caa a22 4500 467901600 CHVBK 20180406152820.0 cr unu---uuuuu 170328e20061101xx s 000 0 eng 10.1007/s00299-006-0190-x doi (NATIONALLICENCE)springer-10.1007/s00299-006-0190-x Agrobacterium tumefaciens -mediated transformation of Rhipsalidopsis gaertneri [Elektronische Daten] [E. A. Al-Ramamneh, S. Sriskandarajah, M. Serek] 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 600mgl−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. Springer-Verlag, 2006 Agrobacterium tumefaciens nationallicence Cactus nationallicence Genetic transformation nationallicence Nutritional stress nationallicence Rhipsalidopsis gaertneri nationallicence Al-Ramamneh E. A. Department of Agricultural Sciences, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark aut Sriskandarajah S. Department of Agricultural Sciences, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark aut Serek M. Department of Agricultural Sciences, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark aut Plant Cell Reports Springer-Verlag 25/11(2006-11-01), 1219-1225 0721-7714 25:11<1219 2006 25 299 https://doi.org/10.1007/s00299-006-0190-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-006-0190-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Al-Ramamneh E. A. Department of Agricultural Sciences, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Sriskandarajah S. Department of Agricultural Sciences, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Serek M. Department of Agricultural Sciences, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, 1871, Frederiksberg C, Copenhagen, Denmark aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/11(2006-11-01), 1219-1225 0721-7714 25:11<1219 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer