TY - JOUR 
A1 - Tomasechová, Jana
AU - Olmos, Antonio
AU - Ruiz-García, Ana B.
AU - Canales, Celia
AU - Mrkvová, Michaela
AU - Glasa, Miroslav
T1 - First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia
Y1 - 2022
SN - 1125-4653 (Print ISSN)

UR - http://hdl.handle.net/20.500.11939/8205
AB - A symptomatic cantaloupe melon plant (Cucumis melo L.) encoded MVU2/21 grown in a garden in Velke Ulany (western Slovakia), showing pronounced chlorotic leaf spots and dwarfed growth was analysed by high-throughput sequencing (HTS). Total RNAs were isolated from apical leaves collected in August 2021. HTS was carried out on an Illumina MiSeq platform and obtained high-quality reads (∼26,4 millions, average length 131 nt) were analyzed using CLC Genomics Workbench v.10.1.1 and Geneious v.2020.2.5. BLASTn analysis of ca. 24,400 generated contigs revealed a complex viral infection involving watermelon mosaic virus (WMV), zucchini yellow mosaic virus (ZYMV) and Cucumis melo endornavirus (CmEV, family Endornaviridae). A 15,075 nt de novo assembled contig (OL957252) corresponded to nearly complete CmEV genome with 97.9% nucleotide identity to NC_029064. Typical genome organisation and CmEV motifs, including three glucosyltransferases (Sabanadzovic et al. 2016), were found in MVU2/21 polyprotein. The MVU2/21 total RNAs were subsequently subjected to RT-PCR using primers reported by Zeng et al. (2020) as well as two newly designed primer pairs: CmE_14443F (5´-TATGCGGTTGACTGGACAGG-3´)/CmE_14931R (5´-TTATGAGCCACAGCGGTCAC-3´) and CmEV_4145F (5´-CTTGCCTTGAAGGTAGATCG-3´)/ CmEV_4634R (5´-GATTGTCCGCACCGTATAAC-3´). Sanger sequencing of the specific 489, 490 and 710 bp PCR products unambiguously confirmed the HTS data. In order to analyse the potential presence of CmEV in Slovakia, additional leaf samples from melons, cucumbers and squashes from four different localities were tested by RT-PCR using the above primers. Five out of 40 samples collected from different locations and hosts, i.e. melon (MVU1/21), cucumber (UVU1/21, UVU2/21), squash (TPE 2/21) and patison (CPTV6) tested positive for CmEV. Partial genomic sequences of all Slovak isolates amplified using CmEV_4145F/4634R primers (OL957253-OL957257) shared high nucleotide identity to MVU2/21 (97.2–100%). CmEV was previously reported outside Europe (Sabanadzovic et al. 2016; da Costa et al. 2019; Zeng et al. 2020). Our work thus extents data on CmEV geographical distribution and host range.
KW - Endornavirus
KW - High-throughput sequencing (HTS)
KW - Detection
KW - Chlorotic leaf spots
KW - H20  Plant diseases
KW - U40  Surveying methods
KW - Cucurbitaceae
KW - Dwarfing (disorder)
KW - Slovakia
LA - en
PB - Springer
ER -