First Report of Cucurbit Aphid-Borne Yellows Virus in Cucurbita pepo and Cucurbita maxima in Slovenia
2019; American Phytopathological Society; Volume: 104; Issue: 2 Linguagem: Inglês
10.1094/pdis-07-19-1524-pdn
ISSN1943-7692
AutoresNataša Mehle, Denis Kutnjak, N. Jakoš, Gabrijel Seljak, Anja Pecman, Sébastien Massart, Maja Ravnikar,
Tópico(s)Plant Pathogenic Bacteria Studies
ResumoHomePlant DiseaseVol. 104, No. 2First Report of Cucurbit Aphid-Borne Yellows Virus in Cucurbita pepo and Cucurbita maxima in Slovenia PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Cucurbit Aphid-Borne Yellows Virus in Cucurbita pepo and Cucurbita maxima in SloveniaN. Mehle, D. Kutnjak, N. Jakoš, G. Seljak, A. Pecman, S. Massart, and M. RavnikarN. Mehle†Corresponding author: N. Mehle; E-mail Address: natasa.mehle@nib.sihttp://orcid.org/0000-0002-3028-0280Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia, D. KutnjakDepartment of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia, N. JakošDepartment of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia, G. SeljakDepartment of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia, A. PecmanDepartment of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, SloveniaJožef Stefan International Postgraduate School, Ljubljana, Slovenia, S. MassartPlant Pathology Laboratory, TERRA-Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium, and M. RavnikarDepartment of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, SloveniaWine Research Centre, University of Nova Gorica, Nova Gorica, Slovenia AffiliationsAuthors and Affiliations N. Mehle1 † D. Kutnjak1 N. Jakoš1 G. Seljak1 A. Pecman1 2 S. Massart3 M. Ravnikar1 4 1Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia 2Jožef Stefan International Postgraduate School, Ljubljana, Slovenia 3Plant Pathology Laboratory, TERRA-Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium 4Wine Research Centre, University of Nova Gorica, Nova Gorica, Slovenia Published Online:18 Dec 2019https://doi.org/10.1094/PDIS-07-19-1524-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat In summer of 2017 and 2018, a survey was conducted in the major cucurbit-growing areas in Slovenia to assess the occurrence of viral diseases. Leaf samples of 20 plants of Cucurbita pepo, three Cucurbita maxima, seven Cucumis melo, one Cucumis sativus, and one Citrullus lanatus showing mosaic, interveinal mottling or yellowing, blisters, and distortions were collected from 11 farms in southwestern Slovenia, seven in northeastern Slovenia, and five in southeastern Slovenia. Total RNA was extracted from leaf tissue using RNeasy plant mini kits (Qiagen, Germantown, MD). Five composite RNA samples were generated by pooling together total RNAs from three to 11 individual samples. Two of those were sent for Illumina small RNA sequencing to SeqMatic (Fremont, CA), two for ribosomal RNA depleted total RNA sequencing at Eurofins Genomics AT (Vienna, Austria), and one for ribosomal RNA depleted total RNA sequencing at Liege University. The sequencing reads (10 to 20 thousand) were analyzed using CLC Genomics Workbench 11 (Qiagen Bioinformatics, Aarhus, Denmark) with the pipeline for plant virus discovery (Pecman et al. 2017) or with Geneious version 10.1.5 (using the plugins Dedupe, SPADES, and blasting the contigs against viral reference database). In four out of five sample pools, sequences of cucurbit aphid-borne yellows virus (CABYV, genus Polerovirus) were detected along with sequences of cucumber mosaic virus (CMV, genus Cucumovirus), watermelon mosaic virus (WMV, genus Potyvirus), and/or zucchini yellow mosaic virus (ZYMV, genus Potyvirus). To confirm the presence of CABYV, each of the 26 individual plants, which were part of the pools of CABYV-positive samples, were tested by reverse transcription PCR using the coat protein (CP) gene-specific primers CE9/CE10 (Juarez et al. 2004). An amplicon of the expected size (600 bp) was generated from samples of 13 plants of C. pepo and from one of the C. maxima samples. All CABYV-positive plants were screened by double-antibody sandwich ELISA for CMV, WMV (both Loewe Biochemica, Germany), and ZYMV (Prime Diagnostics, Wageningen University & Research, the Netherlands). Single infection by CABYV was found in one plant of C. pepo, whereas other 13 plants were coinfected with CMV, WMV, and/or ZYMV. PCR products of the CP gene from all positive samples were purified, Sanger sequenced in both directions, and seven unique sequences were deposited in GenBank (accession nos. MN145451 to MN145457), and subjected to sequence analysis using the BLASTn algorithm (NCBI nt database, 20 June 2019) and MEGA7 software. Sequence comparisons showed 97.3 to 100% nucleotide sequence identity between Slovenian CABYV-positive samples. A maximum likelihood phylogenetic tree obtained based on the alignment of sequences of CABYV isolates from this study and isolates available in the GenBank database revealed that all Slovenian isolates of the virus cluster within the CABYV-Mediterranean group of isolates (Kwak et al. 2018). CABYV is widespread throughout the Mediterranean Basin, but it was found for the first time in Slovenia in this study. We confirmed the presence of CABYV in five different farms in southwestern Slovenia (eight samples) and in six different farms in northeastern Slovenia. Leaf yellowing symptoms were expressed on C. pepo infected by CABYV only. In some of the mixed infected plants, severe symptoms were observed. Further investigation is needed to clarify the significance of CABYV infection.The author(s) declare no conflict of interest.References:Juarez, M., et al. 2004. Plant Dis. 88:907. https://doi.org/10.1094/PDIS.2004.88.8.907A Link, ISI, Google ScholarKwak, H.-R., et al. 2018. Plant Pathol. J. 34:532. Crossref, ISI, Google ScholarPecman, A., et al. 2017. Front. Microbiol. 8:1998. https://doi.org/10.3389/fmicb.2017.01998 Crossref, ISI, Google ScholarThe author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 104, No. 2 February 2020SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionSymptom of maize ear rot caused by Fusarium sporotrichioides (B. B. Wang et al.). Photo credit: C. X. Duan. Systemic symptoms of alfalfa mosaic virus (AMV) isolate CaM on leaves of potato (X. Z. Nie et al.). Photo credit: X. Z. Nie. Metrics Downloaded 2,749 times Article History Issue Date: 31 Jan 2020Published: 18 Dec 2019First Look: 23 Oct 2019Accepted: 11 Oct 2019 Pages: 599-599 Information© 2020 The American Phytopathological SocietyKeywordsvirusesfield cropsepidemiologypathogen detectionThe author(s) declare no conflict of interest.Cited ByCucurbit aphid-borne yellows virus (CABYV) infecting melon and bitter gourd in Java, IndonesiaIOP Conference Series: Earth and Environmental Science, Vol. 948, No. 1Aphid‐borne viruses infecting cultivated watermelon and squash in Spain: Characterization of a variant of cucurbit aphid‐borne yellows virus (CABYV)20 May 2021 | Plant Pathology, Vol. 70, No. 6Occurrence and distribution of viruses infecting Zucchini and Watermelon in Morocco19 October 2020 | Archives of Phytopathology and Plant Protection, Vol. 54, No. 7-8
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