Chrysanthemum (Dendranthema x grandiflora).
This chapter provides reproducible methods for the transformation of Dendranthema x 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 beta-glucoronidase (GUS) reporter and nptII 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.
Teixeira da Silva
Jaime A
JA
Kagawa University, Horticulture Department, Miki-cho, Ikenobe, 761-0795, Kagawa-ken, Japan.
D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics*; D018929:Cell Culture Techniques; D031202:Chrysanthemum / Q000033:anatomy & histology / Q000235:genetics* / Q000502:physiology; D018920:Coculture Techniques; D003470:Culture Media; D017930:Genes, Reporter; D005966:Glucuronidase / Q000032:analysis; D018547:Plant Stems / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology; D016133:Polymerase Chain Reaction; D012038:Regeneration; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
D003470:Culture Media; D005966:Glucuronidase