Transformation Of The ACC Synthase Antisense Gene In Persimmon Fruit

In this study, high-frequency regeneration and transformation systems had been established through selecting the best conduction of regeneration, transformation and antibiotic selection of Uenishiwase leaves in vitro, and ACC gene had been inserted into Uenishiwase. We can increase the storaging function of climacteric fruit by repressing the expression of the ACC synthase enzymes with transgenetic method. The mainresults were as follows,1. Establishing a high efficient and stable regeneration system in Uenishiwase leaf in vitro.In order to found an effective regeneration system in vitro for Uenishiwase, factors of basic medium, different hormone combination, dark treatment and the part of leave was studied. Result shows that MS(l/2N)medium, dark treatment for ten days,the latter of leave was the effective regeneration system.2. Studying effects of antibiotic on the regeneration of Uenishiwase leaves in vitro and the growth of shoots, ensured selecting pressure.It was suitable that 200 mg/L Cef and 20 mg/L Sp used acted as concentration of controlled the propagation of A.g and level of selection in the regeneration experiment of transformation in Uenishiwase, that 200 mg/L Cef and 10 mg/LSp used on growth of resistant shoots.3. Establishing the genetic transformation system and transferring ACC genes into Uenishiwase.Factors that influence percentage of transformation including time of pre-culture, Infection time, concentration of A.g liquid, AS concentration and co-culture were studied. The effective transformation system of Uenishiwase was developed as follow: Leaves pre-cultured in darkness for three days were dipped into Agrobacterium suspension for ten minutes, then were co-cultured in darkness for four day. In this study, we obtained Uenishiwase transgenetic plants carrying ACC gene which identified by PCR .4. Studies on Rooting Conditions of Uenishiwase Persimmon in Vitro.Effects of basal mediums, hormones and dark treatment on rooting in vitro of Uenishiwase persimmon were studied. The results showed that 1/2MS medium was the best basal medium.The best combination of hormones and their concentrations was thatIBAl.Omg/L+IAAOmg/L.Dark treatment in five days improved the rooting in vitro. Rooting of Uenishiwase reached 90% .