OPTIMIZATION OF AGROBACTERIUM-MEDIATED TRANSFORMATION OF SUGARCANE

摘要

Aims: To optimize agrobaterium-mediated transormation in sugarcane. Methods & Results: The embryogenic callus of the cultivar sugarcane cultivar Min-tang 92-649 were used as explant to study the effects on different factors of pre-culture media, pre-culture time, infection time, acetosyringone concentrations and co-culture conditions on transient expression frequency of gus gene. The results showed that condition of optimization of the cultivar Min-tang 92-649 transgened by Agro bacterium-mediated had been pre-cultured for 5 d in pre-culture medium (MS+2.4-D+6-BA0.1mg/L), and then Agrobacterium tumefactions bacterial infection time for 30min, and co-culture at the temperature from 23°C to 25°C. It is also found that 50 mg/L hygromycin would be aneffective concentration to be used for the selection of transformed lines. To be higher in transient expression frequency of gus gene, it is important to add AS 150 mu mol/L to co-culture medium. The presence of the introduced GUS genes in the transgenicplants was confirmed by polymerase chain reaction.