Characterization and application of small RNAs and RNA silencing mechanisms in fungi

Nunes, Cristiano C.; Sailsbery, Joshua; Dean, Ralph A.

doi:10.1016/j.fbr.2011.10.001

Cited by 15 publications

(15 citation statements)

References 68 publications

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“…Indeed, the sRNA pathways appear to be relatively conserved in fungi, and the AGO proteins have been identified in several fungi, particularly in ascomycetes (Nunes et al, 2011). Evolutionary L a n e 1 L a n e 2 L a n e 3 L a n e 4 F I G U R E 4 Colonial morphology of mutants and wild-type (WT).…”

Section: Discussionmentioning

confidence: 99%

“…In plants, many AGOs have been reported to be involved in resistance to different pathogens (Agorio & Vera, 2007;Duan, Fang, & Zhou, 2012;Ellendorff, Fradin, de Jonge, & Thomma, 2009;Mallory & Vaucheret, 2010). Similarly, AGO proteins are also present in the vast majority of filamentous fungi (Hu, Stenlid, Elfstrand, & Olson, 2013;Nunes, Sailsbery, & Dean, 2011). Similarly, AGO proteins are also present in the vast majority of filamentous fungi (Hu, Stenlid, Elfstrand, & Olson, 2013;Nunes, Sailsbery, & Dean, 2011).…”

Section: Introductionmentioning

confidence: 99%

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The distinct roles of Argonaute protein 2 in the growth, stress responses and pathogenicity of the apple tree canker pathogen

Liu

et al. 2017

Forest Pathology

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Valsa mali (V. mali), the causal agent of apple tree Valsa canker, severely damages apple production, a major economic crop in China. To date, our understanding of the molecular mechanisms associated with the pathogenicity of V. mali is still limited. RNA interference participates in various biological processes in multicellular organisms. The Argonaute proteins (AGOs), which are a core component of the RNA interference system, play key roles in vegetable growth, environmental responses and fungal pathogenicity. Previously, transcriptome analysis revealed that the AGO2 gene (VMAGO2) was up-regulated during V. mali infection, suggesting that VMAGO2 plays a potential role in pathogenicity. In this study, we investigated the potential roles of VMAGO2 in the growth, stress responses and pathogenicity of V. mali. VMAGO2 was isolated and found to be orthologous to AGO2 of Neurospora crassa and Fusarium graminearum.Real-time quantitative PCR analysis showed that VMAGO2 expression was 3.4-fold higher than that of the control (mycelium) at 24 hr post-inoculation (hpi). Six positive VMAGO2 mutants were generated using double-joint PCR and PEG-mediated transformation. Deletion of VMAGO2 did not result in any obvious phenotypic change when compared with that of wild-type strain 03-8. Furthermore, the colonial morphology was not obviously affected when the mutants were subjected to osmotic and pH stress treatments. However, the knockout mutants did not grow on PDA with 0.05% H 2 O 2 . More importantly, infection assays showed that the average lesion diameter/ length resulting from mutant infections was 34.8% and 19.8% smaller on apple leaves and twigs, respectively, than those resulting from wild-type infections. All six positive mutants showed a consistent phenotype, and the defects of the mutants were fully complemented by re-introducing the WT VMAGO2 allele. Our results demonstrated that VMAGO2 plays an important role in H 2 O 2 tolerance and the pathogenicity of V. mali.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The distinct roles of Argonaute protein 2 in the growth, stress responses and pathogenicity of the apple tree canker pathogen

Liu

et al. 2017

Forest Pathology

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“…In fungi, several classes of sRNAs, including miRNA-like RNAs, Dicer-independent siRNAs (Lee et al, 2010), qiRNA, esRNA (Lee H.C. et al, 2009; Nicolas et al, 2010; Nunes et al, 2011b; Chen et al, 2015), and tsRNAs (Jochl et al, 2008; Thompson et al, 2008; Thompson and Parker, 2009; Nunes et al, 2011a) have been identified. However, no canonical miRNAs have been reported (Nunes et al, 2011b). In oomycetes, several miRNAs, a variety of siRNAs, and tsRNAs were discovered recently (Vetukuri et al, 2012; Fahlgren et al, 2013; Qutob et al, 2013; Asman et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

The tRNA-Derived Small RNAs Regulate Gene Expression through Triggering Sequence-Specific Degradation of Target Transcripts in the Oomycete Pathogen Phytophthora sojae

Wang

et al. 2016

Front. Plant Sci.

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Along with the well-studied microRNA (miRNA) and small interfering RNA (siRNA) is a new class of transfer RNA-derived small RNA (tsRNA), which has recently been detected in multiple organisms and is implicated in gene regulation. However, while miRNAs and siRNAs are known to repress gene expression through sequence-specific RNA cleavage or translational repression, how tsRNAs regulate gene expression remains unclear. Here we report the identification and functional characterization of tsRNAs in the oomycete pathogen Phytophthora sojae. We show that multiple tRNAs are processed into abundant tsRNAs, which accumulate in a similar developmental stage-specific manner and are negatively correlated with the expression of predicted target genes. Degradome sequencing and 5′ RLM RACE experiments indicate tsRNAs can trigger degradation of target transcripts. Transient expression assays using GUS sensor constructs confirmed the requirement of sequence complementarity in tsRNA-mediated RNA degradation in P. sojae. Our results show that the tsRNA are a class of functional endogenous sRNAs and suggest that tsRNA regulate gene expression through inducing sequence-specific degradation of target RNAs in oomycetes.

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“…The number of argonaute genes identified in filamentous fungal genomes range from zero to as many as eight [20]. The zygomycete M. circinelloides encodes three ago genes, although only ago-1 participates in the RNAi machinery during vegetative growth (Table 1 and Table 2).…”

Section: Discussionmentioning

confidence: 99%

A Single Argonaute Gene Participates in Exogenous and Endogenous RNAi and Controls Cellular Functions in the Basal Fungus Mucor circinelloides

et al. 2013

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The mechanism of RNAi is well described in metazoans where it plays a role in diverse cellular functions. However, although different classes of endogenous small RNAs (esRNAs) have been identified in fungi, their biological roles are poorly described due, in part, to the lack of phenotype of mutants affected in the biogenesis of these esRNAs. Argonaute proteins are one of the key components of the RNAi pathways, in which different members of this protein family participate in the biogenesis of a wide repertoire of esRNAs molecules. Here we identified three argonaute genes of the fungus Mucor circinelloides and investigated their participation in exogenous and endogenous RNAi. We found that only one of the ago genes, ago-1, is involved in RNAi during vegetative growth and is required for both transgene-induced RNA silencing and the accumulation of distinct classes of esRNAs derived from exons (ex-siRNAs). Classes I and II ex-siRNAs bind to Ago-1 to control mRNA accumulation of the target protein coding genes. Class III ex-siRNAs do not specifically bind to Ago-1, but requires this protein for their production, revealing the complexity of the biogenesis pathways of ex-siRNAs. We also show that ago-1 is involved in the response to environmental signals, since vegetative development and autolysis induced by nutritional stress are affected in ago-1 − M. circinelloides mutants. Our results demonstrate that a single Ago protein participates in the production of different classes of esRNAs that are generated through different pathways. They also highlight the role of ex-siRNAs in the regulation of endogenous genes in fungi and expand the range of biological functions modulated by RNAi.

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Characterization and application of small RNAs and RNA silencing mechanisms in fungi

Cited by 15 publications

References 68 publications

The distinct roles of Argonaute protein 2 in the growth, stress responses and pathogenicity of the apple tree canker pathogen

The distinct roles of Argonaute protein 2 in the growth, stress responses and pathogenicity of the apple tree canker pathogen

The tRNA-Derived Small RNAs Regulate Gene Expression through Triggering Sequence-Specific Degradation of Target Transcripts in the Oomycete Pathogen Phytophthora sojae

A Single Argonaute Gene Participates in Exogenous and Endogenous RNAi and Controls Cellular Functions in the Basal Fungus Mucor circinelloides

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