Presence Of dsRNAs Research Articles

GW182 Controls Drosophila Circadian Behavior and PDF-Receptor Signaling

The neuropeptide PDF is crucial for Drosophila circadian behavior: it keeps circadian neurons synchronized. Here, we identify GW182 as a key regulator of PDF signaling. Indeed, GW182 downregulation results in phenotypes similar to those of Pdf and Pdf-receptor (Pdfr) mutants. gw182 genetically interacts with Pdfr and cAMP signaling, which is essential for PDFR function. GW182 mediates miRNA-dependent gene silencing through its interaction with AGO1. Consistently, GW182's AGO1 interaction domain is required for GW182's circadian function. Moreover, our results indicate that GW182 modulates PDFR signaling by silencing the expression of the cAMP phosphodiesterase DUNCE. Importantly, this repression is under photic control, and GW182 activity level--which is limiting in circadian neurons--influences the responses of the circadian neural network to light. We propose that GW182's gene silencing activity functions as a rheostat for PDFR signaling and thus profoundly impacts the circadian neural network and its response to environmental inputs.

A Novel Virus-Like Double-Stranded RNA in an Obligate Biotroph Arbuscular Mycorrhizal Fungus: A Hidden Player in Mycorrhizal Symbiosis

Arbuscular mycorrhizal (AM) fungi form mutualistic associations with most land plants and enhance phosphorus uptake of the host plants. Fungal viruses (mycoviruses) that possess a double-stranded RNA (dsRNA) genome often affect plant-fungal interactions via altering phenotypic expression of their host fungi. The present study demonstrates, for the first time, the presence of dsRNAs, which are highly likely to be mycoviruses, in AM fungi. dsRNA was extracted from mycelia of Glomus sp. strain RF1, purified, and subjected to electrophoresis. The fungus was found to harbor various dsRNA segments that differed in size. Among them, a 4.5-kbp segment was termed Glomus sp. strain RF1 virus-like medium dsRNA (GRF1V-M) and characterized in detail. The GRF1V-M genome segment was 4,557 nucleotides in length and encoded RNA-dependent RNA polymerase and a structural protein. GRF1V-M was phylogenetically distinct and could not be assigned to known genera of mycovirus. The GRF1V-M-free culture line of Glomus sp. strain RF1, which was raised by single-spore isolation, produced twofold greater number of spores and promoted plant growth more efficiently than the GRF1V-M-positive lines. These observations suggest that mycoviruses in AM fungi, at least some of them, have evolved under unique selection pressures and are a biologically active component in the symbiosis.

Induction of systemic resistance by a hypovirulent Rhizoctonia solani isolate in tomato

A polynucleate Rhizoctonia isolate (R3) was analysed for virulence, growth characteristics, enzyme production and presence of dsRNAs. Taxonomic position was assessed morphologically and by anastomosis group (AG) testing and ITS sequence analysis. Results indicated that R3 is a hypovirulent R. solani AG 4. Mechanisms underlying biocontrol towards virulent R. solani and Botrytis cinerea were investigated and plant-mediated resistance was followed using biochemical markers of defence (PR1, laminarinase, chitinase). Control apparently relies on spatial and nutrient competition in soil, and on systemic induced resistance. This is the first report on induction of systemic resistance and of defence markers by a hypovirulent strain of R. solani.

Mycoviruses in Monilinia fructicola

DsRNAs were detected in 36 of 49 Monilinia fructicola isolates from stone fruit orchards in New Zealand. The dsRNA profiles were highly variable, even between isolates from a single tree. Comparison of pathogenicity on detached fruit, in vitro growth rate, and sporulation of 14 isolates showed no obvious correlation with presence of dsRNAs. Partially purified extracts from four isolates were examined for the presence of virus-like particles by transmission electron microscopy. One isolate contained 45 nm isometric particles similar in appearance to totiviruses and partitiviruses. A second isolate contained 200-250 x 25 nm rigid rods similar in appearance to the plant pathogenic tobraviruses and furoviruses. This is the first report of the presence of viral-like agents in the brown rot fungus Monilinia fructicola.

Influence of double-stranded RNAs on growth, sporulation, pathogenicity, and survival of Chalara elegans

Three strains of Chalara elegans from diverse geographical areas that contained multiple (4 or 5) double-stranded RNA fragments were compared with spontaneously derived cultures from these strains that were either partially cured or completely free of dsRNA. In the wild-type strains, presence of the dsRNAs was found to significantly enhance phialospore production and pigmentation of colonies, whereas radial growth and mycelial dry weight accumulation were reduced. The rate and overall percentage of phialospore germination on 1% Noble water agar were also significantly reduced by the presence of the dsRNAs. In two partially cured strains (only one 2.8-kb fragment remaining), pathogenicity to various plant tissues was significantly enhanced when compared with the wild-type strains containing multiple dsRNA. However, survival in field soil was enhanced in one strain and reduced in the other. In the completely cured strain, the loss of multiple dsRNA fragments was associated with enhanced growth, reduced phialospore production, and a complete loss of pathogenicity and capability for survival in soil. These results indicate that the effects of dsRNAs in C. elegans vary with the strain. In general, the presence of multiple dsRNAs in this fungus enhanced sporulation, altered colony morphology, and reduced growth and pathogenicity. However, since the complete loss of dsRNA was found to eliminate pathogenicity and reduce survival, it suggests that some dsRNA fragments in C. elegans may confer an advantage to this soil-borne facultative plant pathogen. Key words: black root rot, soil-borne plant pathogen, Thielaviopsis basicola.

Widespread incidence of double-stranded RNAs of unknown function in rust fungi

Double-stranded (ds) RNAs were found to be present in 33 of 38 collections of rust fungi, representing 15 species from four genera. This finding suggests that dsRNAs occur commonly in rust fungi. Tests were undertaken to determine the role or phenotype associated with the presence of dsRNAs. Comparisons were made between strains of Puccinia sorghi (maize rust) that had different dsRNAs and two fully isogenic strains of Melampsora lini (flax rust), one with no dsRNA and one with at least 13 dsRNAs. It was concluded that dsRNAs had no obvious effect on pathogen virulence or on growth rate as measured by rate of accumulation of leaf chitin or time to pustule eruption. Also it was demonstrated that the presence of dsRNAs had no effect on the interaction that exists between rusts over short distances in the leaf. Key words: dsRNA, rust, function, incidence.