Study On Genetic Transformation Optimization Of Two Indica Rive Varieties

Research were performed on nature embryo of two indica varieties (Oryza sativa L.)Xiangwanxian17,R299. The main result were as follows:1. the best induction condition for Xiangwanxian17and R299is constant temperature with12hour lighting time. after preculture for3days, induction rates of aboved two rice varieties espectly is79%、83%。3days is the optimal preculture time.2.. The result shows:instant transformation of callus of Xiangwanxian17and R299is respectly75.3%、83.2%by vocuum.So vocuum is suitable to aboved two rice viarieties.3.Callus of Xiangwanxian17and R299were vocuumed by agrobacterium tumefa ciens callus whose value is OD600=0.5,the effect is the best.4.during induction period,2,4—D concentration of callus of Xiangwanxian17is2.5mg/L, that of R299is2.0mg/L, in selective stage,the optimium2,4—D concentration to aboved rice varieties is2.5mg/L5. Sugar concentration increase to35mg/L,callus of Xiangwanxian17and R299has better quantity and higher differentiation rate.6. Overall condition,400mg/L carbenicillin (cb)is suitable to Xiangwanxian17and R299.7. When kana concentration increase,plastic rice seedling appeare because of poison effect on callus is strong and result in. Kana is not suitable to Xiangwanxian17and R299.8. Final genetic transformation system is that inducing callus from mature embryo by MS inducing medium with2.5mg/L2,4—D under constant temperature and12h lightness,cutting callus;after pre-culture for3days, contaminated by agrobacterium tumefaciens whose concentration is OD600=0.5for15min by vacuuming,after co-culture for3days,after transferring callus to MS selective medium with50mg/L kana、400mg/L carbenicillin (cb、2.5mg/L2,4—D, and then regeneration、rooting and transplant. Through rebuilding and optimization regeneration system, we successfully get17Xiangwanxian17and24R299transformation rice.