农杆菌介导小麦遗传转化影响因素的研究

选用河南科技学院近年来选育的小麦品种百农160的成熟胚愈伤组织和农杆菌菌株LBA4404,对农杆菌介导遗传转化中农杆菌浓度、感染时间、乙酰丁香酮浓度、头孢霉素质量浓度等几个重要影响因素进行了研究,建立小麦高效转化体系.结果表明.菌液OD<,600>=0.6,侵染时间30min时对愈伤组织的生存和转化最有利;浓度为200...

Factors Affecting Wheat Transformation Mediated by Agrobacterium tume faciens

To establish highly efficient wheat transformation system,factors which influenced wheat transformation including agrobacterium concentration,infection period,and concentration of acetosyringone and cephalosporin were studied with the mature embryo callus of wheat variety Bainong 160 and agrobacterium strain LBA4404.The results showed that the transformation efficiency was clearly improved with agrobacterium density of OD600=0.6 and incubation for 30 min.Acetosyringone with the concentration of 200 μmol/L could significantly enhance the rate of resistant calluses.Considering the inhibition of cephalosporin on agrobacterium cells,it was suggested that 500 mg/L of cephalosporin should be used in the latter stage of antibacterial screening,and then 200 mg/L of cephalosporin should be used in differentiation medium to get resistant calluses of high regeneration efficiency.The transgenic frequency was 2.36% under the optimal conditions.