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An improved chemically inducible gene switch that functions in the monocotyledonous plant sugar cane

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Abstract

Chemically inducible gene switches can provide precise control over gene expression, enabling more specific analyses of gene function and expanding the plant biotechnology toolkit beyond traditional constitutive expression systems. The alc gene expression system is one of the most promising chemically inducible gene switches in plants because of its potential in both fundamental research and commercial biotechnology applications. However, there are no published reports demonstrating that this versatile gene switch is functional in transgenic monocotyledonous plants, which include some of the most important agricultural crops. We found that the original alc gene switch was ineffective in the monocotyledonous plant sugar cane, and describe a modified alc system that is functional in this globally significant crop. A promoter consisting of tandem copies of the ethanol receptor inverted repeat binding site, in combination with a minimal promoter sequence, was sufficient to give enhanced sensitivity and significantly higher levels of ethanol inducible gene expression. A longer CaMV 35S minimal promoter than was used in the original alc gene switch also substantially improved ethanol inducibility. Treating the roots with ethanol effectively induced the modified alc system in sugar cane leaves and stem, while an aerial spray was relatively ineffective. The extension of this chemically inducible gene expression system to sugar cane opens the door to new opportunities for basic research and crop biotechnology.

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Acknowledgments

We thank Michele Yarnall and Rachel Whinna (Syngenta Biotechnology Inc.) for carrying out all of the GUS qELISA analyses, Jamie Huang and Wenling Wang (Syngenta Biotechnology Inc.) for TaqMan analysis, Shujie Dong (Syngenta Biotechnology Inc.) for help with sample deliveries, and Kerry Caffall (Syngenta Biotechnology Inc.) for critical review of the manuscript. The authors also thank the staff at the Queensland Crop Development Facility (Department of Employment, Economic Development and Innovation, Queensland State Government) for their assistance with the growth of transgenic sugar cane. The Syngenta Centre for Sugarcane Biofuels Development is supported by Syngenta, the Queensland University of Technology, Farmacule Bioindustries, and by a grant from the National and International Research Alliances Program of the Queensland State Government. Dr. Mark Harrison is the recipient of a Smart State Fellowship from the Queensland State Government. The Queensland Crop Development Facility is funded by a grant from the Smart State Research Facilities Fund Scheme of the Queensland State Government. The work described in this manuscript is the subject of a patent application by Dr. Mark Kinkema and Dr. Manuel Sainz.

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Author notes

  1. Mark Kinkema

    Present address: AgBiTech, 8 Rocla Court, Glenvale, QLD, 4350, Australia

  2. R. Jason Geijskes & Paulo C. De Lucca

    Present address: Syngenta Proteção de Cultivos Ltda., Rodovia BR 452 km 142, Uberlândia, MG, 38407-049, Brazil

  3. Heather D. Coleman

    Present address: Department of Biology, Syracuse University, 107 College Place, Syracuse, NY, 13244, USA

  4. Anthony Palupe & James L. Dale

    Present address: Centre for Tropical Crops and Biocommodities, Queensland University of Technology, Brisbane, QLD, 4001, Australia

  5. Manuel B. Sainz

    Present address: Syngenta Proteção de Cultivos Ltda., Av Maurílio Biagi 800 9º andar, Ribeirão Prêto, SP, 14020-750, Brazil

Authors and Affiliations

  1. Syngenta Centre for Sugar Cane Biofuels Development, Centre for Tropical Crops and Biocommodities, Queensland University of Technology, Brisbane, QLD, 4001, Australia

    Mark Kinkema, R. Jason Geijskes, Kylie Shand, Heather D. Coleman, Paulo C. De Lucca, Anthony Palupe, Mark D. Harrison, James L. Dale & Manuel B. Sainz

  2. Syngenta Biotechnology Inc., Research Triangle Park, NC, 27709, USA

    Ian Jepson

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Correspondence to Mark Kinkema.

Additional information

Accession numbers: scoalcR (KC189903), scoGUS (KC189904), var4 (KC189905), var5 (KC189906), var8 (KC189907).

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Kinkema, M., Geijskes, R.J., Shand, K. et al. An improved chemically inducible gene switch that functions in the monocotyledonous plant sugar cane. Plant Mol Biol 84, 443–454 (2014). https://doi.org/10.1007/s11103-013-0140-2

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  • DOI: https://doi.org/10.1007/s11103-013-0140-2

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