作　　者： ; ; ;

机构地区： 华南农业大学

出　　处： 《植物生理与分子生物学学报》 2007年第6期517-523,共7页

摘　　要： 通过构建植物表达载体,由农杆菌介导,将望江南核糖体失活蛋白基因cassuin转入烟草。PCR和Southern blot杂交结果证明:外源基因已经以单拷贝整合到烟草基因组内,并且在后代发生遗传分离。RT-PCR和Northern blot杂交结果显示:外源基因可以正常转录。用不同浓度的TMV机械摩擦接种转基因T_1、T_2代各3个自交株系,以非转基因烟草为阴性对照,实验结果表明转基因烟草对TMV表现出不同程度的抗性。 Cassin, the new gene of ribosome-inactivating protein （RIP） isolated from Cassia occidentalis, was inserted into expression vector pBI121 to produce plant expression vector pBI121- cassin （Figs. 1, 2）. pBI121-cassin was introduced into tobacco cultivar ＇K326＇ by the Agrobacteriurm tumefaciens transformation method and more than 100 independent transformants were obtained （Fig. 3）. Southern blot hybridization analysis showed that a single gene locus was inserted into the chromosome of the transgenic tobacco lines （Fig.5） and PCR analysis of segregation population of prog- eny indicated that the inheritance of transgene was dominant in transgenic lines （Fig.4, Table 1）. Resuits of RT-PCR and Northern blot hybridization analysis showed that transgene could be transcribed correctly （Figs.5, 6）. Three self-pollination lines of transgenic T1 and T2 were challenged with TMV at different concentration titers by mechanical inoculation. The transgenic lines exhibited different levels of resistance to TMV with the nontransgenic plants. After both titers of TMV concentration were inoculated, transgenic lines were considered as the highly resistant type with a delay of 4-13 d in development of symptoms and ,10%- 25% of test plants were infected, while nontransgenic control plants were susceptible typical symptoms on the newly emerged leaves （Table 2）. One T2 line, T2-8-2-1, was regarded as an immune type because it did not show any symptoms during 70 d and all plants were shown to be virus free by ELISA tests.

关 键 词： 转基因烟草 核糖体失活蛋白 抗病性评价 烟草花叶病毒

领　　域： [生物学]