Summary
Particle bombardment, or biolistics, is a commonly used method for genetic transformation of plants and other organisms. Millions of DNA-coated metal particles are shot at target cells or tissues using a biolistic device or gene gun. The DNA elutes off the particles that lodge inside the cells, and a portion may be stably incorporated in the host chromosomes. A protocol for the generation of transgenic grapevines via biolistic transformation of embryogenic cell suspension cultures is detailed in this chapter. In a typical experiment, transient gene expression averaged nearly 8000 “hits” per bombarded plate. Five months after bombardment, there were nearly five putative transgenic embryos per bombarded plate. About half of the embryos were regenerated into confirmed transgenic plants. The basic bombardment procedures described are applicable to a wide range of plant genotypes, especially those for which embryogenic cell cultures are available. All users of particle bombardment technology will find numerous useful tips to maximize the success of transformation.
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References
Sanford, J. C., Klein, T. M, Wolf, E. D., and Allen, N. (1987). Delivery of substances into cells and tissues using a particle bombardment process. Particulate Sci. Technol. 5, 27–37.
Sanford, J. C. (2000) The development of the biolistic process. In Vitro Cell. Dev. Biol. Plant 36, 303–308.
Kikkert, J. R. (1993) The Biolistic® PDS-1000/He device. Plant Cell Tiss. Org. Cult. 33, 221–226.
Southgate, E. M., Davey, M. R., Power, J. B., and Marchant, R. (1995). Factors affecting the genetic engineering of plants by microprojectile bombardment. Biotechnol. Adv. 13, 631–651.
Taylor, N. J. and Fauquet, C. M. (2002) Microparticle bombardment as a tool in plant science and agricultural biotechnology. DNA Cell Biol. 21, 963–977.
McCabe, D. and Christou, P. (1993) Direct DNA transfer using electric discharge particle acceleration (ACCELL™ technology). Plant Cell Tiss. Org. Cult. 33, 227–236.
Sanford, J. C., Smith, F. D., and Russell, J. A. (1993) Optimizing the biolistic process for different biological applications. Methods Enzymol. 217, 483–509.
Sanford, J. C., DeVit, M. J., Russell, J. A., et al. (1991) An improved, helium-driven biolistic device. Technique 3, 3–16.
Perl, A., Lotan, O., Abu-Abied, M., and Holland, D. (1996) Establishment of an Agrobacterium-mediated transformation system for grape (Vitis vinifera L.): The role of antioxidants during grape-Agrobacterium interactions. Nat. Biotechnol. 14, 624–628.
Francois, I. E. J. A., Broekaert, W. F., and Cammue, B. P. A. (2002) Different approaches for multi-transgene-stacking in plants. Plant Sci. 163, 281–295.
Fu, X., Duc, L. T., Fontana, S., et al. (2000) Linear transgene constructs lacking vector backbone sequences generate low-copy-number transgenic plants with simple integration patterns. Transgen. Res. 9, 11–19.
Srivastava, V. and Ow, D. (2001) Biolistic mediated site-specific integration in rice. Mol. Breed. 8, 345–350.
Vidal, J. R., Kikkert, J. R., Wallace, P. G., and Reisch, B. I. (2003) High-efficiency biolistic co-transformation and regeneration of ‘Chardonnay’ (Vitis vinifera L.) containing npt-II and antimicrobial peptide genes. Plant Cell Rep. 22, 252–260.
Mauro, M. C., Toutain, S., Walter, B., et al. (1995) High efficiency regeneration of grapevine plants transformed with the GFLV coat protein gene. Plant Sci. 112, 97–106.
Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plantarum 15, 473–497.
Lloyd, G. and McCown, B. (1980) Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Int. Plant Prop. Soc. Proc. 30, 421–427.
Russell, J. A., Roy, M. K., and Sanford, J. C. (1992) Major improvements in biolistic transformation of suspension-cultured tobacco cells. In Vitro Cell. Dev. Biol. 28P, 97–105.
Vain, P., McMullen, M. D., and Finer, J. J. (1993) Osmotic treatment enhances particle bombardment-mediated transient and stable transformation of maize. Plant Cell Rep. 12, 84–88.
Finer, J. J. and McMullen, M. D. (1991) Transformation of soybean via particle bombardment of embryogenic suspension culture tissue. In Vitro Cell. Dev. Biol. 27P, 17–182.
Russell, J. A., Roy, M. K., and Sanford, J. C. (1992) Physical trauma and tungsten toxicity reduce the efficiency of biolistic transformation. Plant Physiol. 98, 1050–1056.
Sawant, S. S., Singh, P. K., and Tuli, R. (2000) Pretreatment of microprojectiles to improve the delivery of DNA in plant transformation. BioTechniques 29, 246–248
Birch, R. G. and Franks, T. (1991) Development and optimisation of microprojectile systems for plant genetic transformation. Aust. J. Plant Physiol. 18, 453–469.
Smith, F. D., Harpending, P. R., and Sanford, J. C. (1992) Biolistic transformation of prokaryotes: factors that affect biolistic transformation of very small cells. J. Gen. Microbiol. 138, 239–248.
Martinelli, L. and Mandolino, G. (1994) Genetic transformation and regeneration of transgenic plants in grapevine (Vitis rupestris S.). Theor. Appl. Genet. 88, 621–628.
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Kikkert, J.R., Vidal, J.R., Reisch, B.I. (2005). Stable Transformation of Plant Cells by Particle Bombardment/Biolistics. In: Peña, L. (eds) Transgenic Plants: Methods and Protocols. Methods in Molecular Biology™, vol 286. Humana Press. https://doi.org/10.1385/1-59259-827-7:061
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DOI: https://doi.org/10.1385/1-59259-827-7:061
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Print ISBN: 978-1-58829-263-6
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