【摘要】 病毒诱导基因沉默是利用RNA介导病毒防卫机制的一项技术.构建含有目的基因片段的人工改造病毒载体,通过农杆菌侵染导致植物内源目的基因沉默.为建立病毒诱导基因沉默体系,选用烟草脆裂病毒(TRV)和烟草为实验材料.构建了八氢番茄红素去饱和酶基因(PDS)的基因沉默病毒载体,病毒载体侵染结果显示目的基因PDS沉默导致烟草幼苗出现光漂白现象.采用RT-PCR的方法检测目的基因PDS的沉默效果,结果显示PDS基因mRNA被显著降解.该体系的建立有利于将来对植物基因进行高通量功能分析.更多还原

【Abstract】 Virus-introduced gene silencing is a technique that exploits the RNA-mediated antiviral mechanism. The modified virus vector carrying the fragment of a target gene were constructed and then inserted to silence the target gene within host plants by Agrobacterium-meditated transformation. The study chose tobacco rattle virus (TRV) and tobacco plants to construct the gene-silencing vector carrying the fragment of LePDS gene and the infection of the vector indicated that the silencing of the target gene PDS resulted in photo-bleached young tobacco plants. RT-PCR analysis was employed to test the effect of silencing the target gene PDS, which show that the gene PDS was significantly degraded. This gene-silencing system will benefit the high-flux analysis of plant genes in the future.更多还原