Reaction of various explants of a Chrysanthemum morifolium cultivar to plant growth regulators in vitro

Document Type : Full Paper


1 Former Ph.D. Student, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Assistant Professor, Science and Research Branch, Islamic Azad University, Tehran, Iran

3 Associate Professor, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

4 Associate Professor, Department of Biotechnology, Faculty of Energy Engineering and New Technology, Shahid Beheshti University, Tehran, Iran


Economic importance of Chrysanthemum among the other ornamental species has resulted in performing various investigations in tissue culture optimization of this plant, worldwide. In the current experiment, leaf, petiole, petal and stem TCL explants, from Chrysanthenum morifolium cv. Reagan Elite Salmon were cultured in MS medium supplemented with different combinations and concentrations of NAA, BAP and TDZ. Regeneration percentage,number of regenerated shoots per explants and regeneration type were investigated in each treatment. Results indicated significant superiority of shoot regeneration percentage from petiole explants (80.5 percent), while minimum regeneration was achieved from the petal explants (37.2 percent). Shoot regeneration type from different explants in the treatments containing only BAP, was direct and in the treatments containing TDZ and NAA was indirect. Reactions of the explants to the PGR treatments, considering shoot regeneration percentage and average number of regenerated shoots per explants, were so different. As among the treatments consisting combinations of NAA plus each of the cytokinins, in T12 (4.5 mg/l BAP + 1 mg/l NAA), petioles and petals represented maximum shoot regeneration percentage and number, while stem TCLs showed minimum rates of regeneration.


  1. Anderson, N. O., Ascher, P. D., Widmer, R. E. & Luby, J. J. (1990). Rapid generation cycling of Chrysanthemum using laboratory seed development and embryo rescue techniques. Journal of American Society of Horticultural Sciences, 115(2), 329-336.
  2. Annadana, S., Rademaker, W., Ramanna, M., Udayakumar, M. & de Jong, J. (2000). Response of stem explants to screening and explant source as a basis for methodical advancing of regeneration protocols for Chrysanthemum. Plant Cell, Tissue and Organ Culture, 62, 47-55.
  3. Barakat, M. N., Abdel Fattah, R. S., Badr, M. & EI-Torky, M. G. (2010). In vitro culture and plant regeneration derived from ray florets of Chrysanthemum morifolium. African Journal of Biotechnology, 9(8), 1151-1158.
  4. Chagas, E. A., Fraguas, C. B., da Silva, E. F., Pasqual, M. & Mendonca, V. (2004). In vitro multiplication of chrysanthemum ‘White polaris’. Revista- Brasileira-de-Agrociencia, 10(1), 123-126.
  5. De Jong, J., van Wordragen, M. F. & Rademaker, W. (1990). Early transformation events in Dendranthema grandiflora. In: Proceedings of EUCARPIA (Section Ornamentals): Integration of in vitro techniques in ornamental plant breeding, Wageningen, pp. 156-161.
  6. Floraholland, Facts and Figures. (2013). Cut flower: FloraHolland departments Finance and MiMa (Market Information & Market Analysis), International Statistics, Flowers and Plants (Bloemen en Planten AIPH), from 2460310/ Kengetallen-EN-2013.pdf
  7. Floraholland, Facts and Figures. (2014). Cut flower: FloraHolland departments Finance and MiMa (Market Information & Market Analysis), International Statistics, Flowers and Plants (Bloemen en Planten AIPH), from https: //www. / media/ 4213130/ floraholland _Kengetallen 2014_Engels.pdf.
  8. Himstedt, J. P. & Jacobsen, H. J. (2001). Shoot regeneration from stem and leaf explants of Chrysanthemum. Acta Horticulturae, 560, 421-424.
  9. Hobbie, L. J. (1998). Auxin: Molecular genetic approaches in Arabidopsis. Plant Physiology and Biochemistry, 36, 91-102. 
  10. Hodson de Jaramillo, E., Forero, A., Cancino, G., Moreno, A. M., Monsalve, L. E. & Acero, W. (2008). In vitro regeneration of three chrysanthemum (Dendrathema grandiflora) varieties via organogenesis and somatic embryogenesis. Universitas Scientiarum, 13(2), 118-127.
  11. Ji, A., Geng, X., Zhang, Y., Yang, H. & Wu. G. (2011). Advances in somatic embryogenesis research of horticultural plants. American Journal of Plant Sciences, 2, 727-732.
  12. Karim, M. Z., Amin, M. N., Azad, M. A. K., Begum, F., Rahman, M. M., Ahmad, S. & Alam, R. (2003). In vitro shoot multiplication of Chrysanthemum morifolium as affected by sucrose, agar and pH. Biotechnology, 2(2), 115-120.
  13. Kaul, V., Miller, R. B., Hutchinson, J. F. & Richards, D. (1990). Shoot regeneration from stem and leaf explants of Dendranthema grandiflora Tzvelev (syn. Chrysanthemum morifolium Ramat.). Plant Cell, Tissue and Organ Culture, 21, 21-30.
  14. Khalili, S., Jafarkhani Kermani, M., Azadi, P. & Kalatejari, S. (2014). Investigating effects of different factors on regeneration efficiency of two commercial cultivars of Chrysanthemum grandiflorum. In: Proceedings of 1st National Ornamental Plants Congress, 21-22 Oct 2014, Karaj, Iran, p. 43. (in Farsi)
  15. Lim, K. B., Kwon, S. J., Lee, S. I., Hwang, Y. J. & Naing,  A. H. (2012). Influence of genotype, explant source, and gelling agent on in vitro shoot regeneration of Chrysanthemum. Horticulture, Environment, and Biotechnology, 53(4), 329-335.
  16. Lindiro, C., Kahia, J., Asiimwe, T., Mushimiyimana, I., Waweru, B., Kouassi, M., Koffi, E., Kone, S. & Sallah, P. Y. (2013). In vitro regeneration of pyrethrum (Chrysanthemum cinerariaefolium) plantlets from nodal explants of in vitro raised plantlets. International Journal of Application or innovation in Engineering & management, 2(7), 207-213.
  17. Malik, S. K., Chadhury, R. & Kalia, R. K. (2005). Rapid in vitro multiplication and conservation of Garcinia indica: A tropical medicinal tree species. Scientia Horticulturae, 106, 539-553.
  18. Mandal, A. K. A. & Datta, S. K. (2005). Direct somatic embryogenesis and plant regeneration from ray florets of Chrysanthemum. Biologia Plantarum, 49(1), 29-33.
  19. Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Plant Physiology, 15, 473-497.
  20. Nahid, J. S., Shyamali, S. & Kazumi, H. (2007). High frequency shoot regeneration from petal explants of Chrysanthemum morifolium Ramat. in vitro. Pakistan Journal of Biological Sciences, 10(19), 3356-3361.
  21. Naing, A. H., Jeon, S. M., Han J. S., Lim, S. H., Lim, K. B. L. & Kim, C. K. (2014). Factors influencing in vitro shoot regeneration from leaf segments of Chrysanthemum. Comptes Rendus Biologies, 337, 383-390.
  22. Park, S. H., Kim, G. H. & Jeong, B. R. (2007). Adventitious shoot regeneration from cultured petal explants of Chrysanthemum. Horticulture, Environment, and Biotechnology, 48(6), 387-392.
  23. Rout, G. R. & Das, P. (1997). Recent trends in the biotechnology of Chrysanthemum: A critical review. Scientia Horticulturae, 69, 239-257.
  24. Song, J. Y., Mattson, N. S., & Jeong, B. R. (2011). Efficiency of shoot regeneration from leaf, stem, petiole and petal explants of six cultivars of Chrysanthemum morifolium. Plant Cell, Tissue and Organ Culture, 107, 295-304.
  25. Teixeira da Silva, J. A. (2003a). Filter paper type affects morphogenic programs and buffers the phytotoxic effect of antibiotics in chrysanthemum and tobacco thin cell layers. Horticultural Science, 38(7), 1403-1407.
  26. Teixeira da Silva, J. A. (2003b). Chrysanthemum: Advances in tissue culture, cryopreservation, postharvest technology, genetics and transgenic biotechnology. Biotechnology Advances, 21, 715-766.
  27. Waseem, K., Khan, M. Q., Jaskani, J., Jilani, M. S. & Khan, M. S. (2009). Effect of different auxins on the regeneration capability of Chrysanthemum leaf discs. International Journal of Agriculture & Biology, 11, 468-472.
  28. Zalewska, M., Miler, N. & Wenda-Piesik, A. (2010). Effect of in vitro topophysis on the growth, development, and rooting of Chrysanthemum explants (Chrysanthemum ´ grandiflorum/ Ramat./ Kitam). Journal of Horticultural Science & Biotechnology, 85(4), 362-366.
  29. Zalewska, M., Tymoszuk, A. & Miler, N. (2011a). New chrysanthemum cultivars as a result of in vitro mutagenesis with the application of different explant types. Acta Scientiarum Polonorum, Hortorum Cultus, 10(2), 109-123.
  30. Zalewska, M., Lema-Rumińska, J., Miler, N., Gruszka M. & Dąbal, W. (2011b). Induction of adventitious shoot regeneration in Chrysanthemum as affected by the season. In Vitro Cellular & Developmental Biology-Plant, 47, 375-378.