Production of transgenic sugarcane (Saccharum officinarum L.) plants by intact cell electroporation

Summary We describe an efficient procedure for genetic transformation of commercial sugarcane varieties POJ 2878 and Ja 60-5. The transformation protocol is based on electroporation of a plasmid conferring GUS activity into cell clusters isolated from embryogenic calli. Six to eight weeks after electroporation, Ja 60-5 plants regenerated from electroporated tissues were tested and confirmed to be transgenic using histochemical glucuronidase and Southern hybridization analysis. Electroporation of intact cells is an efficient and reproducible method for sugarcane transformation and may also be useful for transformation of other plants.Abhrevations GUS -glucuronidase - CAT chloramphenicol acetyl transferase - PCV packed cell volume - PCR polymerase chain reaction - DTT dithiotreitol - Hepes N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - NOS nopaline synthase - 4-MUG 4-methylumbelliferyl -D-glucuronide