The Preliminary Study Of Ion Beam Assisting Agrobacterium-mediated Transforming Soybean Mature Embryos With Gene OXO

Soybean is an important food crop and oil crop, it is the main source of the vegetable protein and edible oil all over the world. Sclerotina sclerotiorum is one of soybean diseases, it can severely impair both the output and quality of soybean. So, breeding of disease-resistant soybean becomes our aim of research.Many pathomycetes harm plants by secreting oxalic acid(OA), but oxalate oxidase(OXO) can break OA down into H2O2 and CO2, relieve its toxicity to delay invasion by plantpathogenicfungi, it plays an important role in defense of plants, resistance to salt stress and processes in pants.But gene pool of soybean is limited, we have not find resistant cultivar of soybean, and breeding needs too long time. For these reasons, Disease-resistant soybean breeding becomes our important research objective. Because of the traditional breeding ways take long time but with limited potential, and the rapid technology development of molecular biology and gene engineering, obtaining new breeds through gene engineering methods has became a speedy and efficienct way. Consequently, regulating the content of oxalate oxidase level through gene engineering methods provides a new method for the research on soybean disease-resistant yield. Transforming mediate by ion beam was brought up by Yu Zengling in 1989, it can avoid the problem that regeneration system difficult of soybean, and facilitate materials, it breaks the problem of traditional method is limited by season. It can be operated in quantity, can improve the efficiency of soybean resistance breedingWe connected the OXO gene and the GFP gene, constructed a plant expression vector carrying the coding sequence of oxalate oxidase gene OXO and genetic transformated soybean:1. Estabishment a soil cultivates system of soybean mature seed Embryos.Compared with some factors which can affect the soil cultivates system, to explore a suitable way for plant cultivation method based on ion-beam-mediated transgene.2. Construction of plant expression vectorThe OXO gene was cloned into the multiple cloning sites of pCAMBIA3304. Subsequently, the pCAMBIA3304-OXO was successfully introduced into A. tumefaciens LBA4404. 3. The foundation of ion beam -assist A. tumefaciens-mediated transformation system.The Agrobacterium tumfaciens contains plasmid pCAMBIA3304-OXO was dipped the soybean mature seed Embryos assisted by ion beam, after screening, Gus assay and PCR amplification, we have got 4 plants which shows positive.