Regeneration of transformed verbena (Verbena x hybrida) by Agrobacterium tumefaciens.
Verbena (Verbena x hybrida), an important floricultural species, was successfully regenerated from stem segments on Murashige and Skoog's basal medium supplemented with thidiazuron and indole-3-acetic acid. A transformation system was developed using cvs. Temari Scarlet, Temari Sakura, Tapien Rose and TP-P2. Agrobacterium tumefaciens strain Agl0 harboring the sGFP gene was infected into stem segments. Transformation efficiency was improved by evaluating and manipulating the age of the plant material, the concentration of kanamycin in the medium during selection, and the length of the culture period in the dark. After 2-3 months of culture on the selection medium, GFP-positive shoots were obtained in all four of the cultivars tested. These shoots were successfully acclimated and set flowers within 2-3 months in a greenhouse. GFP was expressed in all of the organs including the floral parts. Stable genomic transformation was confirmed by Southern blot analysis. No morphological differences were observed between the transformed plants and their host plants.
Tamura
M
M
Institute for Advanced Technology, Suntory Ltd, 1-1-1 Wakayamadai, Shimamoto-cho, Osaka 618-8503, Japan. Yoshikazu_Tanaka@suntory.co.jp
Togami
J
J
Ishiguro
K
K
Nakamura
N
N
Katsumoto
Y
Y
Suzuki
K
K
Kusumi
T
T
Tanaka
Y
Y
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000254:growth & development; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D003433:Crosses, Genetic; D003624:Darkness; D005838:Genotype; D049452:Green Fluorescent Proteins; D006823:Hybrid Vigor / Q000235:genetics; D006824:Hybridization, Genetic; D007210:Indoleacetic Acids / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009280:Naphthaleneacetic Acids / Q000494:pharmacology; D010671:Phenylurea Compounds / Q000494:pharmacology; D018520:Plant Shoots / Q000235:genetics / Q000502:physiology; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D012038:Regeneration; D013395:Sucrose / Q000494:pharmacology; D013830:Thiadiazoles; D013997:Time Factors; D014170:Transformation, Genetic; D032412:Verbena / Q000235:genetics*
D000900:Anti-Bacterial Agents; D007210:Indoleacetic Acids; D008164:Luminescent Proteins; D009280:Naphthaleneacetic Acids; D010671:Phenylurea Compounds; D013830:Thiadiazoles; C016785:thidiazuron; D049452:Green Fluorescent Proteins; D013395:Sucrose; C030737:indoleacetic acid