Lin Yang-Yang, Wu Shui-Han, Chen Jie, Yi Xian-Gui, Wang Xian-Rong, Li Meng
Co-Innovation Center for Sustainable Forestry in Southern China, College of Life Sciences, Nanjing Forestry University, Nanjing 210037, China.
Int J Mol Sci. 2025 Apr 10;26(8):3588. doi: 10.3390/ijms26083588.
, a unique germplasm resource native to China, exhibits significant ornamental value due to its short juvenile phase, early flowering period, abundant flowers, and elegant tree form. However, the lack of an efficient regeneration and genetic transformation system has hindered its genetic improvement and wider application. In this study, we focused on optimizing the tissue culture conditions for and establishing an -mediated transient genetic transformation system. We first determined the optimal medium compositions for different stages of tissue culture, including seed germination, callus induction, adventitious bud differentiation, and rooting. For seed germination, the optimal medium was MS supplemented with 200 mg/L GA3 and 4 mg/L 6-BA. For callus induction, the best medium was MS containing 2.00 mg/L 6-BA, 1.00 mg/L NAA, and 200 mg/L VC. Adventitious bud differentiation was favored on MS medium with 1.00 mg/L 6-BA, 0.10 mg/L NAA, and 200 mg/L VC, while rooting was optimal on 3/4 MS medium supplemented with 0.50 mg/L NAA. Subsequently, we established an -mediated transient genetic transformation system using stem segments of as explants. Through orthogonal experiments, we identified the optimal conditions for genetic transformation as pre-cultivation for 2 days, an concentration of OD = 0.6, an infection time of 30 min, and co-cultivation for 3 days. Under these conditions, the transient genetic transformation efficiency reached 10.42%, as confirmed by PCR and fluorescence detection. This study provides a reliable transient genetic transformation system for , facilitating further functional gene analysis and genetic improvement of this valuable ornamental species.
[物种名称]是中国特有的种质资源，因其幼年期短、开花期早、花量大且树形优美而具有重要的观赏价值。然而，缺乏高效的再生和遗传转化体系阻碍了其遗传改良和更广泛的应用。在本研究中，我们专注于优化[物种名称]的组织培养条件并建立农杆菌介导的瞬时遗传转化体系。我们首先确定了组织培养不同阶段的最佳培养基组成，包括种子萌发、愈伤组织诱导、不定芽分化和生根。对于种子萌发，最佳培养基是添加200 mg/L GA3和4 mg/L 6 - BA的MS培养基。对于愈伤组织诱导，最佳培养基是含有2.00 mg/L 6 - BA、1.00 mg/L NAA和200 mg/L VC的MS培养基。不定芽分化在含有1.00 mg/L 6 - BA、0.10 mg/L NAA和200 mg/L VC的MS培养基上较为有利，而生根在添加0.50 mg/L NAA的3/4 MS培养基上最为适宜。随后，我们以[物种名称]的茎段为外植体建立了农杆菌介导的瞬时遗传转化体系。通过正交实验，我们确定了遗传转化的最佳条件为预培养2天、农杆菌浓度OD = 0.6、侵染时间30分钟和共培养3天。在这些条件下，经PCR和[荧光检测方法]检测证实，瞬时遗传转化效率达到10.42%。本研究为[物种名称]提供了可靠的瞬时遗传转化体系，有助于对这种有价值的观赏物种进行进一步的功能基因分析和遗传改良。
