Brassica napus is an important oilseed crop that offers a considerable amount of biomass for global vegetable oil production. The establishment of an efficient genetic transformation system with a convenient transgenic-positive screening method is of great importance for gene functional analysis and molecular breeding. However, to our knowledge, there are few of the aforementioned systems available for efficient application in B. napus. Based on the well-established genetic transformation system in B. napus, five vectors carrying the red fluorescence protein encoding gene from Discosoma sp. (DsRed) were constructed and integrated into rapeseed via Agrobacterium-mediated hypocotyl transformation. An average of 59.1% tissues were marked with red fluorescence by the visual screening method in tissue culture medium, 96.1% of which, on average, were amplified with the objective genes from eight different rapeseed varieties. In addition, the final transgenic-positive efficiency of the rooted plantlets reached up to 90.7% from red fluorescence marked tissues, which was much higher than that in previous reports. Additionally, visual screening could be applicable to seedlings via integration of DsRed, including seed coats, roots, hypocotyls and cotyledons during seed germination. These results indicate that the highly efficient genetic transformation system combined with the transgenic-positive visual screening method helps to conveniently and efficiently obtain transgenic-positive rapeseed plantlets. A rapid, convenient and highly efficient method was developed to obtain transgenic plants, which can help to obtain the largest proportion of transgene-positive regenerated plantlets, thereby avoiding a long period of plant regeneration. The results of this study will benefit gene functional studies especially in high-throughput molecular biology research.

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一种方便、快速、高效的甘蓝型油菜转基因株系建立方法

甘蓝型油菜是一种重要的油料作物，为全球植物油生产提供大量生物质。建立高效的遗传转化体系和便捷的转基因阳性筛选方法对于基因功能分析和分子育种具有重要意义。然而，据我们所知，上述系统很少可用于在甘蓝型油菜中有效应用。基于甘蓝型油菜中成熟的遗传转化系统，五个载体携带来自Discosoma sp.的红色荧光蛋白编码基因。（DsRed）通过农杆菌介导的下胚轴转化构建并整合到油菜籽中。组织培养基中目视筛选法平均有59.1%的组织出现红色荧光，其中平均有96.1%的组织扩增出8个不同油菜品种的目的基因。此外，红色荧光标记组织生根植株最终转基因阳性率高达90.7%，远高于以往报道。此外，通过红色荧光蛋白整合，视觉筛选可适用于幼苗，包括种子萌发过程中的种皮、根、下胚轴和子叶。这些结果表明，高效的遗传转化系统结合转基因阳性视觉筛选方法有助于方便、高效地获得转基因阳性油菜苗。开发了一种快速、方便、高效的转基因植物获得方法，有助于获得最大比例的转基因阳性再生植株，从而避免植物再生周期过长。这项研究的结果将有利于基因功能研究，特别是高通量分子生物学研究。