Establishment of an Agrobacterium-mediated transformation system for grape (Vitis vinifera L.): The role of antioxidants during grape–Agrobacterium interactions

Abstract

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.

Agrobacterium-mediated transformation of sorghum: factors that affect transformation efficiency

“…Infection solutions with less than 1.0 x 10 8 cfu/mL reduced embryo death to acceptable levels, but no GUS activity was observed. A similar necrotic process elicited by Agrobacterium has been reported in grape (Perl et al, 1996;Pu and Goodman, 1992) and Catharanthus roseus cells (Garner et al 1996). In grapes it was assumed that necrogenesis was due to a hypersensitive response of the grape cells to the Agrobacterium (Perl et al, 1996).…”

“…A similar necrotic process elicited by Agrobacterium has been reported in grape (Perl et al, 1996;Pu and Goodman, 1992) and Catharanthus roseus cells (Garner et al 1996). In grapes it was assumed that necrogenesis was due to a hypersensitive response of the grape cells to the Agrobacterium (Perl et al, 1996). The addition of PVPP or PVPP+DTT to the inoculation, co-cultivation or post-co-cultivation media slightly reduced browning of immature embryos, but did not improve embryo survival rate after co-cultivation.…”

“…Following co-cultivation the embryos were cultured for seven days on I8+C medium or I8+C medium supplemented with 0.5% PVPP and covered with sufficient liquid I8+C medium (i.e. I8+C medium without agarose) containing 0.2% (w/v) DTT to cover the embryos (Perl et al, 1996).…”

Identification and Characterization of a Fruit-Specific, Thaumatin-Like Protein That Accumulates at Very High Levels in Conjunction with the Onset of Sugar Accumulation and Berry Softening in Grapes

“…Progress in this area is likely to require severa1 lines of research. With the discovery that a TL protein is synthesized at a specific time of fruit development, and with the recent development of transformation systems for grape (Mauro et al, 1995;Perl et al, 1996;Scorza et al, 1996), important clues to the functions of VVTLl may now be gained from attenuation of its expression in grapes with parallel studies in other ripening fruits in which TL proteins have been shown to accumulate to high levels.…”

Agrobacterium-mediated genetic transformation of a tropical elite maize line

“…Cysteine, dithiothreitol (DTT), polyvinyl pyrrolidone, and ascorbic acid (Perl et al 1996, Olhoft and Somers 2001, Paz et al 2004, Frame et al 2006, Mishra et al 2014) are antioxidants that have been used to increase the potential of Agrobacterium to infect tissues by inhibiting the antimicrobial exudates produced by plant cells during the infection and cocultivation stages and/or by preventing the occurrence of necrosis in the plant tissue (Gupta 2010).…”