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The RNAi machinery in Colletotrichum higginsianum has a role in antiviral defense

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Colletotrichum higginsianum IMI 349063

PROVIDER: GSE62708 | GEO | 2016/04/28

SECONDARY ACCESSION(S): PRJNA264842

REPOSITORIES: GEO

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The RNAi machinery in Colletotrichum higginsianum has a role in antiviral defense [RNAi_mutants_RNAseq]

Project description:Here we describe the identification and regulation of a novel dsRNA virus in Colletotrichum higginsianum. High throughput sequencing of small RNAs and strand-specific RNA-seq was performed on single gene knock-out mutants created for each RNAi component gene: rdr1, rdr2, rdr3, dcl1, dcl2, ago1, and ago2, and the double mutant: ∆dcl1∆dcl2. De novo assembly of the ∆dcl1 RNA-seq data identified two contigs that represented the forward and reverse strands of an uncharacterized dsRNA virus, designated here as Colletotrichum higginsianum non-segmented dsRNA virus 1 (ChNRV1). We found increased presence of the viral RNA in the RNA-seq datasets of the ∆dcl1, ∆dcl1dcl2, and ∆ago1 strains, suggesting that these genes are required for control of the virus. We show that viral small RNAs co-immunoprecipitate with a 6xFLAG-3xHIS-tagged AGO1 protein by sequencing the small RNAs from immunoprecipitated fractions. Additionally, analyses of the small RNA datasets from the RNAi mutants revealed control of the virus through small RNA-mediated silencing required both AGO1 and DCL1.

2016-04-28 | GSE62707 | GEO

The RNAi machinery in Colletotrichum higginsianum has a role in antiviral defense [AGO_IP_smallRNA]

2016-04-28 | GSE62706 | GEO

The RNAi machinery in Colletotrichum higginsianum has a role in antiviral defense [RNAi_mutants_smallRNA]

2016-04-28 | GSE62705 | GEO

The RNAi machinery in Colletotrichum higginsianum has a role in antiviral defense

Project description:The RNAi machinery in Colletotrichum higginsianum has a role in antiviral defense

| PRJNA264842 | ENA

Small RNA-Based Antiviral Defense in the Phytopathogenic Fungus Colletotrichum higginsianum.

Project description:Even though the fungal kingdom contains more than 3 million species, little is known about the biological roles of RNA silencing in fungi. The Colletotrichum genus comprises fungal species that are pathogenic for a wide range of crop species worldwide. To investigate the role of RNA silencing in the ascomycete fungus Colletotrichum higginsianum, knock-out mutants affecting genes for three RNA-dependent RNA polymerase (RDR), two Dicer-like (DCL), and two Argonaute (AGO) proteins were generated by targeted gene replacement. No effects were observed on vegetative growth for any mutant strain when grown on complex or minimal media. However, ?dcl1, ?dcl1?dcl2 double mutant, and ?ago1 strains showed severe defects in conidiation and conidia morphology. Total RNA transcripts and small RNA populations were analyzed in parental and mutant strains. The greatest effects on both RNA populations was observed in the ?dcl1, ?dcl1?dcl2, and ?ago1 strains, in which a previously uncharacterized dsRNA mycovirus [termed Colletotrichum higginsianum non-segmented dsRNA virus 1 (ChNRV1)] was derepressed. Phylogenetic analyses clearly showed a close relationship between ChNRV1 and members of the segmented Partitiviridae family, despite the non-segmented nature of the genome. Immunoprecipitation of small RNAs associated with AGO1 showed abundant loading of 5'U-containing viral siRNA. C. higginsianum parental and ?dcl1 mutant strains cured of ChNRV1 revealed that the conidiation and spore morphology defects were primarily caused by ChNRV1. Based on these results, RNA silencing involving ChDCL1 and ChAGO1 in C. higginsianum is proposed to function as an antiviral mechanism.

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PARP9 and DTX3L in Antiviral Host Defense

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2015-09-01 | E-GEOD-61421 | biostudies-arrayexpress

RNAi triggered by specialized machinery silences developmental genes and retrotransposons

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2012-11-26 | E-GEOD-41643 | biostudies-arrayexpress

PARP9 and DTX3L in Antiviral Host Defense

Project description:This SuperSeries is composed of the SubSeries listed below.

2015-09-01 | GSE61421 | GEO

RNAi triggered by specialized machinery silences developmental genes and retrotransposons

Project description:This SuperSeries is composed of the SubSeries listed below.