Efficient plant regeneration and Agrobacterium -mediated transformation of Begonia semperflorens-cultorum
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RESEARCH NOTE
Efficient plant regeneration and Agrobacterium‑mediated transformation of Begonia semperflorens‑cultorum Sakiko Hirutani1 · Kazuki Shimomae1 · Akira Yaguchi1 · Dong Poh Chin1,2 · Masahiro Mii1,2 · Tomoko Igawa1 Received: 30 December 2019 / Accepted: 26 May 2020 © Springer Nature B.V. 2020
Abstract 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. Key message Production of transgenic plants through Agrobacterium-mediated transformation using an effective system for plant regeneration from leaf segment of Begonia semperflorens-cultorum was achieved. Keywords Begonia semperflorens-cultorum · Plant regeneration · Agrobacterium-mediated transformation B. semperflorens-cultorum, known as wax begonia, is widely cultivated because of the robust adaptability to the environment, vigorous growth habit and constant blooming all year Communicated by T. Winkelmann. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11240-020-01858-7) contains supplementary material, which is available to authorized users. * Masahiro Mii [email protected]‑u.jp * Tomoko Igawa tigawa@chiba‑u.jp 1
Graduate School of Horticulture, Chiba University, 648 Matsudo, Matsudo, Chiba 271‑8510, Japan
Present Address: Center for Environment, Health and Field Sciences, Chiba University, 6‑2‑1 Kashiwanoha, Kashiwa, Chiba 277‑0882, Japan
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round (Tebbitt 2005). B. semperflorens-cultorum is categorized as a cultivar group of Begonia, which was derived from interspecific hybridization between B. cucullata and B. schmidtiana. Numerous cultivars have been developed by intra-specific crossings and hybridizations with sever
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