The success of in vitro plant regeneration and the competence of genetic transformation greatly depends on the genotype of the species of interest. In previous work, we developed a method for the efficient Agrobacterium-mediated genetic transformation via organogenesis of V. vinifera cultivar Thompson Seedless, by using meristematic bulk (MB) as starting tissue. In this study, we applied this method for the regeneration and transformation of MBs obtained from the Italian cultivar Ciliegiolo and two of the commonly used Vitis rootstocks, 110 Richter and Kober 5BB, in comparison with Thompson Seedless. The A. tumefaciens strain EHA105, harbouring pK7WG2 binary vector, was used for the transformation trials, which allowed selection through the enhanced-green fluorescent protein (eGFP) and the neomycin phosphotransferase (nptII) gene. Putative transformed tissues and/or shoots were identified by either a screening based on the eGFP expression alone or its use in combination with kanamycin in the medium. MBs obtained from Thompson Seedless showed the highest regeneration and transformation cell competence, which subsequently allowed the recovery of stably transformed plants. Ciliegiolo, 110 Richter, and Kober 5BB, produced actively growing transgenic calli showing eGFP fluorescence, more consistently on selective media, but had no regenerative competence.

体外植物再生的成功和遗传转化的能力在很大程度上取决于感兴趣物种的基因型。在之前的工作中，我们开发了一种通过V的器官发生进行有效农杆菌介导的遗传转化的方法。vinifera品种 Thompson Seedless，使用分生组织 (MB) 作为起始组织。在这项研究中，我们应用这种方法对从意大利品种 Ciliegiolo 和两种常用葡萄砧木 110 Richter 和 Koher 5BB 获得的 MB 进行再生和转化，并与 Thompson Seedless 进行比较。A. _ _ 含有 pK7WG2 双元载体的根癌菌株EHA105 用于转化试验，该试验允许通过增强型绿色荧光蛋白 (eGFP) 和新霉素磷酸转移酶 ( nptII ) 基因进行选择。通过单独基于 eGFP 表达的筛选或将其与培养基中的卡那霉素组合使用来鉴定推定的转化组织和/或芽。从 Thompson Seedless 获得的 MB 显示出最高的再生和转化细胞能力，随后可以恢复稳定转化的植物。Ciliegiolo、110 Richter 和Kober 5BB 产生了生长活跃的转基因愈伤组织，显示出eGFP 荧光，在选择性培养基上更加一致，但没有再生能力。