MORPHOGENETIC PATHWAYS FROM Narcissus L. ‘CARLTON’ IN VITRO CULTURES OF PC STAGE FLOWER BUD EXPLANTS ACCORDING TO CYTOKININ AND AUXIN RATIOS
Małgorzata MalikUniversity of Agriculture in Krakow
Anna BachUniversity of Agriculture in Krakow
The effect of the concentration and proportion of growth regulators in an initial medium on Narcissus L. ‘Carlton’ morphogenesis in cultures of Pc stage flower bud explants was investigated. Ovary and flower stem explants were cultured on solid Murashige and Skoog (MS) media with cytokinin – 6-Benzyladenine (BA) or Zeatin (ZEA) (0.5–50 μM) – in combination with auxin – 4-Amino-3,5,6-trichloropicolinic acid (Picloram), 2,4-Dichlorophenoxyacetic acid (2,4-D) or α-Naphthaleneacetic acid (NAA) (1–50 μM). The ratios of cytokinin and auxin in the initial medium determined the morphogenetic pathways and the level of cytokinin proved to be a critical factor. Somatic embryogenesis was achieved for a cytokinin and auxin ratio of 1:1–10, bulb organogenesis – for a 1:0.4–1 ratio and rhizogenesis – for a 1:20–100 ratio. The minimum concentration of cytokinin in the initial medium for somatic embryo and bulb formation was 5 μM. A lower level of cytokinin (0.5 μM) resulted in rhizogenesis. Considering the effect of auxin, Picloram stimulated the initiation of somatic embryos and the 2,4-D enhanced embryo maturation and conversion.
Keywords:callus, growth regulators, organogenesis, somatic embryogenesis
Bronsema, F.B.F., Van Oostveen, W.J.F., Van Lammeren, A.A.M. (2001). Influence of 2,4-D, TIBA and 3,5-D on the growth response of cultured maize embryos. Plant Cell Tiss. Org. Cult., 65, 45–56.
Chen, Z. (1982). A preliminary investigation on the propagation of Narcissus tazetta var. Chinensis Roem by tissue culture. J. Fujian Agricult. Col., 1, 9–13.
Chen, L., Zhu, X., Gu, L., Wu, J. (2005). Efficient callus induction and plant regeneration from anther of Chinese narcissus (Narcissus tazetta L. var. chinensis Roem). Plant Cell Rep., 24, 401–407.
El Tahchy, A., Bordage, S., Ptak, A., Dupire, F., Barre, E., Guillou, C., Henry, M., Chapleur, Y., Laurain-Mattar, D. (2011). Effects of sucrose and plant growth regulators on acetylcholinesterase inhibitory activity of alkaloids accumulated in shoot cultures of Amaryllidaceae. Plant Cell Tiss. Org. Cult., 106, 381–390.
Gude, H., Dijkema, M.H.G.E. (1997). Somatic embryogenesis in tulip. Acta Hortic., 430, 275–300.
Guohua, M. (1998). Effects of cytokinins and auxins on cassava shoot organogenesis and somatic embryogenesis from somatic embryo explants. Plant Cell Tiss. Org. Cult., 54, 1–7.
Hussey, G. (1982). In vitro propagation of Narcissus. Ann. Bot., 49, 707–719.
Jimenez, V.M., Bangerth, F. (2001). Hormonal status of maize initial explants and of the embryogenic and non-embryogenic callus cultures derived from them as related to morphogenesis in vitro. Plant Sci., 160, 247–257.
Kozak, D. (1991). Shoots regeneration from various parts of Narcissus cv. Carlton through tissue culture. Pr. Inst. Sadown. Kwiac., ser. B 16, 41–47.
Kumar, A., Sood, A., Palni, L. M. S., Gupta, A. K. (1999). In vitro propagation of Gladiolus hybridus Hort.: Synergistic effect of heat shock and sucrose on morphogenesis. Micropropagation of gladiolus. Plant Cell Tiss. Org. Cult., 57, 105–112.
Lilien-Kipnis, H., Azizbekova, N., Ziv, M. (1994). Scaled-up proliferation and regeneration of Nerine in liquid cultures Part II. Ontogeny of somatic embryos and bulblet regeneration. Plant Cell Tiss. Org. Cult., 39, 117–123.
Malik, M. (2008). Comparison of different liquid/solid culture systems in the production of somatic embryos from Narcissus L. ovary explants. Plant Cell Tiss. Org. Cult., 94, 337–345.
Mujib, A., Banerjee, S., Maqsood, M., Ghosh, P.D. (2013). Somatic embryogenesis of some member ornamental genera of Amaryllidaceae and allied families: the similarities and differences. Open Hortic. J., 6, 9–18.
Murashige, T., Skoog, F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol. Plant., 15, 473–479.
Ooi, L.S., Wang, H., Ng, T.B., Ooi, V.E. (1998). Isolation and characterization of mannosebinding lectin from leaves of Chinese daffodil Narcissus tazetta. Biochem. Cell Biol., 76, 601–608.
Ptak, A., Bach, A. (2007). Somatic embryogenesis in tulip (Tulipa gesneriana L.) flower stem cultures. In Vitro Cell Dev. Biol.-Plant, 43, 35–39.
Ramirez, A.E., Birnbaum, E.H. (2001). Induction and characterization of callus in Haloxylon aphyllum (Minkw.) Iljin. Proceedings IV International Symposium on In Vitro Culture & Horticulture Breeding. Acta Hortic., 560, 465–468.
Rocha, D.I., Monte-Bello, C.C., Dornelas, M.C. (2015). Alternative induction of de novo shoot organogenesis or somatic embryogenesis from in vitro cultures of mature zygotic embryos of passion fruit (Passiflora edulis Sims) is modulated by the ratio between auxin and cytokinin in the medium. Plant Cell Tiss. Org. Cult., 120, 1087–1098.
Sage, D.O., Lynn, J., Hammatt, N. (2000). Somatic embryogenesis in Narcissus pseudonarcissus cvs. Golden Harvest and St. Keverne. Plant Sci., 150, 209–216.
Seabrook, J.E.A., Cumming, B.G., Dionne, L.A. (1976). The in vitro induction of adventitious shoot and root apices on Narcissus (daffodil and narcissus) cultivar tissue. Can. J. Bot., 54, 814–819.
Selles, M., Bergonon, S., Viladomat, F., Bastida, J., Codina, C. (1997). Effect of sucrose on growth and galanthamine production in shoot-clump cultures of Narcissus confusus in liquidshake medium. Plant Cell Tiss. Org. Cult., 49, 129–136.
Selles, M., Viladomat, F., Bastida, J., Codina, C. (1999). Callus induction, somatic embryogenesis and organogenesis in Narcissus confusus: correlation between the state of differentiation and the content of galanthamine and related alkaloids. Plant Cell Rep., 18, 646–651.
Sochacki, D., Orlikowska, T. (1997). Inicjacja kultur narcyza z różnych eksplantatów. Zesz. Nauk. AR w Krakowie, nr 318. Sesja naukowa, 50, 199–202.
Ulrich, M.R., Davies, F.T.Jr., Koh, Y.C., Duray, S.A., Egilla, J.N. (1999). Micropropagation of Crinum ‘Ellen Bosanquet’ by tri-scales. Sci. Hortic., 82, 95–102.
Wawrosch, C., Malla, P.R., Kopp, B. (2001). Clonal propagation of Lilium nepalense D.Don, a threatened medicinal plant of Nepal. Plant Cell Rep., 20, 285–288.
Yoshida, K., Komae, K. (1994). Cell wall break down and enhancement of cellulase activity during the emergence of callus from rice roots in the presence of 2,4-D. Current tissue in plant molecular and cellular biology. Kluwer Academic Publ., 457–462.
Zimmerman, J.L. (1993). Somatic embryogenesis: a model for early development in higher plants. Plant Cell, 5, 1411–1423.
Ziv, M., Kahany, S., Lilien-Kipnis, H., 1994. Scaled-up proliferation and regeneration of Nerine in liquid cultures Part I. The induction and maintenance of proliferating meristematic clusters by paclobutrazol in bioreactors. Plant Cell Tiss. Org. Cult., 39, 109–115.
Articles are made available under the conditions CC BY 4.0 (until 2020 under the conditions CC BY-NC-ND 4.0).
Submission of the paper implies that it has not been published previously, that it is not under consideration for publication elsewhere.
The author signs a statement of the originality of the work, the contribution of individuals, and source of funding.