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Cereal Genomics pp 219–234Cite as

Sorghum Genetic Transformation by Particle Bombardment

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1099))

Abstract

Particle bombardment transformation describes the acceleration of high-velocity microparticles coated with exotic genes through the plant-protective cell walls, in order for the introduced genes to be integrated into the host genome. This technique has proven to be an effective and versatile approach towards plant genetic modification in preceding decades. Particle bombardment has been successfully applied to cereals including rice, maize, wheat, barley, and sorghum. Historically, sorghum has been considered as one of the most recalcitrant major crops with regard to successful genetic transformation; however, tremendous progress has been made in recent years. Transformation efficiency by particle bombardment has now improved from approximately 1 % to in excess of 20 % utilizing an optimized tissue culture and DNA delivery system. The protocol described in this chapter routinely generates transformants at 10–25 % efficiency within sorghum genotype Tx430. The process generally takes 11–16 weeks from initiation of immature embryos to planting of transformants. This protocol covers the operation of both the Bio-Rad PDS-1000/He System and particle inflow gun. Three factors are crucial to an efficient particle bombardment transformation system: (1) an efficient tissue culture system, (2) a highly efficient DNA delivery system, and (3) an effective selection strategy.

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Acknowledgements

We are thankful to the Australian Research Council (ARC) and Pacific Seeds Ltd. Pty. for funding the Linkage project LP0883808. We are grateful to Sharon Beth Williams, Siti Atiqah Zainul Alam, Azelah Mustapha, and Yue Sun for their efforts during the editing process.

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Authors and Affiliations

  1. School of Agriculture and Food Sciences, The University of Queensland, Brisbane, QLD, Australia

    Guoquan Liu & Bradley C. Campbell

  2. School of Agriculture and Food Sciences, The University of Queensland, St. Lucia, QLD, Australia

    Ian D. Godwin

Authors
  1. Guoquan Liu
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  2. Bradley C. Campbell
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  3. Ian D. Godwin
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Editors and Affiliations

  1. Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St. Lucia, Queensland, Australia

    Robert J. Henry

  2. Centre for Nutrition and Food Science, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, Lismore, New South Wales, Australia

    Agnelo Furtado

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Liu, G., Campbell, B.C., Godwin, I.D. (2014). Sorghum Genetic Transformation by Particle Bombardment. In: Henry, R., Furtado, A. (eds) Cereal Genomics. Methods in Molecular Biology, vol 1099. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-715-0_18

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  • DOI: https://doi.org/10.1007/978-1-62703-715-0_18

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-714-3

  • Online ISBN: 978-1-62703-715-0

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