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Green fluorescent protein as a visual selection marker for papaya (Carica papaya L.) transformation

  • Genetic Transformation and Hybridization
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Abstract

Chemical-based selection for plant transformation is associated with a number of real and perceived problems that might be avoided through visual selection. We have used green fluorescent protein (GFP), as a visual selectable marker to produce transformed papaya (Carica papaya) plants following microprojectile bombardment of embryogenic callus. GFP selection reduced the selection time from 3 months on a geneticin (G418) antibiotic-containing medium to 3–4 weeks. Moreover, GFP selection increased the number of transformed papaya plants by five-to eightfold compared to selection in the presence of antibiotics. Overall, the use of GFP for selecting transgenic papaya lines improved our throughput for transformation by 15- to 24-fold while avoiding the drawbacks associated with the use of antibiotic resistance-based selection markers.

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Fig. 1
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Fig. 3a–e
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Fig. 5a, b

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Abbreviations

BA::

Benzyladenine

2, 4-D::

2,4-Dichlorophenoxyacetic acid

GFP::

Green fluorescent protein

IBA::

Indole-3-butyric acid

NAA::

α-Naphthaleneacetic acid

MS::

Murashige and Skoog plant culture medium

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Acknowledgements

This research was supported in part by a US Department of Agriculture ARS cooperative agreement No. CA58-5320-3-460 with the Hawaii Agriculture Research Center. The authors would like to thank Dr. Mingli Wang, HARC, for providing the pML202 construct, CAMBIA, Canberra, Australia, for providing the pCAMBIA1303 construct, and Dr. Henrik Albert, USDA, ARS, for thoughtful discussions and for technical assistance with the fluorescence microscopy.

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Authors and Affiliations

  1. Hawaii Agriculture Research Center, 99-193 Aiea Heights Drive, Aiea, Hawaii, 96701, USA

    Y. J. Zhu & R. Agbayani

  2. USDA-ARS, PBARC, 99-193 Aiea Heights Drive, Aiea, Hawaii , 96701, USA

    P. H. Moore

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  1. Y. J. Zhu
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  2. R. Agbayani
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  3. P. H. Moore
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Correspondence to Y. J. Zhu.

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Communicated by R.J. Rose

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Zhu, Y.J., Agbayani, R. & Moore, P.H. Green fluorescent protein as a visual selection marker for papaya (Carica papaya L.) transformation. Plant Cell Rep 22, 660–667 (2004). https://doi.org/10.1007/s00299-004-0755-5

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  • DOI: https://doi.org/10.1007/s00299-004-0755-5

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