First Report of Grapevine Pinot gris Virus on Grapevines in Armenia

Grapevine Pinot gris virus (GPGV) is a member of the genus Trichovirus in the family Betaflexiviridae. It was first described on Italian grapevine (Vitis vinifera L.) cultivar Pinot gris, in which it provoked a disease known as grapevine leaf mottling and deformation (GLMD) (Giampetruzzi et al. 2012; Saldarelli et al. 2017). Studies on GPGV revealed a widespread occurrence of GPGV in many wine-producing countries in the world (Eichmeier et al. 2018) and, more recently, in Canada (Poojari et al. 2016), Australia (Wu and Habili 2017), Brazil (Fajardo et al. 2017), Chile (Zamorano et al. 2019), Pakistan (Rasool et al. 2017) and the United Kingdom (Silva et al. 2018). In 2018, 17 grapevine cultivars (Ararati, Arevabuyr, Arevar, Armenia, Eraskheni, Garan Dmak, Itsaptuk, Kishmish White, Masis, Meghrabuyr, Mskhali, Rkatsiteli, Shahumyani, Tokun, Vani, Vardaguyn Yerevani, and Voskehat) were selected in economically important Armenian vineyards of Yerevan city and Merzdavan village for GPGV detection. The grapevines were selected based on possible GLMD symptoms. The plants were tested by reverse transcription polymerase chain reaction for GPGV presence as described by Eichmeier et al. (2018). GPGV was detected in cultivars Ararati, Arevabuyr, Eraskheni, Garan Dmak, Itsaptuk, and Mskhali based on the amplification of gene fragments specific to the MP-CP and RdRp genome regions (Eichmeier et al. 2018). The derived sequences were submitted into GenBank under accession numbers MN186552 to MN186556 (MP-CP) and MN186557 to MN186561 (RdRp). To obtain more genome sequences from a selected GLMD symptomatic isolate (cv. Eraskheni), small RNA high-throughput sequencing (HTS) was used. Total RNA was extracted from cambial scrapings using TRIzol Reagent (TFS, Waltham, MA) and used for constructing a cDNA library (TruSeq Small RNA Library Prep Kit, Illumina, San Diego, CA) followed by sequencing (1 × 36 bp) on an Illumina MiniSeq sequencer using a MiniSeq High Output Reagent Kit (75 cycles) (Illumina). Bioinformatic evaluation was done as described by Eichmeier et al. (2019). HTS revealed the presence of GPGV (one contig: length, 5,535 bp; number of mapped reads, 299; mean read coverage, 1.08; genome coverage, 76.25%; identity with ref. seq. NC_015782, 99.06%) in mixed infection with Grapevine fleck virus, Grapevine leafroll-associated virus 1, Grapevine fanleaf virus, and Grapevine virus A in the sample. The genome sequence of GPGV (cv. Eraskheni) was deposited under accession number MN186562. Our preliminary data showed a high prevalence of GPGV, roughly 35%, in Armenia. The genetic diversity and the impact of this virus on grapevine in Armenia should be studied further.