GENETIC-TRANSFORMATION OF EUSTOMA-GRANDIFLORUM GRISEB BY MICROPROJECTILE BOMBARDMENT

Foreign DNA was introduced into cell suspension cultures and leaf tissue of Eustoma grandiflorum Griseb. (lisianthus) by microprojectile bombardment. For this purpose a low-cost bombardment device that uses a helium flux to accelerate microprojectiles was built. When cell suspensions were used, an average of 4.1 Kan resistant calli were recovered per shot after 4 months' cultivation on selective medium. Most of the Kan resistant plants regenerated from calli were positive to GUS assay. Both the nptII and gus genes were successfully amplified from alkali-treated leaves of putative transgenic plants by PCR analysis. Transgenic plants were not recovered from bombarded leaves. Considering the host range specificity of Agrobacterium, and the response of the species to plant regeneration from suspension culture, microprojectile bombardment is, at present, the most efficient procedure for genetic transformation of lisianthus.