Abstract
Transient expression of electroporation-mediated DNA uptake was monitored in callus-derived protoplasts of two asparagus (Asparagus offcinalis L.) genotypes by measuring the GUS activity. The level of expression and the viability of the protoplasts were influenced by the voltage and duration of the electric pulse. An increased plasmid DNA concentration and the presence of polyethylene glycol (PEG) in the electroporation medium enhanced the transient expression level. A considerable increase in GUS activity was observed in the presence of both PEG and heat-shock treatments than with PEG treatment alone. An optimal level of GUS activity was obtained after electroporation with a capacitive discharge of 500 V/cm and 94 ms duration. The two genotypes differed in their responses in vitro and also showed variable levels of transient expression. The present technique was suitable to obtain transgenic plants, as histochemical GUS assay revealed GUS activity in the protoplast-derived microcolonies as well as in callus tissues.
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