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  • Author or Editor: R.L. Emershad x
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Immature grape embryos from early ripening genotypes of Vitis vinifera were successfully cultured in vitro on Difco orchid agar or a modified White's agar medium. Germination was increased in vitro for five genotypes from 0%, 7%, 11%, 12%, and 16% in vivo to 15%, 24%, 23%, 34%, and 24%, respectively. Subculturing embryos onto liquid culture from seeds that failed to germinate on agar also was possible. Differences in germination rates, as affected by pollen, were significant. This method will allow accelerated development of early ripening cultivars by allowing breeders to use such genotypes as females, as well as males.

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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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