Organogenesis in a Broad Spectrum of Grape Genotypes and <i>Agrobacterium</i>-Mediated Transformation of the Podarok Magaracha Grapevine Cultivar

Abstract

We present data on the ability for organogenesis in 22 genotypes of grapevine and developed a direct organogenesis protocol for the cultivar Podarok Magaracha and the rootstock Kober 5BB. The protocol does not require replacement of culture media and growth regulators, and the duration is 11 weeks. The cultivation of explants occurs on modified MS medium with the addition of 2.0 mg L−1 benzyladenine and indole-3-butyric acid (0.15 mg L−1 for the rootstock Kober 5BB or 0.05 mg L−1 for the cultivar Podarok Magaracha). The direct organogenesis protocol consists of three time periods: (1) culturing explants for 2 weeks in dark conditions for meristematic bulk tissue, (2) followed by 4 weeks of cultivation in light conditions for regeneration, and (3) 5 weeks of cultivation in dark conditions for shoot elongation. Based on this protocol, conditions for the Agrobacterium-mediated transformation of the Podarok Magaracha cultivar were developed with an efficiency of 2.0% transgenic plants per 100 explants. Two stably transformed lines with integration into the genome of the pBin35SGFP plasmid construction, confirmed by Southern blotting, were obtained.

Keywords

• <i>Agrobacterium</i>-mediated transformation
• benzyladenine
• callus
• grapevine
• <i>Vitis</i>
• elongation