Carnation (Dianthus caryophylus L.)
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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/L thidiazuron (TDZ), 0.1 mg/L α-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 µM acetosyringone (AS). After cocultivation, the explants were cut into eight radial segments and subcultured onto selection
medium until transformed shoots regenerated from the explants.
Affiliation(s): (3) Department of Horticulture, Faculty of Agricultural production, Maejo University, Chiangmai, 50290, Thailand
(4) Department of Horticulture Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa 761-0795, Japan
(4) Department of Horticulture Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa 761-0795, Japan
Book Title: Agrobacterium Protocols Volume 2
Series: Methods in Molecular Biology | Volume: 344 | Pub. Date: Sep-01-2006 | Page Range: 311-320 | DOI: 10.1385/1-59745-131-2:311
Subject: Plant Sciences
Key Words: Agrobacterium - genetic transformation - carnation (Dianthus caryophylus L.) - adventitious shoot regeneration - node explants - gus (β-glucuronidase) -
nptII (neomycin phosphotransferase) - AS (acetosyringone)
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