Abstract
Protoplasts were isolated from rice callus and lettuce cotyledons. Their fusion process was studied by light and electron microscopy. Lettuce protoplast fusion could be easily produced by either of polyethylene glycol and electric methods. The transformation of fusion bodies during the fusion process was more easily observed by using the electric method than chemical one. After electric pulse application, the fusion of lettuce protoplasts was finished within about one hr, judging from their externals. On the other hand, the fusion of rice protoplasts occurred in a very short time after electric pulse application. Fusion was performed between lettuce protoplasts and rice protoplasts. Rice protoplasts contain no chloroplasts and lettuce protoplasts contain them. Therefore, both protoplasts could be easily distinguished under a light microscope. When the fusion between rice and lettuce protoplasts was induced by electric pulse application, chloroplasts in lettuce protoplasts moved into rice protoplasts within about one hr. Electron micrographs showed that the components from both protoplasts was located separately in the fusion bodies. Big rice fusion bodies produced by fusion of rice protoplasts contained some round-shaped particles. Their size was almost the same as that of the original protoplasts. These results indicate that it take a long time until the cytoplasm of rice protoplasts mix with the cytoplasm of lettuce protoplasts in the fusion body.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
Binding, H., Krumbiegel-Schroeren, G., and Nehls, R. (1986) ‘Protoplast fusion and early development of fusants’, in J. Reinert and H. Binding (eds), Results and Problems in Cell Differentiation 12, Differentiation of Protoplasts and Transformed Plant Cell, Springer-Verlag Berling Heidelberg, pp. 37 – 66.
Burgess, J., and Fleming, E.N. (1974) ‘Ultrastructural studies of the aggregation and fusion of plant protoplasts’, Planta 118, 183–193.
Chen, L. B. (1989) ‘Fluorescent labeling of mitochondria’ in Y.-L. Wang, and D. Taylor (eds) Method in Cell Biology vol. 29, Fluorescence Microscopy of Living Cells in Culture, Part A, Fluorescent Analogs, Labeling Cells, and Basic Microscopy, Academic Press, San Diego, pp. 103–123.
Cocking, E. C. (1960) ‘A method for the isolation of plant protoplasts and vacuoles’, Nature 187, 962–963.
Davey, M. R., Pearce, N., and Cocking, E. C. (1980) ‘Fusion of legume root nodule protoplasts with non-legume protoplasts: Ultrastructural evidence for the functional activity of Rhizobiumbacteroids in a heterokaryotic cytoplasm’, Z. Pflanzenphysiol. 99, 435–447.
Draget, K. I., Myhre, S., Evjen, K., and Østgaard, K. (1988) ‘Plant protoplast immobilized in calcium alginate; a simple method of preparing fragile cell for transmission electron microscopy’, Stain Tech. 63, 159–164.
Evans, D. A., Bravo, J. E., and Gleba, Y. Y. (1983) ‘Somatic hybridization: Fusion methods, recovery of hybrids, and genetic analysis’, Intern. Rev. Cytol. Suppl. 16, 143–159.
Fowke, L. C. (1980) ‘Applications of protoplasts to the study of plant cells’, Intern. Rev. Cytol. 68, 9–51.
Gahan, P. B. (1984) ‘Plant Histochemistry and Cytochemistry’, Academic Press, London.
Kanai, R., and Edwards, G. E. (1973) Purification of enzymatically isolated mesophyll protoplasts from C3, C4 and crassulacean acid metabolism pints using a aqueous dextran polyethylene glycol two-phase system. Plant Physiol. 52, 484–490.
Küster, E. (1909) ‘Über die Verschmelzung nackter Protoplasten’, Ber. Dtsch. Bot. Ges. 27, 589–598.
Lynch, P. T., Isaac S., and Collin, H. A. (1989) ‘Electrofusion of protoplasts from celery (Apium graveolens L.) with protoplasts from the filamentous fungus Aspergillus nidulans’, Planta 178, 207–214.
Maeda, E, and Maeda, K. (1987) ‘Ultrastructural studies of leaf hydathodes. I. Wheat (Triticum aestivum) leaf tips’, Japan. Jour. Crop Sci. 56, 641–651.
Nagata, T., and Takebe, I. (1971) ‘Plating of isolated tobacco mesophyll protoplasts on agar medium’, Planta 99, 12–20.
Otsuki, Y., and Takebe, I. (1969) ‘Isolation of intact mesophyll cells and their protoplasts from higher plants’, Plant Cell Physiol. 10, 917–921.
Rennie, P. J., Weber, G., Constabel,, F., and Fowke, L. C. (1980) ‘Dedifferentiation of chloroplasts in interspecific and homospecific protoplast fusion products’, Protoplasma 103, 253–262.
Schieder, O., and Vasil, I. K. (1980) ’Protoplast fusion and somatic hybridization’, Intern. Rev. Cytol. Suppl. IIB, 21–46.
Senda, M., Takada, J., Abe, S., and Nakamura, T. (1979) ‘Induction of cell fusion of plant protoplasts by electrical stimulation’, Plant Cell Physiol. 20, 1441–1443.
Shahin, E. A., and Shepard, J. F. (1980) ‘Cassava mesophyll protoplsts; isolation, prolifiration, and shoot formation’, Plant Sci. Lett. 17, 459–465.
Sumardi, I., Taniguchi, T., and Maeda, E. (1987) ‘Effect of enzyme solutions on the isolation of protoplasts from rice, lettuce, sugarcane and coffee leaves’, Rep. Tokai Br. Crop Sci. Soc. Japan 104, 7–13.
Takebe, I., Labib, G., and Melchers, G. (1971) ‘Regeneration whole plants from isolated mesophyll protoplasts of tobacco’, Naturwissenschften 58, 318–320.
Takebe, I., Otsuki, Y., and Aoki, S. (1968) ‘Isolation of tobacco mesophyll cells in intact and active state’, Plant Cell Physiol. 9, 115–124.
Taniguchi, T., and Maeda, E. (1988) ?Fluorescence microscopic observations of chloroplasts in plant protoplasts?, Rep. Tokai Br. Crop Sci. Soc. Japan 106, 15–16.
Taniguchi, T., Sanada, Y., and Maeda,-.E. (1987) ?Effect of coconut water and coconut cream on organ development in calli of rice and lettuce?, Rep. Tokai Br. Crop Sci. Soc. Japan 103, 25–29.
Taniguchi, T., Sato, T., and Maeda, E. (1989) ‘Observation of plant plant protoplasts by various light-microscopes’, Rep. Tokai Br. Crop Soc. Japan 107, 33–38.
Thomas, M. R., and Rose, R. J. (1988) ‘Enrichment for Nicotiana heterokaryons after protoplast fusion and subsequent growth in agarose microdrops’, Planta 175, 396–402.
Togawa, Y., Toda, K., Takayama, S., Miura, Y., Mochisuki, T., and Iwasaki, I. (1987) ‘Cell fusion by electric fields’, Shmadzu Rev. 44, 17–28.
Wu, F. S. (1987) ‘Localization of mitochondria in plant cells by vital staining with rhodamine 123’, Planta 171, 346–357.
Zachrisson, A., and Bornman, C. H. (1986) ‘Electromanipulation of plant protoplasts’, Physiol. Plant. 67, 507–516.
Zimmermann, U., and Scheurich, P. (1981) ‘High frequency fusion of plant protoplasts by electrical fields’, Planta 151, 26–32.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1990 Kluwer Academic Publishers
About this chapter
Cite this chapter
Taniguchi, T., Sato, T., Maeda, K., Maeda, E. (1990). Microscopic Observations of Fusion Process of Rice and Lettuce Protoplasts. In: Sangwan, R.S., Sangwan-Norreel, B.S. (eds) The Impact of Biotechnology on Agriculture. Current Plant Science and Biotechnology in Agriculture, vol 8. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0587-0_18
Download citation
DOI: https://doi.org/10.1007/978-94-009-0587-0_18
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-6752-2
Online ISBN: 978-94-009-0587-0
eBook Packages: Springer Book Archive