Search

Tra1 is an essential coactivator protein of the yeast SAGA and NuA4 acetyltransferase complexes that regulate gene expression through multiple mechanisms including the acetylation of histone proteins. Tra1 is a pseudokinase of the PIKK family characterized by a C-terminal PI3K domain with no known kinase activity. However, mutations of specific arginine residues to glutamine in the PI3K domains (an allele termed tra1Q3) result in reduced growth and increased sensitivity to multiple stresses. In the opportunistic fungal pathogen Candida albicans , the tra1Q3 allele reduces pathogenicity and increases sensitivity to the echinocandin antifungal drug caspofungin, which disrupts the fungal cell wall. Here, we found that compromised Tra1 function, in contrast to what is seen with caspofungin, increases tolerance to the azole class of antifungal drugs, which inhibits ergosterol synthesis. In C. albicans , tra1Q3 increases the expression of genes linked to azole resistance, such as ERG11 and CDR1. CDR1 encodes a multidrug ABC transporter associated with efflux of multiple xenobiotics, including azoles. Consequently, cells carrying tra1Q3 show reduced intracellular accumulation of fluconazole. In contrast, a tra1Q3 Saccharomyces cerevisiae strain displayed opposite phenotypes: decreased tolerance to azole, decreased expression of the efflux pump PDR5 , and increased intracellular accumulation of fluconazole. Therefore, our data provide evidence that Tra1 differentially regulates the antifungal response across yeast species. [ABSTRACT FROM AUTHOR]

To complete its parasitic lifecycle, Salmacisia buchloëana , a biotrophic fungus, manipulates reproductive organ development, meristem determinacy, and resource allocation in its dioecious plant host, buffalograss (Bouteloua dactyloides ; Poaceae). To gain insight into S. buchloëana's ability to manipulate its host, we sequenced and assembled the 20.1 Mb genome of S. buchloëana into 22 chromosome-level pseudomolecules. Phylogenetic analysis suggests that S. buchloëana is nested within the genus Tilletia and diverged from Tilletia caries and Tilletia walkeri ∼ 40 MYA. We find that S. buchloëana contains a novel chromosome arm with no syntenic relationship to other publicly available Tilletia genomes, and that genes on the novel arm are upregulated upon infection, suggesting that this unique chromosomal segment may have played a critical role in S. buchloëana's evolution and host specificity. Salmacisia buchloëana has one of the largest fractions of serine peptidases (1.53% of the proteome) and one of the highest GC contents (62.3%) in all classified fungi. Analysis of codon base composition indicated that GC content is controlled more by selective constraints than directional mutation, and that S. buchloëana has a unique bias for the serine codon UCG. Finally, we identify 3 inteins within the S. buchloëana genome, 2 of which are located in a gene often used in fungal taxonomy. The genomic and transcriptomic resources generated here will aid plant pathologists and breeders by providing insight into the extracellular components contributing to sex determination in dioecious grasses. [ABSTRACT FROM AUTHOR]

White-nose disease (WND), caused by the psychrophilic fungus Pseudogymnoascus destructans, represents one of the greatest threats for North American hibernating bats. Research on molecular data has significantly advanced our knowledge of various aspects of the disease, yet more studies are needed regarding patterns of P. destructans genetic diversity distribution. In the present study, we investigate three sites within the native range of the fungus in detail: two natural hibernacula (karst caves) in Bulgaria, south-eastern Europe and one artificial hibernaculum (disused cellar) in Germany, northern Europe, where we conducted intensive surveys between 2014 and 2019. Using 18 microsatellite and two mating type markers, we describe how P. destructans genetic diversity is distributed between and within sites, the latter including differentiation across years and seasons of sampling; across sampling locations within the site; and between bats and hibernaculum walls. We found significant genetic differentiation between hibernacula, but we could not detect any significant differentiation within hibernacula, based on the variables examined. This indicates that most of the pathogen's movement occurs within sites. Genotypic richness of P. destructans varied between sites within the same order of magnitude, being approximately two times higher in the natural caves (Bulgaria) compared to the disused cellar (Germany). Within all sites, the pathogen's genotypic richness was higher in samples collected from hibernaculum walls than in samples collected from bats, which corresponds with the hypothesis that hibernacula walls represent the environmental reservoir of the fungus. Multiple pathogen genotypes were commonly isolated from a single bat (i.e. from the same swab sample) in all study sites, which might be important to consider when studying disease progression. [ABSTRACT FROM AUTHOR]

Background: Cryptococcosis and cryptococcal meningitis, caused by Cryptococcus neoformans infections, lead to approximately 180,000 deaths per year, primarily in developing countries. Individuals with compromised immune systems, e.g., due to HIV infection (AIDS) or chemotherapy, are particularly vulnerable. Conventional treatment options are often limited and can cause severe side effects. Therefore, this study aimed to investigate the antifungal effect of insect-derived proline-rich antimicrobial peptides (PrAMPs) against C. neoformans. These peptides are known for their low toxicity and their high efficacy in murine infection models, making them a promising alternative for treatment. Results: A preliminary screening of the minimal inhibitory concentrations (MICs) of 20 AMPs, including the well-known PrAMPs Onc112, Api137, and Chex1Arg20 as well as the cathelicidin CRAMP against the C. neoformans strains 1841, H99, and KN99α revealed promising results, with MICs as low as 1.6 μmol/L. Subsequent investigations of selected peptides, determining their influence on fungal colonyforming units, confirmed their strong activity. The antifungal activity was affected by factors such as peptide net charge and sequence, with stronger effects at higher net charges probably due to better intracellular uptake confirmed by confocal laser scanning microscopy. Inactive scrambled peptides suggest a specific intracellular target, although scanning electron microscopy showed that PrAMPs also damaged the cell exterior for a low proportion of the cells. Possible pore formation could facilitate entry into the cytosol. [ABSTRACT FROM AUTHOR]

In February 2023 in Japan, seedlings of wax gourd [Benincasa hispida (Thunb.) Cogn.] in seedling boxes developed water-soaked rot on hypocotyls typical of damping off and eventually died. A Pythium sp. was isolated from the hypocotyls and was identified as Pythium aphanidermatum based on morphological characteristics and DNA sequence data. The same symptoms developed after seedlings were inoculated with the isolated fungus, and the same fungus was reisolated from the seedlings. This is the first report of damping off in wax gourd seedlings caused by P. aphanidermatum in seedling boxes in Japan. [ABSTRACT FROM AUTHOR]

The recent emergence of the fungus Cryptostroma corticale in Seattle, Washington, USA is concerning because of its invasive status in Europe, where it causes sooty bark disease of maples (SBD), killing both forest and urban trees. Additionally, the fruiting bodies (stromata) of C. corticale produce allergenic spores that can impact human health. Therefore, its presence in Washington has the potential to impact the state's populace and trees, including the native bigleaf maple, Acer macrophyllum. To better determine the distribution of C. corticale in western Washington, A. macrophyllum was surveyed on 50 Washington State Park properties. Bark samples were collected from trees with fruiting bodies resembling those of C. corticale and cores were removed from mature asymptomatic trees. The presence of C. corticale was confirmed using nested PCR. Acer macrophyllum with fruiting bodies of C. corticale were well distributed throughout western Washington, with an 89% detection rate at surveyed Park properties containing A. macrophyllum. Core samples indicated that latent infections appeared well distributed and common throughout western Washington on mature A. macrophyllum, with a 91% detection rate at surveyed Park properties containing A. macrophyllum and a 68% detection rate on cored trees. Signs of SBD and evidence of latent infections of C. corticale were widespread at surveyed properties. However, signs of SBD were rare on individual trees and the overall condition of A. macrophyllum was considered vigorous at the majority of Park properties. Awareness of C. corticale, SBD, and their likelihood to be present on a property containing A. macrophyllum in western Washington should help inform management actions and demonstrate the need for future research. [ABSTRACT FROM AUTHOR]

Acacia crassicarpa is a tropical tree species native to Australia, West Papua and Papua New Guinea, which has been widely used to establish plantations in the lowland humid tropics of Sumatra and Kalimantan. These trees, able to grow on sites having relatively poor nutrition, have been relatively free of serious disease problems. A rust disease infecting the phyllodes of A. crassicarpa has recently been encountered in plantations in various areas of Indonesia and Malaysia where they are not native. In this study, the rust was collected and identified as Endoraecium violae‐faustiae using DNA sequence analysis based on the internal transcribed spacer (ITS) region of the rDNA. This is the first record of the rust outside its native range. Damage at present appears to be relatively mild but the pathogen could become important in the future. [ABSTRACT FROM AUTHOR]

Abstract Social insects are prone to pathogen infection because of high exposure rates from social interactions. However, it remains unclear whether queens have enhanced pathogen resistance, because reproduction is largely confined to queens. Here, we used a natural host–pathogen system, the subterranean termite Reticulitermes chinensis and the entomopathogenic fungus Metarhizium anisopliae, to investigate the differences in allogrooming, locomotion, and immune gene expression between queens and workers against pathogen infection. We found that fungal infection significantly reduced survival in both queens and workers. Infected queens received significantly more grooming time from sanitary nestmates than infected workers, but they returned much less grooming time to sanitary nestmates than infected workers. Infection resulted in a reduction in the average locomotion speed and distance of queens but had no effect on worker locomotion. Infection resulted in upregulated expression of two immune genes (termicin and transferrin), two antioxidant genes (CAT and SOD), and phosphate genes CYP450 in queens but not in workers. Our results indicated that eusocial termites evolved strategies that prioritize the reproductive castes' welfare in defending against the pathogen infection to ensure continued reproduction and colony persistence.

ABSTRACTAlthough fungi have been important model organisms for solving genetic, molecular, and ecological problems, recently, they are also becoming an important source of infectious disease. Despite their high medical burden, fungal pathogens are understudied, and relative to other pathogenic microbes, less is known about how their gene functions contribute to disease. This is due, in part, to a lack of powerful genetic tools to study these organisms. In turn, this has resulted in inappropriate treatments and diagnostics and poor disease management. There are a variety of reasons genetic studies were challenging in pathogenic fungi, but in recent years, most of them have been overcome or advances have been made to circumvent these barriers. In this minireview, we highlight how recent advances in genetic studies in fungal pathogens have resulted in the discovery of important biology and potential new antifungals and have created the tools to comprehensively study these important pathogens.

ABSTRACTCandida albicans, one of the most prevalent human fungal pathogens, causes diverse diseases extending from superficial infections to deadly systemic mycoses. Currently, only three major classes of antifungal drugs are available to treat systemic infections: azoles, polyenes, and echinocandins. Alarmingly, the efficacy of these antifungals against C. albicans is hindered both by basal tolerance toward the drugs and the development of resistance mechanisms such as alterations of the drug’s target, modulation of stress responses, and overexpression of efflux pumps. Thus, the need to identify novel antifungal strategies is dire. To address this challenge, we screened 3,049 structurally-diverse compounds from the Boston University Center for Molecular Discovery (BU-CMD) chemical library against a C. albicans clinical isolate and identified 17 molecules that inhibited C. albicans growth by >80% relative to controls. Among the most potent compounds were CMLD013360, CMLD012661, and CMLD012693, molecules representing two distinct chemical scaffolds, including 3-hydroxyquinolinones and a xanthone natural product. Based on structural insights, CMLD013360, CMLD012661, and CMLD012693 were hypothesized to exert antifungal activity through metal chelation. Follow-up investigations revealed all three compounds exerted antifungal activity against non-albicans Candida, including Candida auris and Candida glabrata, with the xanthone natural product CMLD013360 also displaying activity against the pathogenic mould Aspergillus fumigatus. Media supplementation with metallonutrients, namely ferric or ferrous iron, rescued C. albicans growth, confirming these compounds act as metal chelators. Thus, this work identifies and characterizes two chemical scaffolds that chelate iron to inhibit the growth of the clinically relevant fungal pathogen C. albicansIMPORTANCEThe worldwide incidence of invasive fungal infections is increasing at an alarming rate. Systemic candidiasis caused by the opportunistic pathogen Candida albicans is the most common cause of life-threatening fungal infection. However, due to the limited number of antifungal drug classes available and the rise of antifungal resistance, an urgent need exists for the identification of novel treatments. By screening a compound collection from the Boston University Center for Molecular Discovery (BU-CMD), we identified three compounds representing two distinct chemical scaffolds that displayed activity against C. albicans. Follow-up analyses confirmed these molecules were also active against other pathogenic fungal species including Candida auris and Aspergillus fumigatus. Finally, we determined that these compounds inhibit the growth of C. albicans in culture through iron chelation. Overall, this observation describes two novel chemical scaffolds with antifungal activity against diverse fungal pathogens.

The fungal pathogen Zymoseptoria tritici is the causal agent of Septoria tritici blotch (STB), a major wheat disease in Western Europe. Microorganisms inhabiting wheat leaves might act as beneficial, biocontrol, or facilitating agents that could limit or stimulate the development of Z. tritici. Improving our understanding of microbial communities in the wheat phyllosphere would lead to new insights into STB management. This article provides fungal and bacterial metabarcoding datasets obtained by sampling wheat leaves with and without symptoms caused by Z. tritici. Tissues were sampled from three commercial wheat varieties on three sampling dates during a cropping season. Weeds around wheat fields were sampled as well. In total, more than 450 leaf samples were collected. The pathogen Z. tritici was quantified using quantitative PCR. We provide the raw metabarcoding datasets, the amplicon sequence variant tables obtained after bioinformatic processing, the metadata associated with each sample (sampling date, wheat variety and tissue health condition), a preliminary descriptive analysis of the data, and the code used for bioinformatic and descriptive statistical analysis. [Graphic: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Red leaf blotch is a severe soybean disease caused by the select agent fungal pathogen Coniothyrium glycines. With little information about the pathogen, no molecular detection methods are available. Here, we report genome assemblies of five isolates, including reference isolates. These datasets provide a valuable resource for ongoing studies that include marker development, diagnostics, and population diversity. [Figure: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

The conjunctiva has immune-responsive properties to protect the eye from infections. Its innate immune system reacts against external pathogens, such as fungi. The complement factor C5a is an important contributor to the initial immune response. It is known that activation of transient-receptor-potential-vanilloid 1 (TRPV1) and TRP-melastatin 8 (TRPM8) channels is involved in different immune reactions and inflammation in the human body. The aim of this study was to determine if C5a and mucor racemosus e voluminae cellulae (MR) modulate Ca2+-signaling through changes in TRPs activity in human conjunctival epithelial cells (HCjECs). Furthermore, crosstalk was examined between C5a and MR in mediating calcium regulation. Intracellular Ca2+-concentration ([Ca2+]i) was measured by fluorescence calcium imaging, and whole-cell currents were recorded using the planar-patch-clamp technique. MR was used as a purified extract. Application of C5a (0.05–50 ng/mL) increased both [Ca2+]i and whole-cell currents, which were suppressed by either the TRPV1-blocker AMG 9810 or the TRPM8-blocker AMTB (both 20 µM). The N-terminal peptide C5L2p (20–50 ng/mL) blocked rises in [Ca2+]i induced by C5a. Moreover, the MR-induced rise in Ca2+-influx was suppressed by AMG 9810 and AMTB, as well as 0.05 ng/mL C5a. In conclusion, crosstalk between C5a and MR controls human conjunctival cell function through modulating interactions between TRPV1 and TRPM8 channel activity.

The plethora of genome sequences produced in the postgenomic age has not resolved many of our most pressing biological questions. Correlating gene expression with an interrogatable and easily observable characteristic such as the surrogate phenotype conferred by a reporter gene is a valuable approach to gaining insight into gene function. Many reporters including lacZ, amdS, and the fluorescent proteins mRuby3 and mNeonGreen have been used across all manners of organisms. Described here is an investigation into the creation of a robust, synthetic, fusion reporter system for Cryptococcus neoformans that combines some of the most useful fluorophores available in this system with the versatility of the counter-selectable nature of amdS. The reporters generated include multiple composition and orientation variants, all of which were investigated for differences in expression. Evaluation of known promoters from the TEF1 and GAL7 genes was undertaken, elucidating novel expression tendencies of these biologically relevant C. neoformans regulators of transcription. Smaller than lacZ but providing multiple useful surrogate phenotypes for interrogation, the fusion ORF serves as a superior whole-cell assay compared to traditional systems. Ultimately, the work described here bolsters the array of relevant genetic tools that may be employed in furthering manipulation and understanding of the WHO fungal priority group pathogen C. neoformans.

Fusarium head blight (FHB), caused by the hemibiotrophic fungus Fusarium graminearum, is one of the major threats to global wheat productivity. A wheat pore-forming toxin-like (PFT) protein was previously reported to underlie Fhb1, the most widely used quantitative trait locus in FHB breeding programs worldwide. In the present work, wheat PFT was ectopically expressed in the model dicot plant Arabidopsis. Heterologous expression of wheat PFT in Arabidopsis provided a broad-spectrum quantitative resistance to fungal pathogens including F. graminearum, Colletotrichum higginsianum, Sclerotinia sclerotiorum, and Botrytis cinerea. However, there was no resistance to bacterial or oomycete pathogens Pseudomonas syringae and Phytophthora capsici, respectively in the transgenic Arabidopsis plants. To explore the reason for the resistance response to, exclusively, the fungal pathogens, purified PFT protein was hybridized to a glycan microarray having 300 different types of carbohydrate monomers and oligomers. It was found that PFT specifically hybridized with chitin monomer, N-acetyl glucosamine (GlcNAc), which is present in fungal cell walls but not in bacteria or oomycete species. This exclusive recognition of chitin may be responsible for the specificity of PFT-mediated resistance to fungal pathogens. Transfer of the atypical quantitative resistance of wheat PFT to a dicot system highlights its potential utility in designing broad-spectrum resistance in diverse host plants. [Graphic: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

BackgroundCryptococcosis and cryptococcal meningitis, caused by Cryptococcus neoformans infections, lead to approximately 180,000 deaths per year, primarily in developing countries. Individuals with compromised immune systems, e.g., due to HIV infection (AIDS) or chemotherapy, are particularly vulnerable. Conventional treatment options are often limited and can cause severe side effects. Therefore, this study aimed to investigate the antifungal effect of insect-derived proline-rich antimicrobial peptides (PrAMPs) against C. neoformans. These peptides are known for their low toxicity and their high efficacy in murine infection models, making them a promising alternative for treatment.ResultsA preliminary screening of the minimal inhibitory concentrations (MICs) of 20 AMPs, including the well-known PrAMPs Onc112, Api137, and Chex1Arg20 as well as the cathelicidin CRAMP against the C. neoformans strains 1841, H99, and KN99α revealed promising results, with MICs as low as 1.6 μmol/L. Subsequent investigations of selected peptides, determining their influence on fungal colony-forming units, confirmed their strong activity. The antifungal activity was affected by factors such as peptide net charge and sequence, with stronger effects at higher net charges probably due to better intracellular uptake confirmed by confocal laser scanning microscopy. Inactive scrambled peptides suggest a specific intracellular target, although scanning electron microscopy showed that PrAMPs also damaged the cell exterior for a low proportion of the cells. Possible pore formation could facilitate entry into the cytosol.

Ailanthus altissima is among the most invasive woody species worldwide, outcompeting native trees. The fungus Verticillium nonalfalfae (VN) is promising for A. altissima biocontrol, and its effects on the host have been studied via visual assessment in a range of host species. However, little research was performed to address fungal effects on the physiological processes of non-target woody plants. We investigated the occurrence of visual and non-visually recognisable perturbations of VN infection on potted vines to evaluate the potential risks of the biocontrol pathogen on viticulture. Eighteen four-years-old Vitis vinifera (cultivar Grüner Veltliner grafted on Kober 5BB) potted plants were inoculated with VN conidial suspension of the fungus (F), while nine plants were treated with sterile water (C, control). Disease symptoms and physiological parameters were monitored throughout the experiment (seven evaluation dates), while leaf water potential, leaf mass per area (LMA) and biomass were measured at the end of the study when plant tissue was sampled for re-isolation of the fungus. In our trial, inoculations with VN induced characteristic wilting symptoms only in Ailanthus (used as side control of the inoculum), while vines remained asymptomatic, thus indicating a high degree of host specificity of VN. Limited or no impact was detected on the physiology of the non-target V. vinifera. Furthermore, the LMA and biomass measured in the two experimental groups were not different. Although fungal colonisation induced vascular discolouration in both species, the fungus could only be re-isolated from dying Ailanthus but not from vine tissue. Results suggest that V. vinifera cv Grüner Veltliner is resistant to the applied VN isolate. However, the susceptibility and physiology of additional grapevine cultivars, as well as other native woody species to VN, should be studied before promoting large-scale use of the biocontrol agent.

ABSTRACTInnate immune cells, especially neutrophils, play key roles in protecting against infection by Candida albicans and other fungal pathogens. A distinct aspect of neutrophils compared to other phagocytes is that they make much higher levels of myeloperoxidase, which converts H2O2 generated during the oxidative burst into highly reactive hypochlorous acid (HOCl). The effects of HOCl on C. albicans are not well defined, so comparative studies were used to show that HOCl kills C. albicans at lower doses than H2O2 and in a manner that correlates with permeabilization of the plasma membrane. Mutants with defects in plasma membrane organization and antioxidant pathways showed differential sensitivity to these oxidants, consistent with their distinct chemical properties. Transcriptional responses to HOCl and H2O2 were also quite different. Although they induced a common set of genes, most differentially regulated genes were unique. Testing the roles of genes that were predicted to be important for counteracting the types of thiol oxidation that are preferentially caused by HOCl showed that Mxr1 (methionine-S-sulfoxide reductase), Srx1 (sulfiredoxin that reduces cysteine-sulfinic acid groups), and Trx1 (thioredoxin that acts to reduce disulfide bonds) all promoted resistance to HOCl. Altogether, these results suggest that an effective way to help promote the killing of C. albicans by neutrophils would be to perturb the C. albicans plasma membrane, rather than targeting one specific antioxidant pathway.IMPORTANCEHypochlorous acid (HOCl), commonly known as bleach, is generated during the respiratory burst by phagocytes and is a key weapon used to attack Candida albicans and other microbial pathogens. However, the effects of hypochlorous acid on C. albicans have been less well studied than H2O2, a different type of oxidant produced by phagocytes. HOCl kills C. albicans more effectively than H2O2 and results in disruption of the plasma membrane. HOCl induced a very different transcriptional response than H2O2, and there were significant differences in the susceptibility of mutant strains of C. albicans to these oxidants. Altogether, these results indicate that HOCl has distinct effects on cells that could be targeted in novel therapeutic strategies to enhance the killing of C. albicans and other pathogens.

The insect pathogenic fungus, Ascosphaera apis, is the causative agent of honeybee chalk brood disease. Amylases are secreted by many plant pathogenic fungi to access host nutrients through the metabolism of starch, and the identification of new amylases can have important biotechnological applications. Production of amylase by A. apis in submerged culture was optimized using the response surface method (RSM). Media composition was modeled using Box–Behnken design (BBD) at three levels of three variables, and the model was experimentally validated to predict amylase activity (R2 = 0.9528). Amylase activity was highest (45.28 ± 1.16 U/mL, mean ± SE) in media composed of 46 g/L maltose and1.51 g/L CaCl2 at a pH of 6.6, where total activity was ~11-fold greater as compared to standard basal media. The enzyme was purified to homogeneity with a 2.5% yield and 14-fold purification. The purified enzyme had a molecular weight of 75 kDa and was thermostable and active in a broad pH range (> 80% activity at a pH range of 7–10), with optimal activity at 55 °C and pH = 7.5. Kinetic analyses revealed a Km of 6.22 mmol/L and a Vmax of 4.21 μmol/mL·min using soluble starch as the substrate. Activity was significantly stimulated by Fe2+ and completely inhibited by Cu2+, Mn2+, and Ba2+ (10 mM). Ethanol and chloroform (10% v/v) also caused significant levels of inhibition. The purified amylase essentially exhibited activity only on hydrolyzed soluble starch, producing mainly glucose and maltose, indicating that it is an endo-amylase (α-amylase). Amylase activity peaked at 99.38 U/mL fermented in a 3.7 L-bioreactor (2.15-fold greater than what was observed in flask cultures). These data provide a strategy for optimizing the production of enzymes from fungi and provide insight into the α-amylase of A. apis. [ABSTRACT FROM AUTHOR]

This study presents the first report of a Fusarium wilt in Cymbopogon citratus in Portugal. Fusarium is one of the most prominent genera of plant pathogens due to its wide range of hosts and mycotoxin production. The Fusarium fujikuroi species complex (FFSC) encompasses several threatening known plant pathogens. Cymbopogon citratus is a broadly distributed aromatic and medicinal plant rich in bioactive volatiles, which are relevant to several industries. The primary goal of this study was to identify and characterize the Fusarium fujikuroi species putatively responsible for the observed wilt. This report displays symptomatologic, cultural, morphologic, genetic, and cytogenomic characteristics associated with this fungus and disease. The cultural features included flat, white-colored colonies with filiform margins. Additionally, these colonies displayed abundant cottony aerial mycelia at the upper surface and orange-violet color at the lower surface. On Carnation Leaf-Piece Agar, septate fusoid macroconidia were present, displaying a flattened tapering toward the basal part and a number of septa ranging from one to four. The comparison between the amplified and aligned ITS sequences revealed 100% similarity between the isolate and the FFSC. Finally, a flow cytometry assay revealed an estimated genome size of 29.9 Mbp. This finding contrasts with other known pathogens from the Fusarium fujikuroi species complex. Ultimately, novel pathogens might be uncovered by exploring the mycobiome of diseased Cymbopogon citratus plants. [ABSTRACT FROM AUTHOR]

Calonectria pseudoreteaudii is an important causal agent of Eucalyptus leaf blight in southern China. This pathogen causes Eucalyptus tree disease across numerous regions in southern China. In addition to diseased leaves, C. pseudoreteaudii has occasionally been isolated from soil in Eucalyptus plantations. The aim of this study was to clarify whether C. pseudoreteaudii causing Eucalyptus leaf blight in China is mainly clonally reproduced and to determine the potential spreading mechanism of C. pseudoreteaudii between diseased leaves and soil. To this end, 10 polymorphic microsatellite markers were analyzed to detect the genetic diversity of 97 C. pseudoreteaudii isolates from diseased leaves and soil in a Eucalyptus plantation in Guangxi Zhuang Autonomous Region, southern China. The analysis showed that the genetic diversity of the isolates from both the diseased leaves and soil was high. However, the gene and genotype diversity of the C. pseudoreteaudii isolates from diseased leaves were higher than those of the isolates from the soil. Moreover, all genotypes detected in the isolates from the soil were also found in the isolates from the diseased leaves. Structural analyses did not show clear population structures related to the population substrates of the diseased leaves or soil, and molecular variance analyses indicated that no significant genetic differentiation existed between the diseased leaf and soil populations. These results suggest that C. pseudoreteaudii in soil spreads from diseased leaves, and that an asexual cycle is the primary reproductive mode in both diseased leaf and soil populations. This is the first study on the genetic diversity and population structure of C. pseudoreteaudii. The high genetic diversity and spread pathways of this pathogen may pose challenges in controlling the disease. C. pseudoreteaudii from both diseased leaves and soils in Eucalyptus plantations needs to be carefully monitored for disease control and management. [ABSTRACT FROM AUTHOR]

Sorghum is the fifth most important cereal crop in the world. There is a high potential for sorghum production in Indonesia because of its drought tolerance and wide adaptability. Sorghum cultivation is constrained in Indonesia by pests and diseases. One such disease is tar-spot caused by the Phyllachora sp., an obligate fungal pathogen. Tar spot disease of sorghum has not been widely reported. This study reports the finding of Phyllachora species based on the morphology and molecular identification of tar spot on leaves of sorghum plants of Latu Monca and Latu Kaca local varieties.The pathogen was identified as Phyllachora species. This is the first report of tar spots of sorghum caused by Phyllachora in West Lombok Regency, West Nusa Tenggara Province, Indonesia. [ABSTRACT FROM AUTHOR]

The soil fungal community plays an important role in forest ecosystems and is crucially influenced by forest secondary succession. However, the driving factors of fungal community and function during temperate forest succession and their potential impact on succession processes remain poorly understood. In this study, we investigated the dynamics of the soil fungal community in three temperate forest secondary successional stages (shrublands, coniferous forests, and deciduous broad‐leaved forests) using high‐throughput DNA sequencing coupled with functional prediction via the FUNGuild database. We found that fungal community richness, α‐diversity, and evenness decreased significantly during the succession process. Soil available phosphorus and nitrate nitrogen decreased significantly after initial succession occurred, and redundancy analysis showed that both were significant predictors of soil fungal community structure. Among functional groups, fungal saprotrophs and pathotrophs represented by plant pathogens were significantly enriched in the early‐successional stage, while fungal symbiotrophs represented by ectomycorrhiza were significantly increased in the late‐successional stage. The abundance of both saprotroph and pathotroph fungal guilds was positively correlated with soil nitrate nitrogen and available phosphorus content. Ectomycorrhizal fungi were negatively correlated with nitrate nitrogen and available phosphorus content and positively correlated with ammonium nitrogen content. These results indicate that the dynamics of fungal community and function reflected the changes in nitrogen and phosphorus availability caused by the secondary succession in temperate forests. The fungal plant pathogen accumulated in the early‐successional stage and ectomycorrhizal fungi accumulated in the late‐successional stage may have a potential role in promoting forest succession. These findings contribute to a better understanding of the response of soil fungal communities to secondary forest succession and highlight the importance of fungal communities during the successional process. [ABSTRACT FROM AUTHOR]

The Eucalyptus plantation industry in Indonesia has expanded rapidly during the last few decades. During routine nursery disease surveys, symptoms of a leaf and shoot blight disease were detected on Eucalyptus mother plants. Isolates were obtained from symptomatic tissues and identified using DNA sequence analyses. Phylogenetic analyses showed that the isolates were those of Quambalaria eucalypti. Pathogenicity tests were conducted with isolates of Q. eucalypti on clones of E. pellita and E. grandis × E. pellita hybrids. These resulted in symptoms similar to those observed on naturally infected plants. Eucalyptus genotypes tested showed variation in their susceptibility, highlighting the potential to select and breed for resistance and thus to manage future outbreaks of the disease. This is the first report of the pathogen in Indonesia as well as in Southeast Asia. [ABSTRACT FROM AUTHOR]

Infectious diseases caused by invasive, environmentally persistent fungal pathogens have increasingly endangered global biodiversity, yet disease management remains a major conservation challenge. A prominent example is white‐nose syndrome (WNS), a disease caused by the invasive fungal pathogen Pseudogymnoascus destructans (Pd) that has devastated populations of multiple North American bat species, but for which few effective management tools exist. Here, we propose that strategies to delay environmental transmission of Pd during early winter could limit WNS disease effects across winter, benefitting bats. We used a small captive experiment and a multi‐year field trial on wild, free‐ranging bats to assess an environmental control strategy to manage Pd within its environmental reservoir in Pennsylvania, USA, where the pathogen has become endemic. The strategy centers on the application of Polyethylene Glycol 8000 (PEG) to roost substrates in summer, prior to bat hibernation, as a means to disrupt environmental transmission to bats in early winter. In the captive experiment, environmental transmission of Pd to immunologically naïve little brown myotis (Myotis lucifugus) occurred from roost substrates inoculated with Pd, but the application of PEG to these substrates effectively blocked this transmission. In the field trial, Pd load and infection extent both declined substantially in free‐ranging M. lucifugus after treatment relative to controls, with declines exceeding effects of inter‐site and inter‐annual variation. Pathogen prevalence and load also declined substantially after PEG treatment in big brown bats (Eptesicus fuscus). No negative effects of PEG treatment were observed in body condition or colony counts of bats or in the microbial community. Together, these results are consistent with effective environmental control of Pd and reduced WNS disease effects in bats within contaminated hibernacula. The results also highlight the potential of carefully designed environmental control strategies for managing environmentally persistent pathogens. [ABSTRACT FROM AUTHOR]

In August 2017, severe powdery mildew was observed on Potentilla indica in Minhe, Qinghai province, China. The identification of fungal species was performed by morphological study using an optical microscope and scanning electron microscope (SEM), supplemented by sequence analysis of the internal transcribed spacer (ITS) region of rDNA. Based on a combination of morphology, ITS sequence, and host, the powdery mildew on Potentilla indica was identified as Erysiphe thuemenii, a species belonging to the E. aquilegiae complex. This is the first report of E. thuemenii causing powdery mildew on a new host species, P. indica, in China and also worldwide. [ABSTRACT FROM AUTHOR]

Background: Pinellia ternata (Thunb.), a perennial herbal plant in the Araceae family, has great medicinal value and market demand. In August 2020, an outbreak of severe leaf spot blight disease resulted in a huge yield loss of P. ternata. It is necessary to isolate and identify the pathogens that cause spot blight on P. ternata. Methods: In this study, we isolated and identified the pathogens by fulfilling Koch’s postulates. Disease samples with typical spot blight symptoms were collected and pathogens were isolated from the diseased tissues. The pathogen was identified based on its biological characteristics and molecular analysis of internal transcribed (rDNA-ITS) and large subunit (LSU) sequences. Phylogenetic tree were constructed using MEGA7 software and pathogenicity tests were performed using in vivo inoculation. Finally, the pathogen was recovered and identified from the inoculated plants. Results: Based on Koch’s postulates, we identified the pathogen causing spot blight on P. ternata as Stagonosporopsis cucurbitacearum. To our knowledge, this is the first study to explore spot blight on P. ternata caused by S. cucurbitacearum in China. [ABSTRACT FROM AUTHOR]

Pomegranate, a pivotal fruit that is well recognized globally and a rapidly emerging crop in the southeastern United States and other subtropical regions, faces a formidable challenge from Colletotrichum spp., a fungal pathogen causing anthracnose fruit rot, which leads to severe to complete premature fruit drop. The development and use of disease-resistant cultivars are considered the most cost-effective and sustainable approach to managing this disease. Identifying sources of resistance is essential for developing new cultivars with improved resistance to this disease. This project aimed to expand the scope of evaluation through a 2-year field study in central Florida, examining fruit from 35 cultivars from diverse origins using both artificial inoculation at the petal dehiscent stage and natural infection. Lesion size on the fruit was measured during the growing season in a field setting. Subsequently, seven cultivars were selected for further testing by inoculating detached mature fruit and measuring lesion size to confirm observed resistance and determine the correlation between resistance observed in planta in the field and on detached fruit in the laboratory. The field study revealed significant genetic differences among pomegranate cultivars in susceptibility to naturally occurring and induced anthracnose fruit rot and classified cultivars into five resistance or susceptibility classes. Five cultivars that originated from different regions of the world, including 'Azadi', showed consistent resistance to anthracnose fruit rot in the field. Resistance remained strong on detached mature fruit. A strong positive correlation existed between resistance levels on in-planta fruit and on detached mature fruit, suggesting a possible simple, efficient approach to screening breeding populations for anthracnose fruit rot resistance in pomegranate. These findings represent an important step toward developing new anthracnose-resistant cultivars and understanding and improving disease resistance in this increasingly important fruit crop in the world. [ABSTRACT FROM AUTHOR]

Soybean is one of the most important crops grown worldwide and accounting for significant global trade including transgenic soybean. The crop is attacked by several seed-borne fungal pathogens and some of them are of quarantine concern for India. Keeping in view of the risks associated with movement of soybean seeds, sensitive and reliable molecular diagnostics have been developed for precise and simultaneous detection of three pathogens of quarantine concern for India namely, Diaporthe phaseolorum (stem blight), D. longicolla (seed decay), Peronospora manshurica (downy mildew), along with Macrophomina phaseolina causing dry root rot. The targeted pathogens after isolation from imported transgenic and non-transgenic soybean seeds were identified. Quadruplex and qPCR assays were developed targeting the sequences of different genes such as Histone-3 for detection of D. longicolla and M. phaseolina. The markers DlHisF2&R2 and MpHisF1&R1 produced 265 and 309 bp amplicons for D. longicolla and M. phaseolina, respectively. Actin gene based marker DpActF1&R2 was developed for D. phaseolorum which provided 113 bp amplicon whereas, COX2 based marker PmCoxF2&R2 was developed for P. manshurica with amplified product of 152 bp. During qPCR analysis, these markers proved highly specific and sensitive for detection of these pathogens up to 0.1 pg of template DNA. Quadruplex PCR protocol was also developed by combining these specific markers which could distinguish all the targeted pathogens simultaneously in a single reaction. The developed diagnostic protocols are extremely valuable for quarantine clearance and to ensure the safe transboundary exchange and healthy conservation of germplasm in the National Genebank. [ABSTRACT FROM AUTHOR]

Candida auris is a human fungal pathogen classified as an urgent threat to the delivery of health care due to its extensive antimicrobial resistance and the high mortality rates associated with invasive infections. Global outbreaks have occurred in health care facilities, particularly, long-term care hospitals and nursing homes. Skin is the primary site of colonization for C. auris. To accelerate research studies, we developed microbiome sequencing protocols, including amplicon and metagenomic sequencing, directly from patient samples at health care facilities with ongoing C. auris outbreaks. We characterized the skin mycobiome with a database optimized to classify Candida species and C. auris to the clade level. While Malassezia species were the predominant skin-associated fungi, nursing home residents also harbored Candida species, including C. albicans, and C. parapsilosis. Amplicon sequencing was concordant with culturing studies to identify C. auris-colonized patients and provided further resolution that distinct clades of C. auris are colonizing facilities in New York and Illinois. Shotgun metagenomic sequencing from a clinical sample with a high fungal bioburden generated a skin-associated profile of the C. auris genome. Future larger scale clinical studies are warranted to more systematically investigate the effects of commensal microbes and patient risk factors on the colonization and transmission of C. auris. IMPORTANCE Candida auris is a human pathogen of high concern due to its extensive antifungal drug resistance and high mortality rates associated with invasive infections. Candida auris skin colonization and persistence on environmental surfaces make this pathogen difficult to control once it enters a health care facility. Residents in long-term care hospitals and nursing homes are especially vulnerable. In this study, we developed microbiome sequencing protocols directly from surveillance samples, including amplicon and metagenomic sequencing, demonstrating concordance between sequencing results and culturing.

Fungal mitochondrial genomes encode genes involved in crucial cellular processes, such as oxidative phosphorylation and mitochondrial translation, and the molecule has been used as a molecular marker for population genetics studies. Coccidioides immitis and C. posadasii are endemic fungal pathogens that cause coccidioidomycosis in arid regions across both American continents. To date, approximately 150 Coccidioides isolates have been sequenced to infer patterns of variation in nuclear genomes. However, less attention has been given to the mitochondrial genomes of Coccidioides. In this report, we describe the assembly and annotation of mitochondrial reference genomes for two representative strains of C. posadasii and C. immitis, as well as assess population variation among 77 selected genomes. The sizes of the circular-mapping molecules are 68.2 Kb in C. immitis and 75.1 Kb in C. posadasii. We identify 14 mitochondrial protein-coding genes common to most fungal mitochondria, which are largely syntenic across different populations and species of Coccidioides. Both Coccidioides species are characterized by a large number of group I and II introns, harboring twice the number of elements as compared to closely related Onygenales. The introns contain complete or truncated ORFs with high similarity to homing endonucleases of the LAGLIDADG and GIY-YIG families. Phylogenetic comparisons of mitochondrial and nuclear genomes show extensive phylogenetic discordance suggesting that the evolution of the two types of genetic material is not identical. This work represents the first assessment of mitochondrial genomes among isolates of both species of Coccidioides, and provides a foundation for future functional work.

Fungal mitochondrial genomes encode genes involved in crucial cellular processes, such as oxidative phosphorylation and mitochondrial translation, and the molecule has been used as a molecular marker for population genetics studies. Coccidioides immitis and C. posadasii are endemic fungal pathogens that cause coccidioidomycosis in arid regions across both American continents. To date, approximately 150 Coccidioides isolates have been sequenced to infer patterns of variation of nuclear genomes. However, less attention has been given to the mitochondrial genomes of Coccidioides. In this report, we describe the assembly and annotation of mitochondrial reference genomes for two representative strains of C. posadasii and C. immitis, as well as assess population variation among 77 selected genomes. The sizes of the circular-mapping molecules are 68.2 Kb in C. immitis and 75.1 Kb in C. posadasii. We identify fourteen mitochondrial protein-coding genes common to most fungal mitochondria, which are largely syntenic across different populations and species of Coccidioides. Both Coccidioides species are characterized by a large number of group I and II introns, harboring twice the number of elements as compared to closely related Onygenales. The introns contain complete or truncated ORFs with high similarity to homing endonucleases of the LAGLIDADG and GIY-YIG families. Phylogenetic comparisons of mitochondrial and nuclear genomes show extensive phylogenetic discordance suggesting that the evolution of the two types of genetic material is not identical. This work represents the first assessment of mitochondrial genomes among isolates of both species of Coccidioides, and provides a foundation for future functional work.

Candida auris poses a serious threat to infection control and patient care since it can produce invasive infections that have a high fatality rate, has been linked to outbreaks in hospital environments, and is typically resistant to several antifungal medications. Since its first description in 2009, six clades have been described. The emerging fungal pathogen possesses adhesins that allow it to adhere to host tissues and medical devices, can form biofilms, produces various hydrolytic enzymes, employs several strategies to evade host immune responses, and exhibits high genetic diversity, which may contribute to its ability to adapt to different environmental conditions and evade host defenses. C. auris is very resistant to various disinfectants and may be difficult to detect.

Abstract The soil fungal community plays an important role in forest ecosystems and is crucially influenced by forest secondary succession. However, the driving factors of fungal community and function during temperate forest succession and their potential impact on succession processes remain poorly understood. In this study, we investigated the dynamics of the soil fungal community in three temperate forest secondary successional stages (shrublands, coniferous forests, and deciduous broad‐leaved forests) using high‐throughput DNA sequencing coupled with functional prediction via the FUNGuild database. We found that fungal community richness, α‐diversity, and evenness decreased significantly during the succession process. Soil available phosphorus and nitrate nitrogen decreased significantly after initial succession occurred, and redundancy analysis showed that both were significant predictors of soil fungal community structure. Among functional groups, fungal saprotrophs and pathotrophs represented by plant pathogens were significantly enriched in the early‐successional stage, while fungal symbiotrophs represented by ectomycorrhiza were significantly increased in the late‐successional stage. The abundance of both saprotroph and pathotroph fungal guilds was positively correlated with soil nitrate nitrogen and available phosphorus content. Ectomycorrhizal fungi were negatively correlated with nitrate nitrogen and available phosphorus content and positively correlated with ammonium nitrogen content. These results indicate that the dynamics of fungal community and function reflected the changes in nitrogen and phosphorus availability caused by the secondary succession in temperate forests. The fungal plant pathogen accumulated in the early‐successional stage and ectomycorrhizal fungi accumulated in the late‐successional stage may have a potential role in promoting forest succession. These findings contribute to a better understanding of the response of soil fungal communities to secondary forest succession and highlight the importance of fungal communities during the successional process.

Eucalyptus spp. are extensively cultivated in southern China because of their adaptability and versatile timber production. Calonectria leaf blight caused by Calonectria species is considered a major threat to Eucalyptus trees planted in China. The GuangXi Zhuang Autonomous Region is the provincial region with the largest distribution of Eucalyptus plantations in China. The present study aimed to expound the species diversity and pathogenicity of Calonectria isolates obtained from the soil of Eucalyptus plantations in GuangXi. A total of 188 Calonectria isolates were recovered from the soil located close to Eucalyptus trees, and the isolates were identified based on the DNA sequence comparisons of the four partial regions of the translation elongation factor 1-alpha (tef1), β-tubulin (tub2), calmodulin (cmdA), and histone H3 (his3) genes. The isolates were identified as Calonectria aconidialis (74.5%), C. hongkongensis (21.3%), C. pseudoreteaudii (2.1%), C. kyotensis (1.6%), and C. chinensis (0.5%). The inoculation results indicated that 40 isolates representing five Calonectria species were pathogenic to the three Eucalyptus genotypes. Two inoculated experiments consistently showed that the longest lesions were produced by the isolates of C. aconidialis. Some isolates of C. aconidialis, C. hongkongensis, and C. kyotensis produced significantly longer lesions than the positive controls, but not the isolates of C. pseudoreteaudii or C. chinensis. These results indicated that Calonectria isolated from the soil may pose a threat to Eucalyptus plantations. Some Calonectria isolates of the same species differed significantly in their virulence in the tested Eucalyptus genotypes. The resistance of different Eucalyptus genotypes to Calonectria isolates within the same species was inconsistent. The inoculation results in this study suggested that many Calonectria isolates in each species had different levels of pathogenicity, and many Eucalyptus genotypes need to be tested to select disease-resistant Eucalyptus genetic materials in the future. The results of the present study enhance our knowledge of species diversity and the potential damage caused by Calonectria in the soil of Eucalyptus plantations. Our results also provide new insights into the breeding of disease-resistant Eucalyptus genotypes for controlling Calonectria leaf blight in China in the future. [ABSTRACT FROM AUTHOR]

Pomegranate is an important horticultural crop grown in arid and semi-arid regions worldwide. Moreover, due to its widespread nutraceutical and pharmaceutical properties, expansion in its cultivated area has taken place extensively in India which has now become a global leader in its production and export. However, due to changing climatic conditions a plethora of diseases affect its quality and yield. Recently, we observed a typical collar rot occurring on pomegranate plants with 10–25% incidence. The current study was undertaken to identify the causative organism and prove its pathogenicity on pomegranate plants. Therefore, isolation of fungi was carried out using symptomatic tissues. Among the isolated fungi, one of the fungi was recovered up to 80% and, therefore, it was evaluated further. The colony morphology and other microscopic features suggested that it belonged to the genus Calonectria. Homology search using reliable barcode regions, both coding and non-coding, revealed close genetic similarities between the isolate and C. hawksworthii. Pathogenicity tests were performed on pomegranate seedlings, groundnut seedlings (a susceptible host to Calonectria sp.) and 6-month-old pomegranate plants. Artificially inoculated seedlings/plants developed typical symptoms while control plants did not, proving that the isolate was pathogenic. Hence, based on morphological and molecular data we identified the pomegranate collar-rot causal organism as C. hawksworthii, which is a novel record and highlights the expansion in geographical distribution and host range of the fungus. [ABSTRACT FROM AUTHOR]

One of the most harmful diseases that infect maize plants is Fusarium wilt caused by Fusarium oxysporum. The novelty of this study was to investigate the antifungal and plant growth-promoting properties of some rhizobacteria (PGPR). PGPR enhances plant growth and manages plant diseases. In this study, some bacterial isolates were isolated from the rhizosphere of rice plants and selected based on their ability to produce β-1,3 glucanase. The two bacterial isolates that produced higher concentrations of β-1,3 glucanase were selected to complete the study and identified by 16S rDNA sequences as Pseudomonas pseudoalcaligenes with accession number EU921258 and Bacillus pumilus with accession number EU921259. Different biochemical properties of the two bacterial isolates were investigated. Additionally, the effectiveness of P. pseudoalcaligene and B. pumilus against Fusarium oxysporum, which causes maize wilt disease was assessed. The results showed that P. pseudoalcaligenes and B. pumilus, either alone or in combination caused a significant reduction in the percentage of disease severity and an increment in the percentage of protection. In addition, infection with Fusarium caused a significant decrease in the morphological parameters, photosynthetic pigments, and cell membrane stability as compared to healthy control plants. However, lipid peroxidation, osmoprotectant compounds (soluble sugars, glycine betaine, and proline), secondary metabolites (phenol, flavonoid, and lignin content), and the enzymatic antioxidants increased significantly in infected plants as compared to healthy control plants. Furthermore, inoculation with P. pseudoalcaligenes and B. pumilus alone or in a combination caused significantly increased in the morphological parameters, photosynthetic pigment, and cell membrane stability in infected plants. Induction of β-1,3 glucanase gene in maize was not recorded in non-inoculated plants, but detected in inoculated plants with P. pseudoalcaligenes and B. pumilus alone or in combination as 2.4 kb bands on agarose gel electrophoresis. In conclusion, PGPR is regarded as an environmentally friendly method of protecting plants from pathogen infection and increasing plant growth without the use of pesticides, which cause a variety of environmental pollutions. [ABSTRACT FROM AUTHOR]

European canker is one of the most devastating fungal diseases of apple in most temperate regions. The causal agent, Neonectria ditissima, infects trees through wounds in the bark forming cankers that girdle the stem and eventually cause tree death. Timely protection of the trees is challenged by stagnation of symptom expression after infections for a long period of time. The objective of this research is to use a novel TaqMan PCR assay to detect and quantify N. ditissima during the asymptomatic colonization of apple wood. Pruning wounds on branches of the cultivars Elstar and Gala were inoculated with N. ditissima and wood discs were sampled at 2–6, 10–14, and 30–34 mm distance from the inoculation site after 3 hours, 2 weeks, 4 weeks and 8 weeks for the detection and quantification of the pathogen. The TaqMan PCR assay detected N. ditissima in 51% of the inoculated apple tree samples. This was more sensitive than the culturing method detecting N. ditissima in 11% of the samples. An accumulation of N. ditissima DNA up to 34 mm distance from the inoculation site was observed without development of visible symptoms. To our knowledge this is the first time colonization of N. ditissima was detected and quantified in the absence of symptoms of European canker. The implications of this research are discussed. [ABSTRACT FROM AUTHOR]

The present study employed microRNA (miRNA) sequencing and metabolome profiling of Trichoderma parental strains and fusants during normal growth and interaction with the phytopathogen Sclerotium rolfsii Sacc. In‐vitro antagonism indicated that abiotic stress‐tolerant Tricho‐fusant FU21 was examined as a potent biocontroller with mycoparasitic action after 10 days. During interaction with the test pathogen, the most abundant uprising intracellular metabolite was recognized as l‐proline, which corresponds to held‐down l‐alanine, associated with arginine and proline metabolism, biosynthesis of secondary metabolites, and nitrogen metabolism linked to predicted genes controlled by miRNAs viz., cel‐miR‐8210‐3p, hsa‐miR‐3613‐5p, and mml‐miR‐7174‐3p. The miRNAs‐ mml‐miR‐320c and mmu‐miR‐6980‐5p were found to be associated with phenylpropanoid biosynthesis, transcription factors, and signal transduction pathways, respectively, and were ascertained downregulated in potent FU21_IB compared with FU21_CB. The amino benzoate degradation and T cell receptor signaling pathways were regulated by miRNAs cel‐miR‐8210 and tca‐miR‐3824 as stress tolerance mechanisms of FU21. The intracellular metabolites l‐proline, maleic acid, d‐fructose, Myo‐inositol, arabinitol, d‐xylose, mannitol, and butane were significantly elevated as potential biocontrol and stress‐tolerant constituents associated with miRNA regulatory pathways in potent FU21_IB. A network analysis between regulatory miRNA‐predicted genes and intracellular metabolomics acknowledged possible biocontrol pathways/mechanisms in potent FU21_IB to restrain phytopathogen. [ABSTRACT FROM AUTHOR]

Spot form net blotch, caused by Pyrenophora teres f. maculata, is a significant global disease of barley (Hordeum vulgare). Baudin, a barley cultivar that was until recently extensively grown in Western Australia, was reported as having minor seedling resistance. However, Baudin was highly susceptible to a local isolate, M3, suggesting that this isolate had gained virulence against a major susceptibility gene. M3 causes atypical lesions with pale centers early in the infection, with initial screens of a segregating population indicating that this was determined by a single locus in the Baudin genome. The susceptibility was semidominant in F1 progeny and the susceptibility gene, designated Spm1 (Susceptibility to P. teres f. maculata 1), mapped to a 190-kb section of the resistance gene-rich Mla region of chromosome 1H. Phenotyping with Ptm SP1, a non-M3 pathotype, identified a seedling resistance locus on 2H. Minor gene resistance is generally regarded as potentially durable, but our findings suggest the resistance to spot form net blotch in Baudin is nullified by strong susceptibility conferred by a separate locus on 1H. [ABSTRACT FROM AUTHOR]

Ambiviruses are hybrid infectious elements encoding the hallmark gene of RNA viruses, the RNA-dependent RNA polymerase, and self-cleaving RNA ribozymes found in many viroids. Ambiviruses are thought to be pathogens of fungi, although the majority of reported genomes have been identified in metatranscriptomes. Here, we present a comprehensive screen for ambiviruses in more than 46,500 fungal transcriptomes from the Sequence Read Archive (SRA). Our data-driven virus discovery approach identified more than 2,500 ambiviral sequences across the kingdom Fungi with a striking expansion in members of the phylum Basidiomycota representing the most complex fungal organisms. Our study unveils a large diversity of unknown ambiviruses with as little as 27% protein sequence identity to known members and sheds new light on the evolution of this distinct class of infectious agents with RNA genomes. No evidence for the presence of ambiviruses in human microbiomes was obtained from a comprehensive screen of respective metatranscriptomes available in the SRA. [ABSTRACT FROM AUTHOR]

Coccidioidomycosis, or Valley fever, is caused by two species of dimorphic fungi. Based on molecular phylogenetic evidence, the genus Coccidioides contains two reciprocally monophyletic species: C. immitis and C. posadasii. However, phenotypic variation between species has not been deeply investigated. We therefore explored differences in growth rate under various conditions. A collection of 39 C. posadasii and 46 C. immitis isolates, representing the full geographical range of the two species, was screened for mycelial growth rate at 37 °C and 28 °C on solid media. The radial growth rate was measured for 16 days on yeast extract agar. A linear mixed effect model was used to compare the growth rate of C. posadasii and C. immitis at 37 °C and 28 °C, respectively. C. posadasii grew significantly faster at 37 °C, when compared to C. immitis; whereas both species had similar growth rates at 28 °C. These results indicate thermotolerance differs between these two species. As the ecological niche has not been well-described for Coccidioides spp., and disease variability between species has not been shown, the evolutionary pressure underlying the adaptation is unclear. However, this research reveals the first significant phenotypic difference between the two species that directly applies to ecological research.

The pathogenic fungus Phomopsis longicolla causes numerous plant diseases, such as Phomopsis seed decay, pod and stem blight, and stem canker, which seriously affect the yield and quality of soybean production worldwide. Because of a lack of technology for efficient manipulation of genes for functional genomics, understanding of P. longicolla pathogenesis is limited. Here, we developed an efficient polyethylene glycol–mediated protoplast transformation system in P. longicolla that we used to characterize the functions of two genes involved in the cell wall integrity (CWI) pathway of the mitogen-activated protein kinase (MAPK) cascade, including PlMkk1, which encodes MAPK kinase, and its downstream gene PlSlt2, which encodes MAPK. Both gene knockout mutants ΔPlMkk1 and ΔPlSlt2 displayed a reduced growth rate, fragile aerial hyphae, abnormal polarized growth and pigmentation, defects in sporulation, inadequate CWI, enhanced sensitivity to abiotic stress agents, and significant deficiencies in virulence, although there were some differences in degree. The results suggest that PlMkk1 and PlSlt2 are crucial for a series of growth and development processes as well as pathogenicity. The developed transformation system will be a useful tool for additional gene function research and will aid in the elucidation of the pathogenic mechanisms of P. longicolla. [Graphic: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

ABSTRACTDuring a field survey of cultivated Morchella mushroom diseases, Diploöspora longispora and Paecilomyces penicillatus, causal agents of pileus rot or white mould disease were detected, which resulted in up to 80% of yield losses. Multi-locus phylogenic analysis revealed that the fungi were affiliated in a distinct clade in Hypocreales. We further constructed a phylogenetic tree with broader sampling in Hypocreales and estimated the divergence times. The D. longispora and P. penicillatus clades were estimated to have diverged from Hypocreaceae around 129 MYA and Pseudodiploösporeaceae fam. nov is herein proposed to accommodate species in this clade. Two new genera, i.e. Pseudodiploöspora and Zelopaecilomyceswere, were introduced based on morphological characteristics and phylogenic relationships of Diploöspora longispora and Paecilomyces penicillatus, respectively. Five new combinations – Pseudodiploöspora cubensis, P. longispora, P. fungicola, P. zinniae, and Zelopaecilomyces penicillatus – were proposed.

The deployment of plant varieties carrying resistance genes (R) exerts strong selection pressure on pathogen populations. Rapidly evolving avirulence genes (Avr) allow pathogens to escape R‐mediated plant immunity through a variety of mechanisms, leading to virulence. The poplar rust fungus Melampsora larici‐populina is a damaging pathogen of poplars in Europe. It underwent a major adaptive event in 1994, with the breakdown of the poplar RMlp7 resistance gene. Population genomics studies identified a locus in the genome of M. larici‐populina that probably corresponds to the candidate avirulence gene AvrMlp7. Here, to further characterize this effector, we used a population genetics approach on a comprehensive set of 281 individuals recovered throughout a 28‐year period encompassing the resistance breakdown event. Using two dedicated molecular tools, genotyping at the candidate locus highlighted two different alterations of a predominant allele found mainly before the resistance breakdown: a nonsynonymous mutation and a complete deletion of this locus. This results in six diploid genotypes: three genotypes related to the avirulent phenotype and three related to the virulent phenotype. The temporal survey of the candidate locus revealed that both alterations were found in association during the resistance breakdown event. They pre‐existed before the breakdown in a heterozygous state with the predominant allele cited above. Altogether, these results suggest that the association of both alterations at the candidate locus AvrMlp7 drove the poplar rust adaptation to RMlp7‐mediated immunity. This study demonstrates for the first time a case of adaptation from standing genetic variation in rust fungi during a qualitative resistance breakdown. [ABSTRACT FROM AUTHOR]