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Genetic transformation of sweet sorghum

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Abstract

Sweet sorghum has substantial potential as a biofuel feedstock, with advantages in some environments over alternatives such as sugarcane or maize. Gene technologies are likely to be important to achieve yields sufficient for food, fuel and fibre production from available global croplands, but sorghum has proven difficult to transform. Tissue culture recalcitrance and poor reproducibility of transformation protocols remain major challenges for grain sorghum, and there has been no reported success for sweet sorghum. Here we describe a repeatable transformation system for sweet sorghum, based on (1) optimized tissue culture conditions for embryogenic callus production with >90% regenerability in 12-week-old calli, and (2) an effective selection regimen for hygromycin resistance conferred by a Ubi-hpt transgene following particle bombardment. Using this method, we have produced sixteen independent transgenic lines from multiple batches at an overall efficiency of 0.09% transformants per excised immature embryo. Co-expression frequency of a non-selected luciferase reporter was 62.5%. Transgene integration and expression were confirmed in T0 and T1 plants by Southern analysis and luciferase assays. This success using the major international sweet sorghum cultivar Ramada provides a foundation for molecular improvement of sweet sorghum through the use of transgenes. Factors likely to be important for success with other sweet sorghum cultivars are identified.

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Acknowledgments

We thank Mona Singh, Alam Cheng, Amanda Johnson for outstanding technical assistance. This research was supported through a collaboration between CSR Sugar Limited and The University of Queensland, including a grant under the Australian Government Renewable Energy Development Initiative. We thank Queensland DPI&F and Pacific Seeds for supplying seed material to conduct this research.

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Authors and Affiliations

  1. Botany Department/BIOL, The University of Queensland, Brisbane, 4072, Australia

    Anshu Raghuwanshi & Robert G. Birch

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  1. Anshu Raghuwanshi
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  2. Robert G. Birch
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Correspondence to Robert G. Birch.

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Communicated by P. Lakshmanan.

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Raghuwanshi, A., Birch, R.G. Genetic transformation of sweet sorghum. Plant Cell Rep 29, 997–1005 (2010). https://doi.org/10.1007/s00299-010-0885-x

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  • DOI: https://doi.org/10.1007/s00299-010-0885-x

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