Optimization Of Soybean Tissue Culture System And EPSPS Genetic Transformation

Through genetic engineering herbicide resistance gene into the current production is the use of soybean varieties for cultivation of herbicide-resistant varieties available breeding material. The successful genetic transformation depends on good receptor system. Currently, soybean tissue culture still have many problems. This study was designed to improve the soybean tissue culture and transformation system to alleviate the low frequency of regeneration and genetic transformation of soybean problem, while increasing the survival rate of regenerated plantlets. Using agrobacterium- mediated transformation method of soybean cotyledons node, for the receptor Hei Nong 37 and Beijing Xiaolidou, has established a relatively perfect system for genetic transformation. Experiments using cotyledonary node method to genetically modified Xiao Lidou, has got 225 resistant plants and 38 seeds.Studies show that useing chlorine for soybean seeds sterile.Using 300mL sealed cup, with 20mL sodium hypochlorite solution and 2mL concentrated hydrochloric acid, sterilization time of five hours is best. Five hours can guarantee sterilization, and not significantly lower germination rate of soybeans. Germination medium containing about 2mg/L of BAP can improve the rate of genetic transformation. Experiments have also found the most suitable shoot induction days isabout 7 days culture, conducive to shoot differentiation and elongation.Agrobacterium-mediated soybean cotyledon node gene transfer method, when using glyphosate as the transformation of screening agent, for soybean Heinong 37 and Beijing xiaolidou, selected in the shoot induction stage of the appropriate screening concentration is 10~15mg/L, should not be less than 10mg/L. Soybean rooting medium is generally added IBA, but IBA in rooting medium will induce large callus in the base of shoot. After transplanting often due to Bacterial contamination of death. Dip little IBA solution at the base of buds, generally buds can take root healthy and improved survival rate.