Complete Sequence, Genome Organization and Molecular Detection of Grapevine Line Pattern Virus, a New Putative Anulavirus Infecting Grapevine

Abstract

Grapevine line pattern virus (GLPV) was first described 30 years ago in Hungary. The lack of its genomic sequences and of an available antiserum made its detection impossible in other parts of the world. Three different high-throughput sequencing (HTS) protocols applied on a GLPV-infected vine allowed the construction of the full genome sequence of this virus. It includes three RNA segments, encoding four proteins: methyltransferase-helicase (1a), RNA-dependent RNA polymerase (2a), movement protein (3a) and coat protein (3b). The obtained sequences were used to design specific primers for its detection by RT-PCR and Northern blot hybridization, respectively. These diagnostic methods were used to test the presence of GLPV in graft-inoculated plants and in 220 grapevine accessions of different Mediterranean origins. The three RNAs-encoding proteins of GLPV shared a very high amino acid identity with those of hop yellow virus, a tentative member of the Anulavirus genus, leaving no doubt that both are two isolates of the same viral species. A circular RNA originating from the RNA2 was found, for which an alternative silencing suppressor role is hypothesized. Further investigation is needed to determine this possibility and also the host range and pathological significance of the virus.

Keywords

• grapevine
• virus
• line pattern
• anulavirus
• small RNA
• HTS