Bioactive beads-mediated transformation of rice with large DNA fragments containing Aegilops tauschii genes

Plant Cell Reports - Tập 28 - Trang 759-768 - 2009
Naoki Wada1, Shin’ichiro Kajiyama1, Yukio Akiyama1,2, Shigeki Kawakami1, Daisuke No1, Susumu Uchiyama1, Motoyasu Otani3, Takiko Shimada3,4, Naoko Nose5, Go Suzuki5, Yasuhiko Mukai5, Kiichi Fukui1
1Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Japan
2National Institute of Livestock and Grassland Science, Nasushiobara, Japan
3Research Institute of Agricultural Resources, Ishikawa Prefectural University, Ishikawa, Japan
4Department of Plant Sciences, University of Colombo, Colombo, Sri Lanka
5Laboratory of Plant Molecular Genetics, Division of Natural Science, Osaka Kyoiku University, Kashiwara, Japan

Tóm tắt

Transformation with large DNA molecules enables multiple genes to be introduced into plants simultaneously to produce transgenic plants with complex phenotypes. In this study, a large DNA fragment (ca. 100 kb) containing a set of Aegilops tauschii hardness genes was introduced into rice plants using a novel transformation method, called bioactive beads-mediated transformation. Nine transgenic rice plants were obtained and the presence of transgenes in the rice genome was confirmed by PCR and FISH analyses. The results suggested that multiple transgenes were successfully integrated in all transgenic plants. The expression of one of the transgenes, puroindoline b, was confirmed at the mRNA and protein levels in the T2 generation. Our study clearly demonstrates that the bioactive bead method is capable of producing transgenic rice plants carrying large DNA fragments. This method will facilitate the production of useful transgenic plants by introducing multiple genes simultaneously.

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