Transformation of grape (Vitis vinifera L.) zygotic-derived somatic embryos and regeneration of transgenic plants
1995
Scorza, R. | Cordts, J.M. | Ramming, D.W. | Emershad, R.L.
Transgenic grape plants were regenerated from somatic embryos derived from immature zygotic embryos of seedless grape (Vitis vinifera L.) selections. Somatic embryos were bombarded twice with 1 micrometer gold particles using the Biolistic PDS-1000/He device (Bio-Rad Laboratories) and then exposed to Agrobacterium tumefaciens strain C58/Z707 containing the binary plasmid pGA482CG or pCGN73 14. Following cocultivation, secondary embryos were allowed lo proliferate on Emershad/Ramming proliferation (ERP) medium for 6 weeks before selection on ERP medium containg 20-40 microgram/ml kanamycin (kan). Transgenic embryos were identified after 3-5 months under selection and allowed lo germinate and develop into rooted plants on Woody P1ant Medium containing 1 micromolar 6-benzylaminopurine (BAP), 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 kan, positive GUS and PCR assays, and Southern analysis.
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