Fungal Host Research Articles

Human Clinical Isolates of Pathogenic Fungi Are Host to Diverse Mycoviruses.

ABSTRACTFungi host viruses from many families, and next-generation sequencing can be used to discover previously unknown genomes. Some fungus-infecting viruses (mycoviruses) confer hypovirulence on their pathogenic hosts, raising the possibility of therapeutic application in the treatment of fungal diseases. Though all fungi probably host mycoviruses, many human pathogens have none documented, implying the mycoviral catalogue remains at an early stage. Here, we carried out virus discovery on 61 cultures of pathogenic fungi covering 27 genera and at least 56 species. Using next-generation sequencing of total nucleic acids, we found no DNA viruses but did find a surprising RNA virus diversity of 11 genomes from six classified families and two unclassified lineages, including eight genomes likely representing new species. Among these was the first jivivirus detected in a fungal host (Aspergillus lentulus). We separately utilized rolling circle amplification and next-generation sequencing to identify ssDNA viruses specifically. We identified 13 new cressdnaviruses across all libraries, but unlike the RNA viruses, they could not be confirmed by PCR in either the original unamplified samples or freshly amplified nucleic acids. Their distributions among sequencing libraries and inconsistent detection suggest low-level contamination of reagents. This highlights both the importance of validation assays and the risks of viral host prediction on the basis of highly amplified sequencing libraries. Meanwhile, the detected RNA viruses provide a basis for experimentation to characterize possible hypovirulent effects, and hint at a wealth of uncharted viral diversity currently frozen in biobanks.IMPORTANCE Fungal pathogens of humans are a growing global health burden. Viruses of fungi may represent future therapeutic tools, but for many fungal pathogens there are no known viruses. Our study examined the viral content of diverse human-pathogenic fungi in a clinical biobank, identifying numerous viral genomes, including one lineage previously not known to infect fungi.

Pathogenicity, ultra-structural growth and development of green muscardine fungus, Metarhizium anisopliae (Metschnikoff) Sorokin (Ascomycota: Hypocreales) on maize fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae)

BackgroundThe fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) is a devastating pest of maize, difficult to manage using a single pest management tactic. The entomopathogenic fungus (EPF), Metarhizium anisopliae being an eco-friendly component, could be an effective tool for managing this notorious pest owing to its preferred habitat, principally the whorl region.ResultsMetarhizium anisopliae (TNAU-MA-GDU) isolated from FAW was evaluated against 2nd instar larvae of S. frugiperda reared under laboratory conditions. The pathogenicity and infection process were also studied through scanning electron microscope (SEM). The LC50 value for M. anisopliae isolate TNAU-MA-GDU was recorded as 5.8 × 104 spores ml−1 against the second instar larvae of S. frugiperda. SEM observations on the infection process revealed the adherence of the conidia on the head, cuticular ornamentations of the thorax, sensory seta, and legs at 24 h post-infection (hpi). Formation of germ tube and appressoria was observed from 48 hpi with subsequent hyphal development at 72 hpi. Complete networking of mycelium and conidiogenesis observed after 168 hpi.ConclusionAn understanding of fungal–host interactions at the ultramicroscopic levels could reform present tactics for developing hyper-virulent EPF strains.

A highly conserved gene locus in endofungal bacteria codes for the biosynthesis of symbiosis-specific cyclopeptides.

The tight association of the pathogenic fungus Rhizopus microsporus and its toxin-producing, bacterial endosymbionts (Mycetohabitans spp.) is distributed worldwide and has significance for agriculture, food production, and human health. Intriguingly, the endofungal bacteria are essential for the propagation of the fungal host. Yet, little is known about chemical mediators fostering the symbiosis, and universal metabolites that support the mutualistic relationship have remained elusive. Here, we describe the discovery of a complex of specialized metabolites produced by endofungal bacteria under symbiotic conditions. Through full genome sequencing and comparative genomics of eight endofungal symbiont strains from geographically distant regions, we discovered a conserved gene locus (hab) for a nonribosomal peptide synthetase as a unifying trait. Bioinformatics analyses, targeted gene deletions, and chemical profiling uncovered unprecedented depsipeptides (habitasporins) whose structures were fully elucidated. Computational network analysis and labeling experiments granted insight into the biosynthesis of their nonproteinogenic building blocks (pipecolic acid and β-phenylalanine). Deletion of the hab gene locus was shown to impair the ability of the bacteria to enter their fungal host. Our study unveils a common principle of the endosymbiotic lifestyle of Mycetohabitans species and expands the repertoire of characterized chemical mediators of a globally occurring mutualistic association.

A Transfectable Fusagravirus from a Japanese Strain of Cryphonectria carpinicola with Spherical Particles.

A novel dsRNA virus (Cryphonectria carpinicola fusagravirus 1, CcFGV1), isolated from a Japanese strain (JS13) of Cryphonectria carpinicola, was thoroughly characterized. The biological comparison of a set of isogenic CcFGV1-infected and -free (JS13VF) strains indicated asymptomatic infection by CcFGV1. The sequence analysis showed that the virus has a two open reading frame (ORF) genome of 9.6 kbp with the RNA-directed RNA polymerase domain encoded by ORF2. The N-terminal sequencing and peptide mass fingerprinting showed an N-terminally processed or degraded product (150 kDa) of the 5′-proximal ORF1-encoded protein (1462 amino acids) to make up the CcFGV1 spherical particles of ~40 nm in diameter. Interestingly, a portion of CcFGV1 dsRNA co-fractionated with a host protein of 70 kDa. The purified CcFGV1 particles were used to transfect protoplasts of JS13VF as well as the standard strain of an experimental model filamentous fungal host Cryphonectria parasitica. CcFGV1 was confirmed to be associated with asymptomatic infection of both fungi. RNA silencing was shown to target the virus in C. parasitica, resulting in reduced CcFGV1 accumulation by comparing the CcFGV1 content between RNA silencing-competent and -deficient strains. These results indicate the transfectability of spherical particles of a fusagravirus associated with asymptomatic infection.

Seasonal changes in mycophagous insect communities

The phenology of fungal fruiting has changed in the UK over the last 70 years, but whether the associated mycophagous insects are able to exploit 'out of season' fruit bodies is unknown. This study focused on whether fungal baits can be used as a proxy to examine changes in fungal fruiting on insect communities. Using Agaricus bisporus as a bait, mushrooms were placed into two separate woodlands monthly from November 2020 to July 2021. Megaselia rufipes (Phoridae) and Bradysia spp. (Sciaridae) were reared from both wild fungi and fungal baits at different times, making them appropriate species to consider for possible host tracking. Various factors affect an insect's ability to track a fungal host, these include host preference, season, period of fungal fruiting and age of mushroom. Increased fruiting of macrofungi in the future may benefit generalist mycophagous insects, by providing enhanced temporal and spatial resource opportunities. Using fungal baits as a proxy for the effects of climate change on fungal fruiting should be beneficial in uncovering the host preferences of mycophagous insects and may potentially indicate whether mycophagous insects can track fungal hosts across seasons.

Lineage and role in integrative taxonomy of a heterotrophic orchid complex.

Lineage-based species definitions applying coalescent approaches to species delimitation have become increasingly popular. Yet, the application of these methods and the recognition of lineage-only definitions have recently been questioned. Species delimitation criteria that explicitly consider both lineages and evidence for ecological role shifts provide an opportunity to incorporate ecologically meaningful data from multiple sources in studies of species boundaries. Here, such criteria were applied to a problematic group of mycoheterotrophic orchids, the Corallorhiza striata complex, analysing genomic, morphological, phenological, reproductive-mode, niche, and fungal host data. A recently developed method for generating genomic polymorphism data-ISSRseq-demonstrates evidence for four distinct lineages, including a previously unidentified lineage in the Coast Ranges and Cascades of California and Oregon, USA. There is divergence in morphology, phenology, reproductive mode, and fungal associates among the four lineages. Integrative analyses, conducted in population assignment and redundancy analysis frameworks, provide evidence of distinct genomic lineages and a similar pattern of divergence in the extended data, albeit with weaker signal. However, none of the extended data sets fully satisfy the condition of a significant role shift, which requires evidence of fixed differences. The four lineages identified in the current study are recognized at the level of variety, short of comprising different species. This study represents the most comprehensive application of lineage + role to date and illustrates the advantages of such an approach.

High Variability of Fungal Communities Associated with the Functional Tissues and Rhizosphere Soil of Picea abies in the Southern Baltics

Climate change, which leads to higher temperatures, droughts, and storms, is expected to have a strong effect on both health of forest trees and associated biodiversity. The aim of this study was to investigate the diversity and composition of fungal communities associated with the functional tissues and rhizosphere soil of healthy-looking Picea abies to better understand these fungal communities and their potential effect on tree health in the process of climate change. The study sites included 30 P. abies stands, where needles, shoots, roots, and the rhizosphere soil was sampled. DNA was isolated from individual samples, amplified using ITS2 rRNA as a marker and subjected to high-throughput sequencing. The sequence analysis showed the presence of 232,547 high-quality reads, which following clustering were found to represent 2701 non-singleton fungal OTUs. The highest absolute richness of fungal OTUs was in the soil (1895), then in the needles (1049) and shoots (1002), and the lowest was in the roots (641). The overall fungal community was composed of Ascomycota (58.3%), Basidiomycota (37.2%), Zygomycota (2.5%), Chytridiomycota (1.6%), and Glomeromycota (0.4%). The most common fungi based on sequence read abundance were Aspergillus pseudoglaucus (7.9%), Archaeorhizomyces sp. (3.6%), and Rhinocladiella sp. (2.0%). Pathogens were relatively rare, among which the most common were Phacidium lacerum (1.7%), Cyphellophora sessilis (1.4%), and Rhizosphaera kalkhoffii (1.4%). The results showed that the detected diversity of fungal OTUs was generally high, but their relative abundance varied greatly among different study sites, thereby highlighting the complexity of interactions among the host trees, fungi, and local environmental conditions.

Codon Usage Provides Insights into the Adaptive Evolution of Mycoviruses in Their Associated Fungi Host.

Codon usage bias (CUB) could reflect co-evolutionary changes between viruses and hosts in contrast to plant and animal viruses, and the systematic analysis of codon usage among the mycoviruses that infect plant pathogenic fungi is limited. We performed an extensive analysis of codon usage patterns among 98 characterized RNA mycoviruses from eight phytopathogenic fungi. The GC and GC3s contents of mycoviruses have a wide variation from 29.35% to 64.62% and 24.32% to 97.13%, respectively. Mycoviral CUB is weak, and natural selection plays a major role in the formation of mycoviral codon usage pattern. In this study, we demonstrated that the codon usage of mycoviruses is similar to that of some host genes, especially those involved in RNA biosynthetic process and transcription, suggesting that CUB is a potential evolutionary mechanism that mycoviruses adapt to in their hosts.

Advances and challenges in the management of feline sporotrichosis

The domestic cat is the most susceptible host to Sporothrix infection, developing severe clinical forms. Few effective antifungal agents are available for treating feline sporotrichosis, and cases of treatment failure are common. Treatment success depends on cat health status, therapy-related factors, as well as social/economic issues, but it is mainly contingent upon the host–fungus interaction. The owner's adherence is critical and should be reinforced throughout the treatment to increase the chances of a successful outcome. The antifungal agents described for feline sporotrichosis are most often used in monotherapy regimens. Due to cases in which the treatment with itraconazole failed, the use of antifungal agents in combination should be considered to achieve synergy. The combination of itraconazole and potassium iodide represents an important option for the treatment of naïve cats presenting multiple cutaneous lesions, nasal mucosal lesions and/or respiratory signs, as well as for refractory cases. However, the therapeutic options for unsuccessfully treated cases are scarce. Therefore new options are needed, even more taking into account that there are many in vitro potential molecules not available for use in cats yet. More studies are necessary to correlate in vitro antifungal susceptibility tests results and the outcome of cats treated due to sporotrichosis. This review will briefly discuss both the antifungal drugs and treatment protocols used in cats with sporotrichosis, as well as the determinants of treatment failure.

Solenopsora species (Leprocaulaceae) as hosts of lichenicolous fungi

Darmostuk, V., Fačkovcová, Z., Breidy, J., Dokmak, H., van den Boom, P. P. G. & Guttová, A. 2022. Solenopsora-Arten als Wirte flechtenbewohnender Pilze. – Herzogia 35: 115 –130. Wir untersuchten die Thalli von Arten der Flechtengattung Solenopsora (Leprocaulaceae), um mehr über ihr Potenzial als Wirte symptomatischer flechtenbewohnender Pilze zu erfahren. Insgesamt wurden 600 Proben betrachtet, von denen 78 mit flechtenbewohnenden Pilzen infiziert waren. Wir stellten elf Taxa flechtenbewohnender Pilze fest – Arthonia sp., Ceratobasidium bulbillifaciens, Cladosporium sp., Didymocyrtis cladoniicola, Lichenosticta sp., Merismatium cf. peregrinum, Muellerella lichenicola, Muellerella triseptata, Sphaerellothecium griseae, Stigmidium solenopsoricola und Zwackhiomyces solenopsorae, von denen S. griseae und S. solenopsoricola hier als neue Arten beschrieben werden. Das Verhältnis infizierter Proben zur Gesamtprobenmenge (LF/host ratio) beträgt 0,13 für alle betrachteten Proben. Der höchste Wert der LF/host ratio wurde für Solenopsora grisea und S. holophaea gefunden (0,35 bzw. 0,30). Die höchste Diversität flechtenbewohnender Pilze zeigten Solenopsora candicans und S. grisea. Der häufigste flechtenbewohnende Pilz auf S. candicans war Muellerella lichenicola (85 % der infizierten Wirts-Proben) und Sphaerellothecium griseae auf S. grisea (23 % der infizierten Wirts-Proben). Morphologische Merkmale der untersuchten Proben werden beschrieben und taxonomische Bemerkungen gemacht.

ClNOX1/ClNOXR-mediated MAPK and cAMP-PKA signalling pathways and ROS metabolism are involved in Curvularia lunata sexual reproduction and host infection.

NADPH oxidases (NOXs) and hydrogen peroxide (H2 O2 ) are involved in physiological and pathological processes, and cell fate decisions in organisms. However, regulatory mechanism of NOXs and the role of H2 O2 on fungal sexual reproduction and host infection remain largely unexplored. Here, we identified ROS metabolic genes and key signalling genes of MAPK and cAMP-PKA pathways in Curvularia lunata, which were NOX ClNOX1 and ClNOXR, superoxide dismutase ClSOD1 and catalase ClCAT4, redox-regulated transcription factor ClAP1, Ras small GTPases Clg2P, pheromone-response MAPK ClK1 and cAMP-PKA ClSCHA, and characterized the functions of these genes. The results showed that ClNOX1 localized to the plasma membrane. ClNOX1 and ClNOXR were involved in sexual reproduction and host infection via ClNOX1/ClNOXR-derived H2 O2 as well as MAPK and cAMP-PKA signalling pathways. H2 O2 acted as a signalling molecule to regulate sexual reproduction and host infection in C. lunata.

Topical Fungal Infection Induces Shifts in the Gut Microbiota Structure of Brown Planthopper, Nilaparvata lugens (Homoptera: Delphacidae).

Simple SummaryFungal entomopathogens are important natural enemies of insect pests and widely applied for biocontrol. Gut microbiota play important roles in mediating insect physiology and behavior. There is growing evidence that alteration of gut microbial communities due to pathological and environmental exposure can have detrimental impacts on host health and pathogen resistance. Here, we investigated the effects of topical infection with Metarhizium anisopliae fungus on the gut microbial community structure of the brown planthopper (Nilaparvata lugens, BPH), a destructive insect pest of rice. Our results demonstrated dramatic changes of gut bacterial community structure after topical fungal infection in BPH, as indicated by a significant increase in bacterial load, a significant decrease in bacterial community evenness and significant shifts in dominant bacterial abundance at the taxonomic level below the class. The dysbiosis of the gut bacteria might partly be due to the suppression of gut immunity caused by topical fungal infection. Our results highlighted the importance of the gut microbial community in fungal pathogenesis in insects.The brown planthopper (Nilaparvata lugens, BPH) is a destructive insect pest posing a serious threat to rice production. The fungal entomopathogen Metarhizium anisopliae is a promising alternative that can be used for BPH biocontrol. Recent studies have highlighted the significant involvement of gut microbiota in the insect–fungus interactions. In the presented study, we investigated the effects of topical fungal infection on the gut microbial community structure in BPH. Our results revealed that topical infection with M. anisopliae increased the bacterial load and altered the bacterial community structure in the gut of BPH. The relative abundances of the dominant gut bacteria at the order, family and genus level were significantly different between fungus-infected and uninfected groups. At the genus level, the uninfected BPH harbored high proportions of Pantoea and Enterobacter in the gut, whereas the fungus-infected BPH gut was absolutely dominated by Acinetobacter. Moreover, topical fungal infection significantly inhibited the expressions of immune-related genes encoding anti-microbial protein and dual oxidase that were involved in the maintenance of gut microbiota homeostasis, indicating that gut bacteria imbalance might be attributed in part to the suppression of gut immunity caused by fungal pathogen. Our results highlighted the importance of the gut microbial community during interactions between fungal pathogens and insect hosts.

Host-Pathogen Interactions between Metarhizium spp. and Locusts.

The progress in research on the interactions between Metarhizium spp. and locusts has improved our understanding of the interactions between fungal infection and host immunity. A general network of immune responses has been constructed, and the pathways regulating fungal pathogenicity have also been explored in depth. However, there have been no systematic surveys of interaction between Metarhizium spp. and locusts. The pathogenesis of Metarhizium comprises conidial attachment, germination, appressorial formation, and colonization in the body cavity of the host locusts. Meanwhile, the locust resists fungal infection through humoral and cellular immunity. Here, we summarize the crucial pathways that regulate the pathogenesis of Metarhizium and host immune defense. Conidial hydrophobicity is mainly affected by the contents of hydrophobins and chitin. Appressorial formation is regulated by the pathways of MAPKs, cAMP/PKA, and Ca2+/calmodulin. Lipid droplets degradation and secreted enzymes contributed to fungal penetration. The humoral response of locust is coordinated by the Toll pathway and the ecdysone. The regulatory mechanism of hemocyte differentiation and migration is elusive. In addition, behavioral fever and density-dependent population immunity have an impact on the resistance of hosts against fungal infection. This review depicts a prospect to help us understand host–pathogen interactions and provides a foundation for the engineering of entomopathogenic fungi and the discovery of insecticidal targets to control insect pests.

A novel hexa-segmented dsRNA mycovirus confers hypovirulence in the phytopathogenic fungus Diaporthe pseudophoenicicola.

A novel hexa-segmented double-stranded RNA (dsRNA) mycovirus was isolated and characterized from the filamentous phytopathogenic fungus Diaporthe pseudophoenicicola and was named Diaporthe pseudophoenicicola chrysovirus 1 (DpCV1). The full-length cDNAs of dsRNA1-6 were 3335, 3030, 3039, 2980, 963, and 780bp, respectively. Sequence analysis indicated the presence of nine open reading frames (ORFs) in the DpCV1 genome. ORF1 in dsRNA1 putatively encoded the RNA-dependent RNA polymerase (RdRp) and ORF3 in dsRNA2 encoded a capsid protein (CP). The seven remaining ORFs, ORF2 in dsRNA2, ORF4 in dsRNA3, ORF6, seven in dsRNA4, ORF8 in dsRNA5, and ORF9 in dsRNA6, encoded proteins with unknown functions. Phylogenetic analysis revealed that DpCV1 is closely related to members of the cluster I group within the family Chrysoviridae but formed a separate clade. Importantly, all the six segments of DpCV1 were cured successfully through single spore isolation to obtain the isogenic virus-free strains. DpCV1 can confer hypovirulence to the fungal host of Diaporthe pseudophoenicicola. Compared with the virus-free strain, WC02 harbouring the DpCV1 is more sensitive to fungicide prochloraz. Furthermore, the cell wall of DpCV1 infected strain was loose and enlarged. This is the first report of a hexa-segmented tentative chrysovirus in D. pseudophoenicicola.

Regulatory Roles of Histone Modifications in Filamentous Fungal Pathogens.

Filamentous fungal pathogens have evolved diverse strategies to infect a variety of hosts including plants and insects. The dynamic infection process requires rapid and fine-tuning regulation of fungal gene expression programs in response to the changing host environment and defenses. Therefore, transcriptional reprogramming of fungal pathogens is critical for fungal development and pathogenicity. Histone post-translational modification, one of the main mechanisms of epigenetic regulation, has been shown to play an important role in the regulation of gene expressions, and is involved in, e.g., fungal development, infection-related morphogenesis, environmental stress responses, biosynthesis of secondary metabolites, and pathogenicity. This review highlights recent findings and insights into regulatory mechanisms of histone methylation and acetylation in fungal development and pathogenicity, as well as their roles in modulating pathogenic fungi–host interactions.

Hyperparasitic Fungi on Black Mildews (Meliolales, Ascomycota): Hidden Fungal Diversity in the Tropics.

Hyperparasitism on plant-parasitic fungi is a widespread but rarely studied phenomenon. Here, for the first time, we compile in a checklist information provided by peer-reviewed literature for fungi growing on colonies of black mildews (Meliolales, Ascomycota), a species-rich group of tropical and subtropical plant-parasitic microfungi. The checklist contains information on 189 species of contact-biotrophic microfungi in 82 genera. They belong to seven morphological groups: dematiaceous hyphomycetes, moniliaceous hyphomycetes, pycnidioid, perithecioid, catathecioid, and apothecioid fungi. By the fact that species accumulation curves do not reach saturation for any tropical country, it is evident that the knowledge of the diversity of hyperparasitic fungi on Meliolales is incomplete. A network analysis of records of hyperparasitic fungi, their host fungi and host plants shows that genera of hyperparasitic fungi are generalists concerning genera of Meliolales. However, most species of hyperparasitic fungi are restricted to meliolalean hosts. In addition to hyperparasitic fungi, diverse further microorganisms use meliolalean colonies as ecological niche. Systematic positions of most species are unknown because DNA sequence data are lacking for species of fungi hyperparasitic on Meliolales. We discuss the specific challenges of obtaining DNA sequence data from hyperparasitic fungi. In order to better understand the diversity, evolution and biology of hyperparasitic fungi, it is necessary to increase sampling efforts and to undertake further morphological, molecular, and ecological studies.

Coat protein of partitiviruses isolated from mycorrhizal fungi functions as an RNA silencing suppressor in plants and fungi

Orchid seeds depend on colonization by orchid mycorrhizal (OM) fungi for their germination; therefore, the orchids and OM fungi have long maintained a close relationship (e.g., formation of the hyphal mass structure, peloton) during their evolution. In the present study, we isolated new partitiviruses from OM fungi; partitivirus were separately found in different subcultures from the same fungi. Partitiviruses have been believed to lack an RNA silencing suppressor (RSS), which is generally associated with viral pathogenicity, because most partitiviruses isolated so far are latent in both plants and fungi. However, we found that the coat protein (CP) of our partitiviruses indeed had RSS activity, which differed among the virus isolates from OM fungi; one CP showed RSS activity in both plants and fungi, while another CP showed no activity. The family Partitiviridae include viruses isolated from plants and fungi, and it has been suggested that these viruses may occasionally be transmitted between plant and fungal hosts. Given that there are several reports showing that viruses can adapt to nonhost using strong RSS, we here discussed the idea that partitiviruses may be better able to migrate between the orchid and fungus probably through the pelotons formed in the orchid cells, if host RNA silencing is suppressed by partitivirus RSS.

Mode of action of potential biocontrol agents against Fusarium species and Cochliobolus sativus

ABSTRACT The antagonistic activities of three potential biocontrol agents (BCAs), Clonostachys miodochialis, C. rosea, and Minimedusa polyspora, were tested in vitro against the target fungi Fusarium acuminatum, F. avenaceum, F. equiseti, F. graminearum, and Cochliobolus sativus. In vitro dual-culture assay revealed that the BCAs were able to inhibit the mycelial growth of most of the target fungi, with the most common mode of action being mycoparasitism. After 5 d of incubation, depending on the host-parasite interface, biotrophic mycoparasitism was observed via attachment structures on F. avenaceum, F. equiseti, and C. sativus. Haustoria and appressoria were formed by C. miodochialis and M. polyspora on F. avenaceum and C. sativus, respectively. Clamp-like structures were also produced by both BCAs, depending on the host fungus. Clonostachys rosea formed only contact points on its hosts. Coiling structure was observed with C. rosea and M. polyspora occasionally in pure culture but more abundantly in the dual-culture assay. After an additional 3 to 5 d of incubation, the BCAs started damaging their host cells. Asexual fruiting bodies of the BCAs spread necrotrophically on F. acuminatum and F. graminearum and began destroying their mycelia after the initial 5 d of incubation. Furthermore, mycelial dissolution of F. acuminatum by an excreted substance was observed at a distance before direct contact with C. miodochialis. The diffuse metabolite assay revealed that the highest inhibition of the three BCAs was on C. sativus and their least effectiveness was observed on F. graminearum. Overall, these results provide evidence that C. miodochialis, C. rosea, and M. polyspora are potential candidates for biological control of the tested target fungi. This constitutes the first report that these three BCAs are able to establish an initial biotrophic relationship followed by a necrotrophic lifestyle. In addition, C. miodochialis has not previously been reported as a BCA.

Assessing the Old-Growth Dependency of Two Saproxylic Beetle Species in the Southern Appalachian Mountains

AbstractThe southern Appalachian Mountains were intensively logged during the early 1900s, leaving little remaining old-growth forest. Much of the region is now second-growth forest, which may not be suitable to specialist saproxylic species. Moreover, if suitable habitat exists, poorly dispersing species may not be able to colonize it. To investigate this, we assessed the distribution and old-growth dependency of two low-mobility saproxylic beetles in Great Smoky Mountains National Park. Using both field surveys and community science data, we found Megalodacne heros (Say 1823) (Coleoptera: Erotylidae) to be limited to lower elevations regardless of disturbance history, while Phellopsis obcordata (Kirby 1837) (Coleoptera: Zopheridae) was restricted to inside or near old-growth forests. Although trees were generally smaller in second-growth, we detected no habitat limitation for P. obcordata: fungal hosts were present in second-growth areas and the beetle was present on trees as small as 11 cm in diameter. This suggests its distribution is shaped by its low dispersal capability and need for temporal continuity of deadwood habitat, therefore qualifying it as an indicator species. For P. obcordata, old-growth acted as refugia during landscape-wide, anthropogenic disturbances in the early 1900s, though we can draw no conclusions about M. heros from our dataset. The difference in sensitivity to human disturbance displayed between species may be linked to their relative dispersal abilities: P. obcordata is entirely flightless while M. heros is capable of some flight. This study highlights the value of using saproxylic invertebrates with limited dispersal ability for assessing impacts from anthropogenic forest disturbances.

Wild herbivorous mammals (genus Neotoma) host a diverse but transient assemblage of fungi.

Fungi are often overlooked in microbiome research and, as a result, little is known about the mammalian mycobiome. Although frequently detected in vertebrate guts and known to contribute to digestion in some herbivores, whether these eukaryotes are a persistent part of the mammalian gut microbiome remains contentious. To address this question, we sampled fungi from wild woodrats (Neotoma spp.) collected from 25 populations across the southwestern United States. For each animal, we collected a fecal sample in the wild, and then re-sampled the same individual after a month in captivity on a controlled diet. We characterized and quantified fungi using three techniques: ITS metabarcoding, shotgun metagenomics and qPCR. Wild individuals contained diverse fungal assemblages dominated by plant pathogens, widespread molds, and coprophilous taxa primarily in Ascomycota and Mucoromycota. Fungal abundance, diversity and composition differed between individuals, and was primarily influenced by animal geographic origin. Fungal abundance and diversity significantly declined in captivity, indicating that most fungi in wild hosts came from diet and environmental exposure. While this suggests that these mammals lack a persistent gut mycobiome, natural fungal exposure may still impact fungal dispersal and animal health.