Journal of Fungi

Colletotrichum sublineola (Cs), the hemibiotrophic fungus that causes sorghum anthracnose, impacts sorghum grain and biomass crop production worldwide. Although nutrient availability is known to influence development in filamentous fungi, including Colletotrichum species, how in vitro nutrient limitation reprograms the Cs cellular state remains unclear. We cultured Cs on full-strength, half-strength, and one-tenth-strength potato dextrose agar (PDA) to define responses across a nutrient gradient. Nutrient limitation induced a pronounced high-sporulation phenotype, with one-tenth-strength PDA producing the strongest conidiation response, followed by half-strength PDA. To study the underlying molecular programs in each condition, we employed a multiplexed metabolite, protein, and lipid extraction (MPLEx) protocol for global proteomics and metabolomics. Global proteomics resulted in 4,590 protein identifications, including 204 unique to one-tenth-strength PDA. Among them are proteins linked to sporulation, vesicular transport, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, and common in fungal extracellular membrane (CFEM)-domain proteins. Differential abundance and pathway analyses revealed a broad reduction of central carbon and energy metabolism, including glycolysis/gluconeogenesis, pentose phosphate, pyruvate metabolism, and glyoxylate pathways, together with increased ribosome-related processes, cAMP signaling, and cell-surface remodeling in one-tenth-strength PDA conditions. In addition, correlative metabolomics supported selective metabolic depletion and resource reallocation toward stress adaptation, membrane remodeling, and conidiation, supporting proteomics findings. Together, these data support a starvation-adapted Cs developmental state associated with enhanced sporulation, cellular pathway reprogramming, and potential virulence linked preparedness under nutrient-limited growth conditions in vitro. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=101 SRC="FIGDIR/small/724728v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@f6ceb2org.highwire.dtl.DTLVardef@17c4836org.highwire.dtl.DTLVardef@68e995org.highwire.dtl.DTLVardef@1bf3983_HPS_FORMAT_FIGEXP M_FIG C_FIG

Aspergillus fumigatus is a thermotolerant saprobe found in soils and plant debris worldwide and an important pathogen of humans causing two million deaths annually. A. fumigatus makes abundant asexual spores (conidia) which are widely distributed by wind and can be inhaled from the environment. In susceptible individuals inhaled conidia break dormancy, germinate and grow in the lung leading to serious disease. Recent work has shown that conidia made at 37{degrees}C and 50{degrees}C have different morphologies and germination kinetics. While the asexual cycle is well-characterized at 37{degrees}C, much less is known about the asexual cycle at 50{degrees}C. Here, we combine flow cytometry and transcriptomics to track morphology and gene expression in the hyphae, conidiophores and conidia of A. fumigatus during asexual development at 37{degrees}C or 50{degrees}C. We show that the temperature during a narrow time window in late-stage conidiophore development dictates resulting conidial morphology, transcriptional program, and germination kinetics. As expected, conidiation at 37{degrees}C resulted in upregulation of brlA, the master regulator of asexual development, and its downstream targets in conidiophores and conidia. Surprisingly, conidiation at 50{degrees}C resulted in upregulation of MAT1-1, the master regulator of sexual development and its downstream targets in conidiophores and conidia. Our findings suggest that temperature during late conidiophore development transcriptionally primes conidia for asexual, parasexual or sexual development enhancing chances of survival for progeny. Our findings are especially relevant for agricultural compost where a wide gradient of temperatures exists, abundant A. fumigatus has been isolated, and resistance to antifungals is thought to evolve. IMPORTANCEThe human pathogen Aspergillus fumigatus has been found in natural and agricultural environments around the world. Disease is acquired when susceptible individuals inhale airborne asexual spores from the environment, which in agriculture generally includes proximity to compost and plant debris piles. This work shows that the environmental temperature when A. fumigatus spores are made determines the transcriptomes of those spores, priming them for future asexual or sexual development. The survival of asexual and sexual spores is very different at different temperatures, so these results are important for understanding how this pathogen survives in varied hostile environments. In addition, there are very few antifungal drugs with which to treat A. fumigatus infections, and resistance is increasing driven in part by agricultural use of fungicides. These results suggest that higher temperatures during asexual spore formation can lead to increased sexual reproduction and greater chances to evolve antifungal resistance.

The Agaricomycotina accounts for roughly a third of all described fungi. They are important due to their wide range of lifestyles and economic and environmental relevance. Certain agaricomycetes act as lignocellulose degraders, playing a significant role in forest ecosystems and bioremediation processes. These wood-decaying fungi have historically been classified as mostly white- or brown-rot based on their ability to degrade lignin, with white-rot fungi possessing a collection of lignocellulose-degrading enzymes, which are reduced or absent in brown-rot fungi. Here, we sequenced and annotated the genome of the agaricomycete Crepidotus cesatii CBS 511.95 and explored its genome and predicted enzymatic content in a comparative context. The 36.04 Mbp genome is in 235 scaffolds, with 3.34% repeat content and 12,891 predicted genes. We found that the PFAM distributions of identified orthogroups suggested that C. cesatii shows patterns more similar to white-rot fungi compared to brown-rot fungi. Additionally, C. cesatii contained multiple copies of CAZymes CBM1 and AA9 involved in hydrolysis of lignocellulose, similar to white-rot fungi. On the other hand, according to the Conserved Unique Peptide Patterns (CUPP) data for AA2 peroxidases, the key enzymes in lignin degradation, C. cesatii is more similar to brown-rot fungi. Based on our analyses we predict that C. cesatii is another representation of the continuum of wood decaying modes between white and brown rot fungi combining genetic features of both types of fungi.

Background: Invasive pulmonary aspergillosis (IPA) is a serious infection in critically ill patients. Galactomannan detection in endotracheal aspirates (ETA) has emerged as a promising non-invasive diagnostic method. This study evaluates the supportive diagnostic value of ETA galactomannan in ICU patients suspected to have IPA. Methods: We conducted a prospective observational cohort study over two years, enrolling 120 patients in the medicine ICU at a tertiary care centre in India (January 2022 to October 2023). Patients aged over 14 years on mechanical ventilation for >48 hours meeting the entry criteria of the BM-AspICU algorithm were included. ETA galactomannan was measured and correlated with IPA classification. Results: Of 120 patients, 37% (n=44) had probable IPA and 63% (n=76) were classified as colonisers or possible IPA. The optimal ETA galactomannan cut-off was 1.097, yielding sensitivity 72.73% (95% CI 57.2 - 85.0%), specificity 84.2% (95% CI 74.4 - 90.7%), PLR 4.86, NLR 0.35, and AUC 0.844 Conclusion: ETA galactomannan supports IPA diagnosis with favourable sensitivity and specificity. However, given the limitations of clinical scoring-based reference standards and the potential plateau in colonizer reduction at higher cut-offs, it should be integrated into a comprehensive diagnostic approach incorporating clinical, radiological, and microbiological criteria.

Autophagy is a critical cellular process regulated by Atg proteins, yet its modulation by redox-active compounds and iron remains incompletely understood. Here, we investigated the effects of dithiothreitol (DTT) and iron on autophagy and on AaAtg4 protease activity in the plant-pathogenic fungus Alternaria alternata. Using GFP-tagged AaAtg8, fluorescence microscopy and proteolysis assays revealed that DTT markedly enhanced autophagic vacuole formation and GFP release, indicating increased autophagic flux. Western blot analyses confirmed that DTT promoted AaAtg8 lipidation, while co-treatment with hydrogen peroxide (H2O2) suppressed this modification. AaAtg4 was constitutively active and could process AaAtg8 regardless of DTT supplementation, whereas moderate DTT concentrations elevated AaAtg4 protein abundance and phosphorylation. Bimolecular fluorescence complementation assays demonstrated that DTT, but not iron, facilitated AaAtg4-AaAtg8 interactions and vacuolar localization, whereas H2O2 counteracted these effects. Notably, combined DTT and H2O2 sustained autophagy at a low but stable level, suggesting a redox balance in autophagic regulation. Iron supplementation selectively destabilized AaAtg8 and modulated AaAtg4 phosphorylation in a concentration-dependent manner, without altering autophagy or protease activity. Collectively, these findings demonstrate that DTT enhances autophagy primarily by promoting AaAtg8 lipidation, AaAtg4 phosphorylation, and AaAtg4-AaAtg8 complex formation, while exerting minimal influence on AaAtg4 protease activity. In contrast, ion regulates autophagy flux through its effects on AaAtg4 phosphorylation and AaAtg8 stability, without significantly altering AaAtg4 protease activity, AaAtg8 lipidation, or AaAtg4-AaAtg8 interactions. Together, this work underscores the intricate interplay between redox signaling, nutrient cues, and autophagy regulation in A. alternata. IMPORTANCEThis study provides critical new insights into how redox-active compounds and iron modulate autophagy in the plant-pathogenic fungus Alternaria alternata, a pathogen of agricultural relevance. By dissecting the distinct roles of DTT, hydrogen peroxide, and iron in regulating AaAtg8 lipidation, AaAtg4 phosphorylation, and AaAtg4-AaAtg8 interactions, our findings reveal that autophagy is not simply a constitutive process but is finely tuned by redox balance and nutrient cues. This work advances the fundamental understanding of autophagy regulation in filamentous fungi, highlights the interplay between oxidative stress and protease activity, and establishes a framework for exploring how environmental factors shape fungal pathogenicity. Ultimately, these insights may inform novel strategies to mitigate crop fungal diseases by targeting autophagic pathways.

Botrytis cinerea is a necrotrophic plant pathogen with an extremely wide host range. During invasion, the fungus induces rapid host cell death and proliferates in the necrotic tissue. The mechanisms of host killing are still incompletely understood, they involve secretion of lytic enzymes, phytotoxic metabolites and cell death inducing proteins (CDIPs). We have previously shown that the sequential knockout of up to 12 CDIPs leads to a substantial reduction of virulence of B. cinerea mutants. In this study, we have identified additional CDIPs and generated an extended mutant series culminating in a 29x mutant which is deficient in most currently known CDIPs and two phytotoxic metabolites. These mutants are strongly reduced in infection, but still induced necrosis and grey mould symptoms, demonstrating that additional determinants of host killing remain unidentified. Overexpression of the highly phytotoxic Nep1 in a 22-fold CDIP mutant failed to increase its virulence. Reevaluation of several CDIPs previously described as virulence factors revealed for most tested CDIPs no major contribution to pathogenesis. Together with the observation that none of the CDIPs are specific to B. cinerea, our data question a particular role of CDIPs for necrotrophic pathogenesis. In contrast, generation of a mutant lacking all six predicted endo-polygalacturonases confirmed their major but not exclusive role for tissue degradation and infection.

IntroductionLomentospora prolificans is a pathogenic filamentous fungus that causes disease primarily in people with severely compromised immune systems. It is pan-resistant to antifungal drugs, but the mechanism of its resistance to amphotericin B (AMB) is unknown. ObjectivesWe aimed to investigate the mechanism of resistance to AMB of L. prolificans. MethodsThe AMB susceptibility of L. prolificans protoplasts was measured using broth microdilution. L. prolificans, either intact, homogenized or fractionated was incubated with AMB in broth. The same activity was carried out with Aspergillus fumigatus as a control. This broth was then used to prepare microdilution plates with Saccharomyces cerevisiae to determine the activity of the conditioned AMB. ResultsAMB was 16-fold more effective in inhibiting the growth of L. prolificans protoplasts than conidia, but only two-fold more effective against A. fumigatus protoplasts than conidia. Incubation of L. prolificans hyphae with AMB in media diminished drug activity to a much greater extent than A. fumigatus, with 8-fold greater fungal mass of the latter required to achieve the effect of the former. Homogenization and fractionization of L. prolificans revealed that the factor inhibiting AMB activity was soluble with a mass >100 kda. DNase, trypsin, proteinase K, amyloglucosidase, SDS and 0.22 m had no effect on the AMB resistance factor, while treatment with urea, acetonitrile inactivated it. ConclusionWe report a different mechanism for AMB resistance based on the existence of a substance residing in the L. prolificans cell wall that can eliminate the antifungal activity of AMB.

Ubiquitin is a posttranslational modifier that is conserved among eukaryotes. Ubiquitination alters stability and folding of cellular proteins. Deubiquitinases (DUBs) reverse ubiquitination and often function as part of protein complexes. There are 32 predicted DUB-encoding genes present in the soil-borne phytopathogenic fungus Verticillium dahliae. Nuclear ubiquitin-specific protease 3 (Usp3) is a member of the Spt-Ada-Gcn5 acetyltransferase (SAGA) complex, whereas Usp1 is predicted to associate with the COP9 signalosome (CSN), which controls specificities of cellular E3 ubiquitin ligase activities. A proteomics approach using biotin capture and identification (BioID) supports that Usp3 regulates gene expression beyond the transcription level. Western experiments showed a dysregulation in ubiquitinated cellular proteins in corresponding deletion strains. Usp3 and Usp1 are both required for fungal development. They regulate microsclerotia formation based on different environmental cues and provide redundant functions in controlling conidiation. Absence of both corresponding genes resulted in significant impairment of conidiospore formation, which is required for fungal propagation within the plant vascular system. This paralysed spreading ability reduced virulence on tomato plants (Solanum lycopersicum). In summary, V. dahliae responds to environmental cues by Usp3- and Usp1-mediated adjustment of gene expression and protein stability. This is important for key developmental processes of the V. dahliae disease cycle and its virulence towards the host plant. Author summaryUbiquitination and deubiquitination of proteins enable cells to rapidly react to environmental cues and adjust protein stabilities and subsequently transcriptomic profiles. Usp3 is a nuclear deubiquitinase subunit of the Spt-Ada-Gcn5 acetyltransferase (SAGA) transcriptional coactivator complex. Usp1 is predicted to be associated with the COP9 signalosome that regulates substrate specificities of the ubiquitin-proteasome system. BioID experiments suggest that other SAGA complex subunits, histone proteins, spliceosomal proteins, ribosomal proteins, a protein that tackles transcriptionally stalled RNAPII, and a protein that degrades mRNA with premature stop codons locate proximal to Usp3 within the cell. Deletion of USP3 led to the dysregulation of protein ubiquitination. A single deletion of USP1 did not significantly change ubiquitination profiles, however, a double deletion of USP1/3 significantly affected the ubiquitin-proteasome system. The altered ubiquitination profile correlated with a dysregulation of key developmental processes. Microsclerotia formation was decoupled from environmental cues in the {Delta}USP3 strain, whereas an additional deletion of USP1 reconnected it in a media-dependent manner. USP3 and USP1 contribute to a common governing process in conidiation, and the defect in spreading of the {Delta}USP1/3 strain is reflected by a significant reduction in plant pathogenicity.

Colletotrichum siamense is a predominant causal agent of anthracnose in rubber tree and numerous economically important crops, causing severe yield losses worldwide. Conidial germination represents a critical early step for successful infection, while the high-osmolarity glycerol (HOG) MAPK pathway and ergosterol biosynthesis individually govern fungal development, stress adaptation and fungicide responses. However, the molecular crosstalk between these two modules remains largely elusive in phytopathogenic fungi. Here, we identified CsErg5B, a sterol C-22 desaturase homolog, as a direct target of the HOG- regulated transcription factor CsAtf1 in C. siamense. CsErg5B was indispensable for ergosterol biosynthesis, conidial germination, appressorium formation, and full virulence. The {Delta}CsErg5B mutant showed increased conidiation but severely impaired germination, and exhibited elevated resistance to fludioxonil while hypersensitivity to azole fungicides. Epistasis analysis using the {Delta}CsErg5B/{Delta}CsCyp51G1 double mutant - where CsCyp51G1 serves as another downstream target of CsAtf1 - revealed that CsErg5B functions as the predominant downstream effector of CsAtf1 in modulating conidial development and fludioxonil sensitivity. Furthermore, overexpression of CsErg5B significantly rescued the defects in conidial germination and fludioxonil sensitivity in both {Delta}CsAtf1 and {Delta}CsPbs2 mutants. Taken together, our findings uncover a HOG MAPK - CsAtf1 - CsErg5B regulatory axis that connects HOG MAPK signaling to ergosterol homeostasis, thereby governing conidial germination and fungicide sensitivity in C. siamense. This study provides novel insights into the regulatory network underlying fungal development and fungicide response, and offers promising molecular targets for the integrated management of plant anthracnose.

Ergosterol has multiple functions in filamentous fungi and yeasts, although only a part of the functions seems to be understood. An antifungal peptide, theonellamide A (TNM-A) induces drastic morphological changes in fission yeast cells by targeting plasma membrane ergosterol. TNM-A induces overproduction and ectopic accumulation of cell wall glucan at both growing tips and septum through a yet unknown mechanism. Here we show that TNM-A treatment causes accumulation of 1,3-{beta}-glucan at cell-polarity sites, not by increased activity of 1,3-{beta}-glucan synthase, but by an increased, persistent localization of the glucan synthase enzymes. Screening based on subcellular localization of proteins at periphery or polarity sites suggested the involvement of the Rho family GTPase Cdc42. In agreement, TNM-A induced both activation of Cdc42 and enhancement of membrane trafficking of glucan synthase enzymes. In conclusion, our chemical genetics analyses using TNM-A suggest that membrane ergosterol regulates the activity of Cdc42, which further regulates the localization of glucan synthases and cell wall biosynthesis. Highlights (four sentences)- Thenoellamide A (TNM-A) induces an ectopic overproduction of cell wall glucan. - TNM-A treatment causes increased, persistent localization of glucan synthases at the cell tips and septum. - TNM-A activates Cdc42 and upregulates membrane trafficking of glucan synthases. - Ergosterol is involved in proper activation/inactivation of Cdc42.

Understanding how host-microbiome interactions influence tree disease is critical for understanding forest resilience. Here, we present foliar microbiome ITS2 metabarcoding transcriptomic datasets from Pinus sylvestris to investigate susceptibility to Dothistroma needle blight (DNB), a globally important foliar disease caused by Dothistroma septosporum. We hypothesised that host genotype shapes foliar microbial communities and their interactions, thereby influencing disease outcomes. Samples were collected from a progeny-provenance field trial in the south of Scotland representing a broad spectrum of disease susceptibilities. The dataset comprises ITS2 metabarcoding samples from 200 genotypes across three timepoints and RNAseq samples from 48 genotypes across two timepoints. Sampling captured key stages of pathogen exposure and disease progression. Both standardised and bespoke protocols were used for nucleotide extraction, sequencing, and quality control, including multiple negative and positive controls. These datasets, available in the European Nucleotide Archive (project accession PRJEB88228), enable analysis of temporal dynamics in foliar fungal communities, host-microbiome transcriptional responses, and genotype-dependent variation in disease susceptibility.

The pathogenic fungus Trichosporon asahii causes severe mycoses in immunocompromised hosts, such as neutropenic patients. In Cryptococcus neoformans, the unfolded protein response (UPR) sensor Ire1 induces hxl1 mRNA splicing and contributes to stress responses and virulence. The function of Ire1-triggered hxl1 mRNA splicing in stress tolerance and virulence of T. asahii, however, remains unclear. Here, we demonstrated that ire1- and hxl1 gene-deficient T. asahii mutants are sensitive to dithiothreitol (DTT), an inducer of endoplasmic reticulum stress, and exhibit reduced virulence in a silkworm infection model. DTT treatment induced hxl1 mRNA splicing in the wild-type strain, whereas ire1 gene-deficient mutants did not undergo hxl1 mRNA splicing. The ire1 gene-deficient mutants were more sensitive than the parent strain to DTT, H2O2, Congo red, and SDS, and showed reduced virulence in silkworms. Similarly, hxl1 gene-deficient mutants exhibited increased sensitivity to these stressors and reduced virulence. Both the ire1 gene-deficient and hxl1 gene-deficient mutants showed decreased expression of reactive oxygen species-detoxifying related genes CAT2, SOD1, and SOD2, compared with the parent strain. Together, these findings suggest that Ire1-triggered hxl1 mRNA splicing contributes to stress resistance and virulence in T. asahii.

Fungal plant pathogens can be affected by the bacteria they interact with in their environment, yet the characterization of these interactions beyond direct antagonism is lacking, especially in the case of endohyphal bacteria (EHB). Though limited in characterized examples, EHB can alter disease severity of their fungal host, providing either a potential tool or target for control. We screened isolates of Fusarium oxysporum f. sp. niveum (FON), an important soil-borne watermelon pathogen, using 16S PCR and fluorescence in situ hybridization microscopy to identify novel EHB. A symbiont of FON AS124 was identified to be a Paenibacillus sp. through genome sequencing and average nucleotide identity. To begin characterizing this relationship, we conducted watermelon infection assays using FON cured of its symbiont, the native association, and a coinoculation of fungi and bacteria. Disease severity was reduced in watermelon seedlings inoculated with the native association, though not in the coinoculation, and Paenibacillus sp. CB74 did not alone promote plant growth or inhibit fungal growth. This study shows an important functional outcome, reduced disease, for a novel symbiosis between FON and Paenibacillus sp. CB74, setting up further investigation into the mechanisms behind this outcome and the application of this interaction. ImportanceFungi pose a challenge in both the field and hospital as antifungal resistance rises and chemical control is increasingly scrutinized. In plant pathogenic fungi, endohyphal bacteria may present alternative targets or mechanisms of fungal control. These relationships are observed across diverse groups of fungi and bacteria, though few have been studied to the point of understanding impact. To contribute to the small but growing catalog of known endofungal bacterial relationships, we identified a novel symbiosis and began characterizing its functional outcomes with plant infection assays. The identified bacterial symbiont does alter disease severity of the fungal host offering a new system for both application and study of fungal pathogenesis.

The fungal cell wall is populated by proteins (CWPs), mostly uncharacterized, that show an atypical evolutionary behavior. Most CWPs are glycosylphosphatidylinositol(GPI)-proteins, followed by proteins with internal repeats (PIR), and non-covalently attached proteins that harbor carbohydrate binding domains (CBM). Several structural CWPs are initially bound to the same wall carbohydrates, but either covalently or non-covalently. However, it is not clear whether they work in the same way and if they are subjected to the same evolutionary constraints. In Neurospora crassa, CWPs ACW-1 (NCU08936) and NCW-3 (NCU07817) bind to {beta}-1,3-glucans through a GPI anchor or a predicted CBM-52 domain, respectively. Here, the evolutionary trajectories and functional roles of both CWPs were analyzed. Both proteins localized primarily to distal septa and hyphal wall surfaces. Morphological characterization and stress cell wall assays suggested that both proteins contribute to cell wall integrity, but NCW-3 likely plays a more prominent role. ACW-1 and NCW-3 homologues were predominantly identified in Ascomycota. ACW-1 displayed a broader distribution than NCW-3, whose homologues were largely restricted to Sordariales. Despite these differences, both protein families exhibited similar moderate global conservation and signatures of purifying selection within shared taxa. Nevertheless, a divergence gradient was identified within ACW-1, related to its tandem leucine-rich repeat (LRR) regions. A similar local accumulation of evolutionary change was not observed within NCW-3. These findings suggested that distinct CWP architectures can accommodate different patterns of sequence diversification despite sharing similar global evolutionary change.

Fungal-animal mutualisms remain significantly understudied, yet they represent some of the most successful partnerships known in nature. Fungal farming ambrosia beetles cultivate a consortium of fungal partners that include obligate filamentous members and yeasts. These fungi are maintained in highly specialized insect organs, termed mycangia, and are cultivated as food along the beetle galleries elaborated within host trees. Here we identify two previously described filamentous species, Raffaelea arxii and R. fusca, and the yeast, Ambrosiozyma monospora, as well as two new filamentous fungal species, Neocosmospora affinis and Graphium ambrosium, and two novel yeasts, designated Alloascoidea xylebori and Wickerhamomyces ambrosius, from beetle gallery walls and ambrosia beetle mycangia, using a protocol that minimizes biases in recovery by removal of a commonly used ethanol wash. To meet Koch-like postulates, we further demonstrate that all seven fungal species were individually competent at colonizing aposymbiotic X. affinis mycangia, thus demonstrating each as a viable mycangial mutualist. These data highlight methodological considerations that overcome previous limitations in mycangial content characterization, resulting in the discovery of new ambrosia beetle fungal partners. We further validate the fungal-animal mutualism by demonstrating specific colonization of the mycangial organ by potential fungal partners.

Chia (Salvia hispanica L.) is an emerging crop in Bangladesh valued for its medicinal properties and economic significance. In March 2024, target spot-like symptoms were observed in an experimental chia field (24.75{degrees} N, 90.50{degrees} E) at Bangladesh Agricultural University in Mymensingh, Bangladesh with disease incidence ranging from 23% to 47% across approximately 0.25 ha. Initially appearing as brick-red spots, these symptoms developed into target-shaped concentric rings, affecting leaves, stems, and inflorescences. A total of 24 fungal isolates were recovered from infected tissue; two representative isolates (BGECh-3 and BGECh-4) were randomly selected for details characterization. Pathogen identity was established through morphological traits, multilocus phylogenetic analysis of internal transcribed spacer (ITS) and elongation factor 1-alpha (EF-1) genes sequence, and pathogenicity confirmation through Kochs postulates, collectively identifying the causal agent as Corynespora cassiicola. The isolates demonstrated a broad host range, successfully infecting brinjal, chili, bottle gourd, country bean, tomato, and soybean. In vitro fungicide sensitivity assays with seven commercial fungicides showed that both isolates were highly sensitive to Goldzim (50% carbendazim), which completely inhibited mycelial growth at 10 {micro}g mL-{superscript 1}. Conza (10% Hexaconazole) and Amister top (18.2% azoxystrobin + 11.4% difenoconazole) reduced growth by up to 85% and 67%, respectively at equal concentration. Other fungicides showed comparatively lower efficacy even at higher concentrations. This study represents the first report of target spot disease of chia caused by C. cassiicola in Bangladesh and provides insights for effective disease management strategies.

Aphanomyces euteiches, the causative agent of Aphanomyces root rot (ARR), is of major concern for pea and other legume crops globally. This oomycete pathogen causes substantial decreases in crop yields, is unaffected by most fungicides, and persists in the soil for many years via its resilient oospores. Given the significance of pea crops in sustainable agriculture, namely the ability to fix nitrogen and act as a sustainable protein source, solutions to ARR are of high importance. We used RNA-seq in a novel strain of Pseudomonas donghuensis to identify two biosynthetic gene clusters under GacA/S control that are involved in producing bioactive molecules capable of inhibiting A. euteiches. Based on similarity to other reported clusters in Pseudomonas, the first is predicted to encode for a pseudoiodinine compound, while the second is predicted to produce the siderophore 7-hydroxytropolone. Individual knockouts of each cluster showed loss of inhibitory action of P. donghuensis NRC29 against A, euteiches in vivo. This is the first report highlighting the potential of P. donghuensis and the products of the two identified biosynthetic pathways as biocontrol agents for A. euteiches. Further investigations into the efficacy of P. donghuensis NRC29 and its metabolites in inhibiting A. euteiches in field trials will be of high value in developing sustainable strategies for ARR mitigation. ImportanceModern fungicidal treatments for control of root rot in pulse crops are ineffective for control of A. euteiches, leaving limited strategies for management of A. euteiches infected fields. We describe a novel P. donghuensis strain with potential for biocontrol against this persistent pathogen. Given the economic value of peas and other pulses globally, further work into harnessing the bioactive metabolites produced by this strain into a practical in-field treatment will be valuable.

BackgroundSegatella copri is characterised as a prominent glycolytic plant-based fiber utiliser within the human gut. My recent work has introduced a new interpretation of the positive impacts of plant-based polysaccharides on S. copri, as a significant negative relationship between S. copri and Blautia spp. was identified. The high rate of electron donor consumption by Blautia spp. indicated that competition for the electron donors, formate, ferredoxin and fumarate, could be the route of the negative relationship but would also explain the positive relationship with plant-based polysaccharides, as they are products of fiber fermentation intestinally. MethodsForty two genomes of S. copri were annotated via Prokka to identify alternative PEP pathways. Phylogenetics allowed effectively classification of the S. copri isolates into species and subspecies clusters. The sequence homology of nucleotides and proteins were analysed against the control, S. copri DSM 18205, to determine the level of conservation in the alternative phosphoenolpyruvate synthesis pathways. ResultsEnolase (eno) was not identified in the S. copri strains; JCM 13468, LKV-178-WT-2C, RHA03, RHA01 and RHA02, and the whole genome phylogenetic grouping of these strains, has proposed the existence of an eno(-) subspecies of S. copri. This work furthered this idea by identifying alternative PEP pathways from formate, ferredoxin and fumarate, which were the most conserved in the eno(-) S. copri genomes. ConclusionThis work has provided rationale to why enolase may not be present within the eno(-) S. copri isolates and have shown that these alternative PEP synthesis pathways could negate the requirement of enolase in cells and may be factor in evolution of S. copri metabolism.

During mycological surveys in Yunnan Province, China, specimens of a fungus producing massive, upright stromata up to 50 cm high and individually 2.2 Kg in weight were sampled. Through an integrative taxonomic approach combining detailed morphology, multilocus phylogeny (ITS, LSU, RPB2, TUB2), and phylogenomic analyses, this fungus is proposed as the new species Dianjunus rex gen. et sp. nov., the type of the new family Dianjunaceae (Xylariales). Phylogenetic analyses robustly place Dianjunaceae as a distinct sister clade to Graphostromataceae. Divergence time estimation dates the origin of this family to the early Paleocene (~65 Mya), coinciding with the post-K-Pg extinction period, when an estimated 75% of all plant and animal species went extinct, and a significant ecological reorganization of life on earth happened. The stromata of D. rex represent the largest fructifications documented within the Ascomycota, significantly expanding the known morphological range of the Xylariales. The study provides a comprehensive description, including a nodulisporium-like anamorph with periconiella-like branching patterns, and discusses the taxon's phylogenetic placement, and distinctive morphology. This discovery highlights the unexplored fungal diversity in East Asian forests.

Background Erythema nodosum leprosum (ENL) is a severe inflammatory complication of lepromatous leprosy characterised by recurrent inflammatory episodes often requiring prolonged immunosuppression. The severity of ENL can be quantified using the validated and reliable ENLIST ENL Severity Scale (EESS). The longitudinal course of ENL and how it is captured using standardised severity measures has not been well described. We prospectively evaluated the changes in ENL severity over time using the EESS in a randomised clinical trial. Methods We conducted a post-hoc analysis of participants enrolled in the Methotrexate and Prednisolone Study in ENL, an international multicentre randomised controlled trial conducted in Ethiopia, India, Indonesia, and Nepal. Adults with severe ENL (EESS score [&ge;]9) were followed for 60 weeks with repeated EESS assessments. Longitudinal trajectories were analysed using mixed-effects regression models. Item-level analyses characterised the clinical phenotype captured by the scale. Associations between EESS score, prednisolone exposure, and dermatology-specific health-related quality of life measured using the Dermatology Life Quality Index (DLQI) were examined. Findings A total of 135 participants contributed 1,958 EESS assessments. Mean EESS declined rapidly during the first four weeks of treatment (-2.10 points/week; 95% CI -2.36 to -1.84; p<0.001), increased modestly during reduction in corticosteroid dose (weeks 4-20), and gradually declined thereafter. Severe ENL (EESS score [&ge;]9) occurred in 20.6% of visits and was characterised primarily by pain and cutaneous inflammatory manifestations. Participants who required additional prednisolone had persistently higher EESS scores and showed limited improvement compared with those who did not receive additional prednisolone. Longitudinal EESS scores were strongly correlated with the DLQI score (Spearmans {rho}=0.75; p<0.001). Conclusion The EESS captures clinically meaningful changes in ENL severity, aligns with treatment decisions, and reflects patient-reported severity over time. These findings support the use of the EESS as a robust tool for monitoring ENL severity in both clinical research and routine care.