Summary
Agitated layers of liquid medium were created on platform shakers in jars with 25–30 ml of medium (similar to conventional agar culture) rotating at 90 rpm. Thin films were scaled up in larger rectangular vessels on tilted shelves that periodically rock. In jars of liquid medium with a density of 180 explants per liter, multiplication rates of Hota tokudama var. ‘Newberry Gold’ were optimal with a media sucrose concentration of 5% [both with and without 1 μM benzyladenine (BA)]. Endogenous levels of soluble sugars were directly related to the concentration of sucrose in the medium. Three Hosta cultivars (‘Striptease’, ‘Minuteman’, and ‘Stiletto’) with plant densities of 40–200 explants per liter of medium were tested in larger, agitated, thin-film vessels in media with 5% sucrose and directly compared to agar medium. Higher rates of multiplication were observed in liquid than agar with the magnitude of the difference dependent on explant density. Pooled results for the three varieties with 200 explants per liter showed multiplication rates of 1.7x and 2.3x for agar and thin-film liquid, respectively. At 40 explants per liter, the multiplication rate was increased to 2.1x for agar and 3.4x for thin-film liquid. Sugar uptake was greater in liquid than agar and was greater in the higher densities, with the magnitude of the effect dependent on plant variety. Increased vessel size in the liquid, thin-film system and greater sugar uptake allowed more, larger plants to be harvested. Alocasia macrorrhizos was cultured in growth medium containing 1μM BA and 5% sucrose with plant densities in the range of 33–330 explants per liter. Dry weight and multiplication rate were greater in the liquid system than agar with the magnitude of the difference dependent on plant density. With approximately 165 explants per liter, and greater at the initiation of culture, plant density limited growth in both agar and liquid thin-film systems. In a multiplication medium (3 μM BA and 3 μM ancymidol) plant size was reduced by 50% and 60% (fresh weight) in liquid and agar, respectively. Initial density in the range of 165–330 explants per liter did not limit growth with the smaller plants in liquid or semisolid multiplication medium. Sugar uptake was greater in liquid than agar. While ample sugar was present in media for growth at any density on agar, sugar depletion was limiting growth at highest densities with the larger plants in liquid growth medium. In semisolid agar medium, sugar uptake by plants was more rapid than diffusion across the agar medium, resulting in non-equilibrium conditions following the culture cycle. In agitated, liquid medium, a greater transfer of sugars to plant tissue was related to accelerated growth.
Similar content being viewed by others
References
Adelberg, J. Efficiency in thin-film liquid systems for micropropagation of Hosta. Plant Cell Tiss. Organ Cult. (in press); 2004.
Adelberg, J.; Kroggel, M.; Toler, J. Greenhouse and nursery growth of micropropagated Hostas from liquid culture. HortTechnology 10:754–757; 2000.
Adelberg, J.; Simpson, E. Intermittent immersion vessel apparatus and process for plant propagation. US patent pending, pub. no. US 2002/0155595 A1; 2002.
Adelberg, J.; Toler, J. Comparison of agar and thin-film liquid systems for micropropagation of ornamental Alocasia and Colocasia. HortScience (in press); 2004.
Etienne, H.; Berthouly, M. Temporary immersion systems in plant micropropagation. Plant Cell Tiss. Organ Cult. 69:215–231; 2002.
Gollagunta, V.; Adelberg, J.; Riek, J.; Rajapakse, N. Sucrose concentration in liquid media affects soluble carbohydrates, biomass and storage quality of micropropagated Hosta. Plant Cell Tiss. Organ Cult. (in press); 2004.
Levin, R.; Tanny, G. Bioreactors as a low cost option for tissue culture. In: Ahloowalia, B., ed. Low cost options in tissue culture for developing countries. IAEA TecDoc. Vienna: International Atomic Energy Agency; 2004 (in press).
Miller, W.; Langhans, R. Carbohydrate changes of Easter lily during growth in normal and reduced irradiance environments. J. Am. Soc. Hort. Sci. 114:310–315; 1989.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Smith, M. A. L.; Spomer, L. Vessels, gels, liquid media and support systems. In: Aitken-Christie, J.; Kozai, T.; Smith, M. A. L., eds. Automation and environmental control in plant tissue culture. Dordecht: Kluwer Academic Publishers; 1995:371–405.
Williams, R. R. The chemical microenvironment. In: Aitken-Christie, J.; Kozai, T.; Smith, M. A. L., eds. Automation and environmental control in plant tissue culture. Dordecht: Kluwer Academic Publishers; 1995:406–440.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Adelberg, J. SIVB 2003 Congress Symposium Proceeding: Plant Growth and Sugar Utilization in an Agitated, Thin-Film Liquid System for Micropropagation. In Vitro Cell.Dev.Biol.-Plant 40, 245–250 (2004). https://doi.org/10.1079/IVP2003511
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1079/IVP2003511