The Full-length Genome Sequence of a Novel Mitovirus From the Causal Agent Botrosphaeria Dothidea of Pear Ring Rot Disease

The Full-length Genome Sequence of a Novel Mitovirus From the Causal Agent Botrosphaeria Dothidea of Pear Ring Rot Disease | 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 The Full-length Genome Sequence of a Novel Mitovirus From the Causal Agent Botrosphaeria Dothidea of Pear Ring Rot Disease Qi Zou, Yunjing Gao, Qiong Wang, Yuekun Yang, Fang Wang, Ni Hong, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-431413/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 02 Aug, 2021 Read the published version in Archives of Virology → Version 1 posted 3 You are reading this latest preprint version Abstract Here, we describe a novel mycovirus, Botryosphaeria dothidea mitovirus 2 (tentatively designated as BdMV2), isolated from Botryosphaeria dothidea FJ strain causing pear ring rot disease in Fujian Province, China. The complete genome nucleotide sequence of BdMV2 is 2538 nt in length and contains a single 2070 nt open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) of 689 amino acid (aa) using fungal mitochondrial genetic code. BLASTp analysis revealed that the RdRp of BdMV2 shares 28.91%–69.36% (query sequence coverage more than 90%) sequence identity to those of members in the genus Mitovirus , and the closest similarity is 69.36% and 68.79% with the corresponding protein aa sequences of Rhizoctonia solani mitovirus 10 and Macrophomina phaseolina mitovirus 4, respectively. Phylogenetic analysis based on the RdRp aa sequences further revealed that BdMV2 is a newly member in the genus Mitovirus of the family Narnaviridae . To our knowledge, BdMV2 is thus a novel mitovirus from the causal agent B. dothidea of pear ring rot disease. Virology Botryosphaeria dothidea mitovirus 2 Botryosphaeria dothidea rot disease genome sequence novel mitovirus Figures Figure 1 Figure 2 Background Botryosphaeria dothidea is the important and widespread trunk pathogen, which causes pear ring rot disease in the pear production regions in China. Botryosphaeria dothidea with an extensive host range not only harms pear trees by causing severe symptoms such as stem wart and canker, branch dieback and fruit rot, but also other fruit trees including apple, Jujube and grapevine [1–6]. The ring rot disease is becoming increasingly serious and distributes throughout all pear producing areas at present, which seriously affects the yield and quality of pear to further hind the healthy development of pear industry [6–7]. It is urgent to find safe and effective measure to biological control of pear ring rot diseases. As mycoviruses can infect and proliferate in fungi and some mycoviruses present hypovirulence-associated traits, they are potential biological control agents for the control of these fungal disease. Indeed, many mycoviruses with potential biocontrol value have been found and identified from pathogenic fungi infecting fruit trees [8]. Therefore, it is particularly important to find and identify mycoviruses in Botryosphaeria dothidea strains casing pear ring rot disease. To identify and characterize mycovirus, high through-put sequencing techniques and subsequent bioinformatics have been shown as a very effective approach [9–11]. So far, eleven mycoviruses have been identified in B. dothidea strains and characterized. In our lab, Botryosphaeria dothidea chrysovirus 1 (BdCV1), Bipolaris maydis botybirnavirus 1 strain BdEW220 (BmRV1-BdEW220), Botryosphaeria dothidea RNA virus 1 (BdRV1) involved in hypovirulence [12–14], Botryosphaeria dothidea partitivirus 1 (BdPV1), Botryosphaeria dothidea victorivirus 1 (BdVV1), Botryosphaeria dothidea victorivirus 2 (BdVV2) and Botryosphaeria dothidea botourmiavirus 1 (BdBOV-1) with no obvious effect on host, have been identified in B. dothidea strains causing pear ring rot disease [12, 15–17]. Botryosphaeria dothidea bipartite mycovirus 1 (BdBMV1), Botryosphaeria dothidea mitovirus 1 (BdMV1), Botryosphaeria dothidea fusarivirus 1 (BdFV1) and BdCV1-G1 have been also identified in B. dothidea strains isolated from Kerria japonica , pepper and apple hosts, respectively [18–21]. The Narnaviridae genome has positive single strand RNA (+ssRNA) with approximately 2.2–3.6 kb in length and encodes only one RNA-dependent RNA polymerase (RdRp). The Narnaviridae family is divided into two genera, Narnavirus and Mitovirus (Mitochondrial viruses), which replicate in the cytoplasm and mitochondria, respectively. The mycoviruses in Narnaviridae exist widely in fungi, but it is not clear if they have any effect on all kinds of fungi [22–23]. Here, we report the identification of a novel mitovirus isolated from B. dothidea strain, the causal pathogen of pear ring rot disease. We further determined its +ssRNA full-length genome sequence and molecular characteristics. This mycovirus was tentatively designated as Botryosphaeria dothidea mitovirus 2 (BdMV2). Provenance and sequencing of BdMV2 B. dothidea strain FJ was isolated from pear stem bark exhibiting wart symptoms in Fujian Province of China, in 2010, and the hypha blocks of FJ strain were stored in 25% glycerin solution at -80℃. The mycelium was grown on potato dextrose agar (PDA) plates overlaid with a layer of cellophane at 25 ℃ in the dark. The mycelium of B. dothidea strain FJ and other Valsa spp strains were together collected to be used for Illumina sequencing (Beckman company, Beijing, China). The obtained high-quality clean reads were assembled contigs, which were analyzed to search mycoviruses using Blastx. Among these contigs, Contig1603 with 2.5 kb in length shares the highest sequence similarity with 67.14% in comparison to that of Macrophomina phaseolina mitovirus 4. Total RNA was extracted by Trizol method (Aidlab Biotechnologies Co., Ltd, Beijing, China) as template for synthesize cDNA. Finally, the putative mycoviral genome intermediate sequence was determined by RT-PCR and obtained 2042 nt by sequencing in B. dothidea strain FJ using the designed primers based on Contig1603 sequence (Supplementary Table 1). The dsRNA was extracted from the cultured hypha by the column separation method [ 24 ]. The adaptor RACE-OLIGO was ligated to the 3’ terminus of each strand of dsRNA, which were used for template to reversely transcribed cDNA using Oligo REV primer as described previously [ 25 ]. The terminal sequence amplification was performed by nested PCR using the primers designed based on obtained viral sequence and sequence-specific adaptor primers of O5RACE-2 and O5RACE-3, respectively (Supplementary Table 1). The obtained PCR product was purified, linked to pMD18-T vector (Takara, Dalian, China), and transformed into Escherichia coli Top10. For each PCR product, at least three independent identified positive clones were sequenced in both directions (Sangon Biotech Co., Ltd, Shanghai, China). Sequences obtained from clones were assembled using DNAMAN. The viral ORF was predicted using the ORF finder online website of https://www.ncbi.nlm.nih.gov/orffinder . Mycoviral sequences search was performed using NCBI database. Multiple mycoviral sequences were aligned using CLUSTAL X [ 26 ]. Phylogenetic analysis was conducted by using NJ method with 1000 replicates of MEGA 7.0, and RdRp domain motif sequences were analyzed using the GeneDoc software [ 27 – 28 ]. The secondary structures of BdMV2 genome terminus were predicated by online RNAfold software. Sequence properties The colony morphology of B. dothidea strain FJ was shown, which exhibited normal mycelia and the mycelia could grow to cover the whole plate in 5 days (Fig. 1 A). RT-PCR and RNA-ligase-mediated rapid amplification of cDNA ends (RLM-RACE) in combination with cloning and sequence assembly were used to obtain the BdMV2 full-length genome sequence from B. dothidea strain FJ. Its complete genome sequence was deposited in the NCBI GenBank database (Acc. No. MW553911), which was 2538 nt in length with a CG content of 47.24% for the genome. BdMV2 contains a single big ORF with AUG as the initiation condon at positions 176–178 nt and UAA as the termination condon at positions 2243–2245 nt of the genome by fungal mitochondrial codon usage. The ORF (2,070 nt in length) potentially encoded 689 aa RdRp protein with predicated molecular weight of 77.35 kDa. The 5’ and 3’ untranslated regions (UTRs) are 175 and 293 nt long, respectively (Fig. 1 B). Using the RNAfold program, two potential stem-loop structures in the 5’ and 3’ UTRs and a potential panhandle structure of the BdMV2 positive-sense strand UTRs were predicted, respectively (Fig. 1 C), which showed typical characteristic of reported members of the genus Mitovirus [ 22 ]. The BdMV2 contains a conserved RdRp domain at positions 164–495 aa of the genome (Mitovirus RNA-dependent RNA polymeras; pfam05919) by the Conserved Domain Database (CDD) search of NCBI. Multiple sequence alignment analysis of RdRp domain from BdMV2 with those members in Mitovirus showed that BdMV2 RdRp domain contains all six conserved motifs (I to VI), which are characteristics of members in the Mitovirus (Fig. 2 A). A BLASTp comparison of the RdRp sequence of BdMV2 showed the highest similarity of 69.36% with that of RsMV10, and had identity 28.91–68.79% (query coverage more than 90%) to those of members in the genus Mitovirus (Supplementary Table 2). To determine the taxonomic status of BdMV2, phylogenetic analysis was performed using the RdRp aa sequences encoded by BdMV2 and other selected mycoviruses in Mitovirus and Narnavirus of the family Narnaviridae . The results showed that all mitoviruses were classified into two clades of clade I and clade II. It revealed that BdMV2 existed in a separate branch, which was closely clustered in the same subclade with those of RsMV10 and MpMV4 belonging to clade I in Mitovirus genus, distinct from members of the genus Narnavirus (Fig. 2 B). In conclusion, based on its genome organization, sequence alignment and phylogenetic analysis, BdMV2 is a novel mitovirus in the B. dothidea pathogen causing pear ring rot disease. Declarations Acknowledgements This research was supported by the National Natural Science Foundation of China (31972321), the Earmarked Fund for Pear Modern Agro-industry Technology Research System (CARS-29-15) and the National Key Research and Development Program of China (2017YFD0201100). Compliance with ethical standards Conflict of interest All authors declare that they have no any conflicts of interest in this manuscript. Ethical approval This article does not contain any experiments with human participants or animals performed by any of the authors. References Marsberg A, Kemler M, Jami F, Nagel JH, Postma-Smidt A, Naidoo S, Wingfield MJ, Crous PW, Spatafora JW, Hesse CN, Robbertse B, Slippers B (2017) Botryosphaeria dothidea : a latent pathogen of global importance to woody plant health. Mol Plant Pathol 18(4):477–488 Ran L, Zhang M, Shen HJPD (2018) First report of Red Rot of Jujube fruit caused by Botryosphaeria dothidea in China. Plant Dis 102(7):1458 Slippers B, Wingfield MJ (2007) Botryosphaeriaceae as endophytes and latent pathogens of woody plants: diversity, ecology and impact. Fungal Biol Rev 21:90–106 Tang W, Ding Z, Zhou ZQ, Wang YZ, Guo LY (2012) Phylogenetic and Pathogenic analyses show that the causal agent of apple ring rot in China is Botryosphaeria dothidea . Plant Dis 96(4):486–496 Yan JY, Xie Y, Zhang W, Wang Y, Liu JK, Hyde KD, Seem RC, Zhang GZ, Wang ZY, Yao SW, Bai XJ, Dissanayake AJ (2013) Species of Botryosphaeriaceae involved in grapevine dieback in China. Fungal Divers 61(1):221–236 Zhai LF, Zhang MX, Lv G, Chen XR, Jia NN, Hong N, Wang GP (2014) Biological and molecular characterization of four Botryosphaeria species isolated from pear plants showing ring rot and stem canker in China. Plant Dis 98:716–726 Al-Haq MI, Seo Y, Oshita S, Kawagoe Y (2002) Disinfection effects of electrolyzed oxidizing water on suppressing fruit rot of pear caused by Botryosphaeria berengeriana. Food Res Int 35(7):657–664 Chiba S, Salaipeth L, Sasaki A, Kanemastu S, Suzuki N (2009) A novel bipartite dsRNA mycovirus from the white root rot fungus Roselia necatrix: molecular and biological characterization, taxonomic consideration, and potential for biological control. J Virol 83(24):128801–128812 Ghabrial SA, Castón JR, Jiang D, Nibert ML, Suzuki N (2015) 50-plus years of fungal viruses. Virology 479:356–368 Lefkowitz EJ, Dempsey DM, Hendrickson RC, Orton RJ, Siddell SG, Smith DB (2018) Virus taxonomy: the database of the International Committee on Taxonomy of Viruses (ICTV). Nucleic Acids Res 46(D1): D708–D717 Marzano SYL, Nelson BD, Ajayi-Oyetunde O, Bradley CA, Hughes TJ, Hartman GL, Eastburn DM, Domier LL (2016) Identification of diverse mycoviruses through metatranscriptomics characterization of the viromes of five major fungal plant pathogens.J Virol 90(15):6846–6863 Wang LP, Jiang JJ, Wang YF, Hong N, Zhang FP, Xu WX, Wang GP (2014) Hypovirulence ofthe phytopathogenic fungus Botryosphaeria dothidea: association with a coinfecting chrysovirus and a partitivirus. J Virol88(13):7517–7527 Zhai L, Xiang J, Zhang M, Fu M, Yang Z, Hong N, Wang G (2016) Characterization of a noveldouble-stranded RNA mycovirus conferring hypovirulence fromthe phytopathogenic fungus Botryosphaeria dothidea. Virology493:75–85 Zhai L, Yang M, Zhang M, Hong N, Wang G (2019) Characterization of aBotybirnavirus conferring hypovirulence in the phytopathogenicfungus Botryosphaeria dothidea. Viruses 11(3):266 Zhai L, Hong N, Zhang M, Wang G (2015) Complete dsRNA sequenceof a novel victorivirus isolated from the pear stem wart fungusBotryosphaeria dothidea. Arch Virol 160(2):613–616 Yang M, Zhai L, Xiao F, Guo Y, Fu M, Hong N, Wang G (2019) Characterization of a novel victorivirus isolated from the phytopathogenic fungus Botryosphaeria dothidea. Arch Virol 164(6):1609–1617 Yang M, Zhou X, Zhai L, Xiao F, Hong N, Wang G (2020) Molecular characterization of a novel mycovirus infecting the phytopathogenic fungus Botryosphaeria dothidea. Arch Virol 165(7):1667–1670 Liu W, Hai D, Mu F, Yu X, Zhao Y, He B, Xie J, Jiang D, Liu H (2020) Molecular characterization of a novel fusarivirus infecting the plant pathogenic fungus Botryosphaeria dothidea. Arch Virol 165(4):1033–1037 Liu H, Wang H, Zhou Q (2021) A novel mycovirus isolated from the plant-pathogenic fungus Botryosphaeria dothidea. Arch Virol 166:1267–1272 Ding Z, Zhou T, Guo LY (2017) Characterization of a novel strainof Botryosphaeria dothidea chrysovirus 1 from the apple white rotpathogen Botryosphaeria dothidea. Arch Virol 162(7):2097–2102 Liu H, Liu M, Zhu H, Zhong J, Liao X, Zhou Q (2021) Molecular characterization of a novel mitovirus from the plant-pathogenic fungus Botryosphaeria dothidea. Arch Virol 166(2):633–637 Hillman BI, Cai G (2013) The family Narnaviridae: simplest of RNA viruses. Adv Virus Res 86:149–176 Hillman BI, Esteban R (2011) Narnaviridae. In: King AMQ, Adams MJ, Carstens EB, Lefkowitz EJ (eds) Virus taxonomy: classification and nomenclature of viruses: Ninth Report of the International Committee on Taxonomy of Viruses. Elsevier, San Diego, pp 1055–1060 Yang F, Hong N, Wang GP. Mycovirus dsRNA extraction kit and its application: ZL201310072994.3. 2015–06–03 Liu H, Fu Y, Jiang D, Li G, Xie J, Peng Y, Yi X, Ghabrial SA (2009) A novel mycovirus that is related to the human pathogen hepatitis E virus and rubi-like viruses. J. Virol 83:1981–1991. Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23(21):2947–2948 Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729 Nicholas KB (1997) GeneDoc: analysis and visualization of genetic variation. Embnew news 4:14 Supplementary Files supplememtaryfile1.doc supplemantaryfile2.doc BdMV2genomesequence.docx Cite Share Download PDF Status: Published Journal Publication published 02 Aug, 2021 Read the published version in Archives of Virology → Version 1 posted Reviewers invited by journal 20 Apr, 2021 Editorial decision: Major Revision 16 Apr, 2021 First submitted to journal 15 Apr, 2021 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-431413","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":22788099,"identity":"e4903934-0ac2-4d0f-bdd9-12f54d0972dd","order_by":0,"name":"Qi Zou","email":"","orcid":"","institution":"Huazhong Agriculture University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Qi","middleName":"","lastName":"Zou","suffix":""},{"id":22788100,"identity":"e4c29c1f-6218-47d0-a297-53cbd2e621d9","order_by":1,"name":"Yunjing Gao","email":"","orcid":"","institution":"Huazhong Agriculture University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yunjing","middleName":"","lastName":"Gao","suffix":""},{"id":22788101,"identity":"b28692e8-6aaa-4f03-b3e0-b8a931d7ebeb","order_by":2,"name":"Qiong Wang","email":"","orcid":"","institution":"Huazhong Agriculture University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Qiong","middleName":"","lastName":"Wang","suffix":""},{"id":22788102,"identity":"20efe094-4849-4838-9922-432102de93d9","order_by":3,"name":"Yuekun Yang","email":"","orcid":"","institution":"Huazhong Agriculture University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yuekun","middleName":"","lastName":"Yang","suffix":""},{"id":22788103,"identity":"b8eb2f73-a136-4ab6-b9de-8cc15b0484f1","order_by":4,"name":"Fang Wang","email":"","orcid":"","institution":"Huazhong Agriculture University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Fang","middleName":"","lastName":"Wang","suffix":""},{"id":22788104,"identity":"adfcb899-5f4b-4a7d-9ccb-34776f5f20b6","order_by":5,"name":"Ni Hong","email":"","orcid":"","institution":"Huazhong Agriculture University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ni","middleName":"","lastName":"Hong","suffix":""},{"id":22788105,"identity":"4c2d3d4c-b2be-46da-8bb4-acd8973d9a36","order_by":6,"name":"Guoping Wang","email":"","orcid":"","institution":"Huazhong Agriculture University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Guoping","middleName":"","lastName":"Wang","suffix":""},{"id":22788106,"identity":"8c7402d8-234e-4efc-8977-55d69c61996f","order_by":7,"name":"Liping Wang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA8ElEQVRIiWNgGAWjYDACdgaGAx8qGBjY2BnYoEIJBLQwMzA+nHEGqIWZBC3MxrxtYAaRWgwO8x6T5p23TZ4PqOXhj5rDDPzsOQYMP3fg08KXJjl3223DNmYGdgOJY4cZJHveGDD2nsGtxewwj5nE2223GYFa2CQM2A4zGNzIMWBmbCOghXfObXuwloR/hxnsidBibMjbcDsRrOVgG9AWCQJa7A/zJT6ccex2chtQmWRjXzqPxJlnBQd78WiRbO89cOBDzW3b+e3NxyR/fLOW429P3vjgJx4tDAw8MAZjA5x7AJ8GJC2jYBSMglEwCnAAAA/9Sv8xM7XzAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-9095-8682","institution":"Huazhong Agriculture University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Liping","middleName":"","lastName":"Wang","suffix":""}],"badges":[],"createdAt":"2021-04-17 04:52:54","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-431413/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-431413/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00705-021-05189-6","type":"published","date":"2021-08-02T15:10:25+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":8449767,"identity":"97aedf4d-429a-49d6-918f-e98393d18ab5","added_by":"auto","created_at":"2021-04-26 14:39:18","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":166709,"visible":true,"origin":"","legend":"(A) Colony morphology of B. dothidea strain FJ in PDA medium at 25 °C for 5d. (B) Schematic representation of the BdMV2 genome organization. The single ORF and UTRs are indicated by an open bar and black lines, respectively. The blue bar represented the RdRp conserved domain. (C) The putative secondary structures of the 5’ and 3’ termini and panhandle structure of BdMV2 UTRs.","description":"","filename":"Fig1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-431413/v1/b9d360f1fdf214c4edeb5fe9.jpg"},{"id":8449928,"identity":"a9db717d-dc59-4b5d-93a6-0afc865b5122","added_by":"auto","created_at":"2021-04-26 14:42:18","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":434905,"visible":true,"origin":"","legend":"(A) Sequence alignment of BdMV2 RdRp motifs with those of selected mycoviruses in the genus Mitovirus. Horizontal black lines above the alignment sequences indicate the conserved six motifs from I to VI. Shaded areas represented identical aa residues. (B) Phylogenetic tree analysis of BdMV2 in this study and selected mitoviuses and narnaviruses members specified as viral names followed by GenBank accession numbers were shown, based on the RdRp aa sequences using the neighbor-joining (NJ) method with 1000 bootstrap replicates. Bootstrap values with \u003e50% are shown at the branches. The scale bar indicates a genetic distance of 0.2 aa substitutions per site. A red star indicates the newly mycovirus of BdMV2. ","description":"","filename":"Fig2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-431413/v1/699ebf4b969d3666913f78e2.jpg"},{"id":15672305,"identity":"56f3220b-88dc-4727-971a-6ba2f5640c0a","added_by":"auto","created_at":"2021-11-18 14:11:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":405404,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-431413/v1/fd32732d-62da-4d7e-be2f-c6dbb8131389.pdf"},{"id":8450227,"identity":"b4c21371-910b-44eb-a32e-4cb0657e3472","added_by":"auto","created_at":"2021-04-26 14:45:18","extension":"doc","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":37888,"visible":true,"origin":"","legend":"","description":"","filename":"supplememtaryfile1.doc","url":"https://assets-eu.researchsquare.com/files/rs-431413/v1/989ccf808021b6ee2b330fe4.doc"},{"id":8449931,"identity":"e49dea63-a84d-4294-8a3f-03042fd9383f","added_by":"auto","created_at":"2021-04-26 14:42:18","extension":"doc","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":38400,"visible":true,"origin":"","legend":"","description":"","filename":"supplemantaryfile2.doc","url":"https://assets-eu.researchsquare.com/files/rs-431413/v1/20ff22ca3153626701b8b60c.doc"},{"id":8449929,"identity":"2f461de3-e603-4d48-bbc9-7d3dbe5b12da","added_by":"auto","created_at":"2021-04-26 14:42:18","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":14222,"visible":true,"origin":"","legend":"","description":"","filename":"BdMV2genomesequence.docx","url":"https://assets-eu.researchsquare.com/files/rs-431413/v1/1dca735ec40a75184321b7d7.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003eThe Full-length Genome Sequence of a Novel Mitovirus From the Causal Agent \u003cem\u003eBotrosphaeria Dothidea\u003c/em\u003e of Pear Ring Rot Disease\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003e\u003cem\u003eBotryosphaeria dothidea\u003c/em\u003e is the important and widespread trunk pathogen, which causes pear ring rot disease in the pear production regions in China. \u003cem\u003eBotryosphaeria dothidea\u003c/em\u003e with an extensive host range not only harms pear trees by causing severe symptoms such as stem wart and canker, branch dieback and fruit rot, but also other fruit trees including apple, Jujube and grapevine [1\u0026ndash;6]. The ring rot disease is becoming increasingly serious and distributes throughout all pear producing areas at present, which seriously affects the yield and quality of pear to further hind the healthy development of pear industry [6\u0026ndash;7]. It is urgent to find safe and effective measure to biological control of pear ring rot diseases. As mycoviruses can infect and proliferate in fungi and some mycoviruses present hypovirulence-associated traits, they are potential biological control agents for the control of these fungal disease. Indeed, many mycoviruses with potential biocontrol value have been found and identified from pathogenic fungi infecting fruit trees [8]. Therefore, it is particularly important to find and identify mycoviruses in \u003cem\u003eBotryosphaeria dothidea\u003c/em\u003e strains casing pear ring rot disease. To identify and characterize mycovirus, high through-put sequencing techniques and subsequent bioinformatics have been shown as a very effective approach [9\u0026ndash;11]. So far, eleven mycoviruses have been identified in \u003cem\u003eB. dothidea \u003c/em\u003estrains and characterized. In our lab, Botryosphaeria dothidea chrysovirus 1 (BdCV1), Bipolaris maydis botybirnavirus 1 strain BdEW220 (BmRV1-BdEW220), Botryosphaeria\u0026nbsp;dothidea\u0026nbsp;RNA virus 1 (BdRV1) involved in hypovirulence [12\u0026ndash;14], Botryosphaeria dothidea partitivirus 1 (BdPV1), Botryosphaeria dothidea victorivirus 1 (BdVV1), Botryosphaeria dothidea victorivirus 2 (BdVV2) and Botryosphaeria\u0026nbsp;dothidea\u0026nbsp;botourmiavirus 1 (BdBOV-1) with no obvious effect on host, have been identified in\u0026nbsp;\u003cem\u003eB. dothidea\u003c/em\u003e\u0026nbsp;strains causing pear ring rot disease [12, 15\u0026ndash;17]. Botryosphaeria\u0026nbsp;dothidea\u0026nbsp;bipartite\u0026nbsp;mycovirus\u0026nbsp;1 (BdBMV1), Botryosphaeria\u0026nbsp;dothidea\u0026nbsp;mitovirus 1 (BdMV1), Botryosphaeria dothidea fusarivirus 1 (BdFV1) and BdCV1-G1 have been also identified in\u0026nbsp;\u003cem\u003eB. dothidea\u003c/em\u003e\u0026nbsp;strains isolated from \u003cem\u003eKerria japonica\u003c/em\u003e, pepper and apple hosts, respectively [18\u0026ndash;21]. The\u003cem\u003e Narnaviridae\u003c/em\u003e genome has positive single strand RNA (+ssRNA) with approximately 2.2\u0026ndash;3.6 kb in length and encodes only one RNA-dependent RNA polymerase (RdRp). The \u003cem\u003eNarnaviridae\u003c/em\u003e family is divided into two genera, \u003cem\u003eNarnavirus\u003c/em\u003e and \u003cem\u003eMitovirus\u003c/em\u003e (Mitochondrial viruses), which replicate in the cytoplasm and mitochondria, respectively. The mycoviruses in \u003cem\u003eNarnaviridae\u003c/em\u003e exist widely in fungi, but it is not clear if they have any effect on all kinds of fungi [22\u0026ndash;23]. Here, we report the identification of a novel mitovirus isolated from \u003cem\u003eB. dothidea\u003c/em\u003e strain, the causal pathogen of pear ring rot disease. We further determined its +ssRNA full-length genome sequence and molecular characteristics. This mycovirus was tentatively designated as Botryosphaeria dothidea mitovirus 2 (BdMV2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProvenance and sequencing of BdMV2\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eB. dothidea\u003c/em\u003e strain FJ was isolated from pear stem bark exhibiting wart symptoms in Fujian Province of China, in 2010, and the hypha blocks of FJ strain were stored in 25% glycerin solution at -80℃. The mycelium was grown on potato dextrose agar (PDA) plates overlaid with a layer of cellophane at 25 ℃ in the dark. The mycelium of \u003cem\u003eB. dothidea\u003c/em\u003e strain FJ and other \u003cem\u003eValsa\u003c/em\u003e spp strains were together collected to be used for Illumina sequencing (Beckman company, Beijing, China). The obtained high-quality clean reads were assembled contigs, which were analyzed to search mycoviruses using Blastx. Among these contigs, Contig1603 with 2.5 kb in length shares the highest sequence similarity with 67.14% in comparison to that of Macrophomina phaseolina mitovirus 4. Total RNA was extracted by Trizol method (Aidlab Biotechnologies Co., Ltd, Beijing, China) as template for synthesize cDNA. Finally, the putative mycoviral genome intermediate sequence was determined by RT-PCR and obtained 2042 nt by sequencing in \u003cem\u003eB. dothidea\u003c/em\u003e strain FJ using the designed primers based on Contig1603 sequence (Supplementary Table\u0026nbsp;1). The dsRNA was extracted from the cultured hypha by the column separation method [\u003cspan class=\"CitationRef\"\u003e24\u003c/span\u003e]. The adaptor RACE-OLIGO was ligated to the 3\u0026rsquo; terminus of each strand of dsRNA, which were used for template to reversely transcribed cDNA using Oligo REV primer as described previously [\u003cspan class=\"CitationRef\"\u003e25\u003c/span\u003e]. The terminal sequence amplification was performed by nested PCR using the primers designed based on obtained viral sequence and sequence-specific adaptor primers of O5RACE-2 and O5RACE-3, respectively (Supplementary Table\u0026nbsp;1). The obtained PCR product was purified, linked to pMD18-T vector (Takara, Dalian, China), and transformed into \u003cem\u003eEscherichia coli\u003c/em\u003e Top10. For each PCR product, at least three independent identified positive clones were sequenced in both directions (Sangon Biotech Co., Ltd, Shanghai, China). Sequences obtained from clones were assembled using DNAMAN. The viral ORF was predicted using the ORF finder online website of \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/orffinder\u003c/span\u003e\u003c/span\u003e. Mycoviral sequences search was performed using NCBI database. Multiple mycoviral sequences were aligned using CLUSTAL X [\u003cspan class=\"CitationRef\"\u003e26\u003c/span\u003e]. Phylogenetic analysis was conducted by using NJ method with 1000 replicates of MEGA 7.0, and RdRp domain motif sequences were analyzed using the GeneDoc software [\u003cspan class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e28\u003c/span\u003e]. The secondary structures of BdMV2 genome terminus were predicated by online RNAfold software.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSequence properties\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe colony morphology of \u003cem\u003eB. dothidea\u003c/em\u003e strain FJ was shown, which exhibited normal mycelia and the mycelia could grow to cover the whole plate in 5 days (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eA). RT-PCR and RNA-ligase-mediated rapid amplification of cDNA ends (RLM-RACE) in combination with cloning and sequence assembly were used to obtain the BdMV2 full-length genome sequence from \u003cem\u003eB. dothidea\u003c/em\u003e strain FJ. Its complete genome sequence was deposited in the NCBI GenBank database (Acc. No. MW553911), which was 2538 nt in length with a CG content of 47.24% for the genome. BdMV2 contains a single big ORF with AUG as the initiation condon at positions 176\u0026ndash;178 nt and UAA as the termination condon at positions 2243\u0026ndash;2245 nt of the genome by fungal mitochondrial codon usage. The ORF (2,070 nt in length) potentially encoded 689 aa RdRp protein with predicated molecular weight of 77.35 kDa. The 5\u0026rsquo; and 3\u0026rsquo; untranslated regions (UTRs) are 175 and 293 nt long, respectively (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB). Using the RNAfold program, two potential stem-loop structures in the 5\u0026rsquo; and 3\u0026rsquo; UTRs and a potential panhandle structure of the BdMV2 positive-sense strand UTRs were predicted, respectively (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eC), which showed typical characteristic of reported members of the genus \u003cem\u003eMitovirus\u003c/em\u003e [\u003cspan class=\"CitationRef\"\u003e22\u003c/span\u003e]. The BdMV2 contains a conserved RdRp domain at positions 164\u0026ndash;495 aa of the genome (Mitovirus RNA-dependent RNA polymeras; pfam05919) by the Conserved Domain Database (CDD) search of NCBI. Multiple sequence alignment analysis of RdRp domain from BdMV2 with those members in \u003cem\u003eMitovirus\u003c/em\u003e showed that BdMV2 RdRp domain contains all six conserved motifs (I to VI), which are characteristics of members in the \u003cem\u003eMitovirus\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eA). A BLASTp comparison of the RdRp sequence of BdMV2 showed the highest similarity of 69.36% with that of RsMV10, and had identity 28.91\u0026ndash;68.79% (query coverage more than 90%) to those of members in the genus \u003cem\u003eMitovirus\u003c/em\u003e (Supplementary Table\u0026nbsp;2). To determine the taxonomic status of BdMV2, phylogenetic analysis was performed using the RdRp aa sequences encoded by BdMV2 and other selected mycoviruses in \u003cem\u003eMitovirus\u003c/em\u003e and \u003cem\u003eNarnavirus\u003c/em\u003e of the family \u003cem\u003eNarnaviridae\u003c/em\u003e. The results showed that all mitoviruses were classified into two clades of clade I and clade II. It revealed that BdMV2 existed in a separate branch, which was closely clustered in the same subclade with those of RsMV10 and MpMV4 belonging to clade I in \u003cem\u003eMitovirus\u003c/em\u003e genus, distinct from members of the genus \u003cem\u003eNarnavirus\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eB). In conclusion, based on its genome organization, sequence alignment and phylogenetic analysis, BdMV2 is a novel mitovirus in the \u003cem\u003eB. dothidea\u003c/em\u003e pathogen causing pear ring rot disease.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by the National Natural Science Foundation of China (31972321), the Earmarked Fund for Pear Modern Agro-industry Technology Research System (CARS-29-15) and the National Key Research and Development Program of China (2017YFD0201100).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompliance with ethical standards\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors declare that they have no any conflicts of interest in this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis article does not contain any experiments with human participants or animals performed by any of the authors.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eMarsberg A, Kemler M, Jami F, Nagel JH, Postma-Smidt A, Naidoo S, Wingfield MJ, Crous PW, Spatafora JW, Hesse CN, Robbertse B, Slippers B (2017) \u003cem\u003eBotryosphaeria dothidea\u003c/em\u003e: a latent pathogen of global importance to woody plant health. Mol Plant Pathol 18(4):477\u0026ndash;488\u003c/li\u003e\n\u003cli\u003eRan L, Zhang M, Shen HJPD (2018) First report of Red Rot of Jujube fruit caused by Botryosphaeria dothidea in China. Plant Dis 102(7):1458\u003c/li\u003e\n\u003cli\u003eSlippers B, Wingfield MJ (2007) Botryosphaeriaceae as endophytes and latent pathogens of woody plants: diversity, ecology and impact. Fungal Biol Rev 21:90\u0026ndash;106\u003c/li\u003e\n\u003cli\u003eTang W, Ding Z, Zhou ZQ, Wang YZ, Guo LY (2012) Phylogenetic and Pathogenic analyses show that the causal agent of apple ring rot in China is \u003cem\u003eBotryosphaeria dothidea\u003c/em\u003e. Plant Dis 96(4):486\u0026ndash;496\u003c/li\u003e\n\u003cli\u003eYan JY, Xie Y, Zhang W, Wang Y, Liu JK, Hyde KD, Seem RC, Zhang GZ, Wang ZY, Yao SW, Bai XJ, Dissanayake AJ (2013) Species of Botryosphaeriaceae involved in grapevine dieback in China. Fungal Divers 61(1):221\u0026ndash;236\u003c/li\u003e\n\u003cli\u003eZhai LF, Zhang MX, Lv G, Chen XR, Jia NN, Hong N, Wang GP (2014) Biological and molecular characterization of four \u003cem\u003eBotryosphaeria\u003c/em\u003e species isolated from pear plants showing ring rot and stem canker in China. Plant Dis 98:716\u0026ndash;726\u003c/li\u003e\n\u003cli\u003eAl-Haq MI, Seo Y, Oshita S, Kawagoe Y (2002) Disinfection effects of electrolyzed oxidizing water on suppressing fruit rot of pear caused by Botryosphaeria berengeriana. Food Res Int 35(7):657\u0026ndash;664\u003c/li\u003e\n\u003cli\u003eChiba S, Salaipeth L, Sasaki A, Kanemastu S, Suzuki N (2009) A novel bipartite dsRNA mycovirus from the white root rot fungus Roselia necatrix: molecular and biological characterization, taxonomic consideration, and potential for biological control. J Virol 83(24):128801\u0026ndash;128812\u003c/li\u003e\n\u003cli\u003eGhabrial SA, Cast\u0026oacute;n JR, Jiang D, Nibert ML, Suzuki N (2015) 50-plus years of fungal viruses. Virology 479:356\u0026ndash;368\u003c/li\u003e\n\u003cli\u003eLefkowitz EJ, Dempsey DM, Hendrickson RC, Orton RJ, Siddell SG, Smith DB (2018) Virus taxonomy: the database of the International Committee on Taxonomy of Viruses (ICTV). Nucleic Acids Res 46(D1): D708\u0026ndash;D717\u003c/li\u003e\n\u003cli\u003eMarzano SYL, Nelson BD, Ajayi-Oyetunde O, Bradley CA, Hughes TJ, Hartman GL, Eastburn DM, Domier LL (2016) Identification of diverse mycoviruses through metatranscriptomics characterization of the viromes of five major fungal plant pathogens.J Virol 90(15):6846\u0026ndash;6863\u003c/li\u003e\n\u003cli\u003eWang LP, Jiang JJ, Wang YF, Hong N, Zhang FP, Xu WX, Wang GP (2014) Hypovirulence ofthe phytopathogenic fungus Botryosphaeria dothidea: association with a coinfecting chrysovirus and a partitivirus. J Virol88(13):7517\u0026ndash;7527\u003c/li\u003e\n\u003cli\u003eZhai L, Xiang J, Zhang M, Fu M, Yang Z, Hong N, Wang G (2016) Characterization of a noveldouble-stranded RNA mycovirus conferring hypovirulence fromthe phytopathogenic fungus Botryosphaeria dothidea. Virology493:75\u0026ndash;85\u003c/li\u003e\n\u003cli\u003eZhai L, Yang M, Zhang M, Hong N, Wang G (2019) Characterization of aBotybirnavirus conferring hypovirulence in the phytopathogenicfungus Botryosphaeria dothidea. Viruses 11(3):266\u003c/li\u003e\n\u003cli\u003eZhai L, Hong N, Zhang M, Wang G (2015) Complete dsRNA sequenceof a novel victorivirus isolated from the pear stem wart fungusBotryosphaeria dothidea. Arch Virol 160(2):613\u0026ndash;616\u003c/li\u003e\n\u003cli\u003eYang M, Zhai L, Xiao F, Guo Y, Fu M, Hong N, Wang G (2019) Characterization of a novel victorivirus isolated from the phytopathogenic fungus Botryosphaeria dothidea. Arch Virol 164(6):1609\u0026ndash;1617\u003c/li\u003e\n\u003cli\u003eYang M, Zhou X, Zhai L, Xiao F, Hong N, Wang G (2020) Molecular characterization of a novel mycovirus infecting the phytopathogenic fungus Botryosphaeria dothidea. Arch Virol 165(7):1667\u0026ndash;1670\u003c/li\u003e\n\u003cli\u003eLiu W, Hai D, Mu F, Yu X, Zhao Y, He B, Xie J, Jiang D, Liu H (2020) Molecular characterization of a novel fusarivirus infecting the plant pathogenic fungus Botryosphaeria dothidea. Arch Virol 165(4):1033\u0026ndash;1037\u003c/li\u003e\n\u003cli\u003eLiu H, Wang H, Zhou Q (2021) A novel mycovirus isolated from the plant-pathogenic fungus Botryosphaeria dothidea. Arch Virol 166:1267\u0026ndash;1272\u003c/li\u003e\n\u003cli\u003eDing Z, Zhou T, Guo LY (2017) Characterization of a novel strainof Botryosphaeria dothidea chrysovirus 1 from the apple white rotpathogen Botryosphaeria dothidea. Arch Virol 162(7):2097\u0026ndash;2102\u003c/li\u003e\n\u003cli\u003eLiu H, Liu M, Zhu H, Zhong J, Liao X, Zhou Q (2021) Molecular characterization of a novel mitovirus from the plant-pathogenic fungus Botryosphaeria dothidea. Arch Virol 166(2):633\u0026ndash;637\u003c/li\u003e\n\u003cli\u003eHillman BI, Cai G (2013) The family Narnaviridae: simplest of RNA viruses. Adv Virus Res 86:149\u0026ndash;176\u003c/li\u003e\n\u003cli\u003eHillman BI, Esteban R (2011) Narnaviridae. In: King AMQ, Adams MJ, Carstens EB, Lefkowitz EJ (eds) Virus taxonomy: classification and nomenclature of viruses: Ninth Report of the International Committee on Taxonomy of Viruses. Elsevier, San Diego, pp 1055\u0026ndash;1060\u003c/li\u003e\n\u003cli\u003eYang F, Hong N, Wang GP. Mycovirus dsRNA extraction kit and its application: ZL201310072994.3. 2015\u0026ndash;06\u0026ndash;03\u003c/li\u003e\n\u003cli\u003eLiu H, Fu Y, Jiang D, Li G, Xie J, Peng Y, Yi X, Ghabrial SA (2009) A novel mycovirus that is related to the human pathogen hepatitis E virus and rubi-like viruses. J. Virol 83:1981\u0026ndash;1991.\u003c/li\u003e\n\u003cli\u003eLarkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23(21):2947\u0026ndash;2948\u003c/li\u003e\n\u003cli\u003eTamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725\u0026ndash;2729\u003c/li\u003e\n\u003cli\u003eNicholas KB (1997) GeneDoc: analysis and visualization of genetic variation. Embnew news 4:14\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"archives-of-virology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"arvi","sideBox":"Learn more about [Archives of Virology](https://www.springer.com/journal/705)","snPcode":"705","submissionUrl":"https://submission.nature.com/new-submission/705/3","title":"Archives of Virology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Botryosphaeria dothidea mitovirus 2, Botryosphaeria dothidea, rot disease, genome sequence, novel mitovirus","lastPublishedDoi":"10.21203/rs.3.rs-431413/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-431413/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHere, we describe a novel mycovirus, Botryosphaeria dothidea mitovirus 2 (tentatively designated as BdMV2), isolated from \u003cem\u003eBotryosphaeria dothidea \u003c/em\u003eFJ strain causing pear ring rot disease in Fujian Province, China. The complete genome nucleotide sequence of BdMV2 is 2538 nt in length and contains a single 2070 nt open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) of 689 amino acid (aa) using fungal mitochondrial genetic code. BLASTp analysis revealed that the RdRp of BdMV2 shares 28.91%–69.36% (query sequence coverage more than 90%) sequence identity to those of members in the genus \u003cem\u003eMitovirus\u003c/em\u003e, and the closest similarity is 69.36% and 68.79% with the corresponding protein aa sequences of Rhizoctonia solani mitovirus 10 and Macrophomina phaseolina mitovirus 4, respectively. Phylogenetic analysis based on the RdRp aa sequences further revealed that BdMV2 is a newly member in the genus \u003cem\u003eMitovirus\u003c/em\u003e of the\u003cem\u003e \u003c/em\u003efamily \u003cem\u003eNarnaviridae\u003c/em\u003e. To our knowledge, BdMV2 is thus a novel mitovirus from the causal agent\u003cem\u003e B. dothidea\u003c/em\u003e of pear ring rot disease.\u003c/p\u003e","manuscriptTitle":"The Full-length Genome Sequence of a Novel Mitovirus From the Causal Agent Botrosphaeria Dothidea of Pear Ring Rot Disease","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-04-26 14:39:16","doi":"10.21203/rs.3.rs-431413/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewersInvited","content":"","date":"2021-04-21T00:00:00+00:00","index":"","fulltext":""},{"type":"decision","content":"Major Revision","date":"2021-04-16T04:12:21+00:00","index":"","fulltext":""},{"type":"submitted","content":"Archives of Virology","date":"2021-04-15T19:53:56+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"archives-of-virology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"arvi","sideBox":"Learn more about [Archives of Virology](https://www.springer.com/journal/705)","snPcode":"705","submissionUrl":"https://submission.nature.com/new-submission/705/3","title":"Archives of Virology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"bfa44094-2b0d-46a4-bb25-aa9075ced344","owner":[],"postedDate":"April 26th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":3887016,"name":"Virology"}],"tags":[],"updatedAt":"2021-08-22T15:30:01+00:00","versionOfRecord":{"articleIdentity":"rs-431413","link":"https://doi.org/10.1007/s00705-021-05189-6","journal":{"identity":"archives-of-virology","isVorOnly":false,"title":"Archives of Virology"},"publishedOn":"2021-08-02 15:10:25","publishedOnDateReadable":"August 2nd, 2021"},"versionCreatedAt":"2021-04-26 14:39:16","video":"","vorDoi":"10.1007/s00705-021-05189-6","vorDoiUrl":"https://doi.org/10.1007/s00705-021-05189-6","workflowStages":[]},"version":"v1","identity":"rs-431413","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-431413","identity":"rs-431413","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}