The Construction And Application Of Soybean Transient And Genetic Transformation

Soybean 【Glycine max (L) Merr.】is one of the foremost crop plants for its appalication inproduction of vegetable proteins, oils, forage, energe, health care products. After the accomplishmentof sequencing soybean genome in2010, functional genomics turns out to be hot points in soybeanstudy. Efficient and simple transformation techonolgy is one of the limiting factors in this field.Transient expression system is proved as a powerful approach for gene functional evidence, whereasstable genetic transformation is a key technology for construction of mutants mutants and acquisitionof transgenic plants, which are indispensable for gene function study. Soybean is known as a typipcaltransformation-recalcitrant plant. Heterogeneous expression system can explain gene function in someextent, but not in in situ elucidation for a native gene.To answer these questions, we evolved a quickand efficient soybean transient transformation system based on a previous report and advanced astable soybean genetic transformation system in this paper. In addition, we tried to develop a novalapproach construction of Ds mutants. The main results are as followed:1. On the basis of previous frameworks, we established Agrobacterium-mediated soybeantransient system using soybean cultivar Tianlong1hycopotyls as explants. Here, we evolved thistransient array with modified conditions to gene extensive studies, including promoter analysis,protein expression, protein subcellular localization, and protein-protein interaction. Our resultssuggested that such a soybean callus array was efficient and adaptable for multiple purposes of genefunctional investigation in soybean genotype-, binary vector-, and Agrobacterium strain-independentpatterns.2. Using soybean cultivar Tianlong1, we established Agrobacterium-mediated cotyledonarynode transformation system, and successfully obtained GmX-RNAi, GmXL1-OX, GmY-RNAi,GmZ-OX transgenic soybean plants. We verified the transgenic plants by glufosinate screening, PCRand qPCR. Phenotyping results suggests that GmX gene was related to soybean flowering and GmYgene to the shape of compound leaves and the length of petioles.3. We constructed two kinds of Ac/Ds inducibale expression vectors: one with enhancers, theother without enhancers. With the genetic transformation approach established above, we obtainedAc/Ds transgenic soybean plants with enhancers. The research result laid a solid foundation forconstruction of soybean insertion nutants.