Abstract
The protocol described below was initially developed by Shimamoto et al. (1989) and has proved to be highly reproducible and efficient in production of fertile transgenic rice plants (for reviews, Shimamoto 1992; Terada and Shimamoto 1993). The method works well for a wide range of japonica varieties and various genes have been transferred using hygromycin resistance gene (hph) as a selectable marker. The novel nurse culture method (Kyozuka et al. 1987) is used for culture of transformed protoplasts.
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.
References
Baba A, Hasazawa S, Shono K (1986) Cultivation of rice protoplasts and their transformation mediated by Agrobacterium spheroplasts. Plant Cell Physiol 27: 463–472
Harms CT, Potrykus I (1978) Fractionation of plant protoplast types by iso-osmotic density gradient centrifugation. Theor Appl Genet 53: 57–63
Kyozuka J, Hayashi Y, Shimamoto K (1987) High frequency plant regeneration from rice protoplasts by novel nurse culture methods. Mol Gen Genet 206: 408–413
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497
Ohira K, Ojima K, Fujiwara A (1973) Studies on the nutrition of rice cell culture 1. A simple, defined medium for rapid growth in suspension culture. Plant Cell Physiol 14: 1113–1121
Shimamoto K (1992) Genetic manipulation of rice: from protoplasts to transgenic plants. Jpn J Genet 67: 273–290
Shimamoto K, Terada R, Izawa T, Fujimoto H (1989) Fertile transgenic rice plants regenerated from transformed protoplasts. Nature 338: 274–276
Terada R, Shimamoto K (1993) Rice as a model cereal for studies of transformation in monocots. J Plant Res Special Issue 3: 201–211
Yoshida S, Forno DA, Cock JH, Gomez KA (1976) Laboratory manual for physiological studies of rice. The International Rice Research Institute, Philippines, pp 61–66
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Wu, C., Shimamoto, K. (1995). Electroporation-Mediated Gene Transfer to Japonica Rice Protoplasts and Regeneration of Transgenic Plants. In: Potrykus, I., Spangenberg, G. (eds) Gene Transfer to Plants. Springer Lab Manual. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-79247-2_11
Download citation
DOI: https://doi.org/10.1007/978-3-642-79247-2_11
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-48967-9
Online ISBN: 978-3-642-79247-2
eBook Packages: Springer Book Archive