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We previously found that Sclerotinia sclerotiorum endornavirus 3 (SsEV3), belonging to the genus\n              <jats:italic>Betaendornavirus<\/jats:italic>\n              within the family\n              <jats:italic>Endornaviridae<\/jats:italic>\n              , confers hypovirulence on the phytopathogenic fungus\n              <jats:italic>Sclerotinia sclerotiorum<\/jats:italic>\n              , but the underlying mechanisms remains unclear. In this study, we found that the SsEV3-infected strain produced fewer sclerotia, failed to form infection cushions on plant hosts, exhibited increased cell vacuolation, and was more sensitive to abiotic stresses. SsEV3 infection evoked transcriptional rewiring in\n              <jats:italic>S. sclerotiorum<\/jats:italic>\n              , affecting genes related to virulence factors for pathogenicity and RNAi pathway for antiviruses. An unknown biological function of gene\n              <jats:italic>Sssnf1<\/jats:italic>\n              was downregulated following SsEV3 infection. Deletion of\n              <jats:italic>Sssnf1<\/jats:italic>\n              impaired infection cushion formation and decreased virulence of\n              <jats:italic>S. sclerotiorum<\/jats:italic>\n              . Five key RNAi-related genes were significantly upregulated, and deletion of\n              <jats:italic>Ssdcl2<\/jats:italic>\n              contributed to SsEV3 accumulation. Additionally, we identified a hypothetical protein encoded by\n              <jats:italic>Sshp1<\/jats:italic>\n              that directly interacts with the RNA-dependent RNA polymerase (RdRp) domain encoded by SsEV3. Although the deletion mutants of\n              <jats:italic>Sshp1<\/jats:italic>\n              exhibited normal colony morphology, they showed higher SsEV3 accumulation and reduced resistance to reactive oxygen species, indicating that this gene\n              <jats:italic>,<\/jats:italic>\n              similar to RNAi-related genes, plays an antiviral role in response to SsEV3 infection and may represent a new antivirus factor. Therefore, examination of the interaction between endornavirus and\n              <jats:italic>S. sclerotiorum<\/jats:italic>\n              provides new insights into the mechanisms of antivirus and virulence in phytopathogenic fungi.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                Hypovirulence-associated mycoviruses have emerged as promising biocontrol agents, and studying their interactions with phytopathogenic fungi helps uncover mechanisms of fungal pathogenesis and antiviral defense. This study provides critical insights into the interaction between\n                <jats:italic>Sclerotinia sclerotiorum<\/jats:italic>\n                and its hypovirulence-associated endornavirus, SsEV3, elucidating the molecular mechanisms underlying mycovirus-induced changes in fungal virulence and antivirus defense. SsEV3 infection not only impairs fungal virulence traits, including infection cushion formation and sclerotial production but also triggers host antiviral responses involving typical RNA interference pathways. New virulence factors, such as\n                <jats:italic>Sssnf1<\/jats:italic>\n                , and antiviral factors, such as\n                <jats:italic>Sshp1<\/jats:italic>\n                , were identified based on the established interaction system between\n                <jats:italic>S. sclerotiorum<\/jats:italic>\n                and endornavirus. These findings deepen our understanding of fungus-mycovirus interactions, highlighting the role of SsEV3 in reducing the virulence of\n                <jats:italic>S. sclerotiorum<\/jats:italic>\n                , and facilitating the development of mycovirus-based biological control strategies.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/mbio.03365-24","type":"journal-article","created":{"date-parts":[[2025,2,19]],"date-time":"2025-02-19T14:00:11Z","timestamp":1739973611000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":6,"title":["Exploring the interaction between endornavirus and\n            <i>Sclerotinia sclerotiorum<\/i>\n            : mechanisms of phytopathogenic fungal virulence and antivirus"],"prefix":"10.1128","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-8760-8181","authenticated-orcid":false,"given":"Fan","family":"Mu","sequence":"first","affiliation":[{"name":"College of Plant Protection, Shanxi Key Laboratory of Integrated Pest Managementin Agriculture, Shanxi Agricultural University","place":["Jinzhong, China"]},{"name":"National Key Laboratory of Agricultural Microbiology, The Provincial Key Lab of Plant Pathology of Hubei Province, College of Plant Science and Technology, Huazhong Agricultural University","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinsheng","family":"Xia","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Shanxi Key Laboratory of Integrated Pest Managementin Agriculture, Shanxi Agricultural University","place":["Jinzhong, 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