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Mycovirus\u2013induced hypovirulence provides a promising strategy for sustainable disease management. In this study, V. dahliae isolates from Xinjiang were screened by virome sequencing and RT\u2013PCR, and strain 121\u201311C\u20131 co\u2013infected with Verticillium dahliae partitivirus 1 (VdPV1) and Verticillium dahliae magoulivirus 1 (VdMoV1) was identified. Virus\u2013free isogenic strains were obtained via single\u2013conidium purification, verifying that both viruses can be vertically transmitted through fungal conidia. Dual\u2013culture assays uncovered a novel, previously unreported horizontal transmission dependency between the two viruses: VdPV1 achieves horizontal transmission only with the assistance of VdMoV1, whereas VdMoV1 can transmit independently or co\u2013transmit with VdPV1. Biological and pathogenicity assays demonstrated that both viruses significantly inhibit mycelial growth, reduce conidial production and attenuate fungal virulence. Notably, dual infection induced a significantly stronger hypovirulent phenotype than single infection. These findings elucidate the transmission dynamics of VdPV1 and VdMoV1, expand the repertoire of hypovirulent mycoviruses in V. dahliae, and provide evidence that viral co\u2013infection enhances fungal attenuation, offering potential applications for biological control of cotton Verticillium wilt.<\/jats:p>","DOI":"10.3390\/v18070795","type":"journal-article","created":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T10:44:02Z","timestamp":1784544242000},"page":"795","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Co\u2013Infecting Mycoviruses VdPV1 and VdMoV1 Attenuate Verticillium dahliae and Are Transmitted Vertically and Horizontally"],"prefix":"10.3390","volume":"18","author":[{"given":"Yifan","family":"Wang","sequence":"first","affiliation":[{"name":"Key Laboratory of Oasis Agricultural Pest Management and Plant Protection Resources Utilization, College of Agriculture, Shihezi University, Shihezi 832003, China"},{"name":"College of Agriculture, Shihezi University, Shihezi 832003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guolong","family":"Gao","sequence":"additional","affiliation":[{"name":"College of Agriculture, Shihezi University, Shihezi 832003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7180-1791","authenticated-orcid":false,"given":"Jiafeng","family":"Huang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Oasis Agricultural Pest Management and Plant Protection Resources Utilization, College of Agriculture, Shihezi University, Shihezi 832003, China"},{"name":"College of Agriculture, Shihezi University, Shihezi 832003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shicheng","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Agriculture, Shihezi University, Shihezi 832003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongyu","family":"Ji","sequence":"additional","affiliation":[{"name":"College of Agriculture, Shihezi University, Shihezi 832003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukun","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Agriculture, Shihezi University, Shihezi 832003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yejuan","family":"Du","sequence":"additional","affiliation":[{"name":"Key Laboratory of Oasis Agricultural Pest Management and Plant Protection Resources Utilization, College of Agriculture, Shihezi University, Shihezi 832003, China"},{"name":"College of Agriculture, Shihezi University, Shihezi 832003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4077","DOI":"10.1007\/s10570-025-06524-9","article-title":"Sustainable strategies for the valorization of cotton and cotton blended waste: Pathways to circular economy","volume":"32","author":"Grover","year":"2025","journal-title":"Cellulose"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"103262","DOI":"10.1016\/j.jrurstud.2024.103262","article-title":"The global cotton trade network reveals a shift in the cotton import center to the global south from 1986 to 2020","volume":"108","author":"Han","year":"2024","journal-title":"J. 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