Herbicide-resistant transgenic creeping bentgrass plants obtained by electroporation using an altered buffer

Modification of an electroporation buffer using Ca(NO3)(2) and elevated pH (9-10) appeared to have a favorable effect on gene transfer to creeping bentgrass (Agrostis palustris Huds.) Penncross protoplasts, resulting in an increase in the transformation frequency of about twofold. Following electroporation with the plasmid pARK22 containing the bar gene, a total of 278 bialaphos-resistant cell colonies were obtained from four experiments. The bi alaphos-resistant regenerants proved to be transgenic by Southern hybridization of the amplified DNA, All the tested transgenic plants showed herbicide (HERBIE) resistance at the field rate of 0.5-1% (vol/vol). Ammonia contents in leaves after spraying with the herbicide increased less in transgenic plants than in untransformed control plants.