Antiserum to recombinant virus coat protein detects Rupestris stem pitting associated virus in grapevines

Rupestris stem pitting (RSP) is the most widespread virus disease of grapevines. The genome of Rupestris stem pitting associated virus (RSPaV), the putative causal agent of RSP, was recently sequenced. Until recently, the only method to diagnose RSP was biological indexing on woody indicator plants, a process that takes 2 to 3 years to complete. This study reports on the production of a polyclonal antiserum to a recombinant coat protein of RSPaV. The antiserum was used effectively to detect RSPaV from various genotypes and tissues of grapevines by Western blot and indirect enzyme-linked immunosorbent assay. Virus antigens were consistently detected in the cambium of dormant canes and in actively growing leaves of grapevines. Moreover, plants of Vitis rupestris 'St. George', the standard biological indicator for RSP, tested positive for RSPaV. The serological methods developed in this study are advantageous as compared with biological indexing because they are more rapid, less expensive, as reliable, and more suitable for assays of a large number of samples.