Fruit Tree Research Institute, 00040 Ciampino Aeroporto, Roma, Italy.
Plant Cell Rep. 1995 Feb;14(5):267-72. doi: 10.1007/BF00232026.
A protocol for Agrobacterium-mediated transformation of leaf pieces from two almond cultivars has been developed. This protocol gave a transformation percentage of 48.6 for the cultivar MN51 and 27.9 for the cultivar 'Supernova'. The Agrobacterium tumefaciens strain LBA 4404 carrying a binary vector plasmid pBinGUSint has been used. A critical step for optimal transformation was the pre-culture period of leaf pieces prior selection on medium containing kanamycin. Both cultivars showed an almost doubled transformation percentage when the selection was started at day 6 compared to day 0. The presence of neomycin phosphotransferase II and ß-glucuronidase genes carried by pBinGUSint was confirmed in transformed expiants both by Southern blotting analysis and enzyme activity assays.
已开发出一种通过叶组织片用根癌农杆菌转化两种巴旦杏品种的方法。该方法对 MN51 品种的转化率为 48.6%，对‘新星’品种的转化率为 27.9%。所用的根癌农杆菌菌株 LBA 4404 携带一个双元载体质粒 pBinGUSint。在含卡那霉素的培养基上进行选择之前，对叶组织片进行预培养是实现最佳转化的关键步骤。与从第 0 天开始选择相比，第 6 天开始选择时，两个品种的转化率几乎增加了一倍。Southern 印迹分析和酶活性测定均证实，pBinGUSint 携带的新霉素磷酸转移酶 II 和β-葡萄糖醛酸酶基因存在于转化的外植体中。
