Abstract: Protoplasts were isolated from suspension cells of a wheat cultivar“Bodallin”.The protoplasts were transformed by electroporation with uptake of PBC1 plasmid,containing both βglucuronidase(GUS)and hygromycin B phosphotransferase(HPT) genes.By means of BTX ECM6000 Electroporation System and ASP buffer,maximal GUS activity of the transformed protoplasts was obtained under the condition of 300 V(750V/cm)and 50 ms (app.1000 μF),when an effective concentration (25 μg/ml) of PBC1 plasmid was employed.After gene transfer via electroporation,transient expression of GUS in the protoplasts cultured for 2-3 days was able to reach a high leve1.In addition,GUS activity could be enhanced to certain level,by the aid of calf DNA used as a carrier DNA during electroporation.Protoplasts electroporated with PBC1 DNA were cultured on KMP medium containing 50-100 μg/ml hygromycin for 3-4 months and selection of hygromycin resistant call was conducted.A total of 15 calli from transformed protoplasts survived in selective cultures on 100 μg/m1,hugromycin a concentration that completely inhibited the growth of non transformed wheat callus.

Key words: Wheat protoplasts, Electroporation, Gene transfer, GUS activity, Hygromycin resistant callus

CLC Number: 

• S512.101