论文部分内容阅读
RNA沉默(RNAi)介导的植物抗病毒研究是近年来引起广泛关注的一项植物抗病毒基因工程策略。马铃薯Y病毒(PVY)在世界范围内引起马铃薯、烟草等重要经济作物的病害,并且造成严重的经济损失。本文以PVY的外壳蛋白(CP)基因为模板扩增300bp和354bp两个片段,正向和反向分别插入植物表达载体pROKⅡ的35S启动子下游,从而构建了以PVY的CP基因为靶标的RNAi植物表达载体pROKY300,转入农杆菌EHA105中,以农杆菌渗入法在本氏烟中瞬时表达与PVYCP同源的hairpin RNA。结果表明,瞬时表达的hairpinRNA有效干涉了PVY侵染。
RNA silencing (RNAi) -mediated plant antiviral research has attracted widespread attention in recent years as an ARV genetic engineering strategy. Potato virus Y (PVY) causes disease worldwide on important cash crops such as potato and tobacco and causes serious economic losses. In this study, two fragments of 300bp and 354bp were amplified using the coat protein (CP) gene of PVY as a template and inserted into the downstream of 35S promoter of plant expression vector pROKII in the forward and reverse directions, respectively. Thus RNAi targeting CP gene of PVY The plant expression vector pROKY300 was transformed into Agrobacterium tumefaciens EHA105 and the hairpin RNA homologous to PVYCP was transiently expressed in N. benthamiana using Agrobacterium tumefaciens. The results showed that the transiently expressed hairpin RNA effectively interfered with PVY infection.