Wheat transformation with glutenin gene

The rheological properties of wheat grains are associated with the composition of the starchy endosperm in high molecular weight (HMW) glutenin proteins. The HMW glutenin 1xDy12 subunit gene was co introduced with the screenable bar and the reporter gus marker genes in the commercial spring wheat Minaret cultivar (cv). The gene of interest and the marker genes were carried by two separated plasmids, the pBS10BH1 and the pAhC25 respectively. Seven days old calli initiated from immature embryos were bombarded by use of the PDS-1000/He device. A number of different bombardment and culture conditions were tested. These parameters were evaluated on the basis of GUS transient expression. Among those a 4 degrees C cold pre-treatment of the spikes before immature embryos were excised, the culture medium incorporating activated charcoal, silver nitrate and glucose yielded higher GUS transient expression rate. The selective agent bialaphos was maintained at various stages during culture from induction of somatic embryogenesis to rooting of regenerated plantlets. 137 bialaphos resistant plants were obtained among which 109 were carried to maturity. Transgenic plants were characterized by PCR, Southern and SDS-PAGE analysis of the glutenin content.