Clone And Transformation Of Sweet Sorghum Sucrose Transporter Protein SUT1Gene

Sweet sorghum is considered one of the most development potential of energy crops. But itsgermplasm resources is deficient and sugar content is low, which is an obstacle to thedevelopment of sweet sorghum industrialization producing alcohol, leading to the industrydifficultly forming large-scale production. It proved that sucrose transporter SUT1has asignificant impact on the amount of storage sugar sweet sorghum, if we make it clear howsucrose transporter SUT1gene plays an role in the interaction of sorghum source database and itsregulatiory mechanism, We will not provide a reference in the su crose transportation anddistribution mechanism of sweet sorghum, but make a theoretical basis for improvement, whichhas the vital significant on the breeding of sweet sorghum with the high sugar content character.In this study, sucrose transporter SUT1was coloned from sweet sorghum by adopting thetechnology of RACE and homologous cloning, SUT1was analyzed by bioinformatics softwares.After the construction of SUT1overexpression vector, SUT1was transformed into sweetsorghum by Agrobacterium mediation to obtain transgenic plant. The botained transgenic plantscan be for improve sorghum stalk sugar content reference. Current results are as follows:1. It is the first time of clone sucrose transporter protein SUT1gene from sweet sorghum,and the result of its bioinformatics anslysis showed that the full length of SUT1cDNA sequencescloned from sweet sorghum was2472bp, including open reading frame (Open reading frame,ORF)1560bp, which encodes519amino acids. The expression vector containing the SUT1gene was constructed successfully by ligating the SUT1gene to the UBI-1300-GFP plantexpression vector constructed in this laboratory.2. Sweet sorghum has established tissue culture regeneration system. M-81E sweet sorghummature seeds as explants, the study found seeds contamination rate was significantly lower evenbasically no pollution, and seed germination was not affected by using75%of sterile ethanolconducting1.5min;0.1%mercuric chloride13min, and then wash3-4times with sterile water.We eventually set callus induction medium for MS+45g/L sucrose+500mg/L caseinhydrolyzate+1.38g/L L-proline+4.0mg/L2,4-D+0.2mg/L KT+1.2%agar. Callusdifferentiation medium is MS+0.5mg/L6-BA+1.0mg/L IAA+500mg/L casein hydrolysate+30mg/L suter+1.2%agar. Although callus differentiation medium was optimized, but thedifferentiation rate is still low. The last set of rooting medium1/2MS+3mg/L IBA+500mg/Lcasein hydrolyzate+15g/L sucrose+1%agar.3. The Agrobacterium-mediated sweet sorghum genetic transformation was conducted withcallus, shoot tip and pollen tube. The obtained transgenic plants were verified, and the results showed that transgenic plants with target gene were obtained by shoot tip and pollen tube but notby callus.