Optimization Of Soybean Cotyledonary-node Genetic Transformation System And Gene IFS2,CHIⅡGenetic Transformation

Soybean (Glycine max (L.) Merr) as an important crop,which provide proteinand vegetable oil for human ’s daily life. It was Plant in China more than5,000years.Soybean is an important food products. Soybean isoflavone is an important secondarymetabolites. It is many effects on the human,such as anti-cancer effects, regulatingblood lipids effects and improve immunity effecst. However, Soybean contains onlysmall amounts of isoflavones. so it’s cannot meet the needs of the people. After yearsof research found that the means of genetic engineering is one of the ways to improveisoflavones. Soybean isoflavone synthase (IFS2) and chalcone isomerase (CHI) arekeys enzymes in the synthesis pathway of soybean isoflavones. In theory those twogenes cloned and transferred into the soybean that can be raised the content of thesoybean isoflavones..In this study, the laboratory has validated the Agrobacterium tumefaciensEHA105and EHA101containing recombinant plasmid pCB35SR1R2GFP-IFS2. Andverifying the Agrobacterium tumefaciens EHA105and EHA101Agrobacteriumtumefaciens containing the recombinant plasmid pCB35SR1R2GFP-CHI Ⅱ.At the same time, optimizing the soycotyledon node transformation system. Inthe paper Jilin47and Wlliams82soybean varieties as materials. By inducing stembud rate and pumping rate of the optimum level to get two varieties of soybeangermination time is one day. Infection by testing three kinds of ways to get three kindsof infection time47best Jilin infection combination of vacuum and1h10min shaker,Wlliams82any infection methods are feasible. By weight of the suspension wasadded Silwet L-77. Get added0.02%Silwet L-77was the best. Way through theprocess of placing a total of explants cultured hypocotyl come back to put more than hypocotyl-oriented culture medium induced bud release rate. By soybean budswhether to keep pumping stem inoculated into the medium when the cotyledon node,retained cotyledon node can increase the pumping rate of the stem. Then experimentJilin47and silver nitrate concentration willams82pumping stem stage, drawn47andAgNo3Jilin optimum concentration willams82of10mg L-1and8mg L-1.The research was supported by10IFS2gene transfer T0generation and threetransfer CHI Ⅱ gene T0generation of transgenic lines10rpm IFS2validate the T2generation plants to leaf painting Glufosinate diluted, bar strip for detection ofsoybean genes leaf protein and PCR detection35s bar gene and the bar gene are threemethods to detect transgenic plants. Results obtained, six lines are positive for T2generation plants. Due to many reasons not detected by CHI Ⅱ gene T0generation.For the six transgenic lines of soybean seed isoflavone content by HPLCtesting. It was concluded that the wild-type soybean seed in Jilin47daidzein, glycitin,genistin transgenic lines were lower than soybeans. Which IFS-1strains daidzein,inosine, soy genistein content of almost three times the wild-type soybeans..