{"title":"Optimized convolutional neural networks for real-time detection and severity assessment of early blight in tomato (Solanum lycopersicum L.)","authors":"Tushar Dhar , Roaf Ahmad Parray , Bishnu Maya Bashyal , Awani Kumar Singh , Parveen Dhanger , Tapan Kumar Khura , Rajeev Kumar , Murtaza Hasan , Md Yeasin","doi":"10.1016/j.fgb.2025.103984","DOIUrl":"10.1016/j.fgb.2025.103984","url":null,"abstract":"<div><div>Early blight, caused by <em>Alternaria alternata</em>, poses a critical challenge to tomato (<em>Solanum lycopersicum L.</em>) production, causing significant yield losses worldwide. Despite advancements in plant disease detection, existing methods often lack the robustness, speed, and accuracy needed for real-time, field-level applications, particularly under variable environmental conditions. This study addresses these gaps by leveraging transfer learning with optimized MobileNet architectures to develop a highly efficient and generalizable detection system. A diverse dataset of 6451 tomato leaf images, encompassing healthy and varying disease severity levels (low, medium, high) under multiple lighting conditions, was curated to improve model performance across real-world scenarios. Four MobileNet variants—MobileNet, MobileNet V2, MobileNet V3 Small, and MobileNet V3 Large—were fine-tuned, with MobileNet V3 Large achieving the highest classification accuracy of 99.88 %, an F1 score of 0.996, and a rapid inference time of 67 milliseconds. These attributes make it ideal for real-time IoT applications, including smartphone-based disease monitoring, automated precision spraying, and smart agricultural systems. To further validate diseased samples, internal transcribed spacer (ITS) sequence analysis confirmed <em>A. alternata</em> with over 98 % similarity to known isolates in the NCBI database. This study bridges critical research gaps by providing a robust, non-destructive, and real-time solution for early blight severity assessment, enabling timely, targeted interventions to mitigate crop losses in precision agriculture.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"178 ","pages":"Article 103984"},"PeriodicalIF":2.4,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143878589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Li Xu, Jamy Schulpen, Ronald P. de Vries, Mao Peng
{"title":"Identification of putative fungal reference genes with stable expression from large RNA-seq datasets","authors":"Li Xu, Jamy Schulpen, Ronald P. de Vries, Mao Peng","doi":"10.1016/j.fgb.2025.103985","DOIUrl":"10.1016/j.fgb.2025.103985","url":null,"abstract":"<div><div>RNA-sequencing (RNA-seq) is the dominant technology for genome-wide transcript quantification in various biological studies. The wide applications of RNA-seq have played an essential role in elucidating complex molecular mechanisms of fungal physiology, and have generated large volumes of related data that are valuable for further bioinformatic mining. In this study, we focus on identifying fungal reference genes from large available transcriptome datasets. In total, 44 candidate reference genes from <em>Aspergillus niger</em> were identified through strict statistical analysis of 332 transcriptomic samples. These candidates cover both newly identified genes and previously reported housekeeping genes and were enriched in several basic cellular pathways, such as genes encoding ubiquitin-conjugating enzyme, 26S proteasome regulatory subunits, vacuolar H<sup>+</sup>-ATPase subunits, mitochondrial import protein and Ras-related GTPase. Moreover, 26 of the newly identified reference genes with a single ortholog in four other fungi showed stable expression patterns across these fungi. Additionally, these new candidates showed more stable expression than the traditionally used reference genes in the tested datasets, such as <em>gapdh</em>, highlighting their potential to improve normalization of RT-qPCR and transcriptome data.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"178 ","pages":"Article 103985"},"PeriodicalIF":2.4,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143869425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Where to grow and where to go","authors":"Marius Kriegler, Satur Herrero, Reinhard Fischer","doi":"10.1016/j.fgb.2025.103983","DOIUrl":"10.1016/j.fgb.2025.103983","url":null,"abstract":"<div><div>Filamentous fungi grow as very elongated tubular cells that extend by membrane extension and cell-wall biosynthesis. Membrane and enzyme delivery depend on secretory vesicles that travel along microtubules, accumulate in a structure called the <em>Spitzenkörper</em> and then move along actin cables towards the apical membrane. Whereas vesicle fusion and membrane insertion are well studied, less is known about the mechanisms with which the zones of vesicle fusion and hence the growth zones are defined. One mechanism by which polarity is established and maintained is the polar localization of <em><u>c</u>ell-<u>e</u>nd <u>m</u>arker <u>p</u>roteins</em> (CEMPs). They form multi-protein complexes with formin as F-actin polymerase. CEMP delivery depends on microtubules, and hence CEMPs coordinate the microtubule with the actin cytoskeleton. Actin filaments capture microtubule ends, and this positive feedback loop quickly establishes active growth sites. However, CEMP complexes are self-limiting, because fusing vesicles disturb local growth zones and Ca<sup>2+</sup> influx pulses lead to F-actin disassembly. This model emerged from studies in <em>Schizosaccharomyces pombe</em> and <em>Aspergillus nidulans</em>. Surprisingly, deletion of CEMP-coding genes is not lethal. <em>S. pombe</em> mutants form T-shaped cells and <em>A. nidulans</em> germlings grow less straight. In comparison, CEMP-mutants had a strong phenotype in <em>Arthrobotrys flagrans</em>, a nematode-trapping fungus, which produces ring-like trapping structures. CEMP-mutants fail to form adhesive rings and instead form sticks. CEMP overexpression caused a hyperbranching phenotype. Hence, CEMPs are involved in polarity maintenance and play critical roles during modulations of polarity. Here, we are going to discuss the functions of CEMPs and their connections to other polarity determinants.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"178 ","pages":"Article 103983"},"PeriodicalIF":2.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143789359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Distribution and dynamics of hyphal organelles","authors":"Barry Bowman","doi":"10.1016/j.fgb.2025.103982","DOIUrl":"10.1016/j.fgb.2025.103982","url":null,"abstract":"<div><div>Filamentous fungi have been very useful organisms for the investigation of organelles in eukaryotic cells. The structure and function of fungal organelles is generally very similar to that observed in animal cells. However, the nature of a “cell” in many filamentous fungi is unusual, because in many of these organisms the filaments are structured as a large syncytium. In the Ascomycota hyphae are typically a very long tube divided into different compartments by an incomplete cell wall called the septum. The pore in the middle of the septum is large enough to allow virtually all organelles to move from one hyphal compartment to another. In this review, I will look at the dynamics of this movement of organelles and describe what we know about how the structure and distribution of organelles varies from one hyphal compartment to another.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"178 ","pages":"Article 103982"},"PeriodicalIF":2.4,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143725237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
J. Alberto Patiño-Medina , David Vargas-Tejeda , Nancy Y. Reyes-Mares , Viridiana Alejandre-Castañeda , Cesar J. Torres-Cortes , Carlos Pérez-Arques , Leon F. Ruiz-Herrera , Joel Ramírez-Emiliano , Sandeep Vellanki , Marco I. Valle-Maldonado , Karla Viridiana Castro-Cerritos , Martha I. Ramirez-Diaz , Soo Chan Lee , Victoriano Garre , Víctor Meza-Carmen
{"title":"Heterotrimeric G-gamma 1 (Gpg1) participates with G-beta 1 (Gpb1) in the induction of hyphal growth and virulence via the PKA pathway in Mucor lusitanicus","authors":"J. Alberto Patiño-Medina , David Vargas-Tejeda , Nancy Y. Reyes-Mares , Viridiana Alejandre-Castañeda , Cesar J. Torres-Cortes , Carlos Pérez-Arques , Leon F. Ruiz-Herrera , Joel Ramírez-Emiliano , Sandeep Vellanki , Marco I. Valle-Maldonado , Karla Viridiana Castro-Cerritos , Martha I. Ramirez-Diaz , Soo Chan Lee , Victoriano Garre , Víctor Meza-Carmen","doi":"10.1016/j.fgb.2025.103974","DOIUrl":"10.1016/j.fgb.2025.103974","url":null,"abstract":"<div><div>Previous work from our lab indicates that the heterotrimeric Gβ subunit 1 (Gpb1) enhances hyphal development and virulence in <em>Mucor lusitanicus</em>. In this study, three Gγ- and two additional Gβ-encoding genes were deleted to identify which ones might have a similar role as Gpb1. Deletion of <em>gpg1</em> reduces hyphal growth, virulence, cyclic adenosine monophosphate (cAMP) levels, and protein kinase A (PKA) activity, similar to <em>gpb1</em> deletion, suggesting that <em>gpg1</em> participates in the same regulatory pathway as <em>gpb1</em>. The defects observed in Δ<em>gpg1</em> or Δ<em>gpb1</em> were suppressed by overexpression of the gene <em>pkaR1</em> encoding the regulatory subunit 1 of PKA, indicating that this pathway is controlled by Gpg1 and Gpb1. Moreover, Δ<em>gpg1</em> and Δ<em>gpb1</em> show a downregulation of the transcription factors <em>tec1</em> and <em>tec2</em>. Furthermore, <em>tec</em>-overexpression in Δ<em>gpg1</em>, Δ<em>gpb1, and</em> Δ<em>pkaR1</em> restores the wild-type phenotype, indicating that both Tec are under control by the Gpb1, Gpg1, and PKA pathway. Moreover, the Δ<em>gpb1</em>/Δ<em>gbg1</em><sup><em>(+)(−)</em></sup> exhibits lower aerobic germination, hyphal growth and downregulates NAD<sup>+</sup>-glutamate dehydrogenases (<em>gdh</em>2a/b), whereas virulence is similar to that of the wild-type (WT) strain. These alterations in Δ<em>gpb1</em>/Δ<em>gbg1</em><sup><em>(+)(−)</em></sup> were reversed by the presence of glutamate during growth, suggesting that NAD<sup>+</sup>-Gdh2 could be under control of these subunits. Compared to the WT and Δ<em>gpb1</em>/Δ<em>gbg1</em><sup><em>(+)(−)</em></sup> strains under aerobic growth, single deletion strains showed lower rhizoferrin levels, respiration and reactive oxygen species levels. Our results suggest that Gpg1 interacts with Gpb1 to positively control the hyphal development and virulence by repressing the PKA pathway, thereby regulating the mitochondrial oxidative metabolism in <em>M. lusitanicus</em>.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"178 ","pages":"Article 103974"},"PeriodicalIF":2.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143574707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Thomas Svoboda , Dominik Niederdöckl-Loibl , Andreas Schüller , Karin Hummel , Sarah Schlosser , Ebrahim Razzazi-Fazeli , Joseph Strauss
{"title":"Locus-specific chromatin proteomics using dCas-guided proximity labelling in Aspergillus nidulans","authors":"Thomas Svoboda , Dominik Niederdöckl-Loibl , Andreas Schüller , Karin Hummel , Sarah Schlosser , Ebrahim Razzazi-Fazeli , Joseph Strauss","doi":"10.1016/j.fgb.2025.103973","DOIUrl":"10.1016/j.fgb.2025.103973","url":null,"abstract":"<div><div>Proximity labelling that uses promiscuous biotin ligases (BirA) fused to a bait protein is a powerful tool to identify protein interaction partners <em>in vivo</em> under different metabolic or developmental conditions. BirA can also be used to determine protein composition and interaction partners at specific chromatin locations when it is fused with enzymatically-disabled Cas9 (dCas9) and then guided to the location of interest by sgRNAs. We adapted this method (called CasID) for fungal cells using the nitrate assimilation gene cluster of <em>A. nidulans</em> as a model locus and estrogen-inducible expression of the dCas9-BirA fusion to improve condition-specific labelling. For method establishment, we first verified the presence of dCas-BirA and a known transcription factor at the nitrate locus by chromatin immunoprecipitation (ChIP). Results show that both dCas-BirA and the AreA transcription factor are present at the locus of interest under the conditions used for biotinylation. We then optimized the CasID procedure for efficient labelling and background reduction using the CasID-sgRNA strain and two control strains, one lacking the sgRNA and another one lacking the whole CasID system. Here we provide proof-of-concept for the suitability of the method by showing that biotinylated proteins are enriched in the CasID strains in comparison to the controls. After background reduction, 32 proteins remained in two independent experiments exclusively enriched in the Cas-ID-sgRNA strain. Among these proteins was NmrA, an AreA-interacting regulator, and we also found several chromatin-associated proteins. Overall, our results demonstrate that CasID is suitable for locus-specific labelling and identification of chromatin-associated proteins and transcription factors in <em>A. nidulans</em>. However, the high background of proteins that are biotinylated out of chromatin context or unspecifically attach to the affinity purification matrix needs to be addressed by implementing a set of rigorous controls. In summary, we herewith provide a detailed protocol for application of the method that proved to be useful for the identification of novel chromatin-associated proteins and their interaction partners at a specific genomic locus in divers metabolic and developmental conditions.</div></div><div><h3>Author summary</h3><div>This study demonstrates that locus-specific proteomics can be carried out by dCas-BirA guided proximity labelling in <em>Aspergillus nidulans.</em> For establishment, we targeted the well-described bidirectional promoter region between <em>niaD</em>, a nitrate reductase, and <em>niiA</em>, a nitrite reductase. At this locus we could test by chromatin immunoprecipitation (ChIP) in combination with qPCR if both, the dCas9-BirA fusion as well as a central transcription factor are at the locus under the conditions of our CasID experiment. After this first control step, we considered that unspecific labelling by dCas-BirA during the t","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"178 ","pages":"Article 103973"},"PeriodicalIF":2.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143574708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Piotr J. Pietras , Monika Chaszczewska-Markowska , Daniel Ghete , Agata Tyczewska , Kamilla Bąkowska-Żywicka
{"title":"Saccharomyces cerevisiae recovery from various mild abiotic stresses: Viability, fitness, and high resolution three-dimensional morphology imaging","authors":"Piotr J. Pietras , Monika Chaszczewska-Markowska , Daniel Ghete , Agata Tyczewska , Kamilla Bąkowska-Żywicka","doi":"10.1016/j.fgb.2025.103975","DOIUrl":"10.1016/j.fgb.2025.103975","url":null,"abstract":"<div><div>Environmental conditions have a huge impact on the development of all living things but are especially important in the case of single-celled organisms such as <em>Saccharomyces cerevisiae</em> that must respond quickly and appropriately to any change. Many molecular mechanisms of response to stress have been identified in yeast, but only a few reports address physiological and morphological changes. To investigate <em>S. cerevisiae</em> recovery from ten mild stress conditions and to describe the viability and fitness, we performed a series of growth analysis experiments. Moreover, label-free live cell imaging of yeast subjected to ten environmental stresses has been achieved using holotomography - a leading-edge high resolution 3D quantitative phase imaging. We determined that recovery times of yeast cultures subjected to hyperosmotic and sugar starvation stresses were the shortest, as were the doubling times. Substantially lower proliferation capacity was recorded in yeast after applying sugar- and AA starvation, and high pH stresses, compared to control. Furthermore, the stationary growth was much shorter after subjecting yeast to hypoosmotic and heat stresses, and much longer after anaerobic and UV stresses. Further, we determined changes in shape, colony formation, cell wall damage, volume, sphericity, protein and lipid contents in yeast cells under stress conditions. The most prominent changes were observed for UV and hyperosmotic stresses. Condluding, stress conditions applied to yest cultures affected them differently, causing detrimental effects to their growth, metabolism, fitness and morphology. Moreover, we have proven that holotomography is excellent for precisely determining morphological changes of single cells.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"178 ","pages":"Article 103975"},"PeriodicalIF":2.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143574710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jieying Zhu , Weiwei Zhang , Yani Fan , Wei Deng , Liao Zhang , Shunxian Wang , Xingzhong Liu , Meichun Xiang
{"title":"Septin AoCDC11 is involved in trap morphogenesis, conidiation, and vegetative growth in carnivorous Arthrobotrys oligospora","authors":"Jieying Zhu , Weiwei Zhang , Yani Fan , Wei Deng , Liao Zhang , Shunxian Wang , Xingzhong Liu , Meichun Xiang","doi":"10.1016/j.fgb.2025.103971","DOIUrl":"10.1016/j.fgb.2025.103971","url":null,"abstract":"<div><div>Septins, a conserved family of cytoskeletal proteins with GTP-binding domains, play key roles in cell polarity, morphogenesis, cytoskeleton organization, and membrane remodeling. The nematode-trapping fungus <em>Arthrobotrys oligospora</em> can capture and kill nematodes using adhesive networks. It has been highlighted the importance of cell polarity, actin organization, and membrane remodeling in the process of trap formation, but the role of septins in adhesive-network forming remains unclear. In this study, we investigated the functions of <em>AoCDC11</em>, an ortholog of <em>Saccharomyces cerevisiae CDC11</em>, through gene disruption and multiphenotypic analysis. Disruption of <em>AoCDC11</em> led to reduced trap production and abnormal trap morphology. Compared to the wild type, <em>ΔAoCDC11</em> mutants significantly reduced trap formation to emerge more vegetative hyphae and produced more incompletely fused adhesive networks (45 % vs. 10 %) by fewer trap loops and septa. Additionally, <em>ΔAoCDC11</em> mutants exhibited a 36 % reduction in hyphal growth and 88 % decrease in conidiation compared to the wild type. Transcriptomic analysis revealed that AoCDC11 regulated genes involved in trap development, including those related to the cell cycle, anatomical structure development, cellular morphogenesis, vesicle transport, and membrane trafficking. These findings suggest that AoCDC11 plays a crucial role in trap morphogenesis, vegetative growth, and conidiation by modulating multiple biological processes. This study expands our understanding of the functions of septins in morphogenesis and survival strategy of nematode-trapping fungi.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"177 ","pages":"Article 103971"},"PeriodicalIF":2.4,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143534417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Functional redundancy and divergence of UDP-glucose 4-epimerases in galactose metabolism and cell wall biosynthesis in Aspergillus nidulans","authors":"Chihiro Kadooka , Shun Yakabe , Daisuke Hira , Taiki Futagami , Masatoshi Goto , Takuji Oka","doi":"10.1016/j.fgb.2025.103972","DOIUrl":"10.1016/j.fgb.2025.103972","url":null,"abstract":"<div><div>Galactose-containing polysaccharides in the cell walls of filamentous fungi are vital for hyphal formation, mycelial aggregation, and adhesion. Uridine diphosphate (UDP)-glucose 4-epimerase, an enzyme capable of reversibly converting UDP-glucose to UDP-galactose, plays a key role in galactose metabolism. This study investigates the functional specialization and overlapping roles of UDP-glucose 4-epimerases, UgeA and UgeB, in <em>Aspergillus nidulans</em>. Enzyme activity assays revealed that UgeA catalyzes the interconversion of UDP-glucose and UDP-galactose, while UgeB facilitates both UDP-glucose/UDP-galactose and UDP-<em>N</em>-acetylglucosamine/UDP-<em>N</em>-acetylgalactosamine interconversions. Both UgeA and UgeB successfully restored growth in a yeast <em>gal10</em> disruptant, indicating their involvement in galactose metabolism <em>in vivo</em>. Additionally, the <em>ugeB</em> disruptant of <em>A. nidulans</em> exhibited growth retardation during galactose metabolism, a defect that was alleviated by complementation with <em>ugeB</em> or multiple-copy expression of <em>ugeA</em>. These findings elucidate the complex interplay between sugar metabolism and cell wall synthesis in filamentous fungi and offer insights for the development of novel antifungal therapies.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"177 ","pages":"Article 103972"},"PeriodicalIF":2.4,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143484618","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alida van Dijk , Andi M. Wilson , Bianke Marx , Bianca Hough , Benedicta Swalarsk-Parry , Lieschen De Vos , Michael J. Wingfield , Brenda D. Wingfield , Emma T. Steenkamp
{"title":"CRISPR-Cas9 genome editing reveals that the Pgs gene of Fusarium circinatum is involved in pathogenicity, growth and sporulation","authors":"Alida van Dijk , Andi M. Wilson , Bianke Marx , Bianca Hough , Benedicta Swalarsk-Parry , Lieschen De Vos , Michael J. Wingfield , Brenda D. Wingfield , Emma T. Steenkamp","doi":"10.1016/j.fgb.2025.103970","DOIUrl":"10.1016/j.fgb.2025.103970","url":null,"abstract":"<div><div><em>Fusarium circinatum,</em> the causal agent of pine pitch canker, is one of the most destructive pathogens of <em>Pinus</em> species worldwide. Infections by this pathogen result in serious mortality of seedlings due to root and root collar disease, and growth reduction in trees due to canker formation and dieback. Although much is known about the population biology, genetics, and genomics of <em>F. circinatum</em>, relatively little is known regarding the molecular basis of pathogenicity in <em>F. circinatum.</em> In this study, a protoplast-based transformation using CRISPR-Cas9-mediated genome editing was utilized to functionally characterize a putative pathogenicity gene in three different strains of the fungus. <em>In silico</em> analyses suggested the gene likely encodes a small secreted protein, and all isolates in which it was deleted displayed significantly reduced vegetative growth and asexual spore production compared to the wild-type isolates. In pathogenicity tests, lesions induced by the deletion mutants on detached <em>Pinus patula</em> branches were significantly shorter than those produced by the wild-types. The putative pathogenicity gene was named <em>Pgs</em> reflecting its role in <u><strong>p</strong></u>athogenicity, <u><strong>g</strong></u>rowth, and <u><strong>s</strong></u>porulation. Future research will seek to explore the molecular mechanisms underlying the mutant phenotypes observed. Overall, this study represents a significant advance in <em>F. circinatum</em> research as the development and application of a Cas9-mediated gene deletion process opens new avenues for functional gene characterization underlying many of the pathogen's biological traits.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"177 ","pages":"Article 103970"},"PeriodicalIF":2.4,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143426204","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}