First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia

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 202...

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Autores principales: Tomasechová, Jana, Olmos, Antonio, Ruiz-García, Ana B., Canales, Celia, Mrkvová, Michaela, Glasa, Miroslav
Formato: Artículo
Lenguaje:Inglés
Publicado: Springer 2022
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Acceso en línea:http://hdl.handle.net/20.500.11939/8205
https://link.springer.com/article/10.1007/s42161-022-01149-4
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author Tomasechová, Jana
Olmos, Antonio
Ruiz-García, Ana B.
Canales, Celia
Mrkvová, Michaela
Glasa, Miroslav
author_browse Canales, Celia
Glasa, Miroslav
Mrkvová, Michaela
Olmos, Antonio
Ruiz-García, Ana B.
Tomasechová, Jana
author_facet Tomasechová, Jana
Olmos, Antonio
Ruiz-García, Ana B.
Canales, Celia
Mrkvová, Michaela
Glasa, Miroslav
author_sort Tomasechová, Jana
collection ReDivia
description 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.
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spelling ReDivia82052025-04-25T14:48:52Z First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia Tomasechová, Jana Olmos, Antonio Ruiz-García, Ana B. Canales, Celia Mrkvová, Michaela Glasa, Miroslav Endornavirus High-throughput sequencing (HTS) Detection Chlorotic leaf spots H20 Plant diseases U40 Surveying methods Cucurbitaceae Dwarfing (disorder) Slovakia 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. 2022-06-07T10:58:37Z 2022-06-07T10:58:37Z 2022 article publishedVersion Tomašechová, J., Olmos, A., Ruiz‑García, A. B., Canales, C., Mrkvová, M. & Glasa, M. (2022). First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia. Journal of Plant Pathology, 104,1179-1180. 1125-4653 (Print ISSN) 2239-7264 (eISSN) http://hdl.handle.net/20.500.11939/8205 10.1007/s42161-022-01149-4 https://link.springer.com/article/10.1007/s42161-022-01149-4 en M. M. and M. G. acknowledge funding from the project "PandemicFood" (ITMS: 313011AVA9) supported by the Integrated Infrastructure Operational Programme funded by the ERDF. Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess Springer electronico
spellingShingle Endornavirus
High-throughput sequencing (HTS)
Detection
Chlorotic leaf spots
H20 Plant diseases
U40 Surveying methods
Cucurbitaceae
Dwarfing (disorder)
Slovakia
Tomasechová, Jana
Olmos, Antonio
Ruiz-García, Ana B.
Canales, Celia
Mrkvová, Michaela
Glasa, Miroslav
First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia
title First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia
title_full First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia
title_fullStr First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia
title_full_unstemmed First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia
title_short First report of Cucumis melo endornavirus infecting Cucurbitaceae plants in Slovakia
title_sort first report of cucumis melo endornavirus infecting cucurbitaceae plants in slovakia
topic Endornavirus
High-throughput sequencing (HTS)
Detection
Chlorotic leaf spots
H20 Plant diseases
U40 Surveying methods
Cucurbitaceae
Dwarfing (disorder)
Slovakia
url http://hdl.handle.net/20.500.11939/8205
https://link.springer.com/article/10.1007/s42161-022-01149-4
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