Abstract
The filamentous fungus Neurospora crassa is used as a model organism for genetics, developmental biology and molecular biology. Remarkably, it is not known to host or to be susceptible to infection with any viruses. Here, we identify diverse RNA viruses in N. crassa and other Neurospora species, and show that N. crassa supports the replication of these viruses as well as some viruses from other fungi. Several encapsidated double-stranded RNA viruses and capsid-less positive-sense single-stranded RNA viruses can be experimentally introduced into N. crassa protoplasts or spheroplasts. This allowed us to examine viral replication and RNAi-mediated antiviral responses in this organism. We show that viral infection upregulates the transcription of RNAi components, and that Dicer proteins (DCL-1, DCL-2) and an Argonaute (QDE-2) participate in suppression of viral replication. Our study thus establishes N. crassa as a model system for the study of host-virus interactions.
Highlights
The filamentous fungus Neurospora crassa is used as a model organism for genetics, developmental biology and molecular biology
A single fusarivirus harbored in the Louisiana isolate JW60, closest to the standard N. crassa strain 74-OR23-1VA, was fully determined by Sanger sequencing as the standard N. crassa fusarivirus (DDBJ/EMBL/GenBank accession #: LC530175), termed Neurospora crassa fusarivirus 1 (NcFV1) (Fig. 2a)
NcFV1-JW60 was used in the subsequent investigation into virus/host interactions, because the standard N. crassa strain singly infected by this virus could readily be obtained
Summary
The filamentous fungus Neurospora crassa is used as a model organism for genetics, developmental biology and molecular biology. Several encapsidated double-stranded RNA viruses and capsid-less positive-sense single-stranded RNA viruses can be experimentally introduced into N. crassa protoplasts or spheroplasts This allowed us to examine viral replication and RNAi-mediated antiviral responses in this organism. Circadian rhythm-based physiological regulation[22], RNA interference (RNAi) post-transcriptional gene silencing[23,24], and DNA methylation-mediated epigenetic control[25] have been pioneered by researchers using N. crassa The advantages of this fungus over other organisms, especially over other filamentous fungi, include the public availability of a number of biological and molecular tools, biological tractability[26], i.e., rapid vegetative growth and ease of sexual mating, shared techniques, a wellannotated genome sequence[27], and single-gene knockout (KO) collection[28] available from the Fungal Genetics Stock Center (FGSC) (http://www.fgsc.net)[29]. The second group is further categorized into two: quelling and qiRNA
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