Phylogenetic Analysis of a Mitovirus in Fusarium oxysporum from Phytolacca dioica in the Canary Islands

Phylogenetic Analysis of a Mitovirus in Fusarium oxysporum from Phytolacca dioica in the Canary Islands | 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 Research Article Phylogenetic Analysis of a Mitovirus in Fusarium oxysporum from Phytolacca dioica in the Canary Islands Laura Arango-Palacio, Juliana Osorio-Marulanda, Juliana Lopez-Jimenez, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8502341/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract We characterized a novel mitovirus from Fusarium oxysporum isolated from Phytolacca dioica (ombú) in the Canary Islands, Spain. Using transcriptome sequencing, we assembled the full-length genome of the novel Fusarium oxysporum mitovirus 2-like (FoMV2-like), which consists of 2,363 nucleotides and has a GC content of 30.3%. The genome harbors a single open reading frame (ORF) encoding a protein of 713 amino acids. Phylogenetic analysis of the RdRp amino acid sequence showed that FoMV2-like clusters closely with Fusarium oxysporum mitovirus 2, suggesting a close evolutionary relationship. DNADiff analysis revealed that the genome of FoMV2-like shares 100% alignment with Fusarium oxysporum mitovirus 2, with an average identity of 87.33%, supporting its classification as a distinct species within the Mitoviridae family. This study expands our understanding of mycoviral diversity in Fusarium oxysporum and contributes to the genomic characterization of mitoviruses. Figures Figure 1 Figure 2 Introduction Mycoviruses infecting plant-pathogenic fungi exhibit great genomic diversity. According to the International Committee on Virus Taxonomy (ICTV) (https://talk.ictvonline.org/taxonomy/), thirty viral families include representatives that infect fungal species [1–3]. The majority of mycoviruses have RNA genomes, which can be positive (+) or negative (-) sense single-stranded RNA (ssRNA), or double-stranded RNA (dsRNA) [4]. Furthermore, in many cases, mycoviruses do not exhibit significant phenotypic effects on their fungal hosts [5]. Among the various mycoviruses, those infecting Fusarium oxysporum are of particular interest due to their potential impact on this cosmopolitan plant pathogen that infects more than 120 species and causes Fusarium wilt in several crops worldwide [7, 8]. Mycoviruses from different viral families have been reported with hypovirulence activity over F. oxysporum [9–12]. Viruses belonging to Mitoviridae family are positive-sense (+) single-stranded (ss) RNA, with genomes ranging from 2.3 to 5 kb [14]. Their genomes contain a single open reading frame (ORF), which encodes an RNA-dependent RNA polymerase (RdRp) [15, 16]. Mitoviruses replicate in fungal mitochondria, do not form virions, and are unencapsidated, generally causing mitochondrial alterations that result in hypovirulence [17–19]. Several members of the genus Mitovirus have been identified infecting different Fusarium species worldwide [20–23]. Within Fusarium oxysporum , several mitoviruses have been described to date, including Fusarium oxysporum f. sp. Dianthi mitovirus 1 (FodMV1) [24], Fusarium oxysporum mitovirus 1 (FoMV1) [11], Fusarium oxysporum mitovirus 2 (FoMV2), isolated from the pathogenic Fusarium oxysporum f. sp. ginseng [25], and Fusarium oxysporum mitovirus 3 (FoMV3), identified in Fusarium oxysporum f. sp. Melonis causal agent of Fusarium wilt in melons [26]. This study aims to contribute to the genomic understanding of mycoviruses in Fusarium oxysporum by investigating a novel mycovirus in a Fusarium isolate and its genomic characteristics. Genome Assembly and Phylogeny of a Mitovirus from Fusarium oxysporum The Fusarium strain was obtained of from Phytolacca dioica (ombú) in the Canary Islands, Spain, and stored as a conidial suspension. Fusarium oxysporum samples were cultured on potato dextrose agar (PDA) plates at 27 °C. To detect the presence of RNA viruses, a cellulose chromatography method was used to enrich the samples of dsRNA molecules, wich are the replication intermediates of most RNA mycoviruses [27]. For dsRNA extraction, Fusarium oxysporum samples were processed following the protocol described by Cañizares et al. (2015). Subsequently, RNA extraction was performed for sequencing and transcriptome analysis by RNA-seq. Mycelial mass was recovered from the culture medium and macerated in liquid nitrogen and immediately suspended in Trizol TM Reagent to stabilize the genetic material and subsequently processed for the extraction of total RNA, following the standard RNA extraction protocol described by the manufacturer (Trizol TM Reagent, Invitrogen; total RNA extraction protocol). RNA-seq was performed using the Illumina NovaSeq 6000 platform, generating paired-end reads of approximately 100 bp from libraries prepared with the TruSeq Stranded Total RNA kit with Ribo-Zero H/M/R Gold. Reads were quality filtered using Cutadapt v3.5 [28], retaining sequences with a minimum quality score of Q30 and a length of at least 70 bases. De novo assembly was performed using Trinity 2.13.2 with default parameters [29]. Putative viral contigs were identified through BLASTN searches against a custom database containing nucleotide and amino acid sequences from fungal mitovirus genomes and fungal virus genomes retrieved from RefSeq and NCBI Virus. This approach allowed the detection of viral sequences corresponding to a mitovirus, which were subsequently extracted using the RdRp amino acid sequence for further phylogenetic analysis. For the phylogenetic analysis of the mitovirus, the amino acid sequence of its RNA-dependent RNA polymerase (RdRp) was aligned with RdRp sequences of representative members of the family Mitoviridae retrieved from RefSeq, as well as with mitovirus sequences previously described in Fusarium hosts. Narnavirus sequences were included as the outgroup. Multiple sequence alignment was performed using the MAFFT program [30, 31]. The phylogenetic tree based on the RdRp protein alignment was carried out using IQ-TREE 3 [32] under the Q.PFAM+F+I+R5 substitution model, and branch support was evaluated using the ultrafast bootstrap method with 5,000 pseudoreplicates [33] to infer evolutionary relationships among mitovirus taxa. Phylogenetic analysis based on RdRp amino acid sequences showed that the Fusarium oxysporum mitovirus 2-like clusters strongly with Fusarium oxysporum mitovirus 2 (FoMV2), supported by high branch support values (UFB = 100). This clustering indicates a close evolutionary relationship between the two viruses, despite their detection in fungal hosts from geographically distinct locations (Figure 4). DNADiff analysis revealed that the genome of FoMV2-like shares 100% alignment with Fusarium oxysporum mitovirus 2, with an average identity of 87.33%, and 93.93% alignment with Fusarium andiyazi mitovirus 2, with an average identity of 82.28%. Based on the species demarcation criteria for mitoviruses outlined by the ICTV, mitovirus isolates with less than 90% amino acid sequence identity in their RdRp are classified as different species. Consequently, FoMV2-like should be regarded as a member of a new species within the family Mitoviridae . The full-length genome sequence of Fusarium oxysporum mitovirus 2-like (FoMV2-like), accession number (ID 3036262) was obtained through transcriptome assembly. The genome is 2,363 nucleotides in length and has a GC content of 30.3%. A single open reading frame (ORF), starting at position 164 and ending at position 2,304 (Figure 2A), is present within the genome. ORF prediction was carried out using ORF Finder. To assess potential secondary structures, the 5'- and 3'-terminal regions of the mitovirus genome (positive strand) were predicted using the RNAfold WebServer. This ORF is translated into a protein of 713 amino acids, with a molecular weight of 83.136 kDa and an isoelectric point of 9.63. The 5'-terminal sequence is predicted to fold into a stable stem-loop structure with a ΔG value of -3.00 kcal/mol, while the 3'-terminal sequence folds into three stem-loop structures with a ΔG value of -7.00 kcal/mol (Figure 2B). The predicted RNA secondary structures of FoMV2-like resemble those reported for other mitoviruses and are characteristic of members of the Mitoviridae family [13]. Declarations Acknowledgments The authors want to thank the Universidad de Antioquia (UdeA) for financing the publication, and the Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora” and the Centro de Investigaciones del Banano “Cenibanano” for encouraging research. All authors declare no conflict of interest about this publication. Conflict of interest: The authors declare no conflict of interest. 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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-8502341","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":573193648,"identity":"e1848f9c-8084-4b4a-a32e-bdf90f25977d","order_by":0,"name":"Laura Arango-Palacio","email":"","orcid":"","institution":"Universidad de Antioquia","correspondingAuthor":false,"prefix":"","firstName":"Laura","middleName":"","lastName":"Arango-Palacio","suffix":""},{"id":573193649,"identity":"d3c0f25d-b51a-4bb1-9977-f4c1bff635f2","order_by":1,"name":"Juliana Osorio-Marulanda","email":"","orcid":"","institution":"Universidad de 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08:09:14","extension":"html","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":71822,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8502341/v1/73ae7170c24e9ed6ec4f9867.html"},{"id":100370916,"identity":"d2d79609-44eb-4f75-8261-d33adc79b6af","added_by":"auto","created_at":"2026-01-16 08:09:00","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":109366,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum likelihood phylogenetic tree based on RdRp amino acid sequences of the studied mitovirus, representative members of the family \u003cem\u003eMitoviridae\u003c/em\u003e, and mitovirus sequences described in \u003cem\u003eFusarium\u003c/em\u003e hosts. Black circles indicate branches with strong support (UFB \u0026gt; 97).\u003c/p\u003e","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8502341/v1/cf063f130251778f5e951483.png"},{"id":100226670,"identity":"579fbb5b-736a-4458-bf4e-3a53be22a4cf","added_by":"auto","created_at":"2026-01-14 10:36:48","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":158189,"visible":true,"origin":"","legend":"\u003cp\u003eGenomic organization and terminal structure of \u003cem\u003eFusarium oxysporum\u003c/em\u003e mitovirus 2-like (FoMV2-like). \u0026nbsp;(A) Schematic diagram of the genome organization of FoMV2-like. (B) Potential secondary structures of the 5’ and 3’ termini of FoMV2-like.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8502341/v1/0803d480ec6011ac866c3ae5.png"},{"id":100733810,"identity":"9c30b343-cfb9-49e6-a3b3-d046bace682d","added_by":"auto","created_at":"2026-01-20 22:00:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":594594,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8502341/v1/62ca5e96-7280-4df3-8558-79ef1f4eab93.pdf"}],"financialInterests":"","formattedTitle":"Phylogenetic Analysis of a Mitovirus in Fusarium oxysporum from Phytolacca dioica in the Canary Islands","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMycoviruses infecting plant-pathogenic fungi exhibit great genomic diversity. According to the International Committee on Virus Taxonomy (ICTV) (https://talk.ictvonline.org/taxonomy/), thirty viral families include representatives that infect fungal species\u003cem\u003e\u0026nbsp;\u003c/em\u003e[1\u0026ndash;3]. The majority of mycoviruses have RNA genomes, which can be positive (+) or negative (-) sense single-stranded RNA (ssRNA), or double-stranded RNA (dsRNA) [4]. Furthermore, in many cases, mycoviruses do not exhibit significant phenotypic effects on their fungal hosts [5].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAmong the various mycoviruses, those infecting\u003cem\u003e\u0026nbsp;Fusarium oxysporum\u0026nbsp;\u003c/em\u003eare of particular interest due to their potential impact on this cosmopolitan plant pathogen that infects more than 120 species and causes \u003cem\u003eFusarium\u003c/em\u003e wilt in several crops worldwide [7, 8]. Mycoviruses from different viral families have been reported with hypovirulence activity over \u003cem\u003eF. oxysporum\u0026nbsp;\u003c/em\u003e[9\u0026ndash;12]. Viruses belonging to \u003cem\u003eMitoviridae\u003c/em\u003e family are positive-sense (+) single-stranded (ss) RNA, with genomes ranging from 2.3 to 5 kb [14]. Their genomes contain a single open reading frame (ORF), which encodes an RNA-dependent RNA polymerase (RdRp) [15, 16].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMitoviruses replicate in fungal mitochondria, do not form virions, and are unencapsidated, generally causing mitochondrial alterations that result in hypovirulence [17\u0026ndash;19]. Several members of the genus \u003cem\u003eMitovirus\u003c/em\u003e have been identified infecting different \u003cem\u003eFusarium\u003c/em\u003e species worldwide\u0026nbsp;[20\u0026ndash;23]. Within \u003cem\u003eFusarium oxysporum\u003c/em\u003e, several mitoviruses have been described to date, including Fusarium oxysporum f. sp. Dianthi mitovirus 1 (FodMV1) [24], Fusarium oxysporum mitovirus 1 (FoMV1) [11], Fusarium oxysporum mitovirus 2 (FoMV2), isolated from the pathogenic \u003cem\u003eFusarium oxysporum\u003c/em\u003e f. sp. \u003cem\u003eginseng\u003c/em\u003e [25], and Fusarium oxysporum mitovirus 3 (FoMV3), identified in \u003cem\u003eFusarium oxysporum\u003c/em\u003e f. sp. \u003cem\u003eMelonis\u003c/em\u003e causal agent of Fusarium wilt in melons [26]. This study aims to contribute to the genomic understanding of mycoviruses in \u003cem\u003eFusarium oxysporum\u0026nbsp;\u003c/em\u003eby investigating a novel mycovirus in a \u003cem\u003eFusarium\u003c/em\u003e isolate and its genomic characteristics.\u003c/p\u003e"},{"header":"Genome Assembly and Phylogeny of a Mitovirus from Fusarium oxysporum","content":"\u003cp\u003eThe\u003cem\u003e\u0026nbsp;Fusarium\u003c/em\u003e strain was obtained of from \u003cem\u003ePhytolacca dioica\u003c/em\u003e (omb\u0026uacute;) in the Canary Islands, Spain, and stored as a conidial suspension. \u003cem\u003eFusarium\u003c/em\u003e \u003cem\u003eoxysporum\u003c/em\u003e samples were cultured on potato dextrose agar (PDA) plates at 27 \u0026deg;C. To detect the presence of RNA viruses, a cellulose chromatography method was used to enrich the samples of dsRNA molecules, wich are the replication intermediates of most RNA mycoviruses [27]. For dsRNA extraction, \u003cem\u003eFusarium oxysporum\u0026nbsp;\u003c/em\u003esamples were processed following the protocol described by Ca\u0026ntilde;izares et al. (2015).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSubsequently, RNA extraction was performed for sequencing and transcriptome analysis by RNA-seq. Mycelial mass was recovered from the culture medium and macerated in liquid nitrogen and immediately suspended in Trizol TM Reagent to stabilize the genetic material and subsequently processed for the extraction of total RNA, following the standard RNA extraction protocol described by the manufacturer (Trizol TM Reagent, Invitrogen; total RNA extraction protocol). RNA-seq was performed using the Illumina NovaSeq 6000 platform, generating paired-end reads of approximately 100 bp from libraries prepared with the TruSeq Stranded Total RNA kit with Ribo-Zero H/M/R Gold. Reads were quality filtered using Cutadapt v3.5 [28], retaining sequences with a minimum quality score of Q30 and a length of at least 70 bases. De novo assembly was performed using Trinity 2.13.2 with default parameters [29]. Putative viral contigs were identified through BLASTN searches against a custom database containing nucleotide and amino acid sequences from fungal mitovirus genomes and fungal virus genomes retrieved from RefSeq and NCBI Virus. This approach allowed the detection of viral sequences corresponding to a mitovirus, which were subsequently extracted using the RdRp amino acid sequence for further phylogenetic analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFor the phylogenetic analysis of the mitovirus, the amino acid sequence of its RNA-dependent RNA polymerase (RdRp) was aligned with RdRp sequences of representative members of the family \u003cem\u003eMitoviridae\u003c/em\u003e retrieved from RefSeq, as well as with mitovirus sequences previously described in \u003cem\u003eFusarium\u003c/em\u003e hosts. Narnavirus sequences were included as the outgroup. Multiple sequence alignment was performed using the MAFFT program [30, 31]. The phylogenetic tree based on the RdRp protein alignment was carried out using IQ-TREE 3 [32] under the Q.PFAM+F+I+R5 substitution model, and branch support was evaluated using the ultrafast bootstrap method with 5,000 pseudoreplicates [33] to infer evolutionary relationships among mitovirus taxa. Phylogenetic analysis based on RdRp amino acid sequences showed that the Fusarium oxysporum mitovirus 2-like clusters strongly with \u003cem\u003eFusarium oxysporum mitovirus 2\u0026nbsp;\u003c/em\u003e(FoMV2), supported by high branch support values (UFB = 100). This clustering indicates a close evolutionary relationship between the two viruses, despite their detection in fungal hosts from geographically distinct locations (Figure 4).\u003c/p\u003e\u003cp\u003eDNADiff analysis revealed that the genome of FoMV2-like shares 100% alignment with Fusarium oxysporum mitovirus 2, with an average identity of 87.33%, and 93.93% alignment with Fusarium andiyazi mitovirus 2, with an average identity of 82.28%. Based on the species demarcation criteria for mitoviruses outlined by the ICTV, mitovirus isolates with less than 90% amino acid sequence identity in their RdRp are classified as different species. Consequently, FoMV2-like should be regarded as a member of a new species within the family \u003cem\u003eMitoviridae\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eThe full-length genome sequence of Fusarium oxysporum mitovirus 2-like (FoMV2-like), accession number (ID 3036262) was obtained through transcriptome assembly. The genome is 2,363 nucleotides in length and has a GC content of 30.3%. A single open reading frame (ORF), starting at position 164 and ending at position 2,304 (Figure 2A), is present within the genome. ORF prediction was carried out using ORF Finder. To assess potential secondary structures, the 5\u0026apos;- and 3\u0026apos;-terminal regions of the mitovirus genome (positive strand) were predicted using the RNAfold WebServer. This ORF is translated into a protein of 713 amino acids, with a molecular weight of 83.136 kDa and an isoelectric point of 9.63. The 5\u0026apos;-terminal sequence is predicted to fold into a stable stem-loop structure with a \u0026Delta;G value of -3.00 kcal/mol, while the 3\u0026apos;-terminal sequence folds into three stem-loop structures with a \u0026Delta;G value of -7.00 kcal/mol (Figure 2B). The predicted RNA secondary structures of FoMV2-like resemble those reported for other mitoviruses and are characteristic of members of the Mitoviridae family [13].\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors want to thank the Universidad de Antioquia (UdeA) for financing the publication, and the Instituto de Hortofruticultura Subtropical y Mediterr\u0026aacute;nea \u0026ldquo;La Mayora\u0026rdquo; and the Centro de Investigaciones del Banano \u0026ldquo;Cenibanano\u0026rdquo; for encouraging research. All authors declare no conflict of interest about this publication.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis article does not contain any studies with human participants or animals performed by any of the authors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed consent:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all participants in the study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eKuhn JH, Adkins S, Brown K, et al (2023) ICTV Virus Taxonomy Profile: Tulasviridae 2023: This article is part of the ICTV Virus Taxonomy Profiles collection. 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Mol Biol Evol 35:518\u0026ndash;522. https://doi.org/10.1093/molbev/msx281\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-8502341/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8502341/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"We characterized a novel mitovirus from Fusarium oxysporum isolated from Phytolacca dioica (ombú) in the Canary Islands, Spain. Using transcriptome sequencing, we assembled the full-length genome of the novel Fusarium oxysporum mitovirus 2-like (FoMV2-like), which consists of 2,363 nucleotides and has a GC content of 30.3%. The genome harbors a single open reading frame (ORF) encoding a protein of 713 amino acids. Phylogenetic analysis of the RdRp amino acid sequence showed that FoMV2-like clusters closely with Fusarium oxysporum mitovirus 2, suggesting a close evolutionary relationship. DNADiff analysis revealed that the genome of FoMV2-like shares 100% alignment with Fusarium oxysporum mitovirus 2, with an average identity of 87.33%, supporting its classification as a distinct species within the Mitoviridae family. This study expands our understanding of mycoviral diversity in Fusarium oxysporum and contributes to the genomic characterization of mitoviruses.","manuscriptTitle":"Phylogenetic Analysis of a Mitovirus in Fusarium oxysporum from Phytolacca dioica in the Canary Islands","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-14 10:36:43","doi":"10.21203/rs.3.rs-8502341/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"8453670c-26d6-4dab-ace7-556e85c26a87","owner":[],"postedDate":"January 14th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-01-20T20:14:15+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-14 10:36:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8502341","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8502341","identity":"rs-8502341","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}