Green fluorescent protein as a reporter system in the transformation of barley cultivars

A cereal transformation vector, pN1473, containing the strong constitutive rice actin promoter Act-1, a multiple cloning site, and the nos terminator, was constructed. Fusion of a plant-optimized gfp gene to Act-1 in pN1473 r esulted in the vector pN1473GFP. To assess the suitability of pN1473, and G FP as a reporter system in barley transformation, two barley cultivars (Bar onesse and Golden Promise) were transformed by microprojectile bombardment. Transient gfp expression in transformed embryogenic callus material was de tectable by fluorescence microscopy less than 12 h after transformation. Th e presence of the gfp gene in callus and regenerated plantlets was confirme d by PCR amplification and DNA gel-blot analysis.