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Here, a novel circular ssDNA virus, Diaporthe pseudophoenicicola DNA virus 1 (DpDV1), was identified from the phytopathogenic fungus Diaporthe pseudophoenicicola. DpDV1 attenuates the pathogenicity of both its original and experimental host fungi, demonstrating its potential as a biocontrol agent against fungal phytopathogens. Remarkably, DpDV1 displays bidirectional plant\u2013fungal transmission capability without inducing observable phenotypic changes in plant hosts, overcoming the limitations imposed by vegetative incompatibility in conventional hypovirulence-based biocontrol approaches. Immunocytochemical localization assays revealed that DpDV1 is localized in the chloroplasts of plant cells, demonstrating its cross-kingdom transmission. Furthermore, we demonstrated that DpDV1 can cross-protect plants against plant viruses, establishing foundations for cross-protection-based management of plant virus diseases. Hence, we report a novel ssDNA mycovirus and provide evidence of mycovirus cross-kingdom transmission between plant and fungi, providing innovative strategies for mycovirus biocontrol of plant fungal and viral diseases.<\/jats:p>","DOI":"10.1093\/plphys\/kiag464","type":"journal-article","created":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T22:23:24Z","timestamp":1782858204000},"source":"Crossref","is-referenced-by-count":0,"title":["Natural cross-kingdom transmission of a novel ssDNA mycovirus confers broad-spectrum resistance to plant diseases"],"prefix":"10.1093","volume":"201","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1061-0674","authenticated-orcid":false,"given":"Siyu","family":"Zhou","sequence":"first","affiliation":[{"name":"National Key Laboratory for Tropical Crop Breeding, School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University , Sanya, Hainan 572025 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinhui","family":"Sun","sequence":"additional","affiliation":[{"name":"National Key Laboratory for Tropical Crop Breeding, School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University , Sanya, Hainan 572025 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1906-8393","authenticated-orcid":false,"given":"Peng","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Ministry of Education for Ecology of Tropical Islands, Key Laboratory of Tropical Animal and Plant Ecology of Hainan Province, Hainan Normal University , Haikou 571158 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-9499-4270","authenticated-orcid":false,"given":"Xuetuan","family":"Lin","sequence":"additional","affiliation":[{"name":"National 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