Photosynthetica 2011, 49(2):259-266 | DOI: 10.1007/s11099-011-0028-9

Acclimatization of micropropagated plantlets induces an antioxidative burst: a case study with Ulmus minor Mill.

M. C. Dias1,*, G. Pinto1, C. Santos1
1 Centre for Environmental and Marine Studies (CESAM) & Department of Biology, University of Aveiro, Aveiro, Portugal

In this article, the effects of increased light intensities on antioxidant metabolism during ex vitro establishment of Ulmus minor micropropagated plants are investigated. Three month old in vitro plants were acclimatized to ex vitro conditions in a climate chamber with two different light intensities, 200 μmol m-2 s-1 (high light, HL) and 100 μmol m-2 s-1 (low light, LL) during 40 days. Immediately after ex vitro transfer, the increase of both malondialdehyde (MDA) and electrolyte leakage in persistent leaves is indicative of oxidative stress. As the acclimatization continues, an upregulation of the superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GR) enzyme activities were also observed. Simultaneously, MDA content and membrane permeability stabilized, suggesting that the antioxidant enzymes decrease the deleterious effects of reactive oxygen species (ROS) generation. Unexpectedly, newly formed leaves presented a different pattern of antioxidative profile, with high levels of MDA and membrane leakage and low antioxidant enzyme activity. Despite these differences, both leaf types looked healthy (e.g. greenish, with no necrotic spots) during the whole acclimatization period. The results indicate that micropropagated U. minor plantlets develop an antioxidant enzyme system after ex vitro transfer and that, in general, LL treatment leads to lower oxidative stress. Moreover, new leaves tolerate higher levels of ROS without the need to activate the antioxidative pathway, which suggests that the environment at which leaves are exposed during its formation determinate their ability to tolerate ROS.

Keywords: antioxidant enzymes; lipid peroxidation; micropropagation; Ulmus minor

Received: December 15, 2010; Accepted: April 20, 2011; Published: June 1, 2011Show citation

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Dias, M.C., Pinto, G., & Santos, C. (2011). Acclimatization of micropropagated plantlets induces an antioxidative burst: a case study with Ulmus minor Mill. Photosynthetica49(2), 259-266. doi: 10.1007/s11099-011-0028-9.
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References

  1. Ali, M.B., Hahn, E-J., Paek, K-Y.: Effects of light intensities on antioxidant enzymes and malondialdehyde content during short-term acclimatization on micropropagated Phalaenopsis plantlet. - Environ. Exp. Bot. 54: 109-120, 2005. Go to original source...
  2. Agarwal, S., Sairam, R.K., Srivastava, G.C., Meena, R.C.: Changes in antioxidant enzymes activity and oxidative stress by abscisic acid and salicylic acid in wheat genotypes. - Biol. Plant. 49: 541-550, 2005. Go to original source...
  3. Apel, K., Hirt, H.: Reactive oxygen species: Metabolism, oxidative stress, and signal transduction. - Annu. Rev. Plant Biol. 55: 373-399, 2004. Go to original source...
  4. Asada, K.: Production and scavenging of reactive oxygen species in chloroplasts and their functions. - Plant Physiol. 141: 391-396, 2006. Go to original source...
  5. Bacelar, E.A., Santos, D.L., Moutinho-Pereira, J.M., Gonçalves, B.C., Ferreira, H.F., Correia, C.M.: Immediate responses and adaptative strategies of three olive cultivars under contrasting water availability regimes: Changes on structure and chemical composition of foliage and oxidative damage. - Plant Sci. 170: 596-605, 2006. Go to original source...
  6. Baťková, P., Pospíšilová, J., Synková, H.: Production of reactive oxygen and development of antioxidative systems during in vitro growth and ex vitro transfer. - Biol. Plant. 52: 413-422, 2008.
  7. Beers, R.F., Sizer, I.W.: A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. - J. Biol. Chem. 195: 133-140, 1952.
  8. Bradford, M.M.: A rapid and sensitive method for the quantiation of microgram quantities of protein utilizing the principle of protein-dye binding. - Anal. Biochem. 72: 248-254, 1976. Go to original source...
  9. Brito, G., Costa, A., Coelho, C., Santos, C.: Large-scale field acclimatization of Olea maderensis micropropagated plants: morphological and physiological survey. - Trees 23: 1019-1031, 2009. Go to original source...
  10. Carvalho, L.C., Amâncio, S.: Effect of ex vitro conditions on growth and acquisition of autotrophic behaviour during the acclimatisation of chestnut regenerated in vitro. - Sci. Hortic. 95: 151-164, 2002a. Go to original source...
  11. Carvalho, L.C., Amâncio, S.: Antioxidant defence system in plantlets transferred from in vitro to ex vitro: effects of increasing light intensity and CO2 concentration. - Plant Sci. 162: 33-40, 2002b. Go to original source...
  12. Conde, P., Loureiro, J., Santos, C.: Somatic embryogenesis and plant regeneration from leaves of Ulmus minor Mill. - Plant Cell Reports 22: 632-639, 2004. Go to original source...
  13. Conde, P., Sousa, A., Costa, A., Santos, C.: A protocol for Ulmus minor Mill. micropropagation and acclimatization. - Plant Cell Tissue Organ Culture 92: 113-119, 2008.
  14. Chen, G.X., Asada, K.: Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the differences in their enzymatic and molecular properties. - Plant Cell Physiol. 30: 987-998, 1989.
  15. Dias, M.C., Pinto, G., Correia, C.M., Moutinho-Pereira, J., Guerra, C.C., Monteiro, C., Santos, C.: Effects of Light Intensity on Photosynthesis during Acclimatization of U. minor. - Proc. XVIIIth Congress FESPB, Valencia 2010.
  16. Dhindsa, R.S., Plumbdhindsa, P., Thorpe, T.A.: Leaf senescence correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. - J. Exp. Bot. 32: 93-101, 1981. Go to original source...
  17. Dunn, C.P.: The Elms-Breeding, Conservation and Disease Management. - Kluwer, Dordrecht 2000.
  18. Estrada-Luna, A.A., Davies, F.T., Egilla, J.N.: Physiological changes and growth of micropropagated chile ancho pepper plantlets during acclimatization and post-acclimatization. - Plant Cell Tissue Organ Culture 66: 17-24, 2001. Go to original source...
  19. Faisal, M., Anis, M.: Changes in photosynthetic activity, pigment composition electrolyte leakage, lipid peroxidation, and antioxidant enzymes during ex vitro establishment of micropropagated Rauvolfia tetraphylla plantlets. - Plant Cell Tiss. Organ Cult. 99: 125-132, 2009. Go to original source...
  20. FAO: State of the World's Forests, 1999.
  21. Foyer, C.H., Lelandais, M., Kunert, K.J.: Photooxidative stress in plants. - Physiol. Plant. 92: 696-717, 1994. Go to original source...
  22. Foyer, C.H., Lopez-Delgado, H., Dat, J.F., Scott, I.M.: Hydrogen peroxide- and glutathione-associated mechanisms of acclimatory stress tolerance and signalling. - Physiol. Plant. 100: 241-254, 1997. Go to original source...
  23. Guan, Q.Z., Guo, Y.H., Sui, X.L., Zhang, Z.X.: Changes on photosynthesis capacity and antioxidant enzymatic systems on micropropagated Zingiber officinale plantlets during their acclimation. - Photosynthetica 46: 193-201, 2008. Go to original source...
  24. Harvengt, L., Meier-Dinkel, A., Dumas, E., Collin, E.: Establishment of a cryopreserved gene bank of European elms. - Can. J. Forest Res. 34: 43-55, 2004. Go to original source...
  25. Hazarika, B.N.: Morpho-physiological disorders in in vitro culture of plants. - Sci. Hortic. 108: 105-120, 2006. Go to original source...
  26. Ishikawa, T., Yoshimura, K., Sakai, K., Tamoi, M., Takeda, T., Shigeoka, S.: Molecular characterization and physiological role of a glyoxysome-bound ascorbate peroxidase from spinach. - Plant Cell Physiol. 39: 23-34, 1998. Go to original source...
  27. Lutts, S., Kinet, J.M., Bouharmont, J.: NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. - Ann. Bot. 78: 389-398, 1996. Go to original source...
  28. Merkle, S.A., Nairn, C.J.: Hardwood tree biotechnology. - In Vitro Cell Dev. Biol. Plant 41: 602-619, 2005. Go to original source...
  29. Miyake, C., Asada, K.: Inactivation mechanism of ascorbate peroxidase at low concentrations of ascorbate; Hydrogen peroxide decomposes compound I of ascorbate peroxidase. - Plant Cell Physiol. 37: 423-430, 1996. Go to original source...
  30. Nakano, Y., Asada, K.: Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. - Plant Cell Physiol. 22: 867-880, 1981.
  31. Osório, M.L., Osório, J., Romano, A.: Chlorophyll fluorescence in micropropagated Rhododendron ponticum subsp. baeticum plants in response to different irradiances. - Biol. Plant. 54: 415-422, 2010. Go to original source...
  32. Ort, D.R., Baker, N.R.: A photoprotective role for O2 as an alternative electron sink in photosynthesis?- Curr. Opin. Plant Biol. 5: 193-198, 2002. Go to original source...
  33. Park, Y.-S.: Implementation of conifer somatic embryogenesis in clonal forestry: technical requirements and deployment considerations. - Ann. Forest Sci. 59: 651-656, 2002. Go to original source...
  34. Pinto, G., Silva, S., Loureiro, J., Costa, A., Dias, M.C., Araújo, C., Neves, L., Santos, C.: Acclimatization of secondary somatic embryos derived plants of Eucalyptus globulus Labill.: An ultrastructural approach. - Trees-Struc. Func. 25: 383-292, 2011. Go to original source...
  35. Pospíšilová, J., Synková, H., Haisel, D., Čatský, J., Wilhelmová, N., Šrámek, F.: Effect of elevated CO2 concentrations on acclimation of tobacco plantlets to ex vitro conditions. - J. Exp. Bot. 50: 119-126, 1999. Go to original source...
  36. Schützendübel, A., Polle, A.: Plant responses to abiotic stresses: heavy metal-induced oxidative stress and protection by mycorrhization. - J. Exp. Bot. 53: 351-1365, 2002. Go to original source...
  37. Smirnoff, N.: Ascorbic acid: metabolism and functions of a multifacetted molecule. - Curr. Opin. Plant Biol. 3: 229-235, 2000. Go to original source...
  38. Sgherri, C.L.M., Loggini, B., Puliga, S., Navari-Izzo, F.: Antioxidant system in Sporobolus stapfianus: Changes in response to desiccation and rehydration. - Phytochemistry 33: 561-565, 1994. Go to original source...
  39. Van Huylenbroeck, J.M., Piqueras, A., Debergh, P.C.: The evolution of photosynthesis capacity and the antioxidant enzymatic system during acclimatization of micropropagated Calathea plants. - Plant Sci. 155: 59-66, 2000. Go to original source...
  40. Zhou, B.Y., Wang, J.H., Guo, Z.F., Tan, H.Q., Zhu, X.C.: A simple colorimetric method for determination of hydrogen peroxide in plant tissues. - Plant Growth Regul. 49: 113-118, 2006. Go to original source...