Improvement of regeneration and transformation systems for Cyclamen persicum using somatic embryo culture
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<i>Cyclamen persicum</i> is one of the most important pot plants in the world. We developed an effective method for somatic embryo-mediated plant regeneration and genetic transformation using <i>Agrobacterium</i>, with somatic embryos as the source of plant material. Numerous somatic embryos were formed from the leaf segments of <i>Cyclamen</i> cv. "Fragrance Mini" 16 weeks after initiation on a 2,4-D- and kinetin-containing medium. The maximum regeneration frequency from the somatic embryos was achieved using a regeneration medium containing 0.1 mg l<sup>−1</sup> benzylaminopurine, 0.01 mg l<sup>−1</sup> α-naphthaleneacetic acid, and 0.2 mg l<sup>−1</sup> gibberellin. After co-cultivating the somatic embryos with <i>Agrobacterium</i> strain LBA4404 (the plasmid vector pIG121Hm) that harbors genes for hygromycin phosphotransferase, neomycin phosphotransferase, and β-glucuronidase, hygromycin-resistant clones were selected on a medium containing 5 mg l<sup>−1</sup> hygromycin. Hygromycin-resistant plantlets were regenerated from 14 hygromycin-resistant embryos. Using this transformation system, 74 independent plants were obtained from 1-ml packed cell volume, approximately 2,000 embryos.
- Plant tissue culture letters
Plant tissue culture letters 25(1), 77-80, 2008-03-01
Japanese Society for Plant Cell and Molecular Biology