STUDIES ON Agrobacterium tumefaciens-MEDIATED GENETIC TRANSFORMATION IN STRAWBERRY

Several factors affecting Agrobacterium tumefaciens-mediated gene transformation of strawberry were studied by using the transient expression of gus gene with introns as criteria.The results showed that:the explants of strawberry leaves were rather sensitive to kanamycin,and the corresponding concentration of 20 mg/L could substantially inhibit the growth of non-transformed tisse and be used to screen the leaves of strawberry after transformation.Timentin at the concentration of 300 mg/L could be used to eliminate Agrobacterium tumefaciens and had minor impact on the growth of the explants.The preculture time of 2-4 days was beneficial to improve transformation frequency.Co-culture for 2-3 days not only had higher gus transient expression but also avoid the over-growth of agrobacterium.The bacterial concentration of OD_(600nm) 0.4 was the best inoculum density and 10 minutes was the best inoculation time.The PCR analysis proved that the target gene gus was successfully integrated into the genome of strawberry.