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Fungi are generally thought to live in host plants with a single lifestyle, being parasitism, commensalism, or mutualism. The former, known as phytopathogenic fungi, cause various plant diseases that result in significant losses every year; while the latter, such as endophytic fungi, can confer fitness to the host plants. It is unclear whether biological factors can modulate the parasitic and mutualistic traits of a fungus. In this study, we isolated and characterized a mycovirus from an endophytic strain of the fungus Pestalotiopsis theae , a pathogen of tea ( Camellia sinensis ). Based on molecular analysis, we tentatively designated the mycovirus as Pestalotiopsis theae chrysovirus-1 (PtCV1), a novel member of the family Chrysoviridae , genus Alphachrysovirus . PtCV1 has four double-stranded (ds) RNAs as its genome, ranging from 0.9 to 3.4 kbp in size, encapsidated in isometric particles. PtCV1 significantly reduced the growth rates of its host fungus in vitro (ANOVA; P -value < 0.001) and abolished its virulence in planta (ANOVA; P -value < 0.001), converting its host fungus to a non-pathogenic endophyte on tea leaves, while PtCV1-free isolates were highly virulent. Moreover, the presence of PtCV1 conferred high resistance to the host plants against the virulent P. theae strains. Here we report a mycovirus that modulates endophytic and phytopathogenic fungal traits and provides an alternative approach to biological control of plant diseases caused by fungi.

Transcription is fundamentally noisy, leading to significant heterogeneity across bacterial populations. Noise is often attributed to burstiness, but the underlying mechanisms and their dependence on the mode of promotor regulation remain unclear. Here, we measure E. coli single cell mRNA levels for two stress responses that depend on bacterial sigma factors with different mode of transcription initiation (σ 70 and σ 54 ). By fitting a stochastic model to the observed mRNA distributions, we show that the transition from low to high expression of the σ 70 -controlled stress response is regulated via the burst size, while that of the σ 54 -controlled stress response is regulated via the burst frequency. Therefore, transcription initiation involving σ 54 differs from other bacterial systems, and yields bursting kinetics characteristic of eukaryotic systems. Transcription noise in bacteria is often attributed to burstiness, but the mechanisms are unclear. Here, the authors show that the transition from low to high expression can be regulated via burst size or burst frequency, depending on the mode of transcription initiation determined by different sigma factors.

Approximately a year ago, when I accepted the offer to act as a Guest Editor for the Special Issue ‘Mycoviruses’ organised by the MDPI journal Viruses, I dared not expect that ‘Mycoviruses’ would include such a large number of manuscripts [...]

To date, viroids have been found to naturally infect only plants, resulting in substantial losses for some crops. Whether viroids or viroid‐like RNAs naturally infect non‐plant hosts remains unknown. Here the existence of a set of exogenous, single‐stranded circular RNAs, ranging in size from 157 to 450 nucleotides, isolated from the fungus Botryosphaeria dothidea and nominated B. dothidea RNAs (BdcRNAs) is reported. BdcRNAs replicate autonomously in the nucleus via a rolling‐circle mechanism following a symmetric pathway. BdcRNA infection induces symptoms, because BdcRNAs can apparently modulate, to different degrees, specific biological traits (e.g., alter morphology, decrease growth rate, attenuate virulence, and increase or decrease tolerance to osmotic and oxidative stress) of the host fungus. Overall, BdcRNAs have genome characteristics similar to those of viroids and exhibit pathogenic effects on fungal hosts. It is proposed that these novel fungus infecting RNAs should be termed mycoviroids. BdcRNA(s) may be considered additional inhabitants at the frontier of life in terms of genomic complexity, and represent a new class of acellular entities endowed with regulatory functions, and novel epigenomic carriers of biological information. A novel class of viroid‐like RNAs, termed as mycoviroids, naturally infects a filamentous fungus Botryosphaeria dothidea responsible for fungal attenuated virulence and growth rate. These mycoviroids range in size between 157 and 450 nucleotides, replicate autonomously in the nucleus via a rolling‐circle replication mechanism following symmetric pathways, and regulate the metabolic pathways of the host fungus.

Mycoviruses are widely distributed among different groups of filamentous fungi. An awareness of infections caused by mycoviruses was highlighted in the 1980s and 1990s, when the impact of these agents on phenotypes of agriculturally and medically important fungi was reported. However, for entomopathogenic fungi, mycovirus research has only expanded significantly in the last 15 years. Due to the agricultural importance of these fungi, reflected in their use at the forefront of biological control strategies, recent studies have extensively described novel viruses and their effects on their hosts in terms of altered morphological, phenotypical, and ecological characteristics. To summarise the historical progress of mycovirology and recent discoveries, here we describe the state of the art in the study of mycoviruses associated with entomopathogenic fungi. We have limited the review to the occurrence of mycoviruses in fungi of the genera , , , , and and have compiled an inventory of the viruses reported to infect these entomopathogenic genera, as well as a comprehensive review of the biological effects described with respect to infection by mycoviruses in fungi that are relevant to the biological control of insects. Finally, we have outlined possible research scenarios in the light of recent discoveries in the field of mycovirology, such as the use of mycoviruses as virulence modulating factors: the main character sought in biological pest control.

Transcription activator RamA is linked to multidrug resistance of Klebsiella pneumoniae through controlling genes that encode efflux pumps (acrA) and porin‐regulating antisense RNA (micF). In bacteria, σ70, together with activators, controls the majority of genes by recruiting RNA polymerase (RNAP) to the promoter regions. RNAP and σ70 form a holoenzyme that recognizes ‐35 and ‐10 promoter DNA consensus sites. Many activators bind upstream from the holoenzyme and can be broadly divided into two classes. RamA acts as a class I activator on acrA and class II activator on micF, respectively. The authors present biochemical and structural data on RamA in complex with RNAP‐σ70 at the two promoters and the data reveal the molecular basis for how RamA assembles and interacts with core RNAP and activates transcription that contributes to antibiotic resistance. Further, comparing with CAP/TAP complexes reveals common and activator‐specific features in activator binding and uncovers distinct roles of the two C‐terminal domains of RNAP α subunit. Transcription activator RamA is linked to multidrug resistance of Klebsiella pneumoniae. This study reveals the molecular basis for how RamA interacts with RNA polymerase (RNAP) and activates transcription that contributes to antibiotic resistance. Further, comparing with catabolite activator protein/thermophilic CAP homolog ( CAP/TAP) complexes reveals common and activator‐specific features in activator bindings and uncovers distinct roles of the two C‐terminal domains of RNAP.

Bacteriophage T7 infects Escherichia coli and evades the host restriction/modification system. The Ocr protein of T7 was shown to exist as a dimer mimicking DNA and to bind to host restriction enzymes, thus preventing the degradation of the viral genome by the host. Here we report that Ocr can also inhibit host transcription by directly binding to bacterial RNA polymerase (RNAP) and competing with the recruitment of RNAP by sigma factors. Using cryo electron microscopy, we determined the structures of Ocr bound to RNAP. The structures show that an Ocr dimer binds to RNAP in the cleft, where key regions of sigma bind and where DNA resides during transcription synthesis, thus providing a structural basis for the transcription inhibition. Our results reveal the versatility of Ocr in interfering with host systems and suggest possible strategies that could be exploited in adopting DNA mimicry as a basis for forming novel antibiotics. Bacteria and viruses have long been fighting amongst themselves. Bacteriophages are a type of virus that invade bacteria; their name literally means ‘bacteria eater’. The bacteriophage T7, for example, infects the common bacteria known as Escherichia coli. Once inside, the virus hijacks the bacterium’s cellular machinery, using it to replicate its own genetic material and make more copies of the virus so it can spread. At the same time, the bacteria have found ways to try and defend themselves, which in turn has led some bacteriophages to develop countermeasures to overcome those defences. Many bacteria, for example, have restriction enzymes which recognise certain sections of the bacteriophage DNA and cut it into fragments. However, the T7 bacteriophage has one well-known protein called Ocr which inhibits restriction enzymes. Ocr does this by mimicking DNA, which led Ye et al. to wonder if it could also interrupt other vital processes in a bacterial cell that involve DNA. Transcription is the first step in a coordinated process that turns the genetic information stored in a cell’s DNA into useful proteins. An enzyme called RNA polymerase decodes the DNA sequence into a go-between molecule called messenger RNA, and it was here that Ye et al. thought Ocr might jump in to interfere. To begin, Ye et al. examined the structure of Ocr when it binds to RNA polymerase using an imaging technique called cryo-electron microscopy. Ocr has been well-studied before, its structure previously described, but not when attached to RNA polymerase. The analysis showed that Ocr gets in the way of specific molecules, called sigma factors, that show RNA polymerase where to start transcription. Ocr binds to RNA polymerase in exactly the same pocket as part of sigma factors do, which is also the place where DNA must be to be decoded to make messenger RNA. Ye et al. then performed experiments to show Ocr interfering with binding to RNA polymerase did indeed disrupt transcription. This means Ocr is quite versatile as it interferes with the RNA polymerase of the bacterial host and its restriction enzymes. Ocr’s strategy of mimicking DNA to interrupt transcription could be adopted as an approach to develop new antibiotics to stop bacterial infections. DNA transcription is an essential cellular process – without it, no cell can replicate and survive – and RNA polymerase is already a validated target for drugs. Following Ocr’s lead could provide a new way to stop infections, if the right drug can be designed to fit.

There was an error in the original publication [...]

Mycoviral infection can either be asymptomatic or have marked effects on fungal hosts, influencing them either positively or negatively. To fully understand the effects of mycovirus infection on the fungal host, transcriptomic profiling of four Beauveria bassiana isolates, including EABb 92/11-Dm that harbors mycoviruses, was performed 48 h following infection of Tenebrio molitor via topical application or injection. Genes that participate in carbohydrate assimilation and transportation, and those essential for fungal survival and oxidative stress tolerance, calcium uptake, and iron uptake, were found to be overexpressed in the virus-infected isolate during the mid-infection stage. Mycotoxin genes encoding bassianolide and oosporein were switched off in all isolates. However, beauvericin, a mycotoxin capable of inducing oxidative stress at the molecular level, was expressed in all four isolates, indicating an important contribution to virulence against T. molitor. These observations suggest that detoxification of immune-related (oxidative) defenses and nutrient scouting, as mediated by these genes, occurs in mid-infection during the internal growth phase. Consequently, we observe a symbiotic relationship between mycovirus and fungus that does not afflict the host; on the contrary, it enhances the expression of key genes leading to a mycovirus-mediated hypervirulence effect.

The entomopathogenic fungus Beauveria bassiana has a wide host range and is used as a biocontrol agent against arthropod pests. Mycoviruses have been described in phytopathogenic fungi while in entomopathogenic fungi their presence has been reported only rarely. Here we show that 21.3% of a collection of B. bassiana isolates sourced from worldwide locations, harbor dsRNA elements. Molecular characterization of these elements revealed the prevalence of mycoviruses belonging to the Partitiviridae and Totiviridae families, the smallest reported virus to date, belonging to the family Narnaviridae, and viruses unassigned to a family or genus. Of particular importance is the discovery of members of a newly proposed family Polymycoviridae in B. bassiana. Polymycoviruses, previously designated as tetramycoviruses, consist of four non-conventionally encapsidated capped dsRNAs. The presence of additional non-homologous genomic segments in B. bassiana polymycoviruses and other fungi illustrates the unprecedented dynamic nature of the viral genome. Finally, a comparison of virus-free and virus-infected isogenic lines derived from an exemplar B. bassiana isolate revealed a mild hypervirulent effect of mycoviruses on the growth of their host isolate and on its pathogenicity against the greater wax moth Galleria mellonella, highlighting for the first time the potential of mycoviruses as enhancers of biocontrol agents.