ELECTROPORATION-MEDIATED TRANSIENT GENE-EXPRESSION IN ISOLATED SCUTELLA OF HORDEUM-VULGARE

Electroporation was used to introduce DNA into excised scutella of imm ature embryos of Hordeum vulgare L. cvs Golden Promise and Delita. Usi ng the firefly luciferase gene as reporter, parameters were analyzed f or high transient gene expression while maintaining tissue viability. Enzymatic wounding was necessary for DNA uptake. The optimized protoco l involves use of linearized DNA and addition of 15% (w/v) polyethylen e glycol at a field strength of 950 V cm(-1) and approximately 56 ms p ulse length. A one-day preculture was required for obtaining callus af ter electroporation. Transient gene expression was further demonstrate d using the beta-glucuronidase gene. Blue spots were detected at the a baxial scutellar surface, indicating that cells competent for somatic embryogenesis are also amenable to transfection by electroporation,