Abstract:

In order to improve the regeneration frequency and the rate of genetic transformation of agrobacterium-mediated callus of mature embryos for maintainer line (YTB) of HL-CMS, indica rice, a series of experimental program were carried out, such as, composition of culture medium, concentration of bacilli, infection method, co-culture methods, callus drying method, concentration of selection agent. The results revealed that both of LB and YEB medium can culture agrobacterium; vacuum treatment can significantly improve the rate of conversion; adding filter paper on co-culture medium is in favor of subsequent sterilization and can improve the conversion efficiency; callus dried properly can significantly improve the conversion efficiency; the appropriate concentration of hygromycin is between 40-50mg.L-1-1; the use of V-type screening method can increase the positive rate of transgenic plants. So, the systematic optimization of culture methods for agrobacterium can improve regeneration frequency of callus of mature embryos for indica rice.