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Agrobacterium-mediated transformation of the dwarf pomegranate (Punica granatum L. var. nana)

  • Genetic Transformation and Hybridization
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Abstract

The dwarf pomegranate (Punica granatum L. var. nana) is a dwarf ornamental plant that has the potential to be the model plant of perennial fruit trees because it bears fruits within 1 year of seedling. We established an Agrobacterium-mediated transformation system for the dwarf pomegranate. Adventitious shoots regenerated from leaf segments were inoculated with A. tumefaciens strain EHA105 harboring the binary vector pBin19-sgfp, which contains neomycin phosphotransferase (npt II) and green fluorescent protein (gfp) gene as a selectable and visual marker, respectively. After co-cultivation, the inoculated adventitious shoots were cut into small pieces to induce regeneration, and then selected on MS medium supplemented with 0.5 μM α-naphthaleneacetic acid (NAA), 5 μM N6-benzyladenine (BA), 0.3% gellan gum, 50 mg/l kanamycin, and 10 mg/l meropenem. Putative transformed shoots were regenerated after 6–8 months of selection. PCR and PCR-Southern blot analysis revealed the integration of the transgene into the plant genome. Transformants bloomed and bore fruits within 3 months of being potted, and the inheritance of the transgene was confirmed in T1 generations. The advantage of the transformation of dwarf pomegranate was shown to be the high transformation rate. The establishment of this transformation system is invaluable for investigating fruit-tree-specific phenomena.

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Acknowledgments

The authors would like to thank Dr. Jen Sheen (Department of Molecular Biology, Massachusetts General Hospital and Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA) for providing the binary plasmid pBin19-sgfp and Zeneca Mogen for providing the A. tumefaciens strain EHA105.

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Author notes

  1. Nagao Matsuta

    Present address: National Agricultural Research Center for Western Region, Zentsuji, Japan

Authors and Affiliations

  1. University of Tsukuba, Tsukuba, Japan

    Shingo Terakami, Sumiko Sugaya & Hiroshi Gemma

  2. University of Tsukuba, National Institute of Fruit Tree Science, Tsukuba, Japan

    Toshiya Yamamoto

  3. National Institute of Fruit Tree Science, Tsukuba, Japan

    Junichi Soejima

Authors
  1. Shingo Terakami
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  2. Nagao Matsuta
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  3. Toshiya Yamamoto
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  4. Sumiko Sugaya
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  5. Hiroshi Gemma
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  6. Junichi Soejima
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Corresponding author

Correspondence to Sumiko Sugaya.

Additional information

Communicated by H. Ebinuma.

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Terakami, S., Matsuta, N., Yamamoto, T. et al. Agrobacterium-mediated transformation of the dwarf pomegranate (Punica granatum L. var. nana). Plant Cell Rep 26, 1243–1251 (2007). https://doi.org/10.1007/s00299-007-0347-2

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  • DOI: https://doi.org/10.1007/s00299-007-0347-2

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