Published: Vol 4, Iss 19, Oct 5, 2014 DOI: 10.21769/BioProtoc.1258 Views: 13262
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Abstract
Like many woody plant species, walnut (Juglans regia) can be difficult to genetically transform and regenerate. However, somatic embryos have been used successfully for over two decades as a target tissue for transformation and regeneration of transgenic walnut plants. Walnut somatic embryos, initiated originally from developing zygotic embryos or anther tissue, will proliferate numerous secondary embryos from single cells in the epidermal layer. These single cells in intact somatic embryos can be efficiently transformed by Agrobacterium tumefaciens (A. tumefaciens). This gene transfer system is most efficient when Agrobacterium binary vector plasmids contain a scorable maker gene (e.g. uidA) and a selectable marker gene (e.g. nptII). This system should be applicable to any crop that undergoes repetitive embryogenesis from single Agrobacterium-susceptible cells. Here we describe the method of transforming somatic embryos in detail so that this technique can be applied to walnut and other woody plant species.
Materials and Reagents
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Acknowledgments
Note that a similar protocol has been described by Leslie et al. (2006) and Dandekar et al. (1989).
References
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Copyright
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
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Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
Category
Plant Science > Plant transformation > Agrobacterium
Plant Science > Plant developmental biology > General
Molecular Biology > DNA > Transformation
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