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Reef-coral proteins as visual, non-destructive reporters for plant transformation

  • Cell Biology and Morphogenesis
  • Published:
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Abstract

Recently, five novel fluorescent proteins have been isolated from non-bioluminescent species of reef-coral organisms and have been made available through ClonTech. They are AmCyan, AsRed, DsRed, ZsGreen and ZsYellow. These proteins are valuable as reporters for transformation because they do not require a substrate or external co-factor to emit fluorescence and can be tested in vivo without destruction of the tissue under study. We have evaluated them in a large range of plants, both monocots and dicots, and our results indicate that they are valuable reporting tools for transformation in a wide variety of crops. We report here their successful expression in wheat, maize, barley, rice, banana, onion, soybean, cotton, tobacco, potato and tomato. Transient expression could be observed as early as 24 h after DNA delivery in some cases, allowing for very clear visualization of individually transformed cells. Stable transgenic events were generated, using mannose, kanamycin or hygromycin selection. Transgenic plants were phenotypically normal, showing a wide range of fluorescence levels, and were fertile. Expression of AmCyan, ZsGreen and AsRed was visible in maize T1 seeds, allowing visual segregation to more than 99% accuracy. The excitation and emission wavelengths of some of these proteins are significantly different; the difference is enough for the simultaneous visualization of cells transformed with more than one of the fluorescent proteins. These proteins will become useful tools for transformation optimization and other studies. The wide variety of plants successfully tested demonstrates that these proteins will potentially find broad use in plant biology.

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Abbreviations

BMS:

Black Mexican sweet maize

CMP :

Cestrum virus promoter

GUS :

β-Glucuronidase

LUC :

Luciferase

nos :

Nopaline synthetase

pmi :

Phosphomannose isomerase

PCR :

Polymerase chain reaction

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Acknowledgements

The authors would like to thank BD Biosciences Clontech for providing the reef-coral fluorescent protein vectors. We would also like to thank the support teams—quality control, media, greenhouse, licensing and legal—at both Jealott's Hill and Research Triangle Park for making this work possible.

Author information

Author notes

  1. A. Wenck

    Present address: BASF Plant Science, 26 Davis Dr, Research Triangle Park, NC 27709, USA

  2. M. Turner

    Present address: Danisco AS, Langebrogade 1, 1001, Copenhagen, Denmark

  3. G. Legris

    Present address: European Development Centre, Dow AgroSciences, 3 Milton Park, Abingdon, OX14 4RN, UK

Authors and Affiliations

  1. Syngenta Biotechnology Inc., 3054 Cornwallis Road, Research Triangle Park, North Carolina, NC 27709, USA

    A. Wenck, M. Dunn, C. Stacy, E. Dunder, E. van Grinsven, R. Khan, M. Sigareva, W. C. Wang, J. Reed, K. Launis, Y.-F. Chang & L. Melchers

  2. Syngenta Jealott's Hill International Research Centre, Bracknell, RG42 6EY, UK

    C. Pugieux, M. Turner, A. Tiozzo, P. Drayton, D. Oliver, H. Trafford, G. Legris, H. Rushton, S. Tayab & D.-F. Chen

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  1. A. Wenck
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  2. C. Pugieux
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  3. M. Turner
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  8. E. van Grinsven
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Corresponding author

Correspondence to A. Wenck.

Additional information

Communicated by M.C. Jordan

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Wenck, A., Pugieux, C., Turner, M. et al. Reef-coral proteins as visual, non-destructive reporters for plant transformation. Plant Cell Rep 22, 244–251 (2003). https://doi.org/10.1007/s00299-003-0690-x

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  • DOI: https://doi.org/10.1007/s00299-003-0690-x

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