摘要
经比较影响根癌农杆菌介导ACC合成酶反义基因转化枣树的各种因素后 ,建立了稳定的转基因实验体系。预培养 2d的枣树嫩梢段和下胚轴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 +5 0 0mg/lCarb上诱导产生不定芽 ,将抗Km不定芽先后在 2 0mg/lKm的生长培养基和 30mg/lKm生根培养基上继续选择 ,诱导成完整植株 ,转化率为 2 % - 4%。经PCR检测和Southern杂交证实有 6株外源基因已整合到了枣树基因组中。
The anti-ACC gene was ransformed into Zixyphus jujuba by a efficient Agrobacterium tumefactions mediated system developed in this study after studying the transformation factors influencing. In this protocol,using Zixyphus jujuba as transformed materials,transgenic jujube were obtained through the anti-ACC gene transformation by Agrobacterium tumefactions. The younger stems which were preculture for for 2d were infected with Agrobacterium tumefactions and then the stems were used to introduce shoots on the medium supplemented with 0.15mg/l NAA,1.75mg/l ZT,10mg/l Km,500mg/lCarb.The Kanamyci-resistant shoots were further selected on the mediums containing 20mg/l Km,on which the shoots grow,reproduce,root and whole plants were obtained. PCR analysis and Southern blot of the transgenic plants indicated that the foreign genes had been integrated into Zixyphus jujuba.
出处
《分子植物育种》
CAS
CSCD
2003年第5期683-686,共4页
Molecular Plant Breeding
基金
国家自然科学基金资助项目(No.30070365)
