Zhang Kai, He Jianjie, Liu Lu, Xie Runda, Qiu Lu, Li Xicheng, Yuan Wenjue, Chen Kang, Yin Yongtai, Kyaw May Me Me, San Aye Aye, Li Shisheng, Tang Xianying, Fu Chunhua, Li Maoteng
1Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074 China.
2Hubei Collaborative Innovation Center for the Characteristic Resources Exploitation of Dabie Mountains, Huanggang Normal University, Huanggang, 438000 China.
Plant Methods. 2020 Mar 30;16:43. doi: 10.1186/s13007-020-00585-6. eCollection 2020.
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 .
Based on the well-established genetic transformation system in , five vectors carrying the red fluorescence protein encoding gene from sp () were constructed and integrated into rapeseed via -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 , 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.
是一种重要的油料作物，为全球植物油生产提供了大量生物质。建立一个具有便捷的转基因阳性筛选方法的高效遗传转化系统对于基因功能分析和分子育种至关重要。然而，据我们所知，很少有上述系统可有效地应用于。
基于已建立的遗传转化系统，构建了五个携带来自sp（）的红色荧光蛋白编码基因的载体，并通过介导的下胚轴转化将其整合到油菜中。在组织培养基中，通过视觉筛选方法平均有59.1%的组织被红色荧光标记，其中平均96.1%的组织从八个不同油菜品种中扩增出目标基因。此外，从红色荧光标记的组织中获得的生根苗的最终转基因阳性效率高达90.7%，这远高于先前报道的效率。此外，通过整合，视觉筛选可应用于幼苗，包括种子萌发期间的种皮、根、下胚轴和子叶。这些结果表明，高效遗传转化系统与转基因阳性视觉筛选方法相结合有助于方便、高效地获得转基因阳性油菜苗。
开发了一种快速、便捷且高效的方法来获得转基因植物，这有助于获得最大比例的转基因阳性再生苗，从而避免长时间的植物再生。本研究结果将有利于基因功能研究，特别是在高通量分子生物学研究中。
