High-throughput virome analyses with various fungi, from cultured or uncultured sources, have led to the discovery of diverse viruses with unique genome structures and even neo-lifestyles. Examples in the former category include splipalmiviruses and ambiviruses. Splipalmiviruses, related to yeast narnaviruses, have multiple positive-sense (+) single-stranded (ss) RNA genomic segments that separately encode the RNA-dependent RNA polymerase motifs, the hallmark of RNA viruses (members of the kingdom Orthornavirae). Ambiviruses appear to have an undivided ssRNA genome of 3∼5 kb with two large open reading frames (ORFs) separated by intergenic regions. Another narna-like virus group has two fully overlapping ORFs on both strands of a genomic segment that span more than 90% of the genome size. New virus lifestyles exhibited by mycoviruses include the yado-kari/yado-nushi nature characterized by the partnership between the (+)ssRNA yadokarivirus and an unrelated dsRNA virus (donor of the capsid for the former) and the hadaka nature of capsidless 10-11 segmented (+)ssRNA accessible by RNase in infected mycelial homogenates. Furthermore, dsRNA polymycoviruses with phylogenetic affinity to (+)ssRNA animal caliciviruses have been shown to be infectious as dsRNA-protein complexes or deproteinized naked dsRNA. Many previous phylogenetic gaps have been filled by recently discovered fungal and other viruses, which have provided interesting evolutionary insights. Phylogenetic analyses and the discovery of natural and experimental cross-kingdom infections suggest that horizontal virus transfer may have occurred and continue to occur between fungi and other kingdoms. Expected final online publication date for the Annual Review of Phytopathology, Volume 60 is August 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Mycovirus Diversity and Evolution Revealed/Inferred from Recent Studies.

Species of Armillaria are distributed globally and include some of the most important pathogens of forest and ornamental trees. Some of them form large long-living clones that are considered as one of the largest organisms on earth and are capable of long-range spore-mediated transfer as well as vegetative spread by drought-resistant hyphal cords called rhizomorphs. However, the virus community infecting these species has remained unknown. In this study we used dsRNA screening and high-throughput sequencing to search for possible virus infections in a collection of Armillaria isolates representing three different species: Armillaria mellea from South Africa, A. borealis from Finland and Russia (Siberia) and A. cepistipes from Finland. Our analysis revealed the presence of both negative-sense RNA viruses and positive-sense RNA viruses, while no dsRNA viruses were detected. The viruses included putative new members of virus families Mymonaviridae, Botourmiaviridae and Virgaviridae and members of a recently discovered virus group tentatively named "ambiviruses" with ambisense bicistronic genomic organization. We demonstrated that Armillaria isolates can be cured of viruses by thermal treatment, which enables the examination of virus effects on host growth and phenotype using isogenic virus-infected and virus-free strains.

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Armillaria root rot fungi host single-stranded RNA viruses.

Here, a novel mycovirus, Botryosphaeria dothidea mitovirus 1 (BdMV1), was isolated from a phytopathogenic fungus, Botryosphaeria dothidea, and its molecular characteristics were determined. BdMV1 has a genome of 2,667 nt that contains a single large open reading frame (ORF) using the fungal mitochondrial genetic code. The ORF encodes an RNA-dependent RNA polymerase (RdRp) of 727 amino acids with a molecular mass of 81.64 kDa. BLASTp analysis revealed that the RdRp domain of BdMV1 has 39.59% and 39.18% sequence identity to Plasmopara viticola associated mitovirus 43 and Setosphaeria turcica mitovirus 1, respectively. Phylogenetic analysis further suggested that BdMV1 is a new member of the genus Mitovirus within the family Mitoviridae. To the best of our knowledge, this is the first report of a mitovirus in B. dothidea.

Molecular characterization of a novel mitovirus from the plant‑pathogenic fungus Botryosphaeria dothidea.

Nidovirales, which accommodates viruses with the largest RNA genomes, includes the notorious coronaviruses; however, the evolutionary route for nidoviruses is not well understood. We have characterized a positive-sense (+) single-stranded (ss) RNA mycovirus, Rhizoctonia solani hypovirus 2 (RsHV2), from the phytopathogenic fungus Rhizoctonia solani. RsHV2 has the largest RNA genome size of 22,219 nucleotides, excluding the poly(A) tail, in all known mycoviruses, and contains two open reading frames (ORF1 and ORF2). ORF1 encodes a protein of 2,009 amino acid (aa) that includes a conserved helicase domain belonging to helicase superfamily I (SFI). In contrast, ORF2 encodes a polyprotein of 4459 aa containing the hallmark genes of hypoviruses. The latter includes a helicase belonging to SFII. Following phylogenetic analysis, the ORF1-encoded helicase (Hel1) unexpectedly clustered in an independent evolutionary branch together with nidovirus helicases, including coronaviruses, and bacteria helicases. Thus, Hel1 presence indicates the occurrence of horizontal gene transfer between viruses and bacteria. These findings also suggest that RsHV2 is most likely a recombinant arising between hypoviruses and nidoviruses.

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Two distant helicases in one mycovirus: evidence of horizontal gene transfer between mycoviruses, coronaviruses and other nidoviruses

Here, we describe a novel mycovirus, Auricularia heimuer fusarivirus 1 (AhFV1), isolated from the edible fungus Auricularia heimuer strain CCMJ1296. The virus has a single-stranded positive-sense [+ssRNA] genome of 7,127 nucleotides containing two overlapping open reading frames (ORFs) and a poly(A) tail. The large ORF1 encodes a polyprotein of 1,637 amino acids (aa) with conserved RNA-dependent RNA polymerase (RdRp) and DEAD-like helicase superfamily (DEXDc) domains. ORF2 encodes a putative 633-aa protein with unknown function. A BLAST search showed that the nucleotide sequence of the AhFV1 genome is 41.28% identical to that of Sclerotium rolfsii fusarivirus 2 and 40.49% identical to that of Sclerotium rolfsii fusarivirus 1. Phylogenetic analysis based on RdRp and helicase (Hel) sequences indicated that AhFV1 is related to unclassified mycoviruses and other fusariviruses. Our data suggest that AhFV1 should be classified as a member of the newly proposed family "Fusariviridae". This is the second virus and the first full genome sequence of a fusarivirus from A. heimuer.

Molecular characterization of a novel fusarivirus infecting the edible fungus Auricularia heimuer

A novel virus, Botryosphaeria dothidea fusarivirus 1 (BdFV1), was isolated from a fungal strain, SDAU11-86 of Botryosphaeria dothidea, and its complete genome sequence was determined. BdFV1 has a single-stranded positive-sense (+ssRNA) genome with 6,179 nucleotides, excluding the poly(A) tail. The genome of BdFV1 contains two putative open reading frames (ORFs). The first ORF encodes a large polyprotein of 1,544 amino acids (aa) with conserved RNA-dependent RNA polymerase and viral helicase domains. The second ORF encodes a putative 481-aa protein with unknown function. Sequence comparisons and phylogenetic analysis suggested that BdFV1 is a novel mycovirus belonging to the newly proposed family “Fusariviridae”. This is the first report of a +ssRNA mycovirus in B. dothidea.

Molecular characterization of a novel fusarivirus infecting the plant-pathogenic fungus Botryosphaeria dothidea

A novel positive-sense single-stranded RNA virus was isolated from strain CCMJ1271 of the fungus Auricularia heimuer, and the complete genome sequence of the virus was determined. Database searching, sequence alignment, and phylogenetic analysis revealed that this fungal virus and some viruses of family Virgaviridae clustered into a single branch of a phylogenetic tree, and we thus tentatively named the virus "Auricularia heimuer mycovirgavirus 1" (AhMV1). The AhMV1 genome consists of 9,934 nucleotides and contains a short poly(A) tail and three open reading frames (ORFs). ORF1 encodes an RNA-dependent RNA polymerase (RdRp), ORF2 encodes a protein that is homologous to movement proteins of plant virgaviruses, and ORF3 encodes a coat protein (CP). AhMV1 is the first virus to be discovered in A. heimuer.

Molecular characteristics of a novel ssRNA virus isolated from Auricularia heimuer in China.

Hypoviruses are positive-sense single-stranded RNA mycovirus that infect filamentous fungi. However, hypoviruses have not been reported in Bipolaris oryzae, an important phytopathogenic fungus in water bamboo and rice. Here, we report the characterization of a novel hypovirus, tentatively named "Bipolaris oryzae hypovirus 1" (BoHV1), isolated from strain ES35 of B. oryzae infecting water bamboo. The complete genome of BoHV1 consists of 13,596 nucleotides and a poly(A) tail at the 3' end. BoHV1 has single open reading frame (ORF) and encodes a putative polyprotein (4,218 amino acids) containing four potential conserved domains for a papain-like protease, a protein of unknown function (DUF3525), RNA-dependent RNA polymerase (RdRp), and helicase. Phylogenetic analysis of the polyprotein, RdRp, and helicase domains suggested that BoHV1 belongs to the genus Hypovirus within the family Hypoviridae. This is the first report of the presence of a hypovirus in the phytopathogenic fungus B. oryzae.

Du Hai

Papers

Molecular characterization of a novel fusarivirus infecting the plant-pathogenic fungus Botryosphaeria dothidea

Two distant helicases in one mycovirus: evidence of horizontal gene transfer between mycoviruses, coronaviruses and other nidoviruses

Molecular characterization of a novel fusarivirus infecting the edible fungus Auricularia heimuer

Molecular characteristics of a novel ssRNA virus isolated from Auricularia heimuer in China.

The complete genome sequence of a novel hypovirus infecting Bipolaris oryzae.