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ISHS Acta Horticulturae 528: VII International Symposium on Grapevine Genetics and Breeding

TRANSFORMATION STUDIES ON VITIS VINIFERA L., VIA AGROBACTERIUM TUMEFACIENS

Authors:   B.-A. Bornhoff, M. Harst, E. Zyprian, R. Töpfer, C. Iannini
Keywords:   Agrobacterium tumefaciens, Vitis vinifera, embryo suspension culture, pH value, transformation
Abstract:
Somatic embryos of Vitis vinifera cv. DORNFELDER obtained from anthers were transformed via Agrobacterium tumefaciens strain LBA 4404 with a plasmid carrying the genes of beta-glucuronidase (GUS) with intron and neomycinphosphotransferase II (NPT). The somatic embryos proliferated in liquid NN69 medium containing kanamycin for selection and cefotaxim for removing the bacteria. Subsequent cultivation of larger embryos was carried out on solid NN69 medium supplemented with kanamycin. Transgenic shoots were selected and could be converted into rooted plantlets on LS medium. Transformation of the foreign genes was verified by beta-glucuronidase assays and PCR amplifications.

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