Dianthus spiculifolius is a perennial herbaceous flower with strong environmental adaptability and is an important ornamental ground cover plant. In this study, seeds of D. spiculifolius were used as explants for callus induction, adventitious bud differentiation, and rooting by adding different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzyl aminopurine (6-BA), and naphthaleneacetic acid (NAA) to Murashige and Skoog medium. The calli generated were co-cultured with Agrobacterium tumefaciens EHA105 containing pBI121-GUS or pBI121-GFP plasmids for 30 min, and transgenic regenerated plants were obtained by kanamycin (30 mg · L⁻¹) screening. RT-PCR confirmed the stable expression of the exogenous GUS and GFP genes in the D. spiculifolius. The β-glucuronidase (GUS) histochemical staining confirmed GUS gene expression in transgenic calli, adventitious buds, and regenerated plants of D. spiculifolius. The green fluorescent protein (GFP) visual analysis showed GFP gene expression in transgenic calli. Furthermore, subcellular localization analysis showed that the three organelle marker proteins were not only successfully expressed but also accurately localized to their corresponding organelles in D. spiculifolius callus cells. These results indicated a successful establishment of a reliable and efficient A. tumefaciens-mediated genetic transformation system, which will contribute to functional gene research and genetic improvement of D. spiculifolius.

石竹是多年生草本植物，具有很强的环境适应性，是重要的观赏地被植物。在这项研究中，将D. spiculifolius的种子用作外植体，用于诱导愈伤组织，不定芽分化和通过添加不同浓度的2,4-二氯苯氧基乙酸（2,4-D），6-苄基氨基嘌呤（6-BA ）和萘乙酸（NAA）加入Murashige和Skoog培养基中。将产生的愈伤组织与含有pBI121-GUS或pBI121-GFP质粒的根癌农杆菌EHA105共培养30分钟，并通过卡那霉素（30 mg·L ^-1）筛选获得转基因的再生植物。RT-PCR证实外源的稳定表达GUS和D. spiculifolius中的GFP基因。的β葡萄糖醛酸酶（GUS）组织化学染色证实GUS转基因愈伤组织，不定芽，和再生的植物基因表达D. spiculifolius。绿色荧光蛋白（GFP）视觉分析显示GFP基因在转基因愈伤组织中表达。此外，亚细胞定位分析表明，三种细胞器标记蛋白不仅可以成功表达，而且还可以准确地定位在S. spiculifolius愈伤组织细胞中相应的细胞器中。这些结果表明成功建立了可靠和有效的根癌农杆菌。介导的遗传转化系统，将为D. spiculifolius的功能基因研究和遗传改良做出贡献。