Petrovic Natasa, Meng Baozhong, Ravnikar Maja, Mavric Irena, Gonsalves Dennis
National Institute of Biology, Vecna pot 111, 1000 Ljubljana, Slovenia.
Department of Plant Pathology, Cornell University, New York State Agricultural Experiment Station, Geneva 14456-0462.
Plant Dis. 2003 May;87(5):510-514. doi: 10.1094/PDIS.2003.87.5.510.
Rupestris stem pitting associated virus (RSPaV), a member of the genus Foveavirus, is associated with the Rupestris stem pitting component of the Rugose wood (RW) disease complex of grapevines. Heretofore, particles of RSPaV have not been visualized. In this work, flexuous rod particles approximately 723 nm in length were detected in the sap of infected grapevines by immunosorbent electron microscopy (ISEM), using a polyclonal antiserum produced to a recombinant coat protein of RSPaV. Particles of RSPaV were detected in tissue culture-, greenhouse-, and field-grown grapevines infected with RSPaV, but not in healthy control plants. Detection of virus particles by ISEM corresponded with detection of RSPaV by Western blot, enzyme-linked immunosorbent assay, and reverse transcription-polymerase chain reaction. Virus particles were decorated with the antibodies specific to RSPaV but not with antibodies to Grapevine virus A or Grapevine virus B, two other viruses believed to be associated with RW. This definitive identification of RSPaV particles will help define the etiology of RW.
岩蔷薇茎痘相关病毒（RSPaV）是凹顶病毒属的成员，与葡萄藤皱木复合病（RW）的岩蔷薇茎痘组分有关。在此之前，RSPaV的病毒粒子尚未被观察到。在这项研究中，通过免疫吸附电子显微镜（ISEM），使用针对RSPaV重组外壳蛋白产生的多克隆抗血清，在受感染葡萄藤的汁液中检测到了长度约为723纳米的弯曲杆状病毒粒子。在感染RSPaV的组织培养、温室和田间种植的葡萄藤中检测到了RSPaV病毒粒子，但在健康对照植株中未检测到。通过ISEM检测病毒粒子与通过蛋白质免疫印迹、酶联免疫吸附测定和逆转录-聚合酶链反应检测RSPaV的结果一致。病毒粒子用针对RSPaV的特异性抗体进行了标记，但未用针对葡萄病毒A或葡萄病毒B（另外两种被认为与RW有关的病毒）的抗体进行标记。对RSPaV病毒粒子的这种明确鉴定将有助于明确RW的病因。
