2000 Volume 69 Issue 6 Pages 345-352
A reliable method for Agrobacterium-mediated transformation of mulberry (Mores alba L.) callus has been developed. Calli initiated from cotyledons, hypocotyls and roots of seedlings were co-cultivated with Agrobacteria in the presence of acetosyringone under acidic pH (pH5.5) at low temperature (20-22°C). Putative transgenic calli were selected by using a medium containing 12.5μg/ml hygromycin. When the calli initiated from hypocotyls and A. tumefaciens strain EHA101/pIG121-Hm were used, the highest transformation frequency, as evaluated by resistance to hygromycin, was obtained. Completely transformed calli were obtained through several subculturings by gradually increasing the concentration of hygromycin in the selection medium. GUS assay and PCR analysis clearly indicated that the obtained hygromycin-resistant calli were transformed with T-DNA from binary vector to mulberry genome.
