Production of Transgenic Plants via Agrobacterium tumefaciens- Mediated Genetic Transformation in Pinus wallichiana (Himalayan Blue Pine)

We established for the first time an Agrobacterium-mediated genetic transformation system for the Himalayan blue pine. Embryogenic tissue derived from vegetative shoot apices of mature trees of Pinus wallichiana were inoculated with A. tumefaciens strain EHA105 fused with a binary vector pBI121. The plasmid pBI121, containing the neomycin phosphotransferase II (nptII) gene providing kanamycin resistance as a selectable marker and the � -glucuronidase (uidA) reporter gene, was used in the transformation studies. GUS activity was used to monitor transient expression of the uidA gene and to further test lines selected on kanamycin-containing medium. The integration of the transgene (nptII) was confirmed by PCR followed by southern and northern blot analyses. These results demonstrated that a stable and enhanced transformation system could be established in P. wallichiana. This provides an opportunity to transfer economically important genes into Himalayan blue pines.