Transformation of Antisense ACS Gene to Banana Mediated by Agrobacterium tumefaciens

A high efficiency and systematic transgenic procedure mediated by Agrobacterium was developed in Musa spp.cv.Tianbaojiao(AAA group).The introduction of antisense ACS gene to banana was conducted,and the optimized parameters used in Agrobacterium-mediated transformation were obtained.The best transient expression of GUS was obtained under the conditions as follows:the thin cross-sections which were not precultured but were pretreated with 0.1 mol/L mannitol for 4 h,and the Agrobacterium suspension was with OD600 value of 1.0 and supplemented with 100 g/L sucrose,the samples were infected for 10~15 mins,and then they were transferred onto the non-selective medium(pH5.8) supplemented with 0.1 mg/L NAA and 1.0 mg/L BA for co-culture for 4 days at 26 ℃ in the dark.After co-culture,the thin cross-sections were selected on MS medium supplemented with 100 mg/L kanamycin and 2 mg/L AgNO3,which could be used to inhibit Agrobacterium contamination and be good for bud differentiation.At last,5 resistant bud lines were maintained for further studies.The GUS histochemical and PCR assays proved that the gus gene had integrated into the genome of the 5 resistant bud lines.