Carnation (Dianthus caryophylus L.).
Carnation is a valuable crop for the cut flower industry and demand for new and improved varieties is growing. However, genetic transformation of carnations is currently limited because of a lack of efficient routine technique. In this chapter, we present an easy and effective protocol for gene transfer to carnation node explants and subsequent adventitious shoot regeneration. For high-adventitious shoot regeneration, node explants from first to third node of 5- to 8-cm long shoots were cultured on Murashige and Skoog (MS) medium, containing 1.0 mg/Lthidiazuron (TDZ), 0.1 mg/L alpha-napthalenoacetic acid (NAA), 20 g/L sucrose, and 2 g/L Gellan gum for 10 d. Then the explants were cut into 8 radial segments and subcultured onto MS medium, containing 1.0 mg/L BA, 0.1 mg/L NAA, 20 g/L sucrose and 2 g/L Gellan Gum. For effective genetic transformation, 3- to 5-d precultured node explants were submerged in an Agrobacerium suspension for 10 min, then cocultivated on filter paper soaked with water and 50 microM acetosyringone (AS). After cocultivation, the explants were cut into eight radial segments and subcultured onto selection medium until transformed shoots regenerated from the explants.
Nontaswatsri
Chalermsri
C
Department of Horticulture, Faculty of Agricultural Production, Maejo University, Chiangmai, 50290, Thailand.
Fukai
Seiichi
S
D000064:Acclimatization; D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015139:Blotting, Southern; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D003470:Culture Media; D018744:DNA, Plant / Q000737:chemistry; D029749:Dianthus / Q000033:anatomy & histology / Q000235:genetics* / Q000502:physiology; D018014:Gene Transfer Techniques; D005966:Glucuronidase / Q000032:analysis; D018520:Plant Shoots / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology; D030821:Plants, Genetically Modified / Q000254:growth & development; D016133:Polymerase Chain Reaction; D012038:Regeneration; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
D000900:Anti-Bacterial Agents; D003470:Culture Media; D018744:DNA, Plant; D005966:Glucuronidase