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Adaptation of an ecdysone-based genetic switch for transgene expression in soybean seeds

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Abstract

Soybean was used as a model for studies of chemical induction of gene expression in seeds. A chimeric transcriptional activator, VGE, driven by the soybean seed glycinin G1 promoter, was used to induce the expression of an ER-targeted GFPKDEL reporter protein upon addition of the chemical ligand, methoxyfenozide. The chemical gene switch activated gene expression under in vitro conditions in somatic cotyledonary embryos and zygotic seed embryos cultured from transgenic soybean plants, as well as in seeds in planta under greenhouse conditions. The efficiency of induction of GFP expression under different growth conditions was strongly influenced by the developmental stage of the seed and availability of the inducer. The formation of ER-derived GFP-containing protein bodies in seed storage parenchyma cells was correlated with the level of induced expression.

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Abbreviations

TSP:

Total soluble protein

MOF:

Methoxyfenozide

ER:

Endoplasmic reticulum

35SCaMV promoter:

Cauliflower mosaic virus 35S RNA promoter

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Acknowledgments

Dr. Eliot Herman is gratefully acknowledged for providing stimulating discussions and intellectual support. We also thank Dr. Howard Berg in the Integrated Microscopy Facility and Kevin Lutke from Tissue Culture Facility at the Danforth Plant Science Center for their kind assistance throughout this study, and Dr. Mala Padidam (Rohm and Haas Company), Dr. Wayne Parrott (University of Georgia) and Dr. Ed Cahoon (University of Nebraska) for providing certain genetics components for the gene switch and expression systems. This work was supported by grants from the USDA/Illinois-Missouri Alliance for Biotechnology, IMBA (TWT; 2006-5) and from The Consortium for Plant Biotechnology Research, Inc. (CPBR, Inc.) (RNB; GO12026-266).

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Author notes

  1. E. G. Semenyuk

    Present address: Department of Medicine, Northwestern University, 676 N. Saint Clair Street, #1400, Chicago, IL, 60611, USA

  2. T. Moravec

    Present address: Institute of Experimental Botany, Na Karlovce 1A, 160 00, Praha 6, Czech Republic

Authors and Affiliations

  1. Donald Danforth Plant Science Center, 975 N. Warson Road, Saint Louis, MO, 63132, USA

    E. G. Semenyuk, M. A. Schmidt, R. N. Beachy, T. Moravec & T. Woodford-Thomas

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  1. E. G. Semenyuk
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  2. M. A. Schmidt
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  3. R. N. Beachy
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  4. T. Moravec
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  5. T. Woodford-Thomas
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Correspondence to T. Woodford-Thomas.

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Semenyuk, E.G., Schmidt, M.A., Beachy, R.N. et al. Adaptation of an ecdysone-based genetic switch for transgene expression in soybean seeds. Transgenic Res 19, 987–999 (2010). https://doi.org/10.1007/s11248-010-9377-6

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  • DOI: https://doi.org/10.1007/s11248-010-9377-6

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