Begonia semperflorens-cultorum, known as wax begonia, is one of the most popular Begonia species in which variable commercial cultivars have been produced. The genetic transformation technique is required for further modification of this species because introduction of some desired traits such as novel flower colors cannot be achieved by conventional breeding. Here we report the procedures of an efficient plant regeneration from leaf segment and production of transgenic plants using Begonia semperflorens-cultorum. Efficient induction of adventitious shoots was achieved when the explants were cultured on MS medium supplemented with 1.5 mg/l thidiazuron (TDZ), 0.5 mg/l α-naphthaleneacetic acid (NAA), 30 g/l sucrose and 2.5 mg/l gellan gum. Agrobacterium tumefaciens strain EHA101 containing the plasmid pIG121-Hm was used for gene transfer. The transient GUS expression was significantly increased when 10 mM MES was added to the co-cultivation medium. By culturing the infected explants under the selection pressure with 10 mg/l hygromycin (Hm), the Hm-resistant independent shoots were obtained at a frequency of 0.78 per explant. The regenerated Hm-resistant plants showed the integration of hygromycin phosphotransferase (HPT) gene into the genome and stable GUS expression, as the proof of genetic transformation. Consequently, the conditions established in this study enabled the transformation of Begonia semperflorens-cultorum.
Production of transgenic plants through Agrobacterium-mediated transformation using an effective system for plant regeneration from leaf segment of Begonia semperflorens-cultorum was achieved.
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This work was supported by Japan Society for the Promotion of Science KAKENHI Grants (JP17K07600 to TI) and the Strategic Priority Research Promotion Program on Phytochemical Plant Molecular Sciences, Chiba University (Japan).
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Communicated by T. Winkelmann.
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Supplementaly Fig. S1 Effects of various concentrations (mg/l) of NAA and TDZ on the frequency of adventitious shoot formation from leaf segments of B. semperfloren-cultorum ‘Ambassador Coral’. In each treatment, 25 explants were tested and the experiment was repeated three times. The data were recorded after 3 months of culture. Results are represented as the means ± standard error. The different letters on each bar show significant differences based on Tukey–Kramer test (P = 0.05) detected using R software. (DOCX 14 kb)
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Hirutani, S., Shimomae, K., Yaguchi, A. et al. Efficient plant regeneration and Agrobacterium-mediated transformation of Begonia semperflorens-cultorum. Plant Cell Tiss Organ Cult 142, 435–440 (2020). https://doi.org/10.1007/s11240-020-01858-7
- Begonia semperflorens-cultorum
- Plant regeneration
- Agrobacterium-mediated transformation