Summary
The use of scaled-up liquid cultures could be an efficient system for mass propagation of Narcissus, as it can greatly reduce the costs involved with manual handling. Induction of hyperhydric meristematic leaf section clusters and proliferation were carried out in an ancymidol (ANC)-containing liquid medium in flasks and disposable presterilized plastic bioreactors. Non-hyperhydric bulblets started to develop from hyperhydric meristematic leaf section clusters after subculture on a 0.8% agar strength medium, and young bulbs formed after 10 mo. in vivo acclimatization with a 98% survival rate. The present study reveals that in Narcissus leaf sections cultured in liquid medium, morphogenetic changes in leaf sections were associated with metabolic changes. The changes in carbohydrate, protein, and water potential of the liquid media and leaf sections were found to be closely related to meristematic center initiation on Narcissus hyperhydric leaf sections. Starch, sucrose, and glucose were significantly higher in the hyperhydric leaf sections cultured in ANC medium. The water potential was signifieantly higher in ANC-treated leaf sections and significantly lower in the medium containing ANC, at the stage shortly before or after hyperhydricity and meristematic centers hegan forming on the leaf sections. A 30kDa protein was found to be present in the hyperhydric leaf sections. Based on the present study, a largescale micropropagation protocol of Narcissus in agar and liquid cultures is proposed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Chen, J.; Ziv, M. The effect of Ancymidol on hyperhydricity, regeneration, starch and antioxidant enzymatic activities in liquid-cultured Narcissus. Plant Cell Rep. 20:22–27; 2001.
Chin, C. K. Promotion of shoot and root formation in asparagus in vitro by ancymidol. HortScience 17:590–591; 1982.
Cleland, R. The control of cell enlargement. In: Jennings, D. H., ed. Society for Experimental Biology Symposium 31. Cambridge: Cambridge University Press; 1977:101–115.
De Hertogh, A. A.; Aung, L. H.; Benschop, M. The tulip: botany, usage, growth, and development. Hort. Rev. 5:45–125; 1983.
De Klerk-Kiebert, Y. M.; Van Der Plas, L. H. W. Relationship of respiratory pathways in soybean cell suspensions to growth of the cells at various glucose concentrations. Plant Cell Tiss. Organ Cult. 4:225–233; 1985.
Hanks, C. R. Effects of growth retardants on bulbil production by Narcissus twin-scales. Ann. Appl. Biol. 110:203–207; 1987.
Levin, R.; Gaba, V.; Tal, B.; Hirsch, S.; Nola, D.; Vasil, I. K. Automated plant tissue culture for mass propagation. Bio/Technology 6:1035–1040; 1988.
Miller, W. B. A review of carbohydrate metabolism in geophytes. Acta Hort. 325:239–249; 1992.
Murashige, T.; Nakano, R. T. The light requirement for shoot initiation in tobacco callus culture. Am. J. Bot. 55:710; 1968.
Murashige, T.; Skoog, F. revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Preil, W. Application of bioreactors in plant propagation. In: Debergh, P. C.; Zimmerman, R. H., eds. Micropropagation—technology and application. Dordrecht: Kluwer Academic Publishers; 1991:425–445.
Schaffer, A. A.; Petreikov, M. Sucrose-to-starch metabolism in tomato fruit undergoing transient starch accumulation. Plant Physiol. 113:739–746; 1997.
Stark, D. M.; Timmerman, K. P.; Barry, G. F.; Preiss, J.; Kishore, G. M. Regulation of the amount of starch in plant tissues by ADP glucose pyrophosphorylase. Science 258:287–292; 1992.
Styer, D. J. Biorcactor technology for plant propagation. In: Henke, R. R.; Hughes, K. W.; Constantin, M. J.; Hollaender, A., eds. Tissue culture in forestry and agriculture. New York: Plenum Press; 1985:117–130.
Takayama, S.; Swedlund, B.; Miwa, Y. Automated propagation of microbulbs of lilies. In: Vasil, I., ed. Scale-up and automation in plant propagation, cell culture and somatic cell genetic of plants, vol. 8. New York: Academic Press; 1990:112–131.
Theron, K. I.; Jacobs, G. Changes in carbohydrate composition of the different bulb components of Nerine bocdenii W. Watson (Amaryllidaceae). J. Am. Soc. Hort. Sci. 121:343–346; 1996.
Thivend, P.; Mercier, C.; Guilbot, A. Determination of starch with glycoamylase. In: Wistler, R. L.; BeMiller, J. N., eds. Methods in carbohydrate chemistry, vol. 6. New York: Academic Press; 1972:100–105.
Thorpe, T. A.; Meier, D. D. Starch metabolism, respiration and shoot formation in tobacco callus cultures. Physiol. Plant. 27:365–369; 1972.
Van Huystee, R. S. Some molecular aspects of plant peroxidase biosynthetic studies. Annu. Rev. Plant Physiol. 38:205–219; 1987.
Vishnevetsky, J.; Zamski, E.; Ziv, M. Carbohydrate metabolism in Nerine sarniensis bulbs developing in liquid culture. Physiol. Plant. 108:361–369; 2000.
Zamski, E.; Grumberger, Y. Short- and long-eared high-yielding hexaploid wheat cultivars: which has unexpressed potential for higher yield? Ann. Bot. 75:501–506; 1995.
Ziv, M. Vitrification. In: Debergh, P. C.; Zimmerman, R. H., eds. Micropropagation—technology and application. Dordrecht: Kluwer Academic Publishers; 1991:45–69.
Ziv, M. The use of growth retardants for the regulation and acclimatization of in vitro plants. In: Karsen, C. M.; Van Loon, L. C.; Vregdenhil, D., eds. Progress in plant growth regulation. Dordrecht: Kluwer Academic Publishers; 1992:809–817.
Ziv, M. In vitro acclimatization. In: Aitken-Christie, J.; Kozai, T.; Smith, M. L., eds. Automation and environmental control in plant tissue culture. Dordrecht: Kluwer Academic Publishers: 1995:493–516.
Ziv, M. Bioreactor technology for plant micropropagation. Hort. Rev. 24:1–30; 2000.
Ziv, M.; Ariel, T. Bud proliferation and plant regeneration in liquid-cultured Philodendron treated with ancymidol and paclobutrazol. Plant Growth Regul. 10:53–57; 1991.
Ziv, M.; Kahany, S.; Lilien-Kipnis, H. Sealed-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. Organ Cult. 39:109–115; 1994.
Ziv, M.; Ronen, G.; Raviv, M. Proliferation of meristematic clusters in disposable presterilized plastic bioreators for large-scale micropropagation of plants. In Vitro Cell. Dev. Biol. Plant 34:152–158; 1998.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chen, J., Ziv, M. Carbohydrate, metabolic, and osmotic changes in scaled-up liquid cultures of Narcissus leaves. In Vitro Cell Dev Biol -Plant 39, 645–650 (2003). https://doi.org/10.1079/IVP2003451
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1079/IVP2003451