First Report of Watermelon Crinkle Leaf-Associated Virus 2 Infecting Watermelon (Citrullus lanatus) in Oklahoma

HomePlant DiseaseVol. 107, No. 3First Report of Watermelon Crinkle Leaf-Associated Virus 2 Infecting Watermelon (Citrullus lanatus) in Oklahoma PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Watermelon Crinkle Leaf-Associated Virus 2 Infecting Watermelon (Citrullus lanatus) in OklahomaClark Gilford and Akhtar AliClark GilfordDepartment of Biological Science, The University of Tulsa, Tulsa, OK 74104 and Akhtar Ali†Corresponding author: A. Ali; E-mail Address: [email protected]https://orcid.org/0000-0003-3474-0253Department of Biological Science, The University of Tulsa, Tulsa, OK 74104AffiliationsAuthors and Affiliations Clark Gilford Akhtar Ali † Department of Biological Science, The University of Tulsa, Tulsa, OK 74104 Published Online:28 Feb 2023https://doi.org/10.1094/PDIS-05-22-1238-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleWatermelon (Citrullus lanatus) is one of the major cucurbit crops in Oklahoma, which is grown on >1,200 acres. It contributes $16 million to the state's economy (NASS 2022; Shrefler et al. 2017). During the 2021 growing season, virus-like symptoms on watermelon plants were observed in a grower's field in Blaine County, Oklahoma. These symptoms included mottling, crinkled leaf edges, and a unique yellow mosaic pattern. Leaf tissue samples were randomly collected from nine plants and total RNA was extracted using Spectrum Plant Total RNA Kit (Sigma-Aldrich). Total RNA from one sample (named BL13) was subjected to high-throughput sequencing (HTS) on the NextSeq 500/550 High-Output kit v2.5 (Illumina, U.S.A.) at the genomic facility of Oklahoma State University. A total of 21,52,786 trimmed pair-end reads were assembled using CLC Genomics Workbench (v12.0.3) (Qiagen) and subjected to BLASTn analysis. Of these, three contigs of 6,626, 1,383, and 1,233 bp (average coverage 20,658×) showed 99% nucleotide (nt) sequence identity with RNA1 of a watermelon crinkle leaf-associated virus (WCLaV-2) isolate from Brazil (LC636073). The HTS data for BL13 samples also revealed three contigs of 263, 567, and 1,066 bp that matched with Cucumis melo cryptic virus isolate CmCV-HLI but was not confirmed by RT-PCR. The presence of WCLaV-2 in the BL13 sample was confirmed by RT-PCR using primers (RNA1-F: 5′-CAAAACGCCCATAAGAATAG-3′ and RNA1-R: 5′-CTCCATTAATGAGAACTAGGAG-3′) that amplified a 1.2-kb segment of the RNA1. The PCR product was confirmed on 1% agarose gel and cloned and three independent recombinant clones Sanger sequenced (Eurofins Genomics, Louisville, KY). The resulting consensus sequence (1.2 kb) (ON453838) was subjected to a BLASTn search against the GenBank databases, resulting in 99.76% nt identity with WCLaV-2 RNA1 (accession no. LC636073). Screening of the remaining eight symptomatic samples by RT-PCR revealed another positive sample (named BL7) for WCLaV-2. Consensus nt sequence (1.2 kb) analysis for the virus isolate from the BL7 sample (accession no. ON453839) shared 99.84% nt identities with WCLaV-2 RNA1 (LC636073). Total RNA from BL13 and BL7 samples, as well as the remaining seven samples, were tested against three major potyviruses (PRSV-W, WMV, and ZYMV) by multiplex RT-PCR (Rajbanshi and Ali 2019). Both BL13 and BL7 samples were negative to all three potyviruses, but the other eight samples were positive to one or two of the potyviruses. WCLaV-2 is a negative-sense ssRNA virus (family Phenuiviridae) and has been recently reported from watermelon in China (Xin et al. 2017) and Brazil (Maeda et al. 2022). In the United States, it has been reported from Florida (Hendrick et al. 2022), Georgia (Adeleke et al. 2022), and Texas (Hernandez et al. 2021). The mode of transmission of WCLaV-2 and its direct effect on watermelon fruit are not known (Maeda et al. 2022; Xin et al. 2017). Our results report for the first time the occurrence of WCLaV-2 infecting watermelon naturally in Oklahoma and show a further potential threat to watermelon production in the state. Further studies are needed to determine the incidence of WCLaV-2 in other counties of Oklahoma.The author(s) declare no conflict of interest.References:Adeleke, I. A., et al. 2022. Plant Dis. 106:2273. https://doi.org/10.1094/PDIS-11-21-2521-PDN Abstract, Google ScholarHendrick, K. E. M., et al. 2022. Plant Dis. 106:339. Link, ISI, Google ScholarHernandez, R. N., et al. 2021. Plant Dis. 105:2025. https://doi.org/10.1094/PDIS-02-21-0249-PDN Link, ISI, Google ScholarMaeda, M. H. K., et al. 2022. Plant Dis. 106:773. https://doi.org/10.1094/pdis-06-21-1325-pdn Link, ISI, Google ScholarNASS. 2022. USDA-National Agricultural Statistics Service. Washington, DC. Google ScholarRajbanshi, N., and Ali, A. 2019. J. Virol. Methods 273:113725. https://doi.org/10.1016/j.jviromet.2019.113725 Crossref, ISI, Google ScholarShrefler, J., et al. 2017. Watermelon Production. HLA-6236. Oklahoma Cooperative Extension Service. Google ScholarXin, M., et al. 2017. Front. Microbiol. 8:1514. https://doi.org/10.3389/fmicb.2017.01514 Crossref, ISI, Google ScholarFunding: This work was supported by the U.S. Department of Agriculture's (USDA) Agricultural Marketing Service through grant no. 0409018230. The Department of Biological Science and Office of Research and Sponsored Programs, The University of Tulsa, Oklahoma, supported publication charges. The contents are solely the responsibility of the authors and do not necessarily represent the official views of the USDA.The author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 107, No. 3 March 2023SubscribeISSN:0191-2917e-ISSN:1943-7692 Download Metrics Article History Issue Date: 3 Apr 2023Published: 28 Feb 2023First Look: 12 Aug 2022Accepted: 11 Aug 2022 Page: 973 Information© 2023 The American Phytopathological SocietyFundingU.S. Department of Agriculture's (USDA) Agricultural Marketing ServiceGrant/Award Number: 0409018230Keywordspathogen detectionvegetablesviruses and viroidsThe author(s) declare no conflict of interest.PDF downloadCited byFirst Report of Watermelon Crinkle Leaf-Associated Virus (WCLaV)-1 and WCLaV-2 Infecting Straightneck Squash in the United StatesA. Abdul Kader Jailani, Fanny B. Iriarte, and Mathews L. Paret17 August 2023 | Plant Disease, Vol. 107, No. 8