Study On Sweet Sorghum Sucrose Synthase Genes’ Clone And Transgenic Of EPSPS Gene

Sweet sorghum can harvest food on one hand, on the other hand it can also obtain stalk with sugar. At the same time, it has drought tolerance and salt tolerance competing with other crops. Tough sweet sorghum has so many advantages, it is still exist some problems. Such as, sugar content of its stem is not that high, negative sensitive to herbicides are not good for reducing production cost, and sensitive to light is not good for high yield two season planting etc. On the purpose of above objective, I have done there parts of experiment. They are cloning sucrose synthase gene and its front regulating sequence of sweet sorghum North sweet three, establishing regeneration and transformation system of North sweet three, and transporting herbicide resistant gene EPSPS into North sweet three. The conclusions are as follows.1.North sweet three’s sequence and analysis of sucrose synthase. By analysis, the result is the whole sequence total encoding802amino acids, and5992bp in length. Through blast, I know that the sucrose synthase sequence between North sweet three and AFRICA WHITE has a similarity of97%, which proves that sucrose synthase gene is conserved.2.Regeneration and transformation system of North sweet three. Different gene-types of sweet sorghum have different kinds of medium. Through doing experiment and analysis data, I know that the beat callus induction medium of North sweet three is MS+2.5mg/L2,4-D+O.lmg/L TDZ, and the best inducing differentiation medium of North sweet three is MS+0.5mg/L NAA+1.0mg/L6-BA.3.Transgenic Herbicide resistant gene EPSPS. I find that if the infection time is too long, the phenomenon of large numbers of callus browning to die would appear during agro-bacterium co-cultivation period. And if it is too short, there would be much more false positive callus in the screening of callus. The callus in disseminated sloshing process which is easy to loose are more likely to survive during co-culture time. The best optimum culture time is three days and two nights. The first week’s best concentration of antibiotics during screening process is50ug/ml kanamycin,75mg/ml hygromycin and500mg/L carbenicillin, and second week is50ug/ml kanamycin,37.5mg/ml hygromycin and250mg/L carbenicillin. I find the culture the bud separately is better.