LeJA2基因沉默表达载体导入番茄转化系统的建立

Establishment of System for Transforming Antisence LeJA2 Gene into Tomato

试验设侵染时间、菌液浓度和侵染温度3个因素,利用根癌农杆菌将LeJA2基因按L9(34)正交表转化番茄。结果表明,当菌液浓度为OD260=0.6、侵染温度40℃、侵染10 min时的转化效果较好,从而建立了较优的LeJA2基因沉默表达载体导入番茄的转化体系,并已得到14株转化再生的番茄植株,经卡那抗性筛选和PCR检测,证明LeJA2-RNA i基因已导入番茄并表达。

Three factors, the infection time, the concentration of bacterial suspension and the infection temperature were analyzed by L9 （3^4） orthogonal design in this experiment. The results showed that the transformation effect was better when the concentration of bacterial suspension was 0.6 of OD260, the infection temperature was 40℃ and the infection time was 10 minutes. So the optimal transformed system for LeJA2 gene into tomato was established, and 14 transgenic tomato plants had been obtained. Through the PCR determination and kanamycin -resistance screening, the LeJA2 -RNAi was proved to be transformed into tomato and expressed in part of transgenic tomato plants.