Nature Biotechnology
14, 624 - 628 (1996)
doi:10.1038/nbt0596-624
Establishment of an Agrobacterium-mediated transformation system for grape (Vitis vinifera L.): The role of antioxidants during grape−Agrobacterium interactionsAvihai Perl*, 1, Ofra Lotan1, M. Abu-Abied1
& D. Holland1
1Department of Fruit Tree Breeding and Molecular Genetics, Agricultural Research Organization, The Volcani Center, P.O. Box 6 Bet-Dagan, 50250 Israel.
*e-mail: vhlperl@volcani.agri.gov.il. Very short exposures of embryogenic calli of Vitis vinifera cv. Superior Seedless grape plants to diluted cultures of Agrobacterium resulted in plant tissue necrosis and subsequent cell death. Antibiotics used for Agrobacterium elimination or as plant selectable markers were not responsible for this necrotic response. Rather, cell death seemed to be oxygen-dependent and correlated with elevated levels of peroxides. Therefore, we studied the effects on necrosis of various combinations of antioxidants during and after grape-Agrobacterium cocultivation. The combination of polyvinylpolypyrrolidone and dithiothreitol was found to improve plant viability. Tissue necrosis was completely inhibited by these antioxidants while Agrobacterium virulence was not effected. These treatments enabled the recovery of stable transgenic grape plants resistant to hygromycin.
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