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High efficiency inducible gene expression system based on activation of a chimeric transcription factor in transgenic pine

  • Genetics and Genomics
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Abstract

Inducible gene expression systems are needed in functional genomics of tree species. A glucocorticoid-inducible gene expression system was established in a gymnosperm species Virginia pine (Pinus virginiana Mill.) through Agrobacterium tumefaciens-mediated genetic transformation. The results demonstrate that expression of the m-gfp5-ER reporter gene was tightly controlled and 0.1 μM of the glucocorticoid hormone triamcinolone was able to induce m-gfp5-ER expression in transgenic cells. Differential expression of gfp in transgenic cells induced by different concentrations of triamcinolone was observed and confirmed by Northern Blot analysis and by quantitative green fluorescence analyses with Laser Scanning Microscopy. In transgenic plantlets, triamcinolone was taken up efficiently by roots. Triamcinolone was able to induce m-gfp5-ER activity throughout the whole plant. The phenotype of transgenic plantlets was not affected 6 weeks after treatment with 0.1–10 μM triamcinolone. However, 6-week inductions with 100 μM triamcinolone caused growth retardation and developmental defects, as well as inhibition of root formation and elongation. With careful selection of transgenic lines, the inducible gene expression presented in this study could be a very valuable alternative for functional identification of novel genes in plants, especially in pine.

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Acknowledgements

The authors are grateful to Dr. P. B. F. Ouwerkerk and Dr. A. H. Meijer (Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, The Netherlands) for the gift of the vector pINDEX3, to Dr. C. N. Stewart (Department of Plant Sciences, University of Tennessee Knoxville, TN 37996, USA) and Dr. J. Haseloff (Department of Plant Sciences, University of Cambridge, UK) for providing us with the m-gfp5-ER constructs, and to Dr. D. Weidner (The Flow Cytometry-Confocal Microscopy Core Facility, the Brody School of Medicine, East Carolina University, NC 27858, USA) for technical assistance with laser scanning microscopy for imaging and quantitative analysis of green fluorescence

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  1. Department of Biology, Howell Science Complex, East Carolina University, Greenville, NC, 27858-4353, USA

    Wei Tang, Ronald J. Newton & Thomas M. Charles

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  1. Wei Tang
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  2. Ronald J. Newton
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  3. Thomas M. Charles
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Correspondence to Wei Tang.

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Communicated by I.S. Chung

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Tang, W., Newton, R.J. & Charles, T.M. High efficiency inducible gene expression system based on activation of a chimeric transcription factor in transgenic pine. Plant Cell Rep 24, 619–628 (2005). https://doi.org/10.1007/s00299-005-0009-1

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  • DOI: https://doi.org/10.1007/s00299-005-0009-1

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