Maletich Galina, Pushin Alexander, Rybalkin Evgeniy, Plugatar Yuri, Dolgov Sergey, Khvatkov Pavel
Federal State Funded Institution of Science "The Labor Red Banner Order Nikita Botanical Gardens-National Scientific Center of the RAS", 298648 Yalta, Russia.
Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, 142290 Puschino, Russia.
Plants (Basel). 2024 Oct 3;13(19):2779. doi: 10.3390/plants13192779.
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 benzyladenine and indole-3-butyric acid (0.15 mg L for the rootstock Kober 5BB or 0.05 mg L 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 -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 plasmid construction, confirmed by Southern blotting, were obtained.
我们展示了22种葡萄基因型的器官发生能力的数据,并为“波达罗克·马加拉查”品种和“考贝5BB”砧木开发了一种直接器官发生方案。该方案无需更换培养基和生长调节剂,持续时间为11周。外植体在添加了2.0毫克/升苄基腺嘌呤和吲哚-3-丁酸(“考贝5BB”砧木为0.15毫克/升,“波达罗克·马加拉查”品种为0.05毫克/升)的改良MS培养基上培养。直接器官发生方案包括三个时间段:(1)将外植体在黑暗条件下培养2周以形成分生组织块,(2)随后在光照条件下培养4周以进行再生,(3)在黑暗条件下培养5周以促进芽伸长。基于此方案,开发了“波达罗克·马加拉查”品种的介导转化条件,每100个外植体产生转基因植株的效率为2.0%。通过Southern杂交确认,获得了两个稳定整合到质粒构建体基因组中的转化株系。