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As a biotrophic fungus, Ustilago maydis, the causal agent of common smut of maize, must establish and maintain a relationship with the host throughout the pathogenic cycle. Functional characterization of the U. maydis transcription factor Zfp1 suggests a role in modulating pathogenic development. Deletion of zfp1 resulted in attenuated pathogenic hyphal growth, reduced infection frequency, arrested disease symptom development, and decreased anthocyanin production. Complementation with Zfp1 constructs lacking conserved domains indicated it may function with and without DNA binding and suggests a role for the Fungal_trans domain in enabling full virulence. RNA‐sequencing revealed the transcription of approximately 30% of U. maydis genes, including those encoding predicted and confirmed effectors, were altered in the zfp1 deletion strain during pathogenic growth. Together, the results suggest that Zfp1 is a Zn(II)2Cys6 transcription factor that is required for the full induction of infection, anthocyanin production, and virulence. [ABSTRACT FROM AUTHOR]

Summary: Reactive oxygen species (ROS) play an important role during host–pathogen interactions and are often an indication of induced host defence responses. In this study, we demonstrate for the first time that Puccinia triticina (Pt) generates ROS, including superoxide, H2O2 and hydroxyl radicals, during wheat infection. Through pharmacological inhibition, we found that ROS are critical for both Pt urediniospore germination and pathogenic development on wheat. A comparative RNA‐Seq analysis of different stages of Pt infection process revealed 291 putative Pt genes associated with the oxidation–reduction process. Thirty‐seven of these genes encode known proteins. The expressions of five Pt genes, including PtNoxA, PtNoxB, PtNoxR, PtCat and PtSod, were subsequently verified using RT‐qPCR analysis. The results show that the expressions of PtNoxA, PtNoxB, PtNoxR, PtCat and PtSod are up‐regulated during urediniospore germination. In comparison, the expressions of PtNoxA, PtNoxB, PtNoxR and PtCat are down‐regulated during wheat infection from 12 to 120 h after inoculation (HAI), whereas the expression of PtSod is up‐regulated with a peak of expression at 120 HAI. We conclude that ROS are critical for the full virulence of Pt and a coordinate down‐regulation of PtNox genes may be important for successful infection in wheat. [ABSTRACT FROM AUTHOR]

Mitigation of emerging infectious diseases that threaten global biodiversity requires an understanding of critical host and pathogen responses to infection. For multihost pathogens where pathogen virulence or host susceptibility is variable, host-pathogen interactions in tolerant species may identify potential avenues for adaptive evolution in recently exposed, susceptible hosts. For example, the fungus Pseudogymnoascus destructans causes white-nose syndrome ( WNS) in hibernating bats and is responsible for catastrophic declines in some species in North America, where it was recently introduced. Bats in Europe and Asia, where the pathogen is endemic, are only mildly affected. Different environmental conditions among Nearctic and Palearctic hibernacula have been proposed as an explanation for variable disease outcomes, but this hypothesis has not been experimentally tested. We report the first controlled, experimental investigation of response to P. destructans in a tolerant, European species of bat (the greater mouse-eared bat, Myotis myotis). We compared body condition, disease outcomes and gene expression in control (sham-exposed) and exposed M. myotis that hibernated under controlled environmental conditions following treatment. Tolerant M. myotis experienced extremely limited fungal growth and did not exhibit symptoms of WNS. However, we detected no differential expression of genes associated with immune response in exposed bats, indicating that immune response does not drive tolerance of P. destructans in late hibernation. Variable responses to P. destructans among bat species cannot be attributed solely to environmental or ecological factors. Instead, our results implicate coevolution with the pathogen, and highlight the dynamic nature of the 'white-nose syndrome transcriptome.' Interspecific variation in response to exposure by the host (and possibly pathogen) emphasizes the importance of context in studies of the bat- WNS system, and robust characterization of genetic responses to exposure in various hosts and the pathogen should precede any attempts to use particular bat species as generalizable 'model hosts.' [ABSTRACT FROM AUTHOR]

The basidiomycete smut fungus Ustilago maydis causes common smut of corn. This disease is spread through the production of teliospores, which are thick-walled dormant structures characterized by low rates of respiration and metabolism. Teliospores are formed when the fungus grows within the plant, and the morphological steps involved in their formation have been described, but the molecular events leading to dormancy are not known. In U. maydis, natural antisense transcripts (NATs) can function to alter gene expression and many NATs have increased levels in the teliospore. One such NAT is as-ssm1 which is complementary to the gene for the mitochondrial seryl-tRNA synthetase ( ssm1), an enzyme important to mitochondrial function. The disruption of ssm1 leads to cell lysis, indicating it is also essential for cellular viability. To assess the function of as-ssm1, it was ectopically expressed in haploid cells, where it is not normally present. This expression led to reductions in growth rate, virulence, mitochondrial membrane potential and oxygen consumption. It also resulted in the formation of as-ssm1/ ssm1 double-stranded RNA and increased ssm1 transcript levels, but no change in Ssm1 protein levels was detected. Together, these findings suggest a role for as-ssm1 in facilitating teliospore dormancy through dsRNA formation and reduction of mitochondrial function. [ABSTRACT FROM AUTHOR]

Ustilago maydis infection of Zea mays leads to the production of thick-walled diploid teliospores that are the dispersal agent for this pathogen. Transcriptome analyses of this model biotrophic basidiomycete fungus identified natural antisense transcripts ( NATs) complementary to 247 open reading frames. The U. maydis NAT cDNAs were fully sequenced and annotated. Strand-specific RT- PCR screens confirmed expression and identified NATs preferentially expressed in the teliospore. Targeted screens revealed four U. maydis NATs that are conserved in a related fungus. Expression of NATs in haploid cells, where they are not naturally occurring, resulted in increased steady-state levels of some complementary mRNAs. The expression of one NAT, as - um02151, in haploid cells resulted in a twofold increase in complementary mRNA levels, the formation of sense-antisense double-stranded RNAs, and unchanged Um02151 protein levels. This led to a model for NAT function in the maintenance and expression of stored teliospore mRNAs. In testing this model by deletion of the regulatory region, it was determined that alteration in NAT expression resulted in decreased pathogenesis in both cob and seedling infections. This annotation and functional analysis supports multiple roles for U. maydis NATs in controlling gene expression and influencing pathogenesis. [ABSTRACT FROM AUTHOR]

Fungi are models for investigating many eukaryotic molecular processes. The identification of natural antisense transcripts (NATs) in fungi led to the discovery of mechanisms for controlling gene expression through transcriptional interference, chromatin remodelling and dsRNA formation. An overview of these mechanisms and the description of specific NAT functions is provided to give context for a broader discussion of fungal NATs. Transcriptome analyses have revealed a large number of NATs in a divergent group of fungi. The timing of NAT expression suggests roles in core life functions, such as responding to the environment and sexual reproduction. The transcriptome studies also uncover a large number of NATs whose functions remain elusive. These could provide novel control of gene expression, targeted responses to stimuli, or other functions. The goal of this review is provide background for this expanding field of research while highlighting opportunities for future discoveries. [ABSTRACT FROM AUTHOR]