Zhai Yuhui, Xie Xinrong, Zhang Liping, Wang Xuefei, Zhang Zixuan, Niu Lixin, Zhang Yanlong
College of Landscape Architecture and Arts, Northwest A&F University, Yangling 712100, China.
Plants (Basel). 2025 Aug 12;14(16):2498. doi: 10.3390/plants14162498.
is an economically significant species serving as an ornamental, medicinal herb, and woody oilseed crop. Gene function elucidation and molecular breeding are hindered by the lack of efficient, stable transformation methods due to tissue culture challenges. To enable year-round functional studies without material constraints, we established a novel transient transformation system mediated by Agrobacterium using in vitro embryo-derived seedlings (TTAES) in . By optimizing embryo germination media, we achieved consistent seedling production. Orthogonal experiments with a GUS reporter identified optimal conditions: OD = 1.0, 200 μM of acetosyringone, six negative-pressure treatments, and 2 h infection. Under this optimized system, maximum transformation efficiency was achieved at 35 days after germination. With this system, we demonstrated its application in investigating transcription factor-mediated regulation of target gene promoters using GUS as a reporter gene. To achieve non-destructive identification of transiently transformed plants, we employed GFP as a reporter gene. Using transient expression of VIGS (knockdown) and 35S constructs (overexpression), we characterized gene functions, thereby confirming the system's effectiveness for functional analysis. This system facilitates the acquisition of plant experimental materials and significantly improves research efficiency for year-round gene function elucidation in .
是一种具有重要经济意义的物种，可作为观赏植物、药用草本植物和木本油料作物。由于组织培养方面的挑战，缺乏高效、稳定的转化方法阻碍了基因功能的阐明和分子育种。为了在不受材料限制的情况下进行全年的功能研究，我们在[具体物种]中建立了一种由农杆菌介导的新型瞬时转化系统，该系统使用体外胚衍生幼苗（TTAES）。通过优化胚萌发培养基，我们实现了一致的幼苗生产。用GUS报告基因进行的正交实验确定了最佳条件：OD = 1.0、200 μM乙酰丁香酮、六次负压处理和2小时感染。在这个优化系统下，萌发后35天达到最大转化效率。利用这个系统，我们以GUS作为报告基因，展示了其在研究转录因子介导的靶基因启动子调控中的应用。为了实现对瞬时转化植物的无损鉴定，我们采用GFP作为报告基因。利用VIGS（基因沉默）和35S构建体（过表达）的瞬时表达，我们对基因功能进行了表征，从而证实了该系统在功能分析中的有效性。该系统有助于获取植物实验材料，并显著提高了[具体物种]全年基因功能阐明的研究效率。
