Affiliation:
1. Nikitsky Botanical Gardens: FGBUN Nikitskij botaniceskij sad Nacional'nyj naucnyj centr Rossijskoj akademii nauk
2. Pushchino Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences: Filial Instituta bioorganiceskoj himii im akademikov M M Semakina i U A Ovcinnikova Rossijskoj akademii nauk
Abstract
Abstract
We present data on the ability to organogenesis in 22 genotypes of grapevine and developed a direct organogenesis protocol for the cultivar Podarok Magaracha and the rootstock Kober 5BB. Protocol does not require replacement of culture media and growth regulators, the duration is 11 weeks. 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 callogenesis, (2) followed by 4 weeks of cultivation in light conditions for regeneration, (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.
Publisher
Research Square Platform LLC