Abstract
Maize (Zea mays L.) protoplasts obtained from Type I and Type II calli from several genotypes were shown to be capable of synthesizing cell walls and forming small clusters of cells. The medium used also supported cluster formation from protoplasts obtained from root tips. The effects of various additions to the medium (such as casein hydrolysate, coconut water, amino acids, sugars, phytohormones, nitrate, calcium, and dimethylsulfoxide as well as pH variations on cellcluster formation were determined. The method of culture (protoplasts plated in agarose or supported in alginate beads in liquid medium) as well as several components of the medium were found to be critical for microcallus formation. Protoplasts obtained from embryogenic Type I callus and cultured in the medium of C. Nitsch and J.P. Nitsch (1967, Planta 72, 355–370) modified by various additions (NN 67-mod medium) were affected most by various sugars, casein hydrolysate, coconut water, and a combination of the auxins napthalene-1-acetic acid (2 mg/l) and 2,4-dichlorophenoxyacetic acid (0.1 mg/l), and the cytokinin N6-benzylaminopurine (0.5 mg/l). Cluster size in the agarose culture system was from 0.1 to 0.5 mm diameter and in the alginate culture system, up to 2.0 mm diameter.
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References
Bui-Dang-Ha, D., Mackenzie, I.A. (1973) The division of protoplasts from Asparagus officinalis L. and their growth and differentiation. Protoplasma 78, 215–221
Carlberg, I., Glimelius, K., Eriksson, T. (1983) Improved culture ability of potato protoplasts by use of activated charcoal. Plant Cell Rep. 2, 223–225
Chin, J.C., Scott, K.J. (1979) A large scale isolation procedure for cereal mesophyll protoplasts. Ann. Bot. 43, 23–32
Chu, C.C., Wang, C.C., Sun, C.S., Hsu, C., Yin, K.C., Chu, C.Y. (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci. Sinica 18, 659–668
Evans, D.A., Bravo, J.E. (1983) Protoplast isolation and culture. In: Handbook of plant cell culture, pp. 124–176, Evans, D.A., Sharp, W.R., Ammirato, P.V., Yamada, Y., eds. Academic Press, New York
Green, C.E. (1982) Somatic embryogenesis and plant regeneration from the friable callus of Zea mays. In: Plant tissue culture 1982 (Proc. V. Int. Congr. Plant Tissue and Cell Culture), pp. 107–108, Fujuwara, A., ed. Japanese Association for Plant Tissue Culture, Tokyo, Japan
Hahne, G., Hoffmann, F. (1984) Dimethyl sulfoxide can initiate cell divisions of arrested callus protoplasts by promoting cortical microtubule assembly. Proc. Natl. Acad. Sci. USA 81, 5449–5453
Hess, D., Leipoldt, G. (1979) Regeneration of shoots and roots from isolated mesophyll protoplasts of Nemesia strumosa. Biochem. Physiol. Pfl. 174, 411–417
Hodges, T.K., Kamo, K.K., Becwar, M.R., Schroll, S. (1985) Regeneration of maize. In: Biotechnology in plant science: revelence to agriculture in the eighties, pp. 15–33, Zaitlin, M., Day, P., Hollaender, A., eds. Academic Press, New York
Imbrie-Milligan, C.W., Hodges, T.K. (1986) Microcallus formation from maize protoplasts prepared from embryogenic callus. Planta 168, 395–401
Johansson, L., Andersson, B., Eriksson, T. (1982) Improvement of anther culture technique: Activated charcoal bound in agar medium in combination with liquid medium and elevated CO2 concentration. Physiol. Plant. 54, 24–30
Kamo, K.K., Becwar, M.R., Hodges, T.K. (1985) Regeneration of Zea mays L. from embryogenic callus. Bot. Gaz. 146, 327–334
Kao, K.N., Michayluk, M.R. (1975) Nutrient requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126, 105–110
King, P.J., Potrykus, I., Thomas, E. (1978) In vitro genetics of cereal: Problems and perspectives. Physiol. Vég. 16, 381–399
Kohlenbach, H.W., Wernicke, W. (1977) Investigations on the inhibitory effect of agar and the function of active carbon in anther culture. Z. Pflanzenphysiol. 86, 463–472
Koop, H.-U., Weber, G., Schweiger, H.-G. (1983) Individual culture of selected single cells and protoplasts of higher plants in microdroplets of defined media. Z. Pflanzenphysiol. 112, 21–34
Landgren, C.R. (1976) The influence of culture conditions on mitotic activity in protoplasts derived from Pisum root cortical explants. Protoplasma 87, 49–69
Linsefors, L., Brodelius, P. (1985) Immobilization of plant protoplasts: Viability studies. Plant Cell Rep. 4, 23–27
Morgan, A., Cocking, E.C. (1982) Plant regeneration from protoplasts of Lycopersicum esculentum Mill. Z. Pflanzenphysiol. 106, 97–104
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol. Plant. 15, 473–497
Nitsch, C., Nitsch, J.P. (1967) The induction of flowering in vitro in stem segments of Plumbago indica L. II. The production of reproductive buds. Planta 72, 355–370
Rao, P.S. (1982) Protoplast culture. In: Experimental embryology of vascular plants, pp. 231–262, Johri, B.M., ed. Springer, Berlin
Scheurich, P., Schnabl, H., Zimmermann, U., Klein, J. (1980) Immobilisation and mechanical support of individual protoplasts. Biochim. Biophys. Acta 598, 645–651
Schnabl, H., Youngman, R.J. (1985) Immobilisation of plant cell protoplasts inhibits enzymic lipid peroxidation. Plant Science 40, 65–69
Schnabl, H., Youngman, R.J., Zimmermann, U. (1983) Maintenance of plant cell membrane integrity and function by the immobilisation of protoplasts in alginate matrices. Planta 158, 392–397
Shahin, E.A. (1983) Totipotency of tomato protoplasts. Theor. Appl. Gen. 69, 235–240
Stuart, R., Street, H.E. (1971) Studies on the growth in culture of plant cells. X. Further studies on the conditioning of culture media by suspensions of Acer pseudoplatanus L. cells. J. Exitl. Bot 22, 96–106
Uchimiya, H., Murashige, T. (1976) Influence of the nutrient medium on the recovery of dividing cells from tobacco protoplasts. Plant Physiol. 57, 424–429
Vasil, V., Vasil, L.K. (1986) Plant regeneration from friable embryogenic callus and cell suspension cultures of Zea mays L. J. Plant Physiol. 124, 399–408
Von Arnold, S., Eriksson, T. (1977) A revised medium for growth of pea mesophyll protoplasts. Physiol. Plant. 39, 257–260
White, P.R. (1963) The cultivation of animal and plant cells. Ronald Press, New York
Widholm, J.M. (1972) The use of fluorescein diacetate and phenosafranin for determining viability of cultured plant cells. Stain Technol. 47, 189–193
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Imbrie-Milligan, C., Kamo, K.K. & Hodges, T.K. Microcallus growth from maize protoplasts. Planta 171, 58–64 (1987). https://doi.org/10.1007/BF00395067
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DOI: https://doi.org/10.1007/BF00395067