49.
    Wimmer, M. A., Muhling, K. H., Lauchli, A. (2003) The interaction between salinity and boron toxicity affects the sub cellular distribution of ions and proteins in wheat leaves. Plant Cell Environ. 26, 1267–1274.CrossRefGoogle Scholar
  50. 50.
    Zhou, R., Zhao, H. (2004) Seasonal pattern of antioxidant enzyme system in the roots of perennial forage grasses grown in alpine habitat, related to freezing tolerance. Physiol. Plant. 121, 399–408.CrossRefGoogle Scholar

Copyright information

© Akadémiai Kiadó, Budapest 2015

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • Mohamed A. K. Shaddad
    • 1
  • Gaber K. Abd El-Baki
    • 2
    Email author
  • Mostafa Doaa
    • 2
  • Rafat Al-Shimaa
    • 2
  1. 1.Botany Department, Faculty of ScienceAssiut UniversityAssiutEgypt
  2. 2.Botany and Microbiology Department, Faculty of ScienceMinia UniversityEl-MiniaEgypt

Personalised recommendations