Improvement of Agrobacterium-mediated transformation efficiency and transgenic plant regeneration of Vitis vinifera L. by optimizing selection regimes and utilizing cryopreserved cell suspensions

• Author(s): Gafny, R.; Hanania, Uri; Li, Ping; Mawassi, M.; Perl, Avihai; Sahar, N.; Sela, I.; Tanne, E.; Wang, Qiaochun;
• Abstract: Various concentrations of kanamycin and paromomycin were examined for their effect on embryogenic cell suspension viability, transformation efficiency, and transgenic plant regeneration. Paromomycin (10-25 mg/1) induced an earlier killing effect on cell suspensions than kanamycin (40-100 mg/l). Transformation efficiency and the number of embryos developed on selection medium were positively correlated with an increase in paromomycin concentrations from 10 to 20 or 25 mg/l. Paromomycin was more effective than kanamycin in selection of transformed cells and induction of embryo development during selection. Although no differences in transformation efficiency were observed between cryopreserved and non-cryopreserved cells, conversions of transformed cells to embryos and plantlets were significantly improved when cryopreserved cells were used as the target material. With the improved transformation procedure proposed, 65-80% of GUS-positive cells and embryos were produced with about 40% of transformed embryos regenerated into plants. Copyright 2004 Elsevier Ireland Ltd. All rights reserved.
• Publication Date: Feb 2005
• Journal: Plant Science (Oxford)