根癌农杆菌介导反义ACC合成酶基因对枣树的转化  

Transformation of Zizyphus jujuba Hetau with Antisense ACC Synthase Gene

作　　者：何业华[1] 熊兴华[2] 林顺权[1] 吴会桃[1] 林良斌[2] 何钢[3] 陈建华[3] 

机构地区：[1]华南农业大学园艺植物生物技术研究所,广东广州510642 [2]作物基因工程湖南省重点实验室,湖南长沙410128 [3]中南林学院生物技术学院,湖南长沙410016

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Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金资助项目(30070365)

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.

关 键 词：枣树 ACC合成酶反义基因 转化 

分 类 号：S665.1[农业科学—果树学]