Stable transformation of Gladiolus by particle gun bombardment of cormels

Abstract

Transgenic plants of Gladiolus were produced following particle bombardment of cormel slices. Plant cells were cotransformed with the gene for phosphinothricin acetyltransferase under control of the cauliflower mosaic virus 35S promoter and the uidA gene coding for β-glucuronidase (GUS) under control of the actin promoter isolated from rice. The optimum concentration for the first selection of transformed plants was 8 mg/l phosphinothricin which resulted in 14% of the bombarded cormel slices regenerating plants that were transformed as confirmed by polymerase chain reaction amplification. Polymerase chain reaction amplification, Southern hybridization and histochemical staining for GUS gene expression on plants after two selective screenings with phosphinothricin indicated that regenerated plants were transformed. Histochemical staining for GUS gene expression showed that the actin promoter resulted in GUS gene expression primarily in callus cells and root meristems. Leaves were typically chimeric for GUS gene expression.