B - Book chapters etc edited externally
Sparks, C. A. and Jones, H. D. 2014. Genetic transformation of wheat via particle bombardment. in: Henry, R. J. and Furtado, A. (ed.) Cereal genomics: methods and protocols. Methods in Molecular Biology, vol 1099 Springer, New York. pp. 201-218
Authors | Sparks, C. A. and Jones, H. D. |
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Editors | Henry, R. J. and Furtado, A. |
Abstract | Since its first invention in the late 1980s the particle gun has evolved from a basic gunpowder driven machine firing tungsten particles to one more refined which uses helium gas as the propellant to launch alternative heavy metal particles such as gold and silver. The simple principle is that DNA-coated microscopic particles (microcarriers) are accelerated at high speed by helium gas within a vacuum and travel at such a velocity as to penetrate target cells. However, the process itself involves a range of parameters which are open to variation: microparticle type and size, gun settings (rupture pressure, target distance, vacuum drawn, etc.), preparation of components (e.g., gold coating), and preparation of plant tissues. Here is presented a method optimized for transformation of wheat immature embryos using the Bio-Rad PDS-1000/He particle gun to deliver gold particles coated with a gene of interest and the selectable marker gene bar at 650 psi rupture pressure. Following bombardment, various tissue culture phases are used to encourage embryogenic callus formation and regeneration of plantlets and subsequent selection using glufosinate ammonium causes suppression of non-transformed tissues, thus assisting the detection of transformed plants. This protocol has been used successfully to generate transgenic plants for a wide range of wheat varieties, both spring and winter bread wheats (T. aestivum L.) and durum wheats (T. turgidum L.). |
Page range | 201-218 |
Year of Publication | 2014 |
Book title | Cereal genomics: methods and protocols. Methods in Molecular Biology, vol 1099 |
Publisher | Springer, New York |
ISBN | 9781627037150 |
Digital Object Identifier (DOI) | https://doi.org/10.1007/978-1-62703-715-0_17 |
Funder project or code | Wheat |
Wheat Transformation | |
Open access | Published as non-open access |
Output status | Published |
Publication dates | |
Online | 13 Oct 2013 |
Copyright license | Publisher copyright |
Permalink - https://repository.rothamsted.ac.uk/item/8qxxq/genetic-transformation-of-wheat-via-particle-bombardment
Publisher's version