Producing Transgenic `Thompson Seedless' Grape (Vitis vinifera L.) Plants

in Journal of the American Society for Horticultural Science
Authors:
R. ScorzaU.S. Department of Agriculture-Agricultural Research Service, Appalachian Fruit Research Station, 45 Wiltshire Road, Kearneysville, WV 25430

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J.M. CordtsU.S. Department of Agriculture-Agricultural Research Service, Appalachian Fruit Research Station, 45 Wiltshire Road, Kearneysville, WV 25430

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D.J. GrayCentral Florida Research and Education Center, Institute of Food and Agricultural Sciences, University of Florida, 5336 University Avenue, Leesburg, FL 34748

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D. GonsalvesDepartment of Plant Pathology, Cornell University, Geneva, NY 14456

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R.L. EmershadU.S. Department of Agriculture-Agricultural Research Service, Horticultural Research Laboratory, 2021 South Peach Avenue, Fresno, CA 93727

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D.W. RammingU.S. Department of Agriculture-Agricultural Research Service, Horticultural Research Laboratory, 2021 South Peach Avenue, Fresno, CA 93727

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Transgenic grape plants were regenerated from somatic embryos derived from leaves of in vitro-grown plants of `Thompson Seedless' grape (Vitis vinifera L.) plants. Somatic embryos were either exposed directly to engineered Agrobacterium tumefaciens or they were bombarded twice with 1-μm gold particles and then exposed to A. tumefaciens. Somatic embryos were transformed with either the lytic peptide Shiva-1 gene or the tomato ringspot virus (TomRSV) coat protein (CP) gene. After cocultivation, secondary embryos proliferated on Emershad/Ramming proliferation (ERP) medium for 6 weeks before selection on ERP medium containing 40 μg·mL-1 kanamycin (kan). Transgenic embryos were identified after 3 to 5 months under selection and allowed to germinate and develop into rooted plants on woody plant medium containing 1 μm 6-benzylaminopurine, 1.5% sucrose, 0.3% activated charcoal, and 0.75% agar. Integration of the foreign genes into these grapevines was verified by growth in the presence of kanamycin (kan), positive β-glucuronidase (GUS) and polymerase chain-reaction (PCR) assays, and Southern analysis.

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