Lin Kai, Lu Li-Xin, Pan Bang-Zhen, Chai Xia, Fu Qian-Tang, Geng Xian-Chen, Mo Yi, Fei Yu-Chong, Xu Jia-Jing, Li Meng, Ni Jun, Xu Zeng-Fu
Guangxi Key Laboratory of Forest Ecology and Conservation, Guangxi Colleges and Universities Key Laboratory for Cultivation and Utilization of Subtropical Forest Plantation, College of Forestry, Guangxi University, Nanning 530004, China.
Key Laboratory of National Forestry and Grassland Administration on Cultivation of Fast-Growing Timber in Central South China, State Key Laboratory for Conservation and Utilization of Subtropical Agro-BioreSources, College of Forestry, Guangxi University, Nanning 530004, China.
Int J Mol Sci. 2025 Mar 11;26(6):2496. doi: 10.3390/ijms26062496.
L., a woody oilseed plant rich in α-linolenic acid, represents a promising source of polyunsaturated fatty acids. However, the lack of an efficient genetic transformation system has significantly hindered gene function research and molecular breeding in . In this study, we developed a highly efficient -mediated hairy root transformation system for via the use of gel in combination with the visually detectable reporter for gene function analysis in roots. The results indicate that the optimal transformation method involves infecting seedlings with gel containing acetosyringone and inducing hairy root formation in perlite. This approach resulted in more than 18.97% of the seedlings producing positive hairy roots overexpressing the gene. Using this genetic transformation system, we successfully overexpressed the antimicrobial peptide-encoding gene in hairy roots, which enhanced the resistance of to . Furthermore, by combining this transformation system with the CRISPR-Cas9 tool, we validated the regulatory role of in the development of root epidermal cells in . Unexpectedly, a 123-bp DNA fragment from the T-DNA region of the Ri plasmid was found to be knocked in to the genome, replacing a 110-bp fragment of at CRISPR-Cas9 induced double-strand DNA breaks. Conclusively, this system provides a powerful tool for gene function research in and provides novel insights into the development of transformation and gene editing systems for other woody plants.
L.是一种富含α-亚麻酸的木本油料植物，是多不饱和脂肪酸的一个有前景的来源。然而，缺乏高效的遗传转化系统严重阻碍了L.的基因功能研究和分子育种。在本研究中，我们通过使用含有可视化检测报告基因的凝胶结合发根农杆菌，开发了一种用于L.的高效发根农杆菌介导的发根转化系统，用于根系基因功能分析。结果表明，最佳转化方法是用含有乙酰丁香酮的凝胶感染L.幼苗，并在珍珠岩中诱导发根形成。这种方法使超过18.97%的幼苗产生过表达该基因的阳性发根。利用该遗传转化系统，我们在发根中成功过表达了编码抗菌肽的基因，增强了L.对[具体病害名称未给出]的抗性。此外，通过将该转化系统与CRISPR-Cas9工具相结合，我们验证了L.中[基因名称未给出]在根表皮细胞发育中的调控作用。意外的是，发现来自发根农杆菌Ri质粒T-DNA区域的一个123 bp DNA片段插入到L.基因组中，在CRISPR-Cas9诱导的双链DNA断裂处取代了L.的一个110 bp片段。总之，该系统为L.的基因功能研究提供了一个强大的工具，并为其他木本植物的转化和基因编辑系统的开发提供了新的见解。
