Abstract
SummaryA successful transformation procedure using Agrobacterium-mediated gene transfer was established for the bacterial wilt-resistant Dianthus species, D. acicularis. Highly regenerable cell suspension cultures derived from calli, which originated from leaf segments, were infected with A. tumefaciens strain EHA101 (pIG121Hm) and co-cultivated for 2 d. After selection on medium with 30 mg l–1 hygromycin, approximately five hygromycin-resistant plants were obtained from 1 g fresh weight of cell culture. For the production of hygromycin-resistant calli, addition of acetosyringone to the co-cultivation medium was indispensable, with an optimum concentration of 200 µM. Hygromycin-resistant calli and plantlets showed stable and uniform GUS expression, and integration of the GUS gene into the plant genome was also confirmed by Southern blot analysis.These transgenic plants had a normal morphology, and no somaclonal variation was observed in their level of ploidy.
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