机构地区: 华南农业大学
出 处: 《湖南农业大学学报(自然科学版)》 2004年第1期33-36,共4页
摘 要: 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. With Zizyphus jujuba as transformed materials ,transgenic jujube is obtaine d through the anti-ACC gene transformation by Agrobacterium tumefactions .Th e younger stems precultured for 1 d are infected with Agrobacterium tumefactio ns and then the stems were used to introduce shoots on the medium supplemented with 0.15 mg/L NAA,1.75 mg/L ZT,10 mg/L Km,500 mg/L Carb.The Kanamyci-resistan t shoots are further selected on the mediums containing 15 mg/L Km,on which the shoots grow,reproduce,root and whole plants are obtained.With the PCR test ,se ven positive plants are selected from those Kanamycin-resistant plants,and the southern hybridization shows that the foreign gene has been integrated into th e six of seven positive plants'genome,and Real time PCR reveals that anti-ACC g ene is expressed in the five of six plants.