The Hibiscus genus, encompassing roughly 300 species across 10 sections, presents a diverse and economically significant range of industrial, ornamental, and medicinal properties. Polyploidy, whether occurring spontaneously or induced through external agents, such as chemicals like colchicine and oryzalin, plays a crucial role in plant breeding. It enhances various attributes including flower size, resilience, and metabolite production. Among the species in this genus, Hibiscus hamabo is noted for its salt tolerance and its attractive yellow flowers in summer and golden-yellow or burnt orange leaves in fall. However, H. hamabo typically has small and sparse flowers, which led to our efforts to enhance its ornamental value through induced polyploidy. In our study, we treated germinating seeds of H. hamabo with three concentrations of colchicine (0, 0.125, and 0.25% v/v) for varying durations (6, 12, or 24 hours). The most effective conversion was achieved with seeds treated with 0.25% v/v colchicine for 24 hours. This treatment produced solid polyploids (4n = 184) and mixed-polyploids (2n 4n). The 4n plants exhibited a 2C-DNA content of 8.50 pg, compared to 4.23 pg in the untreated (2n = 92) plants. We evaluated the impact of induced polyploidy on several morphological traits including leaf color, shape, size, trichome density, and plant height. Significant differences were observed between the polyploid plants and the control plants. Additionally, we explored the cytomolecular analysis of induced polyploidy, particularly focusing on the distribution and organization of rDNA. In 2n plants, one locus of 5S and four loci of 35S rDNA (two major and two minor) were identified. The 5S site is pericentromeric, while one of the major 35S sites is sub-terminal, and the others are at terminal locations. In 4n plants, the number of 5S and 35S sites was exactly duplicated, confirming the polyploidization at the genetic level. Our results proved that colchicine can be used to induce polyploidy in germinating H. hamabo seeds, paving the way for the improvement of this species through this method.
