4.7 Article

Characterization of a New Nepovirus Infecting Grapevine

Journal

PLANT DISEASE
Volume 105, Issue 5, Pages 1432-1439

Publisher

AMER PHYTOPATHOLOGICAL SOC
DOI: 10.1094/PDIS-08-20-1831-RE

Keywords

GNVA; grapevine; grapevine nepovirus A; high-throughput sequencing; Nepovirus; phylogenetic analysis; Secoviridae; virus evolution

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In 2012, a dormant canes of a proprietary wine grape were added to the collection of the University of California-Davis Foundation Plant Services. Research revealed the presence of a mechanically transmissible virus in the grape, leading to the discovery of a novel bipartite nepovirus-like genome. The new virus was classified within the genus Nepovirus based on its biological and molecular characteristics.
In 2012, dormant canes of a proprietary wine grape (Vitis vinifera L.) accession were included in the collection of the University of California-Davis Foundation Plant Services. No virus-like symptoms were elicited when bud chips from propagated own-rooted canes of the accession were graft-inoculated onto a panel of biological indicators. However, chlorotic ringspot symptoms were observed on sap-inoculated Chenopodium amaranticolor Coste & A. Rein and C. quinoa Willd. plants, indicating the presence of a mechanically transmissible virus. Transmission electron microscopy of virus preparations from symptomatic C. quinoa revealed spherical, nonenveloped virions about 27 nm in diameter. Nepovirus-like haplotypes of sequence contigs were detected in both the source grape accession and symptomatic C. quinoa plants via high-throughput sequencing. A novel bipartite nepovirus-like genome was assembled from these contigs, and the termini of each RNA segment were verified by rapid amplification of complementary DNA ends assays. The RNA1 (7,186-nt) of the virus encodes a large polyprotein 1 of 231.1 kDa, and the RNA2 (4,460-nt) encodes a large polyprotein 2 of 148.9 kDa. Each of the polyadenylated RNA segments is flanked by 5'- (RNA1 = 156-nt; RNA2 = 170-nt) and 3'- (RNA1 = 834-nt; RNA2 = 261-nt) untranslated region sequences with >90% identities. Maximum likelihood phylogenetic analyses of the conserved Pro-Pol amino acid sequences revealed the clustering of the new virus within the genus Nepovirus of the family Secoviridae. Considering its biological and molecular characteristics, and based on current taxonomic criteria, we propose that the novel virus, named grapevine nepovirus A, be assigned to the genus Nepovirus.

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