Biologia Plantarum

, Volume 54, Issue 3, pp 509–514 | Cite as

Activity of antioxidant enzyme during in vitro organogenesis in Crocus sativus

Brief Communication


The effect of various hormonal combinations on regeneration of shoots and roots from meristem-derived callus of Crocus sativus L. and activities of antioxidant enzymes have been studied. The most efficient regeneration occurred with 1.0 mg dm−3 1-naphthaleneacetic acid (NAA) + 1.0 mg dm−3 thidiazuron and 1.0 mg dm−3 NAA + 2.0 mg dm−3 kinetin. For sprouting, regenerated shoot were subcultured on Murashige and Skoog medium containing 1.0 mg dm−3 NAA + 1.0 mg dm−3 benzylaminopurine (BAP). Protein content and superoxide dismutase activity decreased in regenerated shoots and roots and increased in sprouting shoots, while catalase (CAT), peroxidase (POX) and polyphenol oxidase (PPO) activities increased during organogenesis and decreased in sprouting shoots. High CAT and PPO activities were detected in regenerated roots, whereas high POX activity was observed in regenerated shoot.

Additional key words

catalase peroxidase polyphenol oxidase protein saffron superoxide dismutase 







ethylenediaminetetraacetic acid






Murashige and Skoog


1-naphthaleneacetic acid


nitroblue tetrazolium


polyacrylamide gel electrophoresis


picloram (4-amino-3,5-trichloropicolinic acid)




polyphenol oxidase




reactive oxygen species


sodium dodecyl sulfate


superoxide dismutase




Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



The financial support of this research was provided by University of Tehran. The authors are grateful to the anonymous reviewers for very helpful and valuable advices.


  1. Aebi, H.: Catalases. — In: Bergmeyer, H.U. (ed.): Methods of Enzymatic Analysis. Vol. 2. Pp. 673–684. Academic Press, New York 1974.Google Scholar
  2. Abeles, F.B., Biles, C.L.: Characterization of peroxidases in lignifying peach fruit endocarp. — Plant Physiol. 95: 269–273, 1991.CrossRefPubMedGoogle Scholar
  3. Agrawal, V., Sardar, P.: In vitro propagation of Cassia angustifolia through leaflet and cotyledon derived calli. — Biol. Plant. 50: 118–122, 2006.CrossRefGoogle Scholar
  4. Basker, D., Negbi, M.: Uses of saffron. — Econ. Bot. 37: 228–236, 1983.Google Scholar
  5. Benson, E.E.: Do free radicals have a role in plant tissue culture recalcitrance? — In Vitro cell. dev. Biol. Plant 36: 163–170, 2000.Google Scholar
  6. Baťková, P., Pospíšilová, J., Synková, H.: Production of reactive oxygen species and development of antioxidative systems during in vitro growth and ex vitro transfer — Biol. Plant. 52: 413–422, 2008.CrossRefGoogle Scholar
  7. Bhagyalakshmi, N.: Factors influencing direct shoot regeneration from ovary explants of saffron. — Plant Cell Tissue Organ Cult. 58: 205–211, 1999.CrossRefGoogle Scholar
  8. Bradford, M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. — Anal. Biochem. 72: 248–254, 1976.CrossRefPubMedGoogle Scholar
  9. Davis, B.J.: Disc electrophoresis. II. Method and application to human serum proteins. — Ann. N.Y. Acad. Sci. 121: 404–427, 1964.CrossRefPubMedGoogle Scholar
  10. Ding, B.Z., Bai, S.H., Wu, Y., Fan, X.P.: Induction of callus and regeneration of plantlets from corm of Crocus sativus L. — Acta bot. sin. 23: 434–440, 1981.Google Scholar
  11. Franck, T., Kevers, C., Gaspar, T.: protective enzymatic systems against activated oxygen species compared in normal and vitrified shoots of Prunus avium L. raised in vitro. — Plant Growth Regul. 16: 253–256, 1995.CrossRefGoogle Scholar
  12. Gaspar, T.: The concept of cancer in in vitro plant cultures and the implication of habituation to hormones and hyperhydricity. — Plant Tissue Cult. Biotechnol. 1: 126–136, 1995.Google Scholar
  13. Giannopolitis, C.N., Ries, S.K.: Superoxide dismutases II. purification and quantitative relationship with water-soluble protein in seedlings. — Plant Physiol. 59: 315–318, 1977.CrossRefPubMedGoogle Scholar
  14. Gupta, S.D., Datta, S.: Antioxidant enzyme activities during in vitro morphogenesis of gladiolus and the effect of application of antioxidants on plant regeneration. — Biol. Plant. 47: 179–183, 2003/4.CrossRefGoogle Scholar
  15. Huetteman, C.A., Preece, J.E.: Thidiazuron: a potent cytokinin for woody plant tissue culture. — Plant Cell Tissue Organ Cult. 33: 105–119, 1993.CrossRefGoogle Scholar
  16. Ilahi, I., Jabeen, M., Firdous, N.: Morphogenesis with saffron tissue culture. — J. Plant Physiol. 128: 227–232, 1987.Google Scholar
  17. Jasska, V.: Isoenzyme diversity and phylogenetic affinities among the Phaseolus beans (Fabaceae). — Plant Syst. Evol. 200: 233–252, 1996.CrossRefGoogle Scholar
  18. Kumar, G.N.M., Knowles, N.R.: Changes in lipid peroxidation and lipolytic and free radical scavenging enzyme activities during aging and sprouting of potato (Solanum tuberosum) seed-tubers. — Plant Physiol. 102: 115–124, 1993.PubMedGoogle Scholar
  19. Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. — Nature 227: 680–685, 1970.CrossRefPubMedGoogle Scholar
  20. Lee, D.L., Kim, Y.S., Lee, C.B.: The inductive responses of the antioxidant enzymes by salt stress in the rice (Oryza sativa L.). — J. Plant Physiol. 158: 737–745, 2001.CrossRefGoogle Scholar
  21. Libik, M., Konieczny, R., Pater, B., Ślesak, I., Miszalski, Z.: Differences in the activities of some antioxidant enzymes and in H2O2 content during rhizogenesis and somatic embryogenesis in callus cultures of the ice plant. — Plant Cell Rep. 23: 834–841, 2005.CrossRefPubMedGoogle Scholar
  22. Molassiotis, A.N., Dimassi, K., Diamantidis, G., Therios, I.: Changes in peroxidases and catalase activity during in vitro rooting. — Biol. Plant. 48: 1–5, 2004.CrossRefGoogle Scholar
  23. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. — Physiol. Plant. 15: 473–497, 1962.CrossRefGoogle Scholar
  24. Piqueras, A., Han, B.H., Escribano, J., Rubio, C., Hellín, E., Fernández, J.A.: Development of cormgenic nodules and microcorms by tissue culture, a new tool for the multiplication and genetic improvement of saffron. — Agronomie 19: 603–610, 1999.CrossRefGoogle Scholar
  25. Radhika, K., Sujatha, M., Nageshwar Rao, T.: Thidiazuron stimulates adventitious shoot regeneration in different sunflower explants. — Biol. Plant. 50: 174–179, 2006.CrossRefGoogle Scholar
  26. Raymond, J., Rakariyatham, N., Azanza, J.L.: Purification and some properties of polyphenoloxidase from sunflower seeds. — Phytochemistry 34: 927–931, 1993.CrossRefGoogle Scholar
  27. Rey, H.Y., Mroginski, L.A.: Somatic embryogenesis and plant regeneration in diploid and triploid Arachis pintoi. — Biol. Plant. 50: 152–155, 2006.CrossRefGoogle Scholar
  28. Sampathu, S.R., Shivashankar, S., Lewis, Y.S.: Saffron (Crocus sativus L.) cultivation, processing, chemistry and standardization. — CRC Food Sci. Nutr. 20: 123–157, 1984.CrossRefGoogle Scholar
  29. Sharma, K.D., Rathour, R., Sharma, R., Goel, S., Sharma, T.R., Singh, B.M.: In vitro development in Crocus sativus. — Biol. Plant. 52: 709–712, 2008.CrossRefGoogle Scholar
  30. Tian, M., Gu, Q., Zhu, M.: The involvement of hydrogen peroxide and antioxidant enzymes in the process of shoot organogenesis of strawberry callus. — Plant Sci. 165: 701–707, 2003.CrossRefGoogle Scholar
  31. Tang, W., Newton, R.J.: Peroxidase and catalase activities are involved in direct adventitious shoot formation induced by thidiazuron in eastern white pine (Pinus strobes L.) zygotic embryos. — Plant Physiol. Biochem. 43: 760–769, 2005.CrossRefPubMedGoogle Scholar
  32. Van Loon, L.C., Geelen, J.L.M.C.: The relation of polyphenol oxidase and peroxidase to symptom expression in tobacco var. “Samsun NN“ after infection with tobacco mosaic virus. — Acta phytopathol. Acad. Sci. hung. 6: 9–20, 1971.Google Scholar
  33. Wang, S.Y., Faust, M.: Changes of fatty acids and sterols in apple buds during the break induced by a plant bioregulator, thidiazuron. — Physiol. Plant. 72: 115–120, 1988.CrossRefGoogle Scholar
  34. Wang, S.Y., Jiao, H. J., Faust, M.: Changes in activities of catalase, peroxidase and polyphenol oxidase in apple buds during bud break induced by thidiazuron. — J. Plant Growth Regul. 10: 33–39, 1991.CrossRefGoogle Scholar
  35. Wendel, J.F., Weeden, N.F.: Visualization and interpretation of plant isozymes. — In: Soltis, D.E., Soltis, P.S. (ed.): Isozymes in Plant Biology. Pp. 4–45. Dioscorides Press, Portland 1989.Google Scholar
  36. Woodbury, W., Spencer, A.K., Stahman, M.A.: An improved procedure using ferricyanide for detecting catalase isozymes. — Anal. Biochem. 44: 301–305, 1971.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Department of Plant Sciences, School of Biology, College of SciencesUniversity of TehranTehranI.R. Iran

Personalised recommendations