Agrobacterium-mediated genetic transformation of date palm (Phoenix dactylifera L.) cultivar "Khalasah" via somatic embryogenesis
DOI:
https://doi.org/10.14719/pst.2015.2.3.119Keywords:
Date palm, Phoenix dactylifera, Agrobacterium tumefaciens, genetic transformation, somatic embryogenesis, kanamycin resistanceAbstract
In present investigation, an efficient Agrobacterium- mediated genetic transformation was successfully carried out for a well known date palm (Phoenix dactylifera L.) cultivar “Khalasah” using matured somatic embryos. Somatic embryogenesis was initiated from offshoot’s shoot tips of date palm cultivar. For genetic transformation, morphologically advanced matured somatic embryos developed on MS medium fortified with TDZ (1.0 mg/l) were co-cultured with A. tumefaciens strain LBA 4404 harboring binary vector pBI 121, containing uidA (GUS) and npt II genes and incubated for 4 days and later it was (somatic embryos) inoculated on germinating and plantlet conversion MS medium supplemented with BAP (0.75 mg/l) + kanamycin (100 mg/l). Prolific shoots developed from putatively transformed matured embryos showed 47.5 % transformation efficiency. A large number of transgenic plants were obtained and later established in black hard plastic bags. A strong GUS activity was detected in the putatively transformed plant leaves by histochemical assay and, the integration of uidA (GUS) and npt II genes into transgenic plants was confirmed by polymerase chain reaction (PCR) and Southern hybridization analysis. The established transformation protocol allows effective and quick regeneration via somatic embryogenesis, and it would be highly valuable for existing date palm orchards and improving their productivity.
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