Efficient Genetic Transformation of Chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) Using Stem Segments.

Abstract

Efficient genetic transformation of chrysanthemum (Dendranthema grandi/lorum (Ramat.) Kitamura) based on the rapid direct regeneration of stem explants is described. The genotype (cv. 1581), which showed a high regeneration potential with no negative effects from Agrobacterium inoculation, was selected for transformation experiments. Explants were cocultivated for 2 days with Agrobacterium tumefaciens strain AGLO containing the binary vector pMOG 410 carrying the NPT 11 and GUS intron genes. After cocultivation, the explants were placed on selection medium containing 10 mg/l kanamycin. About 10 olo of the inoculated explants produced GUS-positive shoots within 65 days of culture. Higher transformation efficiency based on harvested shoots was obtained when the explants were exposed to higher IAA for 7 days after cocultivation. The presence of T-DNA in the plants has been confirmed by PCR analysis.