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Development of Micropropagation System for Apple Breeding from Cotyledon Explants
Van Giap Do,Seonae Kim,Youngsuk Lee,Hun-Joong Kweon 한국원예학회 2021 한국원예학회 학술발표요지 Vol.2021 No.10
Agrobacterium-mediated transformation is one of the well-known methods and proficient use for introducing the interest genes onto the plant cell tissue to obtain transgenic plants for study the function of the interest genes and established new traits through the expression of these genes. In this study, we developed an efficient system for in vitro culture and Agrobacterium-mediated transformation protocol for apples using cotyledons as explants. The apple seed of commercial apple Hongno cultivar was surface disinfected and then removed the peel and the external coat and then culture on embryo culture media for 2 ‒ 3 weeks. The cotyledons were cut down to 2 ‒ 4 slices and be used as explants for in vitro culture and Agrobacterium-mediated transformation. For in vitro shoot regeneration system, a high percentage of callus formation and of adventitious shoot regeneration was observed. Besides that, for the development of the transgenic apple plants, the cotyledon explants were also used for Agrobacterium-mediated transformation. The transgenic plant with the integration of the T-DNA region contained a resistant gene htp (hygromycin phosphotransferase) and report gene GFP (green fluorescent protein) into the apple genome was confirmed by the genomic DNA PCR of hpt gene. The transformed efficiency of transgenic plant regeneration obtained from explants was 15%. An in vitro culture system for development of the whole apple seedling from micropropagation of cotyledon material to produce many progenies apple plants and transgenic apple plants were generated via Agrobacterium-mediated transformation method were obtained. It is useful for apple breeding and elucidates the function of genes of interest in apples.