Rapeseed (Brassica napus) is an important oil crop that supplies a considerable amount of global vegetable oil production. Genetic transformation system is important to gene functional analysis and molecular breeding. Here, an efficient Agrobacterium-mediated transformation protocol using hypocotyl of rapeseed as explants is described. To develop this protocol, we compared several essential factors that would affect the transformation efficiency, such as Agrobacterium strains, selection marker genes, and genotypes of rapeseed. Comparison of different Agrobacterium strains showed that the GV3101 had higher transformation efficiency than that of C58C1 and EHA105. HPTII, NPTII, and RePAT were used as selection marker genes in tissue culture. The results showed that the transformation efficiency was 3.7–4.8%, 2.2–22.5%, and 1.6–5.9% when the hypocotyl of Westar was infected by GV3101 and screened under hygromycin, kanamycin, and basta, respectively. The transformation efficiency of Westar was the highest and ZS11 was the lowest when five different genotypes of rapeseed (Westar, ZS9, ZS11, GY284, and WH3417) were infected by GV3101. Using this protocol, it will take 8–10 weeks to obtain transgenic plants. This protocol has been used to study gene function in several genotypes of rapeseed in our laboratory. These results indicate that it is efficient to obtain transgenic plant of rapeseed using this protocol.

油菜籽（甘蓝型油菜）是一种重要的油料作物，可提供大量的全球植物油产量。遗传转化系统对于基因功能分析和分子育种很重要。在此，描述了一种有效的农杆菌介导的转化方法，该方法使用油菜的下胚轴作为外植体。为了开发此协议，我们比较了一些会影响转化效率的重要因素，例如农杆菌菌株，选择标记基因和油菜籽的基因型。对不同农杆菌菌株的比较表明，GV3101的转化效率高于C58C1和EHA105。HPTII，NPTII和RePAT用作组织培养中的选择标记基因。结果表明，当Westar的下胚轴被GV3101感染并在潮霉素，卡那霉素和巴斯德下筛选时，转化效率分别为3.7–4.8％，2.2–22.5％和1.6–5.9％。当五种不同基因型的油菜籽（Westar，ZS9，ZS11，GY284和WH3417）感染GV3101时，Westar的转化效率最高，ZS11最低。使用此方案，将需要8-10周才能获得转基因植物。在我们的实验室中，该协议已用于研究几种基因型油菜的基因功能。这些结果表明，使用该方案可有效获得油菜的转基因植物。