Artificial Infection of <i>Rosellinia necatrix</i> with Purified Viral Particles of a Member of the Genus <i>Mycoreovirus</i> Reveals Its Uneven Distribution in Single Colonies

Sasaki, Atsuko; Kanematsu, Satoko; Onoue, Mari; Oikawa, Yuri; Nakamura, Hitoshi; Yoshida, Kouji

doi:10.1094/phyto-97-3-0278

Cited by 50 publications

(35 citation statements)

References 28 publications

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“…This fact, and the low levels of RnVV1 replication in EP155, suggested that RnVV1 would be eliminated during repeated subculture. This possibility is supported by previous reports indicating that even in natural hosts, some mycoviruses are distributed unevenly and are easily lost during subculture (15,71). However, RnVV1 was readily transmitted via anastomosis (Fig.…”

Section: Discussionsupporting

confidence: 84%

“…Subsequently, similar techniques were applied for several other mycoviruses belonging to the same and different families such as Reoviridae (15), Partitiviridae (11), and Megabirnaviridae (13) and even for a gemini-like ssDNA virus (14). The present study investigated the transfection of two different fungal hosts, the natural one and an experimental one, with the novel victorivirus RnVV1 belonging to the genus Victorivirus of the family Totiviridae, for which no reproducible, efficient infectivity system has been available hitherto.…”

Section: Discussionmentioning

confidence: 99%

“…However, reproducible transfection protocols with purified particles have been established for several other dsRNA viruses (see, for example, references 11, 12, and13) and also a single-stranded DNA (ssDNA) virus (14). The establishment of transfection protocols has led to better understanding of the etiology of mycovirus infection, exploration of mycovirus-host and virus-virus interactions, and the determination of experimental host ranges (15)(16)(17). These developments encouraged us to test a victorivirus from a filamentous fungus for infectivity as particles.…”

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A Novel Victorivirus from a Phytopathogenic Fungus, Rosellinia necatrix, Is Infectious as Particles and Targeted by RNA Silencing

Chiba

Lin

Kondō

et al. 2013

J Virol

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Japan bA novel victorivirus, termed Rosellinia necatrix victorivirus 1 (RnVV1), was isolated from a plant pathogenic ascomycete, white root rot fungus Rosellinia necatrix, coinfected with a partitivirus. The virus was molecularly and biologically characterized using the natural and experimental hosts (chestnut blight fungus, Cryphonectria parasitica). RnVV1 was shown to have typical molecular victorivirus attributes, including a monopartite double-stranded RNA genome with two open reading frames (ORFs) encoding capsid protein (CP) and RNA-dependent RNA polymerase (RdRp), a UAAUG pentamer presumed to facilitate the coupled termination/reinitiation for translation of the two ORFs, a spherical particle structure ϳ40 nm in diameter, and moderate levels of CP and RdRp sequence identity (34 to 58%) to those of members of the genus Victorivirus within the family Totiviridae. A reproducible transfection system with purified RnVV1 virions was developed for the two distinct fungal hosts. Transfection assay with purified RnVV1 virions combined with virus elimination by hyphal tipping showed that the effects of RnVV1 on the phenotype of the natural host were negligible. Interestingly, comparison of the RNA silencing-competent (standard strain EP155) and -defective (⌬dcl-2) strains of C. parasitica infected with RnVV1 showed that RNA silencing acted against the virus to repress its replication, which was restored by coinfection with hypovirus or transgenic expression of an RNA silencing suppressor, hypovirus p29. Phenotypic changes were observed in the ⌬dcl-2 strain but not in EP155. This is the first reported study on the host range expansion of a Totiviridae member that is targeted by RNA silencing.

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Section: Discussionsupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

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A Novel Victorivirus from a Phytopathogenic Fungus, Rosellinia necatrix, Is Infectious as Particles and Targeted by RNA Silencing

Chiba

Lin

Kondō

et al. 2013

J Virol

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“…It may be infected with several dsRNA mycoviruses (37,38), including partitiviruses (RnPV1 and RnPV2), a mycoreovirus (Rosellinia necatrix mycoreovirus 3; RnMyRV3), a megabirnavirus (Rosellinia necatrix megabirnavirus 1; RnMBV1), a quadrivirus (Rosellinia necatrix quadrivirus 1; RnQV1), a victorivirus (RnVV1), and a novel ssRNA virus (Rosellinia necatrix fusarivirus 1; RnFV1) (28,29,33,34,(39)(40)(41)(42). Among them, RnPV1, RnMyRV3, RnMBV1, RnQV1, and RnVV1 were successfully introduced into the same R. necatrix strain W97, in which RnMyRV3 and RnMBV1 affect mycelial growth and virulence on apples but RnPV1, RnQV1, and RnVV1 were asymptomatic (28,29,34,43,44). Moreover, we showed that the RNA silencing mechanism is present in R. necatrix and is suppressed by RnMyRV3 but not by RnPV1, RnQV1, RnMBV1 (45), or RnVV1 (our unpublished data).…”

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Differential Inductions of RNA Silencing among Encapsidated Double-Stranded RNA Mycoviruses in the White Root Rot Fungus Rosellinia necatrix

Yaegashi

Shimizu

Ito

et al. 2016

J Virol

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RNA silencing acts as a defense mechanism against virus infection in a wide variety of organisms. Here, we investigated inductions of RNA silencing against encapsidated double-stranded RNA (dsRNA) fungal viruses (mycoviruses), including a partitivirus (RnPV1), a quadrivirus (RnQV1), a victorivirus (RnVV1), a mycoreovirus (RnMyRV3), and a megabirnavirus (RnMBV1) in the phytopathogenic fungus Rosellinia necatrix. Expression profiling of RNA silencing-related genes revealed that a dicer-like gene, an Argonaute-like gene, and two RNA-dependent RNA polymerase genes were upregulated by RnMyRV3 or RnMBV1 infection but not by other virus infections or by constitutive expression of dsRNA in R. necatrix. Massive analysis of viral small RNAs (vsRNAs) from the five mycoviruses showed that 19-to 22-nucleotide (nt) vsRNAs were predominant; however, their ability to form duplexes with 3= overhangs and the 5= nucleotide preferences of vsRNAs differed among the five mycoviruses. The abundances of 19-to 22-nt vsRNAs from RnPV1, RnQV1, RnVV1, RnMyRV3, and RnMBV1 were 6.8%, 1.2%, 0.3%, 13.0%, and 24.9%, respectively. Importantly, the vsRNA abundances and accumulation levels of viral RNA were not always correlated, and the origins of the vsRNAs were distinguishable among the five mycoviruses. These data corroborated diverse interactions between encapsidated dsRNA mycoviruses and RNA silencing. Moreover, a green fluorescent protein (GFP)-based sensor assay in R. necatrix revealed that RnMBV1 infection induced silencing of the target sensor gene (GFP gene and the partial RnMBV1 sequence), suggesting that vsRNAs from RnMBV1 activated the RNA-induced silencing complex. Overall, this study provides insights into RNA silencing against encapsidated dsRNA mycoviruses. IMPORTANCEEncapsidated dsRNA fungal viruses (mycoviruses) are believed to replicate inside their virions; therefore, there is a question of whether they induce RNA silencing. Here, we investigated inductions of RNA silencing against encapsidated dsRNA mycoviruses (a partitivirus, a quadrivirus, a victorivirus, a mycoreovirus, and a megabirnavirus) in Rosellinia necatrix. We revealed upregulation of RNA silencing-related genes in R. necatrix infected with a mycoreovirus or a megabirnavirus but not with other viruses, which was consistent with the relatively high abundances of vsRNAs from the two mycoviruses. We also showed common and different molecular features and origins of the vsRNAs from the five mycoviruses. Furthermore, we demonstrated the activation of RNA-induced silencing complex by mycoviruses in R. necatrix. Taken together, our data provide insights into an RNA silencing pathway against encapsidated dsRNA mycoviruses which is differentially induced among encapsidated dsRNA mycoviruses; that is, diverse replication strategies exist among encapsidated dsRNA mycoviruses. RNA silencing is a sequence-specific RNA degradation mechanism that is widely conserved among eukaryotic organisms, including animals, plants, and fungi (1-4). This mechanism is triggered by th...

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“…However, it has to be noted that successful infections with mycoviruses with rigid virus particles have been achieved using mycoviruses from families Reoviridae (Hillman et al, 2004;Sasaki et al, 2007;Kanematsu et al, 2010) and Partitiviridae (Sasaki et al, 2006;Kanematsu et al, 2010). In these studies, infection was achieved by inoculating fungal protoplasts with virus particles.…”

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Effect of Diaporthe RNA virus 1 (DRV1) on growth and pathogenicity of different Diaporthe species

Moleleki

Wingfield

et al. 2011

Eur J Plant Pathol

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Artificial Infection of Rosellinia necatrix with Purified Viral Particles of a Member of the Genus Mycoreovirus Reveals Its Uneven Distribution in Single Colonies

Cited by 50 publications

References 28 publications

A Novel Victorivirus from a Phytopathogenic Fungus, Rosellinia necatrix, Is Infectious as Particles and Targeted by RNA Silencing

A Novel Victorivirus from a Phytopathogenic Fungus, Rosellinia necatrix, Is Infectious as Particles and Targeted by RNA Silencing

Differential Inductions of RNA Silencing among Encapsidated Double-Stranded RNA Mycoviruses in the White Root Rot Fungus Rosellinia necatrix

Effect of Diaporthe RNA virus 1 (DRV1) on growth and pathogenicity of different Diaporthe species

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