Izawati Abang Masli Dayang, Parveez Ghulam Kadir Ahmad, Masani Mat Yunus Abdul
Advanced Biotechnology and Breeding Centre, MPOB, Kajang, Selangor, Malaysia.
Methods Mol Biol. 2012;847:177-88. doi: 10.1007/978-1-61779-558-9_15.
Transgenic oil palm (Elaeis guineensis Jacq.) plantlets are regenerated after Agrobacterium tumefaciens-mediated transformation of embryogenic calli derived from young leaves of oil palm. The calli are transformed with an Agrobacterium strain, LBA4404, harboring the plasmid pUBA, which carries a selectable marker gene (bar) for resistance to the herbicide Basta and is driven by a maize ubiquitin promoter. Modifications of the transformation method, treatment of the target tissues using acetosyringone, exposure to a plasmolysis medium, and physical injury via biolistics are applied. The main reasons for such modifications are to activate the bacterial virulence system and, subsequently, to increase the transformation efficiency. Transgenic oil palm cells are selected and regenerated on a medium containing herbicide Basta. Molecular analyses revealed the presence and integration of the introduced bar gene into the genome of the transformants.
在根癌农杆菌介导下,将油棕幼叶来源的胚性愈伤组织进行转化后,再生出转基因油棕(Elaeis guineensis Jacq.)植株。用携带质粒pUBA的根癌农杆菌菌株LBA4404对愈伤组织进行转化,该质粒带有抗除草剂Basta的选择标记基因(bar),并由玉米泛素启动子驱动。对转化方法进行了改进,使用乙酰丁香酮处理靶组织,使其暴露于质壁分离培养基中,并通过生物弹道学进行物理损伤。进行这些改进的主要原因是激活细菌毒力系统,进而提高转化效率。在含有除草剂Basta的培养基上筛选并再生转基因油棕细胞。分子分析表明,导入的bar基因存在并整合到转化体的基因组中。