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以枣树鲜食品种鸡蛋枣为材料,通过根癌农杆菌介导ACC合成酶反义基因转化,得到了转基因植株.预培养1d的枣树幼嫩茎段经根癌农杆菌感染后共培养3d,转移至分化选择培养基MS(1/2NO3)+0.15mg/LNAA+1.75mg/LZT+10mg/LKm+500mg/LCarb上诱导产生不定芽,将抗Km不定芽先后在15mg/LKm的生长、增殖和生根培养基上继续选择,诱导成完整植株.经PCR检测从抗Km植株中选择到7株阳性植株,进一步经Southern杂交证实有6株外源基因已整合到了枣树基因组中.Real timePCR定量检测表明,ACC合成酶反义基因在5株转基因植株中得到表达.
The jujube tree jujube was taken as the material and transgenic plants were transformed by Agrobacterium tumefaciens-mediated ACC synthase antisense gene transformation.The young stem segments of jujube tree pre-cultured for 1 day were co-cultured for 3 days after Agrobacterium tumefaciens infection, The adventitious buds were induced to differentiate into selective medium MS (1 / 2NO3) + 0.15mg / L NAA + 1.75mg / LZT + 10mg / LKm + 500mg / LCarb and the anti-Km adventitious buds were grown at 15mg / And rooting medium continue to select, induced into a complete plant.From the PCR detection, 7 positive plants were selected from the anti-Km plants, further confirmed by Southern hybridization 6 foreign genes have been integrated into the jujube genome.Real timePCR quantitative The results showed that ACC synthase antisense gene was expressed in five transgenic plants.