Nickel Holger, Kawchuk Lawrence, Twyman Richard M, Zimmermann Sabine, Junghans Holger, Winter Stephan, Fischer Rainer, Prüfer Dirk
Fraunhofer Institute for Molecular Biotechnology and Applied Ecology (IME), Forckenbeckstrasse 6, 52074 Aachen, Germany.
Virus Res. 2008 Sep;136(1-2):140-5. doi: 10.1016/j.virusres.2008.05.001. Epub 2008 Jun 11.
The P1 protein of Potato leafroll virus (PLRV) is thought to play a major role in the replication cycle by promoting the maturation of the genome-linked virion protein VPg. To study the relevance of P1 and its autoproteolytic derivative P1-C25 in the viral life cycle, the V H and V L domains of monoclonal antibody mAbP1-1, raised against the C-terminus of P1, were used to develop a single chain variable fragment antibody scFvP1-1 for expression in plants. The transient expression of scFvP1-1 in tobacco (Nicotiana tabacum) strongly reduced virus accumulation, while transgenic potato (Solanum tuberosum) plants expressing scFvP1-1 showed high levels of resistance following PLRV inoculation by viruliferous aphids. This is the first report that conclusively demonstrates that a PLRV gene product is essential for the completion of the virus life cycle in vivo without genetic alteration of the viral genome. This is also the first time plantibody-mediated resistance has been demonstrated with a luteovirus.
马铃薯卷叶病毒(PLRV)的P1蛋白被认为通过促进基因组连接病毒粒子蛋白VPg的成熟在复制周期中发挥主要作用。为了研究P1及其自蛋白水解衍生物P1-C25在病毒生命周期中的相关性,针对P1 C末端产生的单克隆抗体mAbP1-1的VH和VL结构域被用于开发在植物中表达的单链可变片段抗体scFvP1-1。scFvP1-1在烟草(Nicotiana tabacum)中的瞬时表达强烈降低了病毒积累,而表达scFvP1-1的转基因马铃薯(Solanum tuberosum)植株在接种带毒蚜虫传播的PLRV后表现出高水平抗性。这是第一份确凿证明PLRV基因产物在不改变病毒基因组的情况下对病毒在体内完成生命周期至关重要的报告。这也是首次用黄症病毒证明植物抗体介导的抗性。