A fluorescent antibiotic resistance marker for rapid production of transgenic rice plants

Tetsuko Ochiai-Fukuda, Naoko Takahashi-Ando, Shuichi Ohsato, Tomoko Igawa, Kaori Kadokura, Hiroshi Hamamoto, Masayoshi Nakasako, Toshiaki Kudo, Takehiko Shibata, Isamu Yamaguchi, Makoto Kimura

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Blasticidin S (BS) is an aminoacylnucleoside antibiotic used for the control of rice blast disease. To establish a new cereal transformation system, we constructed a visual marker gene designated gfbsd, encoding an enhanced green fluorescent protein (EGFP) fused to the N-terminus of BS deaminase (BSD). It was cloned into a monocot expression vector and introduced into rice (Oryza sativa L. cv. Nipponbare) calluses by microprojectile bombardment. Three to five weeks after the bombardment, multicellular clusters emitting bright-green EGFP fluorescence were obtained with 10 μg/ml BS, which is not sufficient to completely inhibit the growth of non-transformed tissues. Fluorescent sectors (approximately 2 mm in diameter) excised from the calluses regenerated into transgenic plantlets (approximately 10 cm in height) as early as 51 (average 77 ± 11) days after the bombardment. The visual antibiotic selection was more efficient and required less time than the bialaphos selection with bar. In addition, the small size (1.1 kb) of gfbsd is preferable for construction of transformation vectors. This new marker gene will make a significant contribution in molecular genetic studies of rice plants.

Original languageEnglish
Pages (from-to)521-527
Number of pages7
JournalJournal of Biotechnology
Volume122
Issue number4
DOIs
Publication statusPublished - 2006 Apr 20

Keywords

  • Agricultural antibiotics
  • Blasticidin S resistance
  • Cereal transformation
  • Enhanced green fluorescent protein (EGFP)
  • Fusion reporter/marker gene
  • Genetically modified organisms (GMO)

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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  • Cite this

    Ochiai-Fukuda, T., Takahashi-Ando, N., Ohsato, S., Igawa, T., Kadokura, K., Hamamoto, H., Nakasako, M., Kudo, T., Shibata, T., Yamaguchi, I., & Kimura, M. (2006). A fluorescent antibiotic resistance marker for rapid production of transgenic rice plants. Journal of Biotechnology, 122(4), 521-527. https://doi.org/10.1016/j.jbiotec.2005.09.015