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Improved molecular tools for sugar cane biotechnology

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Abstract

Sugar cane is a major source of food and fuel worldwide. Biotechnology has the potential to improve economically-important traits in sugar cane as well as diversify sugar cane beyond traditional applications such as sucrose production. High levels of transgene expression are key to the success of improving crops through biotechnology. Here we describe new molecular tools that both expand and improve gene expression capabilities in sugar cane. We have identified promoters that can be used to drive high levels of gene expression in the leaf and stem of transgenic sugar cane. One of these promoters, derived from the Cestrum yellow leaf curling virus, drives levels of constitutive transgene expression that are significantly higher than those achieved by the historical benchmark maize polyubiquitin-1 (Zm-Ubi1) promoter. A second promoter, the maize phosphonenolpyruvate carboxylate promoter, was found to be a strong, leaf-preferred promoter that enables levels of expression comparable to Zm-Ubi1 in this organ. Transgene expression was increased approximately 50-fold by gene modification, which included optimising the codon usage of the coding sequence to better suit sugar cane. We also describe a novel dual transcriptional enhancer that increased gene expression from different promoters, boosting expression from Zm-Ubi1 over eightfold. These molecular tools will be extremely valuable for the improvement of sugar cane through biotechnology.

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Acknowledgments

We thank Michele Yarnall and Rachel Whinna (Syngenta Biotechnology, Incorporated) for carrying out all of the GUS qELISA analyses, Jamie Huang and Wenling Wang (Syngenta Biotechnology, Incorporated) for TaqMan analysis, and Shujie Dong (Syngenta Biotechnology, Incorporated) for help with sample deliveries. We also would like to acknowledge Mark Harrison, Robyn Lloyd, Rachael Hassall, Amanda Johnson, and Jan Zhang for their assistance with plant sampling, and Mark Harrison for critical review of the manuscript. We would like to acknowledge a potential conflict of interest in that the author Dr Mark Kinkema has filed a patent application for the dual transcriptional enhancer sequence described in this manuscript.

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Author notes

  1. Anthony Palupe

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

  2. Mark Kinkema

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

  3. Jason Geijskes & Paulo deLucca

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

  4. Heather D. Coleman

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

  5. Manuel Sainz

    Present address: Syngenta Proteção de Cultivos Ltda., Av Maurílio Biagi 800 9° 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, Jason Geijskes, Paulo deLucca, Anthony Palupe, Kylie Shand, Heather D. Coleman, Manuel Sainz & James L. Dale

  2. Centre for Tropical Crops and Biocommodities, Queensland University of Technology, Brisbane, QLD, 4001, Australia

    Anthony Brinin, Brett Williams & James L. Dale

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  1. Mark Kinkema
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Correspondence to Mark Kinkema.

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Kinkema, M., Geijskes, J., deLucca, P. et al. Improved molecular tools for sugar cane biotechnology. Plant Mol Biol 84, 497–508 (2014). https://doi.org/10.1007/s11103-013-0147-8

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