Study Of The ACC Synthase Transgenic And Rooting And Transplantation System Of Jiro

Persimmon is a common fruit, whose production is nearly 90% in the world. Its nutrition is abundant and flavor is unique. It has high edible and medical value, but to be able to soft. A protocal of a reproducible Agrobacterium tumefaciens-mediated transgenic system for the production of resisting soft transgenic plants of persimmon that transmits transgenes of the ACC synthase RNAi gene into progeny have been developed.1. Establishing a high efficient and stable genetic transformation system of Jiro in vitro. In the work, it studied systemicly the transformation conditions. The main results were as follows,1.1 Establishing the proper concentration of antibioticCompared the restraining effect with the different Cef concrete rate, 200 mg/L could have restrained the A.t. Compared the restraining effect to the shoot with the Sp of 20, 30, 40, 50 and 60 mg/L, the proper concentration was 30 mg/L.1.2 The transgenic technic of JiroCutting 0.5 mm~2 from the base of 2 to 3 leaves of the top of Jiro of 3.5 years in vitro from generation to generation, putting with top of the back of leaf on the medium of DKW(1/2N) + ZT 1.0 mg/L + TDZ 0.5 mg/L, dipping the leaves after pre-cultured in darkness for four days into Agrobacterium suspension which was AS 50 mg/L in it for ten minutes, transferring the leaves to the medium of DKW(1/2N) + ZT 1.0 mg/L + TDZ 0.5 mg/L with co-cultured in darkness for three days, then diverting the leaves to the medium of DKW(1/2N) + ZT 1.0 mg/L + TDZ 0.5 mg/L + Cef 200 mg/L with de-cultured in darkness for three days, finally transferring the leaves to the medium of DKW(1/2N) + ZT 1.0 mg/L + TDZ 0.5 mg/L + Cef 200 mg/L + Sp 30 mg/L for selecting in light, once every 20 days, transferring the leaves to the medium of DKW + ZT 1.0 mg/L + Sp 30 mg/L + Cef 200 mg/L after three times, differentiation percentage of leaves was able to get 27%.In this study, we obtained 8 Jiro transgenetic plants carrying ACC synthase RNAi gene which identified by PCR, 1.6% of regenerate shoots. 2 Establishing the rooting and transplantation system of Jiro in vitro.2.1 It selected the high 15 to 25 mm, strong tube culture with 2 to 3 leaves in the medium of DKW + ZT 1.0 mg/L as explants. The explants were curtured in darkness for seven days, then curtured in light of 36μmol/m~2s for thirty days after inserting in the rooting medium of 1/2 MS + IBA 1.0 mg/L. Rooting rate of Jiro was upwards to 95% in the case of above conditions.2.2 Take the above rooting explants from vitro after light of 180μmol/m2s for seven days, cleaning them and disinfecting them, inserting them into vermiculite disinfected in the high temperature. The humidity in the air was upwards to 90%. the temperature was 25℃. The intensity of light was 180μmol/m~2s. Reduced gradually the humidity(upwards to 80%), and enhanced the intensity of light, 180 to 540μmol/m~2s.