Biology subjects

Urban, M.

Publications and source records attributed to Urban, M..

14 recordsLinked to original sources

bioRxiv2026

Comparative analysis of 15 chromosome-scale near T2T assemblies of Brazilian Fusarium graminearum isolates

Fusarium graminearum is the causative agent of Fusarium head blight of wheat, yet genomic resources for Brazilian populations, where it is the dominant species, remain limited to a single reference genome. We report fifteen near telomere-to-telomere (T2T) assemblies of Brazilian F. graminearum isolates generated with Oxford Nanopore sequencing. Each genome was assembled into four nuclear chromosomes, with 118 of 120 telomeres resolved, BUSCO completeness of 99.199.3%, and 14,80214,894 predicted genes per isolate. The genomes are structurally conserved, showing no chromosome-scale rearrangements and uniform transposable element content. Orthology assignment placed 90.4% of genes in the core genome. This suggests that the large phenotypic variance in aggressiveness observed between isolates likely resides in regulatory differences and/or within the small and largely uncharacterised accessory fraction. Ultimately, these assemblies provide a high-resolution resource for the global FHB community and a foundation for comparative analysis of a previously underrepresented population.

bioinformatics↗

bioRxiv2026

The impact of long-read sequencing on fungal genome assemblies: progress and disparity

Fungal genomics has expanded rapidly over the past 30 years, and recently the pace and breath has further quickened for many taxa, although many taxonomic gaps persist. With three decades of rapid growth, fungal genomics now merits a re-examination of its history, progress, and unresolved taxonomic gaps. Here, we review the development of fungal genomics from early efforts such as the Fungal Genome Initiative to current progress driven by third-generation long-read sequencing. We have compiled and summarised publicly available fungal genomes to highlight trends in assembly quality, adoption of long-read technologies, and taxonomic representation. Notably, substantial phylogenetic gaps remain, particularly outside Dikarya, and significant challenges persist for unculturable taxa. This review identifies priorities for the fungal community, including: (1) coordinated efforts to close major taxonomic gaps across the fungal tree of life; (2) improved repository metrics to facilitate identification of high-quality assemblies; and (3) improved and standardised genome annotation which is lacking for most assemblies. Together, these steps will support the development of reliable genomic resources that capture the full breadth of diversity across the fungal kingdom, generating foundational data for comparative genomics, evolutionary biology, functional studies, genetic studies and applied research.

genomics↗

bioRxiv2026

Recognizing dUTPase as a mitotic factor essential for early embryonic development

dUTPase is universally regarded as a metabolic sanitizing enzyme that protects genomes by preventing the incorporation of uracil into DNA. Despite its essentiality across eukaryotes, no function beyond nucleotide sanitization has been demonstrated. Here, we uncover a conserved, non-canonical role for dUTPase as a regulator of mitosis. Using Drosophila and mouse models, we demonstrate that dUTPase loss causes early embryonic lethality characterized by severe mitotic failure that, cannot be rescued by disabling uracil-DNA repair, uncoupling dUTPase essentiality from DNA repair pathways. Mechanistically, dUTPase dynamically associates with the mitotic spindle and centrosomes, and its depletion induces centrosome amplification and chromosome segregation defects. Beyond cell division, dUTPase dosage bidirectionally controls cell migration, linking its mitotic function to cellular behaviors relevant for metastasis. Together, our findings redefine dUTPase as a moonlighting mitotic factor that coordinates centrosome integrity and spindle dynamics, expanding its known repertoire beyond nucleotide metabolism. HighlightsdUTPase deficiency leads to mitotic defects during early embryonic development in both Drosophila and mouse models dUTPase shows dynamic spatiotemporal localization associated with microtubules through mitosis dUTPase is essential for the normal number and intracellular localization of centrosomes dUTPase deficiency counteracts while its overexpression enhances cell migration

molecular biology↗

bioRxiv2026

Cross-View Latent Integration via Nonparametric Gamma Shrinkage Factor Analysis

Factor analysis is a dominant paradigm for multi-omic heterogeneous data, but is challenged by partially redundant signals and noise across views and by an unknown true number of factors. We present CLING, an unsupervised multi-view factor model with hierarchical Bayesian sparsity priors: a product-of-Gammas prior inducing cumulative column-wise shrinkage (increasing with factor index) coupled with a Gamma-Gamma local-precision hierarchy on loadings yielding heavy-tailed marginals. This pairing enables automatic factor selection by adaptively deactivating unsupported factors while retaining active ones during inference, and induces selective sparsity that allows salient loadings to escape shrinkage while collapsing negligible ones. As a fully conjugate hierarchical model, CLING admits a scalable variational inference algorithm for multi-view data. Across synthetic benchmarks and multiomics datasets, CLING recovers more accurate factors and more informative loadings while explaining at least as much variance as competitive multi-view baselines; on glioblastoma gene expression and DNA methylation data, CLING identifies pathways linked to tumor subtype and patient age. Source code: https://github.com/szczurek-lab/CLING.

bioinformatics↗

bioRxiv2025

Acclimation of carbon metabolism to a changing environment across a leaf rosette of Arabidopsis thaliana

O_LIPlants need to efficiently and quickly stabilize photosynthesis and carbon metabolism under changing environmental conditions to prevent irreversible tissue damage C_LIO_LIDifferently developed leaves of Arabidopsis thaliana rosettes are typically homogenized for metabolic analyses which might result in significant over- or underestimation of metabolite dynamics. C_LIO_LIHere, photosynthesis and carbon metabolism were analysed in mature and immature leaves of rosettes of natural Arabidopsis accessions originating from southern and northern Europe. Ambient growth condition at 22 {degrees}C was compared to a combined low temperature/elevated light treatment. C_LIO_LIGradients of Fv/Fm and CO2 assimilation rates across leaf rosettes hinted towards tissue-specific acclimation capacities of photosynthesis and carbon metabolism. C_LIO_LIDynamics of carbohydrates and carboxylic acids were integrated with photosynthetic parameters in a quantitative carbon balance model. Model simulations suggested that mature leaf tissue stabilizes acclimation of carbon metabolism in immature leaf tissue and/or other sink tissue. C_LIO_LIIn conclusion, acclimation capacities of photosynthesis to low temperature and elevated light significantly differ within one leaf rosette of Arabidopsis. This directly affects carbon metabolism which shapes the metabolic acclimation capacity of the whole plant. C_LIO_LIIt is suggested to consider such tissue-specific effects which might otherwise be hidden behind a high variance of experimental data. C_LI

plant biology↗

bioRxiv2025

Chromosome segregation in Escherichia coli involves DNA binding by the MinC-MinD complex

The Escherichia coli Min system, composed of the proteins MinC, MinD, and MinE, is crucial for directing septum formation at mid-cell. MinD has also been shown to bind DNA in a non-sequence-specific manner and to tether plasmid DNA to liposomes. A role for the Min system in chromosome segregation has been proposed, whereby repeated, transient membrane-tethering events mediated by MinD could help prevent backward movements of the duplicated chromosomes once entropic forces weaken. However, the contribution of the other Min proteins to DNA binding has remained unclear. Here, we demonstrate that DNA binding is strongly enhanced when MinD and MinC form a complex, while MinE and FtsZ (at high concentrations) interfere with this binding. We reveal a critical role for the linker connecting MinCs N- and C-terminal domains in both DNA binding and inhibition of FtsZ polymerization. E. coli strains expressing MinC linker variants from the native locus, in combination with the alpha subunit of HU tagged with GFP, display chromosomes that appear more spread throughout the cell. This phenotype could reflect reduced compaction, chromosome segregation defects, or a combination of both. These findings suggest a previously unrecognized role for the MinC linker in DNA binding and provide new insights into a potential physiological role of MinCD-DNA bnding in chromosome organization and/or dynamics.

microbiology↗

bioRxiv2024

The Fusarium graminearum effector protease FgTPP1 suppresses immune responses and facilitates Fusarium Head Blight Disease

Most plant pathogens secrete effector proteins to circumvent host immune responses, thereby promoting pathogen virulence. One such pathogen is the fungus Fusarium graminearum, which causes Fusarium Head Blight (FHB) disease on wheat and barley. Transcriptomic analyses revealed that F. graminearum expresses many candidate effector proteins during early phases of the infection process, some of which are annotated as proteases. However, the contributions of these proteases to virulence remains poorly defined. Here, we characterize a F. graminearum endopeptidase, FgTPP1 (FGSG_11164), that is highly upregulated during wheat spikelet infection and is secreted from fungal cells. To elucidate the potential role of FgTPP1 in F. graminearum virulence, we generated FgTPP1 deletion mutants ({Delta}Fgtpp1) and performed FHB infection assays. While the number of completely bleached spikes infected by F. graminearum wild-type reached 50% of total infected spikes, the number of fully bleached spikes infected by{Delta} Fgtpp1 mutants was 25%, suggesting FgTPP1 contributes to fungal virulence. Transient expression of green fluorescent protein (GFP)-tagged FgTPP1 revealed that FgTPP1 localizes, in part, to chloroplasts and attenuates chitin-mediated activation of mitogen-activated protein kinase (MAPK) signaling, reactive oxygen species production, and cell death induced by an autoactive disease resistance protein when expressed in planta. Notably, the FgTPP1 protein is conserved across the Ascomycota phylum, making it a core effector among ascomycete plant pathogens. These properties make FgTPP1 an ideal candidate for decoy substrate engineering, with the goal of engineering resistance to FHB, and likely other crop diseases caused by ascomycete fungi.

plant biology↗

bioRxiv2024

A conserved fungal Knr4/Smi1 protein is vital for maintaining cell wall integrity and host plant pathogenesis

Filamentous plant pathogenic fungi pose significant threats to global food security, particularly through diseases like Fusarium Head Blight (FHB) and Septoria Tritici Blotch (STB) which affects cereals. With mounting challenges in fungal control and increasing restrictions on fungicide use due to environmental concerns, there is an urgent need for innovative control strategies. Here, we present a comprehensive analysis of the stage-specific infection process of Fusarium graminearum in wheat spikes by generating a dual weighted gene co-expression network (WGCN). Notably, the network contained a mycotoxin-enriched fungal module that exhibited a significant correlation with a detoxification gene-enriched wheat module. This correlation in gene expression was validated through quantitative PCR. By examining a fungal module with genes highly expressed during early symptomless infection, we identified a gene encoding FgKnr4, a protein containing a Knr4/Smi1 disordered domain. Through comprehensive analysis, we confirmed the pivotal role of FgKnr4 in various biological processes, including morphogenesis, growth, cell wall stress tolerance, and pathogenicity. Further studies confirmed the observed phenotypes are partially due to the involvement of FgKnr4 in regulating the fungal cell wall integrity pathway by modulating the phosphorylation of the MAP-kinase MGV1. Orthologues of FgKnr4 are widespread across the fungal kingdom but are absent in other Eukaryotes, suggesting the protein has potential as a promising intervention target. Encouragingly, the restricted growth and highly reduced virulence phenotypes observed for{Delta} Fgknr4 were replicated upon deletion of the orthologous gene in the wheat fungal pathogen Zymoseptoria tritici. Overall, this study demonstrates the utility of an integrated network-level analytical approach to pinpoint genes of high interest to pathogenesis and disease control.

plant biology↗

bioRxiv2023

The trichothecene mycotoxin deoxynivalenol facilitates cell-to-cell invasion during wheat-tissue colonisation by Fusarium graminearum

Fusarium Head Blight (FHB) disease on small grain cereals is primarily caused by the ascomycete fungal pathogen Fusarium graminearum. Infection of floral spike tissues is characterised by the biosynthesis and secretion of potent trichothecene mycotoxins, of which deoxynivalenol (DON) is widely reported due to its negative impacts on grain quality and consumer safety. The TRI5 gene encodes an essential enzyme in the DON biosynthesis pathway and the single gene deletion mutant,{Delta} Tri5, is widely reported to restrict disease progression to the inoculated spikelet. In this study, we present novel bioimaging evidence revealing that DON facilitates the traversal of the cell wall through plasmodesmata, a process essential for successful colonisation of host tissue. Chemical complementation of{Delta} Tri5 did not restore macro- or microscopic phenotypes, indicating that DON secretion is tightly regulated both spatially and temporally. A comparative qualitative and quantitative morphological cellular analysis revealed infections had no impact on plant cell wall thickness. Immuno- labelling of callose at plasmodesmata during infection indicates that DON can increase deposits when applied exogenously, but is reduced when F. graminearum hyphae are present. This study highlights the complexity of the inter-connected roles of mycotoxin production, cell wall architecture and plasmodesmata in this highly specialised interaction.

plant biology↗

bioRxiv2023

Identification and functional characterisation of a locus for target site integration in Fusarium graminearum

BackgroundFusarium Head Blight is a destructive floral disease of different cereal crops. The Ascomycete fungus Fusarium graminearum (Fg) is one of the main causal agents of FHB in wheat and barley. The role(s) in virulence of Fg genes include genetic studies that involve the transformation of the fungus with different expression cassettes. We have observed in several studies where Fg genes functions were characterised that integration of expression cassettes occurred randomly. Random insertion of a cassette may disrupt gene expression and/or protein functions and hence the overall conclusion of the study. Target site integration (TSI) is an approach that consists in identifying a chromosomal region where the cassette can be inserted. The identification of a suitable locus for TSI in Fg would avert the potential risks of ectopic integration. ResultsHere, we identified a highly conserved intergenic region on chromosome 1 suitable for TSI. We named this intergenic region the TSI locus 1. We developed an efficient cloning vector system based on the Golden Gate method to clone different expression cassettes for use in combination with TSI locus 1. We present evidence that integrations in the TSI locus 1 affects neither fungal virulence nor fungal growth under different stress conditions. Integrations at the TSI locus 1 resulted in the expression of different gene fusions. In addition, the activities of Fg native promoters were not altered by integration into the TSI locus 1. We have developed a bespoke bioinformatic pipeline to analyse the existence of ectopic integrations and tandem insertions of the cassette that may occurred during the transformation process. Finally, we established a protocol to study protein secretion in wheat coleoptiles using confocal microscopy and the TSI locus 1. ConclusionThe TSI locus 1 can be used in Fg and potentially other cereal infecting Fusarium species for diverse studies including promoter activity analysis, secretion, protein localisation studies and gene complementation. The bespoke bioinformatic pipeline developed in this work can be an alternative to southern blotting, the gold standard technique to identify ectopic integration and tandem insertions in fungal transformation.

genetics↗

bioRxiv2022

A framework for community curation of interspecies interactions literature

The quantity and complexity of data being generated and published in biology has increased substantially, but few methods exist for capturing knowledge about phenotypes derived from molecular interactions between diverse groups of species, in such a way that is amenable to data-driven biology and research. To improve access to this knowledge, we have constructed a framework for the curation of the scientific literature studying interspecies interactions, using data curated for the Pathogen-Host Interactions Database (PHI-base) as a case study. The framework provides a curation tool, phenotype ontology and controlled vocabularies to curate pathogen-host interaction data (at the level of the host, pathogen, strain, gene and genotype). The concept of a multispecies genotype, the metagenotype, is introduced to facilitate capturing changes in the pathogens disease-causing abilities, and host resistance or susceptibility observed by gene alterations. We report on this framework and describe PHI-Canto, a community curation tool for use by publication authors.

microbiology↗

bioRxiv2021

In vitro evaluation of the effect of mutations in primer binding sites on detection of SARS-CoV-2 by RT-qPCR

A number of RT-qPCR assays for the detection of SARS-CoV-2 have been published and are listed by the WHO as recommended assays. Furthermore, numerous commercial assays with undisclosed primer and probe sequences are on the market. As the SARS-CoV-2 pandemic progresses, the virus accrues mutations, which in some cases - as seen with the B.1.1.7 variant - can outperform and push back other strains of SARS-CoV-2. If mutations occur in primer or probe binding sites, this can impact RT-qPCR results and impede SARS-CoV-2 diagnostics. Here we tested the effect of primer mismatches on RT-qPCR performance in vitro using synthetic mismatch in vitro transcripts. The effects of the mismatches ranged from a shift in ct values from -0.13 to +7.61. Crucially, we found that a mismatch in the forward primer has a more detrimental effect for PCR performance than a mismatch in the reverse primer. Furthermore, we compared the performance of the original Charite RdRP primer set, which has several ambiguities, with a primer version without ambiguities and found that without ambiguities the ct values are ca. 3 ct lower. Finally, we investigated the shift in ct values observed with the Seegene Allplex kit with the B.1.1.7 SARS-CoV-2 variant and found a three-nucleotide mismatch in the forward primer of the N target.

molecular biology↗

bioRxiv2019

The Vertebrate Codex Gene Breaking Protein Trap Library For Genomic Discovery and Disease Modeling Applications

The zebrafish is a powerful model to explore the molecular genetics and expression of the vertebrate genome. The gene break transposon (GBT) is a unique insertional mutagen that reports the expression of the tagged member of the proteome while generating Cre-revertible genetic alleles. This 1000+ locus collection represents novel codex expression data from the illuminated mRFP protein trap, with 36% and 87% of the cloned lines showcasing to our knowledge the first described expression of these genes at day 2 and day 4 of development, respectively. Analyses of 183 molecularly characterized loci indicate a rich mix of genes involved in diverse cellular processes from cell signaling to DNA repair. The mutagenicity of the GBT cassette is very high as assessed using both forward and reverse genetic approaches. Sampling over 150 lines for visible phenotypes after 5dpf shows a similar rate of discovery of embryonic phenotypes as ENU and retroviral mutagenesis. Furthermore, five cloned insertions were in loci with previously described phenotypes; embryos homozygous for each of the corresponding GBT alleles displayed strong loss of function phenotypes comparable to published mutants using other mutagenesis strategies (ryr1b, fras1, tnnt2a, edar and hmcn1). Using molecular assessment after positional cloning, to date nearly all alleles cause at least a 99+% knockdown of the tagged gene. Interestingly, over 35% of the cloned loci represent 68 mutants in zebrafish orthologs of human disease loci, including nervous, cardiovascular, endocrine, digestive, musculoskeletal, immune and integument systems. The GBT protein trapping system enabled the construction of a comprehensive protein codex including novel expression annotation, identifying new functional roles of the vertebrate genome and generating a diverse collection of potential models of human disease.

genomics↗

bioRxiv2019

Elucidating small RNA pathways in Arabidopsis thaliana egg cells

O_LISmall RNA pathway components and small RNA profiles of flowering plant egg cells are largely unexplored, mainly because they are not easily accessible but deeply buried inside the ovary.\nC_LIO_LIWe describe here the utilization of proliferating callus tissue that adopted transcriptome features of Arabidopsis egg cell as a tool to explore small RNA pathway components and small RNA profiles in egg cells. We furthermore complement our studies with mRNA-Seq data from isolated Arabidopsis egg cells and provide data validation by promoter-reporter studies and whole mount in situ hybridization.\nC_LIO_LISequencing of small RNA libraries demonstrate the predominance of TE-derived siRNAs in the egg cell-related callus. TE-features and expression profiles suggest post-transcriptional silencing of activated Gypsy-like LTR retrotransposons, whereas the majority of class II DNA transposons belonging to Copia, CACTA, hAT-like and Mutator superfamilies are subjected to transcriptional silencing.\nC_LIO_LISmall RNA-seq furthermore led to the identification of differentially expressed known and novel miRNAs whose expression in the egg cell was verified by small RNA whole mount in situ hybridization. Both the strong expression of miRNAs in the egg-cell-adjoining synergids and the secretion of miRNAs into the micropyle suggest hitherto undescribed roles for these accessory cells in intercellular communication with the egg cell and the arriving pollen tube.\nC_LIO_LIIn conclusion, our datasets provide valuable and comprehensive resources to study small RNA pathways and small-RNA-mediated epigenetic reprogramming during egg cell differentiation and the onset of plant embryogenesis.\nC_LI

plant biology↗