Ohnuma Mariko, Teramura Hiroshi, Shimada Hiroaki
Department of Biological Science and Technology, Tokyo University of Science, Katsushika, Tokyo 125-8585, Japan.
Plant Biotechnol (Tokyo). 2020 Mar 25;37(1):25-30. doi: 10.5511/plantbiotechnology.19.1209a.
Potato () is one of the important crop plants, and many potato cultivars consist of a tetraploid genome with high heterozygosity. The techniques of transformation and genome editing require plant regeneration. However, no efficient regeneration method has been established except for some specific cultivars, such as 'Sayaka'. In general, it is necessary to determine the adequate concentrations of auxin and cytokinin for plant regeneration. We established an efficient method using a 24-well microplate that easily enabled determination of the concentrations of these plant growth regulators suitable for shoot regeneration. Using this method, the optimal concentrations of these factors were analyzed for two representative potato cultivars, 'Sayaka' and 'Konafubuki'. This analysis revealed there was a large difference in the optimal concentrations between them. Based on this result, a specialized medium for the efficient regeneration of 'Konafubuki' cultivars was proposed. This assay method was also applied for determination of hygromycin sensitivity of these potato cultivars, and it was observed that 'Konafubuki' was rather sensitive to hygromycin. These findings suggested that the selection of a 'Konafubuki' transformant could be achieved using a medium containing a lower amount of hygromycin than that used for 'Sayaka'.
马铃薯（）是重要的农作物之一，许多马铃薯栽培品种具有高度杂合的四倍体基因组。转化和基因组编辑技术需要植株再生。然而，除了一些特定品种，如“Sayaka”外，尚未建立有效的再生方法。一般来说，有必要确定用于植株再生的生长素和细胞分裂素的合适浓度。我们建立了一种使用24孔微孔板的有效方法，该方法能够轻松确定适合芽再生的这些植物生长调节剂的浓度。使用该方法，分析了两个代表性马铃薯品种“Sayaka”和“小并吹雪”的这些因素的最佳浓度。该分析表明它们之间的最佳浓度存在很大差异。基于此结果，提出了一种用于“小并吹雪”品种高效再生的专用培养基。该测定方法还用于确定这些马铃薯品种对潮霉素的敏感性，并且观察到“小并吹雪”对潮霉素相当敏感。这些发现表明，使用比“Sayaka”所用潮霉素含量更低的培养基可以实现“小并吹雪”转化体的筛选。
