Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 1219-1225 |
| Seitenumfang | 7 |
| Fachzeitschrift | Plant cell reports |
| Jahrgang | 25 |
| Ausgabenummer | 11 |
| Publikationsstatus | Veröffentlicht - 24 Juni 2006 |
Abstract
A protocol for Agrobacterium tumefaciens-mediated genetic transformation of Rhipsalidopsis cv. CB5 was developed. Calluses derived from phylloclade explants and sub-cultured onto fresh callus induction medium over a period of 9-12 months were co-cultivated with A. tumefaciens LBA4404. Plasmid constructs carrying the nptII gene, as a selectable marker, and the reporter uidA gene were used. Transformed Rhipsalidopsis calluses with a vigorous growth phenotype were obtained by extended culture on media containing 600 mg l-1 kanamycin. After 9 months of a stringent selection pressure, the removal of kanamycin from the final medium together with the culture of the transformed calluses under nutritional stress led to the formation of several transgenic adventitious shoots. Transformation was confirmed by GUS staining (for uidA gene), ELISA analysis and Southern blot hybridization (for the nptII gene). With this approach, a transformation efficiency of 22.7% was achieved. Overall results described in this study demonstrate that Agrobacterium-mediated transformation is a promising approach for this cactus species.
ASJC Scopus Sachgebiete
- Agrar- und Biowissenschaften (insg.)
- Agronomie und Nutzpflanzenwissenschaften
- Agrar- und Biowissenschaften (insg.)
- Pflanzenkunde
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in: Plant cell reports, Jahrgang 25, Nr. 11, 24.06.2006, S. 1219-1225.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Agrobacterium tumefaciens-mediated transformation of Rhipsalidopsis gaertneri
AU - Al-Ramamneh, E. A.
AU - Sriskandarajah, S.
AU - Serek, M.
N1 - Funding Information: Acknowledgements The project was supported by a Ph.D. grant from Al Balqa Applied University in Jordan, additional grants from University of Hannover, Danish Ministry of Agriculture and Fisheries in Denmark (93s-2466-Å01-01430) and Danish Schlumbergera Growers and Breeders: Gartneriet Thoruplund A/S (Odense), Ro-hdes Gartneri A/S (Kerteminde), Gartneriet PKM ApS (Odense), and Hansson DK (Søndersø).
PY - 2006/6/24
Y1 - 2006/6/24
N2 - A protocol for Agrobacterium tumefaciens-mediated genetic transformation of Rhipsalidopsis cv. CB5 was developed. Calluses derived from phylloclade explants and sub-cultured onto fresh callus induction medium over a period of 9-12 months were co-cultivated with A. tumefaciens LBA4404. Plasmid constructs carrying the nptII gene, as a selectable marker, and the reporter uidA gene were used. Transformed Rhipsalidopsis calluses with a vigorous growth phenotype were obtained by extended culture on media containing 600 mg l-1 kanamycin. After 9 months of a stringent selection pressure, the removal of kanamycin from the final medium together with the culture of the transformed calluses under nutritional stress led to the formation of several transgenic adventitious shoots. Transformation was confirmed by GUS staining (for uidA gene), ELISA analysis and Southern blot hybridization (for the nptII gene). With this approach, a transformation efficiency of 22.7% was achieved. Overall results described in this study demonstrate that Agrobacterium-mediated transformation is a promising approach for this cactus species.
AB - A protocol for Agrobacterium tumefaciens-mediated genetic transformation of Rhipsalidopsis cv. CB5 was developed. Calluses derived from phylloclade explants and sub-cultured onto fresh callus induction medium over a period of 9-12 months were co-cultivated with A. tumefaciens LBA4404. Plasmid constructs carrying the nptII gene, as a selectable marker, and the reporter uidA gene were used. Transformed Rhipsalidopsis calluses with a vigorous growth phenotype were obtained by extended culture on media containing 600 mg l-1 kanamycin. After 9 months of a stringent selection pressure, the removal of kanamycin from the final medium together with the culture of the transformed calluses under nutritional stress led to the formation of several transgenic adventitious shoots. Transformation was confirmed by GUS staining (for uidA gene), ELISA analysis and Southern blot hybridization (for the nptII gene). With this approach, a transformation efficiency of 22.7% was achieved. Overall results described in this study demonstrate that Agrobacterium-mediated transformation is a promising approach for this cactus species.
KW - Agrobacterium tumefaciens
KW - Cactus
KW - Genetic transformation
KW - Nutritional stress
KW - Rhipsalidopsis gaertneri
UR - http://www.scopus.com/inward/record.url?scp=33750353993&partnerID=8YFLogxK
U2 - 10.1007/s00299-006-0190-x
DO - 10.1007/s00299-006-0190-x
M3 - Article
C2 - 16799807
AN - SCOPUS:33750353993
VL - 25
SP - 1219
EP - 1225
JO - Plant cell reports
JF - Plant cell reports
SN - 0721-7714
IS - 11
ER -