First Report of Grapevine Virus H in Grapevine in Greece

Grapevine virus H (GVH) is a member of the genus Vitivirus in family Betaflexiviridae (subfamily Trivirinae, order Tymovirales) that infects grapevine (Candresse et al., 2018). GVH was first identified symptomless an unknown cultivar from Portugal 2018 al. 2018), and since then has been reported only California (Diaz‑Lara 2019). Several vitiviruses have detected Greek vineyards (Avgelis Roubos 2000; Dovas Katis 2003a; 2003b; Panailidou 2019; Lotos 2020), but no information available on presence GVH. In fall 2020, to investigate virome commercial vineyard Assyrtiko northern Greece, composite sample made leaves petioles nine vines exhibiting leafroll disease symptoms. Total RNA extracted according protocol White (2008) subjected rRNA depletion, library construction (TruSeq Stranded kit), high-throughput sequencing (HTS) NovaSeq6000 platform (Illumina Inc.) at Macrogen (Korea). The resulting ~42 million 101-nt paired-end reads were analyzed Geneious Prime assembled de novo contigs local BLASTn search, which revealed 18 infecting viruses viroids, among also GVH-like contig (GeA-9). GeA-9 7,404 nucleotides (nt) long, covering 99.4% full genome shared 98.2 % nt identity with isolate USA (MN716768). To confirm GVH, samples Assyrtiko, used initially produce for HTS analysis, tested individually by RT-PCR, using primers GVH_F_2504 (5'-CTGCTTCGCTGAACATATGC-3') GVH_R_2835 (5'-ATCATTRTGATCGAGAGAGTAGTG-3') amplify 331-nt segment ORF1. five out one these reamplified Sanger sequencing. fragment ORF1 obtained (MW460005) 97.5% identical nucleotide sequence corresponding (GeA-9) analysis it 97.2% sequences USA, respectively (NC_040545 MN716768), confirming samples. This report thus further increasing number known infect expanding geographic locations recorded so far.