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Development of genotype-independent regeneration system for transformation of rice (Oryza sativa ssp. indica)

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Abstract

Rice (Oryza sativa ssp. indica) is an important economic crop in many countries. Although a variety of conventional methods have been developed to improve this plant, manipulation by genetic engineering is still complicated. We have established a system of multiple shoot regeneration from rice shoot apical meristem. By use of MS medium containing 4 mg L−1 thidiazuron (TDZ) multiple shoots were successfully developed directly from the meristem without an intervening callus stage. All rice cultivars tested responded well on the medium and regenerated to plantlets that were readily transferred to soil within 5–8 weeks. The tissue culture system was suitable for Agrobacterium-mediated transformation and different factors affecting transformation efficiency were investigated. Agrobacterium strain EHA105 containing the plasmid pCAMBIA1301 was used. The lowest concentration of hygromycin B in combined with either 250 mg L−1 carbenicillin or 250 mg L−1 cefotaxime to kill the rice shoot apical meristem was 50 mg L−1 and carbenicillin was more effective than cefotaxime. Two-hundred micromolar acetosyringone had no effect on the efficiency of transient expression. Sonication of rice shoot apical meristem for 10 s during bacterial immersion increased transient GUS expression in three-day co-cultivated seedlings. The gus gene was found to be integrated into the genome of the T0 transformant plantlets.

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Abbreviations

BA:

6-Benzylaminopurine

cv:

Cultivar

GUS:

β-Glucuronidase

TDZ:

Thidiazuron

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Acknowledgements

We would like to thank Dr Richard Jefferson for providing the plasmid pCAMBIA1301 and the Pathum Thani Rice Research Center, Thailand, for the rice seeds. This research was supported in part by Thailand Research Fund No. RSA/4/2538; The Faculty of Graduate Studies, Mahidol University; The Department of Agriculture; and The Ministry of Education, Commission of Higher Education, subproject “Graduate Study and Research in Agricultural Biotechnology”, Thailand.

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok, 10400, Thailand

    Nimnara Yookongkaew & Jarunya Narangajavana

  2. Department of Agriculture, Office of Biotechnology Research and Development, Rangsit Ongkarak Road, Klong 6, Thanyaburi, Pathumtani, 12110, Thailand

    Methinee Srivatanakul

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  1. Nimnara Yookongkaew
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  2. Methinee Srivatanakul
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  3. Jarunya Narangajavana
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Correspondence to Jarunya Narangajavana.

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Yookongkaew, N., Srivatanakul, M. & Narangajavana, J. Development of genotype-independent regeneration system for transformation of rice (Oryza sativa ssp. indica). J Plant Res 120, 237–245 (2007). https://doi.org/10.1007/s10265-006-0046-z

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  • DOI: https://doi.org/10.1007/s10265-006-0046-z

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