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Combining Analysis of DNA in a Crude Virion Extraction with the Analysis of RNA from Infected Leaves to Discover New Virus Genomes
Published on: July 27, 2018
Identification and complete genome sequence of a new cheravirus discovered in grapevine
Xudong Fan1, Yuxin Hao1, Fang Ren1
1National Center for Eliminating Viruses from Deciduous Fruit Tree, Research Institute of Pomology, Chinese Academy of Agriculture Sciences, Xingcheng, 125100, Liaoning, China.
Abstract:
High-throughput sequencing (HTS) was applied to identify viruses present in grapevine samples collected in Ningxia, China, that exhibited obvious symptoms of viral disease. A putative novel cheravirus was found through the analysis of sequencing data, and its presence was confirmed by RT-PCR in a Cabernet Sauvignon sample with vein yellowing. The complete genome sequence of this virus was determined by Sanger sequencing of overlapping RT-PCR products and the rapid amplification of cDNA ends (RACE) method. The genome of the putative virus comprises two RNA segments 7,395 and 3,645 nucleotides in length, excluding their poly-A tails. The new virus was tentatively named "grapevine cheravirus" (GCheV). The Pro-Pol region of RNA1 and the CP region of RNA2 of GCheV shared the highest amino acid sequence identity, reaching 72.0% and 48.9%, respectively, with the corresponding sequences of Corymbium villosum cheravirus (CvCV). Phylogenetic analysis revealed that GCheV clustered with CvCV in the genus Cheravirus. These results suggest that GCheV is a new member of that genus. To the best of our knowledge, this is the first report of a virus belonging to the genus Cheravirus in grapevines.
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