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Genetic Modification of Grapevine Embryogenic Cultures

  • S. A. Dhekney
  • S. K. Sessions
  • M. Brungart-Rosenberg
  • C. Claflin
  • Z. T. Li
  • D. J. Gray
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1864)

Abstract

Precision breeding is an approach to grapevine genetic improvement that transfers only specific traits among sexually compatible species via the relatively stable mitotic cell division pathway in order to avoid the significant disruption imposed upon conventional breeding by meiosis. Factors enabling precision breeding include the availability of the Vitis genome sequence combined with highly optimized gene insertion and plant regeneration protocols. A protocol for the production of grapevine embryogenic cultures and their genetic transformation is described. Embryogenic cultures are produced from either leaf or floral explants. Somatic embryos at the cotyledonary stage of development are used for Agrobacterium-mediated transformation. Following co-cultivation with Agrobacterium containing the genes of interest, modified embryos are selected on the basis of anthocyanin pigmentation and antibiotic resistance. Somatic embryos are then germinated to produce modified plants that are hardened and transferred to a greenhouse. The presence of the genes of interest is confirmed by PCR.

Key words

Vitis Somatic embryogenesis Agrobacterium Transgenic plants Transgenes Culture medium Growth regulators Plant tissue culture 

Notes

Acknowledgments

S. A. Dhekney holds the E.A Whitney Endowed Professorship in the Department of Plant Sciences. M. Brungart-Rosenberg and C. Claflin were supported in part by a grant from the National Institute of General Medical Sciences (2P20GM103432) from the National Institute of Health. The research was supported in part by the Wyoming Agricultural Experiment Station and grants from the USDA/USAID US-Egypt Science and Technology Development Funds program (Grant No. 0210-22310-005-24G).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • S. A. Dhekney
    • 1
  • S. K. Sessions
    • 1
  • M. Brungart-Rosenberg
    • 1
  • C. Claflin
    • 2
  • Z. T. Li
    • 3
  • D. J. Gray
    • 4
  1. 1.Department of Plant SciencesSheridan Research and Extension Center, University of WyomingSheridanUSA
  2. 2.Sheridan CollegeSheridanUSA
  3. 3.USDA-ARSKearneysvilleUSA
  4. 4.Mid-Florida Research and Education Center, University of Florida/IFASApopkaUSA

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