根癌农杆菌介导反义ACC合成酶基因对枣树的转化

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／L NAA 1．75mg／L．ZT 10mg／L Km 500mg／L Carb上诱导产生不定芽，将抗Km不定芽先后在15mg／L Km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—time PCR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．

Transformation of Zizyphus jujuba Hetau with Antisense ACC Synthase Gene

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.