Plant regeneration from leaf explants and efficient Agrobacterium-mediated transformation system of chrysanthemum (Dendranthema grandiflorum)

Leaf explants of the chrysanthemum (Dendranthema grandiflorum) were cultured on MS medium supplemented with various concentrations of NAA, IAA, 2,4-D, and BA. The highest frequency of shoot formation was observed on MS medium supplemented with 1.0 mg(.)L(-1) BA and 0.3 mg(.)L(-1) 2,4-D in 'Puma' and with 2.0 mg(.)L(-1) NAA and 0.5 mg(.)L(-1) BA in 'Subangryeok'. The suitable concentrations of kanamycin to select the transformant are 20 mg(.)L(-1) in 'Puma' and 50 mg(.)L(-1) in 'Subangryeok'. In the 'Puma', the calli formed most efficiently in the medium containing kanamycin from leaf discs co-cultivated with A. tumefaciens inoculation at 1:50 dilution ratio for 5 min. There were 700 bp npt gene bands in the two transformants through the PCR detection of npt gene.