Studies On Construction Of Sucrose Phophate Synthesis Gene Anti Expression Vector And Transformation Into Tomato

In this research,anti-TSPS expression vector was constructed and transformed to tomato plant via Agrobacterium-mediated transformation.The main results were as follows:1.The experiments digested tomato fruit sucrose phosphate synthase gene which was cloned to pMD18-T vector with double enzymes Hincâ…¡and XbaI from 1.9 kb long to 830bp long.Antisense expression vector of tomato fruit sucrose phosphate synthase gene was successfully constructed after linking the 830 bp fragment and pROK3 vector which was digested by XbaI and SmaI using T4DNA ligase enzymes.The result showed that TSPS successfully linked to plant expression vector pROK3.2.The experiment concluded that the optimal medium for callus induction and bud differentiation was MS+ZT 2.0 mg/L +IAA 0.5 mg/L,the optimal medium for root induction and sound seedling was 1/2 MS+IBA 0.5 mg/L,the optimal explants for transformation was cotyledon.It was to assure the lived rate of regenerated plant after cultured about 7 days in cultured room and then growed to field.3.Following regeneration system above,tomato cotyledons were transformed via Agrobacterium-mediated transformation and 135 regeneration tomato plants were obtained. Distilling tomato leaves DNA grouping by CTAB method and using PCR analysis,the results showed that two groups had 830 bp lines.Then distilling DNA from identified groups separately,two of them had the aimed line.Southern blot analysis showed that the two plants had blot signal.It was said that Anti-TSPS gene was integrated into tomato genome and transformed rate was 1.4%.4.The experiments about SPS active,sugar and acid contents in genetic tomato showed dramatically lower than CK,it indicated that sucrose metabolized and accumulated were influenced by Anti-TSPS.