Mycovirus Hunting Revealed the Presence of Diverse Viruses in a Single Isolate of the Phytopathogenic Fungus Diplodia seriata From Pakistan

• Published in: Frontiers in cellular and infection microbiology Vol. 12; p. 913619
• Main Authors: Khan, Haris Ahmed, Telengech, Paul, Kondo, Hideki, Bhatti, Muhammad Faraz, Suzuki, Nobuhiro
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 29.06.2022
• Subjects: Ascomycota - virology; Botryosphaeriaceae; Cellular and Infection Microbiology; Diplodia seriata; Fungal Viruses - classification; Fungal Viruses - isolation & purification; Genome, Viral; mycovirome; next-generation sequencing; Pakistan; Phylogeny; phytopathogenic fungi; RNA Viruses - classification; RNA Viruses - isolation & purification; RNA, Double-Stranded - genetics; RNA, Viral - genetics; ssRNA virus; Pakistan; ssRNA virus; next-generation sequencing; Botryosphaeriaceae; virus/virus interaction; dsRNA virus; Diplodia seriata; mycovirome; phytopathogenic fungi
• ISSN: 2235-2988; 2235-2988
• DOI: 10.3389/fcimb.2022.913619

Diplodia seriata in the family Botryosphaeriaceae is a cosmopolitan phytopathogenic fungus and is responsible for causing cankers, fruit rot and leaf spots on economically important plants. In this study, we characterized the virome of a single Pakistani strain (L3) of D. seriata . Several viral-like contig sequences were obtained via a previously conducted next-generation sequencing analysis. Multiple infection of the L3 strain by eight RNA mycoviruses was confirmed through RT-PCR using total RNA samples extracted from this strain; the entire genomes were determined via Sanger sequencing of RT-PCR and RACE clones. A BLAST search and phylogenetic analyses indicated that these eight mycoviruses belong to seven different viral families. Four identified mycoviruses belong to double-stranded RNA viral families, including Polymycoviridae , Chrysoviridae , Totiviridae and Partitiviridae , and the remaining four identified mycoviruses belong to single-stranded RNA viral families, i.e., Botourmiaviridae , and two previously proposed families “ Ambiguiviridae ” and “ Splipalmiviridae ”. Of the eight, five mycoviruses appear to represent new virus species. A morphological comparison of L3 and partially cured strain L3ht1 suggested that one or more of the three viruses belonging to Polymycoviridae , “ Splipalmiviridae ” and “ Ambiguiviridae ” are involved in the irregular colony phenotype of L3. To our knowledge, this is the first report of diverse virome characterization from D. seriata .