Chrysanthemum (Dendranthema grandiflora)

This chapter provides reproducible methods for the transformation of Dendranthema � grandiflora ‘Shuhou-no-chikara’ (standard) and ‘Lineker’ (spray) stem internode thin cell layers (TCLs) and conventional stem internode explants using Agrobacterium strains carrying a binary vector with β-glucoronidase (GUS) reporter and npt II selector genes. Transformation efficiencies are reported at the kanamycin selection (callus formation and plantlet rooting), GUS assay, polymerase chain reaction (PCR), and Southern analysis levels. Notes on regeneration improvement through the use of TCLs, as well as the effective use of sonication for both regeneration and generation of transformants, stimulation of Agro -infection and elimination of Agrobacterium , are included.