Abstract
Agrobacterium-mediated transformation of rice has been routinely performed according to the protocol reported by Hiei et al. (Plant J. 6:271–282, 1994). However, several elite japonica and many indica varieties cannot be efficiently transformed by Agrobacterium system. Also a large number of transformants are required to generate T-DNA insertion and FOX libraries as well as gene-targeting studies. To overcome these challenges, we established a high-efficiency transformation system in rice by cocultivating rice calli with Agrobacterium on filter papers moistened with enriched liquid media instead of using solid media (Ozawa, Plant Sci. 176:522–527, 2009; Ozawa and Takaiwa, Plant Sci. 179:333–337, 2010). In this system, the transformation efficiency of the calli is almost 100% in many varieties.
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References
Hiei, Y., Ohta, S., Komari, T., Kumashiro, T. (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6, 271–282.
Ozawa, K. (2009) Establishment of a high efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.). Plant Sci. 176, 522–527.
Ozawa, K., Takaiwa, F. (2010) Highly efficient Agrobacterium-mediated transformation of suspension-cultured cell clusters of rice (Oryza sativa L.). Plant Sci. 179, 333–337.
Chu, C.C., Wang, C.C., Sun, C.S., Hsu, C., Yin, K.C., Chu, C.Y., Bi, F.Y. (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on nitrogen sources. Sci. Sin. 18, 659–668.
Daigen, M., Kawakami, O., Nagasawa, Y. (2000) Efficient anther culture method of the japonica rice cultivar Koshihikari. Breed. Sci. 50, 197–202.
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497.
Hiei, Y., Komari, T., Kubo, T. (1997) Transformation of rice mediated by Agrobacterium tumefaciens. Plant Mol. Biol. 35, 205–218.
Jefferson, R. (2007) Science as Social Enterprise: The CAMBIA BiOS Initiative. Innovations: Technology, Governance, Globalization. Vol. 1, No. 4, 13–44.
Kawai, K., Kaku, K., Izawa, N., Shimizu, T., Fukuda, A., Tanaka, Y. (2007) A novel mutant acetolactate synthase gene from rice cells, which confers resistance to ALS-inhibiting herbicides. J. Pestic. Sci. 32, 89–98.
Acknowledgments
The author would like to thank Dr. Fumio Takaiwa and Dr. Sakiko Hirose for their advice and encouragement.
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Ozawa, K. (2012). A High-Efficiency Agrobacterium-Mediated Transformation System of Rice (Oryza sativa L.). In: Dunwell, J., Wetten, A. (eds) Transgenic Plants. Methods in Molecular Biology, vol 847. Humana Press. https://doi.org/10.1007/978-1-61779-558-9_5
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DOI: https://doi.org/10.1007/978-1-61779-558-9_5
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