Perl A, Lotan O, Abu-Abied M, Holland D
Department of Fruit Tree Breeding and Molecular Genetics, Volcani Center, Bet-Dagan, Israel.
Nat Biotechnol. 1996 May;14(5):624-8. doi: 10.1038/nbt0596-624.
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.
将酿酒葡萄品种“超级无核”葡萄植株的胚性愈伤组织短暂暴露于稀释的农杆菌培养物中，会导致植物组织坏死及随后的细胞死亡。用于消除农杆菌或作为植物选择标记的抗生素并非这种坏死反应的原因。相反，细胞死亡似乎依赖于氧气，且与过氧化物水平升高相关。因此，我们研究了葡萄与农杆菌共培养期间及之后各种抗氧化剂组合对坏死的影响。发现聚乙烯聚吡咯烷酮和二硫苏糖醇的组合可提高植物活力。这些抗氧化剂完全抑制了组织坏死，而农杆菌的毒性未受影响。这些处理使得能够获得对潮霉素具有抗性的稳定转基因葡萄植株。
