Abstract
A genetic transformation protocol was developed using the transfer of a synthetic CP4 EPSPS transgene, as a conditional positive selectable marker, into commercially relevant zonal pelargoniums using an Agrobacterium tumefaciens strain in combination with a novel step-down glyphosate selection system. The transformation efficiency based on independent T-DNA integration events averaged 1.9 % over 10 experiments. Some 273 independent transformants were produced within an average time of 6 mo from explant inoculation with Agrobacterium to plantlet recovery. For plantlet recovery, three aromatic amino acids were incorporated into the rooting medium to ameliorate the accumulative effects of glyphosate selection. The T-DNA also contained a mutant ethylene receptor (etr1-1) cDNA from Arabidopsis thaliana, under control of the petunia flower-specific, floral-binding protein promoter, to confer ethylene insensitivity. However, delayed flower senescence was not obtained. The transformation protocol provides a reliable method to add herbicide resistance and other traits to zonal pelargonium.
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