Deng Jie, Li Wenyun, Li Xiaomin, Liu Diqiu, Liu Guanze
Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China.
State Key Laboratory of Conservation and Utilization of Bio-Resources in Yunnan, The Key Laboratory of Medicinal Plant Biology of Yunnan Province, National & Local Joint Engineering Research Center on Germplasms Innovation & Utilization of Chinese Medicinal Materials in Southwest China, Yunnan Agricultural University, Kunming 650201, China.
Plants (Basel). 2024 Sep 6;13(17):2509. doi: 10.3390/plants13172509.
The -based transgenic technique is commonly used for gene function validation and molecular breeding. However, it is not suitable for plants with a low regeneration capacity or a low transformation rate, such as (Burk) F.H. Chen and Wilson. In this study, a novel transformation method based on injection in the meristems was developed using and as experimental models. PCR analysis confirmed the successful integration of the reporter gene () into the genome of two experimental models. QRT-PCR and Western blot analysis demonstrated the transcriptional and translational expression of DsRed2. Additionally, laser confocal microscopy confirmed the significant accumulation of the red fluorescent protein in the leaves, stems, and roots of transformed and . Most importantly, in the second year after injection, the specific bright orange fluorescence from DsRed2 expression was observed in the transgenic and plants. This study establishes a fast, efficient, and tissue-culture-independent transgenic technique suitable for plants with a low regeneration capacity or a low transformation rate. This technique may improve the functional genomics of important medicinal and ornamental plants such as and , as well as their molecular breeding.
基于[具体技术名称未给出]的转基因技术常用于基因功能验证和分子育种。然而,它不适用于再生能力低或转化率低的植物,如[具体植物名称未给出](伯克)F.H.陈和[具体植物名称未给出]威尔逊。在本研究中,以[具体植物名称未给出]和[具体植物名称未给出]为实验模型,开发了一种基于分生组织注射的新型[具体植物名称未给出]转化方法。PCR分析证实报告基因[DsRed2]成功整合到两个实验模型的基因组中。QRT-PCR和蛋白质免疫印迹分析证明了DsRed2的转录和翻译表达。此外,激光共聚焦显微镜证实了红色荧光蛋白在转化的[具体植物名称未给出]和[具体植物名称未给出]的叶、茎和根中大量积累。最重要的是,在注射后的第二年,在转基因[具体植物名称未给出]和[具体植物名称未给出]植株中观察到了DsRed2表达产生的特定亮橙色荧光。本研究建立了一种快速、高效且不依赖组织培养的转基因技术,适用于再生能力低或转化率低的植物。该技术可能会改善重要药用和观赏植物如[具体植物名称未给出]和[具体植物名称未给出]的功能基因组学及其分子育种。