Transformation Of Stress Resistance Related Gene GM And FMDV Structure Protein Gene P1 Into Soybean

In this paper, somatic embryogenesis and plant regeneration were induced by higher concentrations of auxins with immature cotyledon of ten varieties from Northeast area in soybean. Correlative factors with prolixferation, germination and regeneration of embryogenic tissue were discussed. The system of tissue culture and regeneration were optimized. Genetic transformation system was established successfully in soybean. The stress resistance-related gene GM (coding the transcription factor DREB )in plasmid pRdGM-200 was transformed into Agrobacterium tumefaciens by using freeze-thaw method and electroporation. These could afford the basic research for soybean transformation via Agrobacterium-mediated. The stress resistance-related gene GM and Foot and Mouth disease virus structure protein gene P1 were transformed into immature cotyledon and somatic embryogenesis of soybean via Agrobacterium-mediated and particle bombardment. The regenerated plantlets were examined by PCR and PCR-southern blot. The results indicated that target gene had been transformed into soybean, and the factors affecting genetic transformation were investigated. The main results were summarized as followed: somatic embryotenesis of ten varieties with immature cotyledon was induced successfully. All varieties generated somatic embryotenesis and the highest frequency reached to 49.8% for somatic embryogenesis. The best explant was immature cotyledon for inducing somatic embryogendsis in soybean. Induction of somatic embryogenesis with high concentration of 2,4-D could produce better results. The frequencies of somatic embryogenesis varied with genotypes of soybean (3.8-49.8%). The frequencies of somatic embryogenesis in five varieties were the highest (13.2-49.8%). The plasmid pRdGM-200 was transformed into Agrobacterium tumefaciens(EHA101,EHA105 and LBA4404) by using freeze-thaw method and electroporation. These could afford the basic research for soybean transformation via Agrobacterium-mediated. Conditions of Agrobacterium tumefaciens transformation with freeze-thaw method were optimized, and the best scheme of Agrobacterium tumefaciens transformation was groped...