Construction of an infectious full-length cDNA clone of a melon yellowing-associated virus from Brazil

Construction of an infectious full-length cDNA clone of a melon yellowing-associated virus from Brazil | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Construction of an infectious full-length cDNA clone of a melon yellowing-associated virus from Brazil B. A. Silva, D. M. T. A. Freitas, C. M. Kauffmann, A. M. V. Batista, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8866350/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 03 May, 2026 Read the published version in Virus Genes → Version 1 posted 13 You are reading this latest preprint version Abstract Melon yellowing-associated virus (MYaV), a member of the genus Carlavirus (family Betaflexiviridae ), is a positive-sense, single-stranded RNA virus with a poly(A) tail. It is thought to be one of causative agents of severe yellowing disease in melon “Amarelão do meloeiro” (in Portugues) in northeastern Brazil and is transmitted by the whitefly Bemisia tabaci . In the field, MYaV frequently occurs in mixed infections with the recombinant form of cucurbit aphid-borne yellows virus (CABYV), contributing to the disease complex of melon severe yellowing disease. Considering its impact, inability of virus transmission by mechanical inoculation and the lack of molecular tools to study this pathogen, we constructed a full-length infectious cDNA clone of MYaV using Gibson Assembly technology. The genome was amplified in two overlapping fragments spanning the 5′ and 3′ regions, cloned in the binary vector pJL89 and subsequently introduced to Agrobacterium tumefaciens strain GV3101. Agroinoculation with this construct revealed that the MYaV clone was infectious in melon, cucumber ( Cucumis sativus ), West Indian gherkin ( Cucumis anguria ), and watermelon ( Citrullus lanatus ). Notably, infections in watermelon and West Indian gherkin were asymptomatic, a finding of epidemiological relevance since these crops may act as unnoticed inoculum sources in the field to melon crops. In contrast, no infection was detected in squash ( Cucurbita maxima ) or zucchini ( Cucurbita pepo ). This infectious clone provides not only a valuable tool to advance studies on MYaV and virus–host interactions but also an essential resource for breeding programs aimed at developing resistant melon varieties. MYaV Cucumis melo Infectious clone Melon yellowing-associated virus Gibson Assembly Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 03 May, 2026 Read the published version in Virus Genes → Version 1 posted Editorial decision: Revision requested 22 Apr, 2026 Reviews received at journal 21 Apr, 2026 Reviews received at journal 14 Apr, 2026 Reviews received at journal 14 Apr, 2026 Reviewers agreed at journal 13 Apr, 2026 Reviewers agreed at journal 13 Apr, 2026 Reviewers agreed at journal 11 Apr, 2026 Reviewers agreed at journal 10 Apr, 2026 Reviewers agreed at journal 10 Apr, 2026 Reviewers invited by journal 09 Apr, 2026 Editor assigned by journal 23 Feb, 2026 Submission checks completed at journal 23 Feb, 2026 First submitted to journal 12 Feb, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8866350","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":623154114,"identity":"4f860150-6cdb-45fe-8520-89ece3cb592e","order_by":0,"name":"B. A. Silva","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"B.","middleName":"A.","lastName":"Silva","suffix":""},{"id":623154116,"identity":"5984d50c-ef0f-4259-9927-8baa0055f9ed","order_by":1,"name":"D. M. T. A. Freitas","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"D.","middleName":"M. T. A.","lastName":"Freitas","suffix":""},{"id":623154125,"identity":"8bfe52d9-4745-4a77-a99e-54d55b560793","order_by":2,"name":"C. M. Kauffmann","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"C.","middleName":"M.","lastName":"Kauffmann","suffix":""},{"id":623154128,"identity":"4dfda27f-2261-4245-aa37-ac378f79ce49","order_by":3,"name":"A. M. V. Batista","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"A.","middleName":"M. V.","lastName":"Batista","suffix":""},{"id":623154131,"identity":"513efe44-b138-4192-921f-99f6a29475f0","order_by":4,"name":"P. S. Queiroz","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"P.","middleName":"S.","lastName":"Queiroz","suffix":""},{"id":623154136,"identity":"499e99a1-9481-4569-8fd1-e8d3d8673526","order_by":5,"name":"S. B. S. Cardenas","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"S.","middleName":"B. S.","lastName":"Cardenas","suffix":""},{"id":623154146,"identity":"cf148379-4f41-4205-9c02-4fe0d8bf86ce","order_by":6,"name":"D. F. Fraga","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"D.","middleName":"F.","lastName":"Fraga","suffix":""},{"id":623154149,"identity":"25984a06-2218-4903-9c45-873fd512c280","order_by":7,"name":"H. B. S. Mota","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"H.","middleName":"B. S.","lastName":"Mota","suffix":""},{"id":623154151,"identity":"66ee9696-f62b-4279-847d-c892dc4536fe","order_by":8,"name":"P. R. M. S. Machado","email":"","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":false,"prefix":"","firstName":"P.","middleName":"R. M. S.","lastName":"Machado","suffix":""},{"id":623154152,"identity":"6f583c86-e083-4a34-b792-82f99c31d176","order_by":9,"name":"T. Nagata","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAApUlEQVRIiWNgGAWjYBADOQYGHgZmopTC1BmTriWxgWgt9uznD3/mbbNL33C79wBz4R5ibOFJZpPmbUvO3XDnXALzjGdEOSyZjZm3jTl3w40cA2aeA8Ro4X/MDHRYfboB8VokkhmADjucQIKWG4/NJOecO24480ZewuEZxGhh7098/OFNWbU8343cg48LiNECBoxsEJpoDUDwhwS1o2AUjIJRMPIAAC87Ml7gEtNdAAAAAElFTkSuQmCC","orcid":"","institution":"Universidade de Brasília","correspondingAuthor":true,"prefix":"","firstName":"T.","middleName":"","lastName":"Nagata","suffix":""}],"badges":[],"createdAt":"2026-02-13 01:23:36","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8866350/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8866350/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11262-026-02240-z","type":"published","date":"2026-05-03T15:58:34+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":108809124,"identity":"fdd1ca97-6e59-4c71-bef5-a6348889666a","added_by":"auto","created_at":"2026-05-08 15:50:07","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":301317,"visible":true,"origin":"","legend":"","description":"","filename":"20260212MYaVinfectiouscloneFinal.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8866350/v1_covered_e41adb0d-fcfd-4c1f-8ea5-43b96bf17dad.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Construction of an infectious full-length cDNA clone of a melon yellowing-associated virus from Brazil","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"virus-genes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"viru","sideBox":"Learn more about [Virus Genes](http://link.springer.com/journal/11262)","snPcode":"11262","submissionUrl":"https://submission.nature.com/new-submission/11262/3","title":"Virus Genes","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"MYaV, Cucumis melo, Infectious clone, Melon yellowing-associated virus, Gibson Assembly","lastPublishedDoi":"10.21203/rs.3.rs-8866350/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8866350/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMelon yellowing-associated virus (MYaV), a member of the genus \u003cem\u003eCarlavirus\u003c/em\u003e (family \u003cem\u003eBetaflexiviridae\u003c/em\u003e), is a positive-sense, single-stranded RNA virus with a poly(A) tail. It is thought to be one of causative agents of severe yellowing disease in melon \u0026ldquo;Amarel\u0026atilde;o do meloeiro\u0026rdquo; (in Portugues) in northeastern Brazil and is transmitted by the whitefly \u003cem\u003eBemisia tabaci\u003c/em\u003e. In the field, MYaV frequently occurs in mixed infections with the recombinant form of cucurbit aphid-borne yellows virus (CABYV), contributing to the disease complex of melon severe yellowing disease. Considering its impact, inability of virus transmission by mechanical inoculation and the lack of molecular tools to study this pathogen, we constructed a full-length infectious cDNA clone of MYaV using Gibson Assembly technology. The genome was amplified in two overlapping fragments spanning the 5\u0026prime; and 3\u0026prime; regions, cloned in the binary vector pJL89 and subsequently introduced to \u003cem\u003eAgrobacterium tumefaciens\u003c/em\u003e strain GV3101. Agroinoculation with this construct revealed that the MYaV clone was infectious in melon, cucumber (\u003cem\u003eCucumis sativus\u003c/em\u003e), West Indian gherkin (\u003cem\u003eCucumis anguria\u003c/em\u003e), and watermelon (\u003cem\u003eCitrullus lanatus\u003c/em\u003e). Notably, infections in watermelon and West Indian gherkin were asymptomatic, a finding of epidemiological relevance since these crops may act as unnoticed inoculum sources in the field to melon crops. In contrast, no infection was detected in squash (\u003cem\u003eCucurbita maxima\u003c/em\u003e) or zucchini (\u003cem\u003eCucurbita pepo\u003c/em\u003e). This infectious clone provides not only a valuable tool to advance studies on MYaV and virus\u0026ndash;host interactions but also an essential resource for breeding programs aimed at developing resistant melon varieties.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e","manuscriptTitle":"Construction of an infectious full-length cDNA clone of a melon yellowing-associated virus from Brazil","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-16 23:24:24","doi":"10.21203/rs.3.rs-8866350/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-22T07:04:38+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-21T23:28:31+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-14T18:09:01+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-14T16:59:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"175825901748877549632665406356029780378","date":"2026-04-13T15:39:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"154075760494245402387605850870235362899","date":"2026-04-13T15:03:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"199438003489605158511621356396603117157","date":"2026-04-11T07:53:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"221585346460673485155492337493611586689","date":"2026-04-10T16:17:27+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"163589302043185722465871493345598631100","date":"2026-04-10T11:48:38+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-09T07:30:51+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-23T15:31:24+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-23T15:30:51+00:00","index":"","fulltext":""},{"type":"submitted","content":"Virus Genes","date":"2026-02-13T01:12:52+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"virus-genes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"viru","sideBox":"Learn more about [Virus Genes](http://link.springer.com/journal/11262)","snPcode":"11262","submissionUrl":"https://submission.nature.com/new-submission/11262/3","title":"Virus Genes","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"c50de22d-11e1-4ebc-95ec-a78a0b7e9122","owner":[],"postedDate":"April 16th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-05-08T15:15:58+00:00","versionOfRecord":{"articleIdentity":"rs-8866350","link":"https://doi.org/10.1007/s11262-026-02240-z","journal":{"identity":"virus-genes","isVorOnly":false,"title":"Virus Genes"},"publishedOn":"2026-05-03 15:58:34","publishedOnDateReadable":"May 3rd, 2026"},"versionCreatedAt":"2026-04-16 23:24:24","video":"","vorDoi":"10.1007/s11262-026-02240-z","vorDoiUrl":"https://doi.org/10.1007/s11262-026-02240-z","workflowStages":[]},"version":"v1","identity":"rs-8866350","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8866350","identity":"rs-8866350","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}