Optimum for Regeneration and Transformation System in Cucumis sativus L.

Cotyledon-nod of 4 days of cucumber cv.gl plant and Agrobacterium tumefaciens GV3101(habored pCAMBIA 2301) were used to optimize the condition for regeneration and transformation system of cucumber.For regeneration,we set grade concentrations of 6-BA and ABA and added proper amount of AgNO3 to get the optimal medium.For genetic transformation,kanamycin sensitivity of cotyledon-nod was tested.The optional parameter of the pre-culture,infection time,co-culture time and the addition of antioxidant were obtained by gus stain.After the optimization,the parameter in transgenic cucumber were investigated by PCR.The results showed that MS + 1.5 mg/L 6-BA + 1.0 mg/L ABA + 2.0 mg/L AgNO3 was the optimal formula,which can induce shooting and a new plantlet can be gotten in 45 days with the formula.The critical concentration of kanamycin was 50 mg/L,and the optimal pre-culture time was 1~2 days,the optimal co-culture time was 3 days after 20~30 min infected by GV3101.The addition of antioxidant enhanced the efficiency distinctly and foreign gene was successful integrated into cucumber genome.