Excision of DNA fragments with the piggyBac system in Chrysanthemum morifolium

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  • Kishi-Kaboshi Mitsuko
    Institute of Vegetable and Floriculture Science, National Agriculture and Food Research Organization
  • Nishizawa-Yokoi Ayako
    Institute of Agrobiological Sciences, National Agriculture and Food Research Organization
  • Mitsuhara Ichiro
    Institute of Agrobiological Sciences, National Agriculture and Food Research Organization
  • Toki Seiichi
    Institute of Agrobiological Sciences, National Agriculture and Food Research Organization Laboratory of Plant Genome Engineering, Department of Plant Life Science, Faculty of Agriculture, Ryukoku University
  • Sasaki Katsutomo
    Institute of Vegetable and Floriculture Science, National Agriculture and Food Research Organization

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  • Excision of DNA fragments with the <i>piggyBac</i> system in <i>Chrysanthemum morifolium</i>

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<p>Chrysanthemum morifolium is one of the most popular ornamental plants in the world. However, as C. morifolium is a segmental hexaploid, self-incompatible, and has a sizable heterologous genome, it is difficult to modify its trait systematically. Genome editing technology is one of the attractive methods for modifying traits systematically. For the commercial use of genetically modified C. morifolium, rigorous stabilization of its quality is essential. This trait stability can be achieved by avoiding further genome modification after suitable trait modification by genome editing. Since C. morifolium is a vegetatively propagated plant, an approach for removing genome editing tools is required. In this study, we attempted to use the piggyBac transposon system to remove specific DNA sequences from the C. morifolium genome. Using the luminescence as a visible marker, we demonstrated that inoculation of Agrobacterium harboring hyperactive piggyBac transposase removes inserted 2.6 kb DNA, which harbors piggyBac recognition sequences, from the modified Eluc sequence.</p>

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