Pub Date : 2025-09-01DOI: 10.1016/j.fgb.2025.104025
Steven D. Harris
A defining feature of filamentous fungi is the ability to form branched hyphae that facilitate the efficient colonization of ecological niches. Two distinct branching patterns have been recognized in fungal hyphae; apical branching and lateral branching. The use of physiological approaches and image analysis has generated sufficient data to enable the development of robust models that predict branching behavior. However, insight into the molecular mechanisms that underlie branching has lagged. The intent of this review is to summarize processes that are known to contribute to the regulation of branching and the morphogenesis of branches. Potential directions for future study will also be suggested. Ultimately, advances in understanding hyphal branching will likely be best achieved using a “design-build-test-learn” approach that combines genetic analyses in model systems with ensemble modeling that generates testable predictions.
{"title":"Duplicating the mold: Branching of fungal hyphae","authors":"Steven D. Harris","doi":"10.1016/j.fgb.2025.104025","DOIUrl":"10.1016/j.fgb.2025.104025","url":null,"abstract":"<div><div>A defining feature of filamentous fungi is the ability to form branched hyphae that facilitate the efficient colonization of ecological niches. Two distinct branching patterns have been recognized in fungal hyphae; apical branching and lateral branching. The use of physiological approaches and image analysis has generated sufficient data to enable the development of robust models that predict branching behavior. However, insight into the molecular mechanisms that underlie branching has lagged. The intent of this review is to summarize processes that are known to contribute to the regulation of branching and the morphogenesis of branches. Potential directions for future study will also be suggested. Ultimately, advances in understanding hyphal branching will likely be best achieved using a “design-build-test-learn” approach that combines genetic analyses in model systems with ensemble modeling that generates testable predictions.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104025"},"PeriodicalIF":2.3,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144932127","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}
Pub Date : 2025-08-26DOI: 10.1016/j.fgb.2025.104024
Xuan Shang , Zhenwei Yang , Guanzu Peng , Yawen Wu , Fei Dou , Jin Liu , Wanjie Li
Accurate quantification of yeast sporulation efficiency is essential for genetic studies, but manual counting remains time-consuming and susceptible to subjective bias. Although deep learning tools like cellpose provide automated solutions, there exists a compelling need for alternative approaches that enable the quantification of spores. Our methodology employs ilastik's texture-feature optimization to reliably segment sporulating mother cells, intentionally avoiding explicit tetrad discrimination to ensure robustness across diverse spore morphologies. Subsequent Fiji-based image processing employs optimized algorithms for accurate spore quantification within cellular boundaries, facilitating automated batch classification of dyads, triads, and tetrads. Quantitative validation demonstrates our pipeline maintains strong concordance with manual counting (93.4 % agreement, ICC = 0.94) alongside a 68 % reduction in processing time (P < 0.001). The pipeline's reliability was further verified in Hsp82 phosphorylation mutants, consistently enables quantification of sporulation efficiency across genetic backgrounds. To balance throughput and precision, our workflow intentionally combines automated image processing (ilastik segmentation, Fiji quantification) with manual quality control checkpoints (segmentation validation). This modular pipeline allows adjustable segmentation parameters, compatibility with alternative nuclear markers, and batch processing of diverse imaging datasets. By combining accessibility with precision, our method provides laboratories a reproducible alternative to fully manual counting while maintaining compatibility with standard microscopy setups.
{"title":"Machine learning driven semi-automated framework for yeast sporulation efficiency quantification using ilastik segmentation and Fiji nuclear enumeration","authors":"Xuan Shang , Zhenwei Yang , Guanzu Peng , Yawen Wu , Fei Dou , Jin Liu , Wanjie Li","doi":"10.1016/j.fgb.2025.104024","DOIUrl":"10.1016/j.fgb.2025.104024","url":null,"abstract":"<div><div>Accurate quantification of yeast sporulation efficiency is essential for genetic studies, but manual counting remains time-consuming and susceptible to subjective bias. Although deep learning tools like cellpose provide automated solutions, there exists a compelling need for alternative approaches that enable the quantification of spores. Our methodology employs ilastik's texture-feature optimization to reliably segment sporulating mother cells, intentionally avoiding explicit tetrad discrimination to ensure robustness across diverse spore morphologies. Subsequent Fiji-based image processing employs optimized algorithms for accurate spore quantification within cellular boundaries, facilitating automated batch classification of dyads, triads, and tetrads. Quantitative validation demonstrates our pipeline maintains strong concordance with manual counting (93.4 % agreement, ICC = 0.94) alongside a 68 % reduction in processing time (<em>P</em> < 0.001). The pipeline's reliability was further verified in Hsp82 phosphorylation mutants, consistently enables quantification of sporulation efficiency across genetic backgrounds. To balance throughput and precision, our workflow intentionally combines automated image processing (ilastik segmentation, Fiji quantification) with manual quality control checkpoints (segmentation validation). This modular pipeline allows adjustable segmentation parameters, compatibility with alternative nuclear markers, and batch processing of diverse imaging datasets. By combining accessibility with precision, our method provides laboratories a reproducible alternative to fully manual counting while maintaining compatibility with standard microscopy setups.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104024"},"PeriodicalIF":2.3,"publicationDate":"2025-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144913658","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}
Pub Date : 2025-07-13DOI: 10.1016/j.fgb.2025.104023
Xinai Liu , Xiuwen Wang , Yueping Yang , Kang Li , Wenrong Liu , Qing Liu , Yuanda Song
As a well-established oleaginous model fungus, Mucor circinelloides has been extensively employed in lipid metabolism research due to its high lipid accumulation capacity and genetic tractability. Metformin hydrochloride, a biguanide drug, exerts significant effects on glucose and lipid metabolism in animal model, however, its effect on lipid metabolism in oleaginous microorganism is unclear. This investigation elucidates the regulatory effects of metformin hydrochloride on lipid biosynthesis in the model oleaginous fungus M. circinelloides. The findings revealed that metformin hydrochloride suppresses fatty acid biosynthesis in M. circinelloides. Addition of 4 g/L metformin hydrochloride to the growth medium of the fungus reduced the total fatty acid content from 29.57 % to 23.27 %, representing a decrease of 21.30 %. Furthermore, metformin hydrochloride significantly upregulated the transcriptional levels of AMPK subunits (such as Snf-α1, Snf-γ1, and Snf-γ3) while suppressing the expression of key genes involved in lipid synthesis, including acl, acc1, and acc2. Our enzymatic assays revealed that metformin hydrochloride treatment markedly inhibited ACC and 6PGDH activities, thereby restricting the availability of both acetyl-CoA precursors and NADPH reducing equivalents required for lipid biosynthesis. This study provided evidence supporting the relationship between metformin hydrochloride and lipid synthesis and validated metformin hydrochloride as a targeted drug for inhibiting lipid synthesis in M. circinelloides.
{"title":"The effect of exogenous addition of metformin hydrochloride on lipid synthesis in Mucor circinelloides WJ11","authors":"Xinai Liu , Xiuwen Wang , Yueping Yang , Kang Li , Wenrong Liu , Qing Liu , Yuanda Song","doi":"10.1016/j.fgb.2025.104023","DOIUrl":"10.1016/j.fgb.2025.104023","url":null,"abstract":"<div><div>As a well-established oleaginous model fungus, <em>Mucor circinelloides</em> has been extensively employed in lipid metabolism research due to its high lipid accumulation capacity and genetic tractability. Metformin hydrochloride, a biguanide drug, exerts significant effects on glucose and lipid metabolism in animal model, however, its effect on lipid metabolism in oleaginous microorganism is unclear. This investigation elucidates the regulatory effects of metformin hydrochloride on lipid biosynthesis in the model oleaginous fungus <em>M. circinelloides</em>. The findings revealed that metformin hydrochloride suppresses fatty acid biosynthesis in <em>M. circinelloides</em>. Addition of 4 g/L metformin hydrochloride to the growth medium of the fungus reduced the total fatty acid content from 29.57 % to 23.27 %, representing a decrease of 21.30 %. Furthermore, metformin hydrochloride significantly upregulated the transcriptional levels of <em>AMPK</em> subunits (such as Snf-α1, Snf-γ1, and Snf-γ3) while suppressing the expression of key genes involved in lipid synthesis, including <em>acl</em>, <em>acc1</em>, and <em>acc2</em>. Our enzymatic assays revealed that metformin hydrochloride treatment markedly inhibited ACC and 6PGDH activities, thereby restricting the availability of both acetyl-CoA precursors and NADPH reducing equivalents required for lipid biosynthesis. This study provided evidence supporting the relationship between metformin hydrochloride and lipid synthesis and validated metformin hydrochloride as a targeted drug for inhibiting lipid synthesis in <em>M. circinelloides</em>.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104023"},"PeriodicalIF":2.4,"publicationDate":"2025-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144644190","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}
Pub Date : 2025-07-02DOI: 10.1016/j.fgb.2025.104021
Yuekun Zou , Shuang Bai , Minna Han , Xin Yang , Xiaoxian Cheng , Jiamin Wu , Zhikuan Xia , Rongya Yang
Cell pyroptosis serves as a critical regulatory mechanism by which host cells respond to fungal infections. However, whether pyroptosis contributes to alterations in fungal pathogenicity during microevolution remains unclear. In this study, we investigated the role of pyroptosis in the pathogenicity of Trichosporon asahii (T. asahii) intraspecies variation, as well as the underlying mechanisms. An in vitro model was established by co-culturing a strain (Ori 2.2174) and a intraspecies variation (Evo 1403.3) of T. asahii with mouse macrophages, while an in vivo model was developed by inoculating immunosuppressed ICR mice with these strains. Survival analysis was performed to assess mortality differences between groups. In vitro experiments revealed that the macrophages infected with Evo 1403.3 exhibited significantly lower lysis rates, pyroptosis levels, and inflammatory cytokine production compared to those infected with Ori 2.2174. Similarly, mice inoculated with Evo 1403.3 showed significantly higher survival rates and reduced fungal burden and lesion areas in organ tissues compared to those infected with Ori 2.2174. Furthermore, pyroptosis mediated by the NLRP3/Caspase-1/GSDMD signaling axis was observed in macrophages infected with both strains, with the extent of pyroptosis positively correlating with strain pathogenicity. Collectively, these findings indicate that cell pyroptosis mediated through the NLRP3/Caspase-1/GSDMD pathway plays a pivotal role in regulating the pathogenicity of T. asahii intraspecies variation.
{"title":"The role of NLRP3/caspase−/GSDMD signal axis mediated cell pyroptosis in the pathogenicity of T. asahii","authors":"Yuekun Zou , Shuang Bai , Minna Han , Xin Yang , Xiaoxian Cheng , Jiamin Wu , Zhikuan Xia , Rongya Yang","doi":"10.1016/j.fgb.2025.104021","DOIUrl":"10.1016/j.fgb.2025.104021","url":null,"abstract":"<div><div>Cell pyroptosis serves as a critical regulatory mechanism by which host cells respond to fungal infections. However, whether pyroptosis contributes to alterations in fungal pathogenicity during microevolution remains unclear. In this study, we investigated the role of pyroptosis in the pathogenicity of <em>Trichosporon asahii</em> (<em>T. asahii</em>) intraspecies variation, as well as the underlying mechanisms. An in vitro model was established by co-culturing a strain (Ori 2.2174) and a intraspecies variation (Evo 1403.3) of <em>T. asahii</em> with mouse macrophages, while an in vivo model was developed by inoculating immunosuppressed ICR mice with these strains. Survival analysis was performed to assess mortality differences between groups. In vitro experiments revealed that the macrophages infected with Evo 1403.3 exhibited significantly lower lysis rates, pyroptosis levels, and inflammatory cytokine production compared to those infected with Ori 2.2174. Similarly, mice inoculated with Evo 1403.3 showed significantly higher survival rates and reduced fungal burden and lesion areas in organ tissues compared to those infected with Ori 2.2174. Furthermore, pyroptosis mediated by the NLRP3/Caspase-1/GSDMD signaling axis was observed in macrophages infected with both strains, with the extent of pyroptosis positively correlating with strain pathogenicity. Collectively, these findings indicate that cell pyroptosis mediated through the NLRP3/Caspase-1/GSDMD pathway plays a pivotal role in regulating the pathogenicity of <em>T. asahii</em> intraspecies variation.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104021"},"PeriodicalIF":2.4,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565522","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}
Pub Date : 2025-06-30DOI: 10.1016/j.fgb.2025.104022
Haiyan Wang , Peiya Chen , Yun Wang , Babar Khan , Ai-Ping Pang , Fu-Gen Wu , Fengming Lin
While endo-β-1,3-glucanases, crucial for hydrolyzing β-1,3-glucan in cell wall, play significant roles in cellular activities of filamentous fungi, research on these enzymes in Trichoderma reesei remains scarce. In this study, we explored the cellular functions and molecular properties of a novel endo-β-glucanase, GLU1, in T. reesei. The mRNA level of glu1 was noticeably upregulated under cellulase-producing condition as compared to non-cellulase-producing condition. Moreover, the deletion of glu1 significantly repressed cellulase production, primarily by reducing the transcript levels of key genes involved in cellulase synthesis during the early stages of fermentation. This deletion further led to weakened cell growth, diminished sporulation, and mutated colony morphology. These suggest that glu1 plays a crucial positive role in cellulase biosynthesis, cell growth, and sporulation. Additionally, the absence of glu1 led to increased β-glucan content within cell wall, making T. reesei more susceptible to Congo red but less sensitive to NaCl. This suggests that GLU1 is involved in modulating the composition and functionality of cell wall. Bioinformatic analyses classified GLU1 within the GH64 family, while the cellular distribution analysis revealed GLU1 was located in the cytoplasm. These experimental data broaden our understanding of the roles and molecular characteristics of endo-β-1,3-glucanases in filamentous fungi.
{"title":"Functional characterization of endo-β-1,3-glucanase in Trichoderma reesei","authors":"Haiyan Wang , Peiya Chen , Yun Wang , Babar Khan , Ai-Ping Pang , Fu-Gen Wu , Fengming Lin","doi":"10.1016/j.fgb.2025.104022","DOIUrl":"10.1016/j.fgb.2025.104022","url":null,"abstract":"<div><div>While <em>endo</em>-β-1,3-glucanases, crucial for hydrolyzing β-1,3-glucan in cell wall, play significant roles in cellular activities of filamentous fungi, research on these enzymes in <em>Trichoderma reesei</em> remains scarce. In this study, we explored the cellular functions and molecular properties of a novel <em>endo</em>-β-glucanase, GLU1, in <em>T</em>. <em>reesei</em>. The mRNA level of <em>glu1</em> was noticeably upregulated under cellulase-producing condition as compared to non-cellulase-producing condition. Moreover, the deletion of <em>glu1</em> significantly repressed cellulase production, primarily by reducing the transcript levels of key genes involved in cellulase synthesis during the early stages of fermentation. This deletion further led to weakened cell growth, diminished sporulation, and mutated colony morphology. These suggest that <em>glu1</em> plays a crucial positive role in cellulase biosynthesis, cell growth, and sporulation. Additionally, the absence of <em>glu1</em> led to increased β-glucan content within cell wall, making <em>T</em>. <em>reesei</em> more susceptible to Congo red but less sensitive to NaCl. This suggests that GLU1 is involved in modulating the composition and functionality of cell wall. Bioinformatic analyses classified GLU1 within the GH64 family, while the cellular distribution analysis revealed GLU1 was located in the cytoplasm. These experimental data broaden our understanding of the roles and molecular characteristics of <em>endo</em>-β-1,3-glucanases in filamentous fungi.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104022"},"PeriodicalIF":2.4,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144555885","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}
Pub Date : 2025-06-27DOI: 10.1016/j.fgb.2025.104020
Guifang Lin , Huakun Zheng , Dal-Hoe Koo , Zonghua Wang , David Cook , Barbara Valent , Sanzhen Liu
Magnaporthe oryzae (syn. Pyricularia oryzae), the causative agent of devastating crop diseases, exhibits remarkable genomic plasticity that contributes to its adaptability and pathogenicity. Individual M. oryzae strains may contain supernumerary mini-chromosomes, which are dispensable and highly repetitive. Here, we explored the stability of two mini-chromosomes of a Lolium strain isolated in the US, TF05–1, in which one mini-chromosome contains sequences nearly identical to the mini-chromosome of the wheat isolate B71 from Bolivia. The discordance of their phylogenetic relationships based on genomic polymorphisms in core chromosomes and polymorphisms in mini-chromosomes indicated horizontal transfer of the mini-chromosome. Contour-clamped homogeneous electric field (CHEF) karyotyping and genome sequencing analysis found variation in numbers and sizes of mini-chromosomes among asexual monoconidial progeny of TF05–1. Optimization of a fluorescence in situ hybridization (FISH) protocol enabled single-cell karyotyping and revelation of drastic cellular variation in numbers of mini-chromosomes. In addition, rearrangement within mini-chromosomes occurred frequently in the TF05–1 progeny. We characterized an intrachromosomal rearrangement presumably mediated by a palindrome repeat. The rearrangement resulted in a 300-kb deletion and a 900-kb duplication. We found that, in contrast to Long Terminal Repeat (LTR) retrotransposons in core chromosomes, LTR retrotransposons in mini-chromosomes were more recently inserted, less methylated, and with higher G + C content. The data indicated that most LTR retrotransposons in mini-chromosomes retain high activity and have yet to be silenced by fungal genome defense mechanisms such as repeat-induced point mutation, which may contribute to highly dynamic mini-chromosome content in fungi.
{"title":"Highly dynamic supernumerary mini-chromosomes in a Magnaporthe oryzae strain contributes to cellular variance of genomic content","authors":"Guifang Lin , Huakun Zheng , Dal-Hoe Koo , Zonghua Wang , David Cook , Barbara Valent , Sanzhen Liu","doi":"10.1016/j.fgb.2025.104020","DOIUrl":"10.1016/j.fgb.2025.104020","url":null,"abstract":"<div><div><em>Magnaporthe oryzae</em> (syn. <em>Pyricularia oryzae</em>), the causative agent of devastating crop diseases, exhibits remarkable genomic plasticity that contributes to its adaptability and pathogenicity. Individual <em>M. oryzae</em> strains may contain supernumerary mini-chromosomes, which are dispensable and highly repetitive. Here, we explored the stability of two mini-chromosomes of a <em>Lolium</em> strain isolated in the US, TF05–1, in which one mini-chromosome contains sequences nearly identical to the mini-chromosome of the wheat isolate B71 from Bolivia. The discordance of their phylogenetic relationships based on genomic polymorphisms in core chromosomes and polymorphisms in mini-chromosomes indicated horizontal transfer of the mini-chromosome. Contour-clamped homogeneous electric field (CHEF) karyotyping and genome sequencing analysis found variation in numbers and sizes of mini-chromosomes among asexual monoconidial progeny of TF05–1. Optimization of a fluorescence in situ hybridization (FISH) protocol enabled single-cell karyotyping and revelation of drastic cellular variation in numbers of mini-chromosomes. In addition, rearrangement within mini-chromosomes occurred frequently in the TF05–1 progeny. We characterized an intrachromosomal rearrangement presumably mediated by a palindrome repeat. The rearrangement resulted in a 300-kb deletion and a 900-kb duplication. We found that, in contrast to Long Terminal Repeat (LTR) retrotransposons in core chromosomes, LTR retrotransposons in mini-chromosomes were more recently inserted, less methylated, and with higher G + C content. The data indicated that most LTR retrotransposons in mini-chromosomes retain high activity and have yet to be silenced by fungal genome defense mechanisms such as repeat-induced point mutation, which may contribute to highly dynamic mini-chromosome content in fungi.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104020"},"PeriodicalIF":2.4,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144517737","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}
Hyper-branching morphology boosts protein production in filamentous fungi as protein secretion occurs predominantly at the actively growing apical hyphal tip. However, mechanisms regulating hyphal morphology in fungi remain poorly characterized. In this paper, we report that the Cys2-His2 Zinc finger transcription factor NsdC negatively regulates both conidiation and hyphal branching in Aspergillus oryzae. The ∆nsdC A. oryzae mutants produced 10-fold more conidia than the wild type (WT) strain and overexpressed BrlA which is the master transcription factor for the conidiation pathway. The mutant also produced hyphae that were more branched in comparison to the WT strain. Analysis of macro-morphology of ∆nsdC mutant strain expressing human lysozyme in protein production medium (5 × DPY) revealed that it formed large mycelial clumps compared to compact pellets in the WT strain. However, lysozyme secretion in the ∆nsdC mutants was comparable to the WT strain, indicating that hyper-branching does not necessarily result in increased protein secretion. Our results also demonstrate that NsdC is a critical regulator of conidiation, hyphal branching and mycelial macro-morphology in A. oryzae.
丝状真菌的超分支形态促进了蛋白质的产生，因为蛋白质的分泌主要发生在活跃生长的顶端菌丝尖端。然而，真菌菌丝形态的调节机制仍不清楚。本文报道了Cys2-His2锌指转录因子NsdC对米曲霉分生和菌丝分支均有负调控作用。∆nsdC A. oryzae突变体产生的分生孢子数是野生型（WT）的10倍，并且过表达分生途径的主要转录因子BrlA。与WT菌株相比，突变体也产生了更多分支的菌丝。表达人溶菌酶的∆nsdC突变菌株在蛋白生产培养基（5 × DPY）中的宏观形态分析显示，与WT菌株的致密球相比，它形成了较大的菌丝团块。然而，∆nsdC突变体的溶菌酶分泌与WT菌株相当，这表明超分支不一定会导致蛋白质分泌增加。我们的研究结果还表明，NsdC是米孢霉分生、菌丝分支和菌丝宏观形态的关键调节因子。
{"title":"The Cys2-His2 zinc finger transcription factor NsdC regulates hyphal branching, conidiation and mycelial pellet morphology in Aspergillus oryzae","authors":"Hui Ting Chu , Naazneen Sofeo , Jasmin Jebarani Jebamony , Prakash Arumugam","doi":"10.1016/j.fgb.2025.104019","DOIUrl":"10.1016/j.fgb.2025.104019","url":null,"abstract":"<div><div>Hyper-branching morphology boosts protein production in filamentous fungi as protein secretion occurs predominantly at the actively growing apical hyphal tip. However, mechanisms regulating hyphal morphology in fungi remain poorly characterized. In this paper, we report that the Cys2-His2 Zinc finger transcription factor NsdC negatively regulates both conidiation and hyphal branching in <em>Aspergillus oryzae.</em> The <em>∆nsdC A. oryzae</em> mutants produced 10-fold more conidia than the wild type (WT) strain and overexpressed BrlA which is the master transcription factor for the conidiation pathway. The mutant also produced hyphae that were more branched in comparison to the WT strain. Analysis of macro-morphology of <em>∆nsdC</em> mutant strain expressing human lysozyme in protein production medium (5 × DPY) revealed that it formed large mycelial clumps compared to compact pellets in the WT strain. However, lysozyme secretion in the ∆<em>nsdC</em> mutants was comparable to the WT strain, indicating that hyper-branching does not necessarily result in increased protein secretion. Our results also demonstrate that NsdC is a critical regulator of conidiation, hyphal branching and mycelial macro-morphology in <em>A. oryzae</em>.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104019"},"PeriodicalIF":2.4,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144491119","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}
Pub Date : 2025-06-20DOI: 10.1016/j.fgb.2025.104017
Andrea P. Angeles-Vázquez , Mirna G. García-Castillo , Gabriela Parra-Olea , Marco T. Solano de la Cruz , M. Delia Basanta
Chytridiomycosis, caused by Batrachochytrium dendrobatidis (Bd), is a major emerging disease contributing to the global decline of amphibians. Phenotypic variations in Bd, such as zoosporangia size and zoospore production, may be linked to differences in pathogenicity among isolates and adaptation to temperatures. Bd has been detected in various regions in Mexico, but its phenotypic variability and relationship to pathogenicity remain unexplored. Here, we examine phenotypic characteristics of Bd such as the growth rate, carrying capacity, zoospore production, and zoosporangia size of ten Bd isolates representing different sublineages within the Bd Global Panzootic Lineage, collected from diverse temperature regimes in Mexico. We found high phenotypic variability between isolates, with growth rate and zoosporangia size influenced by the environmental temperature of their respective sites. Other traits, such as carrying capacity and zoospore production, did not show clear patterns related to environmental temperature. Our findings provide insights into the phenotypic variability of Bd in Mexico and highlight the complexity of pathogen-environment interactions suggesting local adaptations.
{"title":"Influence of environmental temperature on the phenotypic variation of Batrachochytrium dendrobatidis isolates from Mexico","authors":"Andrea P. Angeles-Vázquez , Mirna G. García-Castillo , Gabriela Parra-Olea , Marco T. Solano de la Cruz , M. Delia Basanta","doi":"10.1016/j.fgb.2025.104017","DOIUrl":"10.1016/j.fgb.2025.104017","url":null,"abstract":"<div><div>Chytridiomycosis, caused by <em>Batrachochytrium dendrobatidis</em> (<em>Bd</em>), is a major emerging disease contributing to the global decline of amphibians. Phenotypic variations in <em>Bd</em>, such as zoosporangia size and zoospore production, may be linked to differences in pathogenicity among isolates and adaptation to temperatures. <em>Bd</em> has been detected in various regions in Mexico, but its phenotypic variability and relationship to pathogenicity remain unexplored. Here, we examine phenotypic characteristics of <em>Bd</em> such as the growth rate, carrying capacity, zoospore production, and zoosporangia size of ten <em>Bd</em> isolates representing different sublineages within the <em>Bd</em> Global Panzootic Lineage, collected from diverse temperature regimes in Mexico. We found high phenotypic variability between isolates, with growth rate and zoosporangia size influenced by the environmental temperature of their respective sites. Other traits, such as carrying capacity and zoospore production, did not show clear patterns related to environmental temperature. Our findings provide insights into the phenotypic variability of <em>Bd</em> in Mexico and highlight the complexity of pathogen-environment interactions suggesting local adaptations.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104017"},"PeriodicalIF":2.4,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144369600","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}
Pub Date : 2025-06-20DOI: 10.1016/j.fgb.2025.104018
Berl Oakley , Miguel A. Peñalva
Hyphal tip growth, the primary growth form in fungi, has not yielded its secrets easily or completely, but decades of research in many labs have greatly clarified this fascinating process. In this review, we will summarize progress that has been made in understanding the multiple roles microtubule and microfilament cytoskeletal networks play in tip growth. We will give particular attention to work in Aspergillus nidulans in which these subjects have been studied most intensely, but we will include findings obtained with other organisms where appropriate. Microtubules play a critical role in long range vesicular transport which is powered by the plus end-directed kinesin motor molecules KinA (a type 1 kinesin) and UncA (a type 3 kinesin). The minus end directed motor, dynein, plays an important role in moving kinesins and other cargos away from the hyphal apex so that they can be reused. Actin microfilaments and the motor molecule myosin V play an equally important role, and we will discuss the mechanisms by which microtubule- and actin-dependent transport cooperate to sustain rapid tip growth. With several motors operating in the same cytoplasm, adapter molecules are required to provide the mechanisms by which motors discriminate among cargos. These adapters are being identified and the critical roles of small GTPases are becoming increasingly clear. Endocytosis and exocytosis at the hyphal apex are absolutely required for tip growth and many of the key molecules in these processes have now been identified and their roles clarified. Myosin V is critical for concentrating vesicles carrying vesicular SNAREs at the Spitzenkörper. They then fuse with the apical membrane driven by interaction of vesicular (R-)SNAREs with target (Q-)SNAREs. Localization of the small GTPase RAB11 to the apex is likely a critical marker for the site of exocytosis. Actin patches are the major site of endocytosis, forming a collar near the apex in rapidly growing tip cells, and important progress has been made in understanding the roles of components of actin patches. In total, the machinery for delivering vesicles to the cell apex, the exocytosis machinery and the endocytosis machinery collectively interact to form a tip growth apparatus. Although we celebrate the progress that has been made, we will also point out some of the important remaining questions in this field.
{"title":"The roles of the cytoskeleton in fungal tip growth: Insights from Aspergillus nidulans","authors":"Berl Oakley , Miguel A. Peñalva","doi":"10.1016/j.fgb.2025.104018","DOIUrl":"10.1016/j.fgb.2025.104018","url":null,"abstract":"<div><div>Hyphal tip growth, the primary growth form in fungi, has not yielded its secrets easily or completely, but decades of research in many labs have greatly clarified this fascinating process. In this review, we will summarize progress that has been made in understanding the multiple roles microtubule and microfilament cytoskeletal networks play in tip growth. We will give particular attention to work in <em>Aspergillus nidulans</em> in which these subjects have been studied most intensely, but we will include findings obtained with other organisms where appropriate. Microtubules play a critical role in long range vesicular transport which is powered by the plus end-directed kinesin motor molecules KinA (a type 1 kinesin) and UncA (a type 3 kinesin). The minus end directed motor, dynein, plays an important role in moving kinesins and other cargos away from the hyphal apex so that they can be reused. Actin microfilaments and the motor molecule myosin V play an equally important role, and we will discuss the mechanisms by which microtubule- and actin-dependent transport cooperate to sustain rapid tip growth. With several motors operating in the same cytoplasm, adapter molecules are required to provide the mechanisms by which motors discriminate among cargos. These adapters are being identified and the critical roles of small GTPases are becoming increasingly clear. Endocytosis and exocytosis at the hyphal apex are absolutely required for tip growth and many of the key molecules in these processes have now been identified and their roles clarified. Myosin V is critical for concentrating vesicles carrying vesicular SNAREs at the Spitzenkörper. They then fuse with the apical membrane driven by interaction of vesicular (R-)SNAREs with target (Q-)SNAREs. Localization of the small GTPase RAB11 to the apex is likely a critical marker for the site of exocytosis. Actin patches are the major site of endocytosis, forming a collar near the apex in rapidly growing tip cells, and important progress has been made in understanding the roles of components of actin patches. In total, the machinery for delivering vesicles to the cell apex, the exocytosis machinery and the endocytosis machinery collectively interact to form a tip growth apparatus. Although we celebrate the progress that has been made, we will also point out some of the important remaining questions in this field.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104018"},"PeriodicalIF":2.4,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144364920","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}
Pub Date : 2025-06-11DOI: 10.1016/j.fgb.2025.104016
Oscar Romero , Magalie Galarneau , Samantha Pladwig , Boyan Liu , Sherri Cox , Jennifer Geddes-McAlister
Invasive aspergillosis (IA) is a fungal infection caused by Aspergillus species affecting humans and animals, including birds. Such infections have severe impacts on host health, with the efficacy of current treatment options dwindling against rising rates of antifungal resistance. This scenario represents a critical One Health challenge influenced by climate change at the intersection of animal, human, and environmental health. In this study, we isolated and identified four fungal isolates from infected wildfowl in Southern Ontario, Canada, as Aspergillus fumigatus. Antifungal susceptibility assays against amphotericin B, itraconazole, voriconazole and terbinafine were performed following the Clinical and Laboratory Standards Institute guidelines for filamentous fungi. All strains displayed similar sensitivity to amphotericin B and itraconazole, whereas differences were observed in the response to voriconazole and terbinafine. Next, we performed whole genome sequencing integrated with a comparative genomic analysis to define differences across isolates potentially influencing antifungal susceptibility. As expected, the isolates were phylogenetically similar but demonstrated distinct clustering with A. fumigatus isolate AfB6 mapping closely with the ATCC reference strain compared to the other isolates (i.e., AfB2, AfB8, and AfB7). Notably, single nucleotide polymorphisms (SNPs) were detected across the strains with some correlation between SNPs in antifungal resistance-associated genes and susceptibility profiles; however, antifungal tolerance towards terbinafine was not directly correlated with genetic factors. These data suggest that A. fumigatus isolated from wildfowl with lethal infections from Southern Ontario, Canada, have varying levels of susceptibility to known antifungals and that drivers beyond the anticipated genetic factors influence antifungal response.
{"title":"Aspergillus fumigatus isolated from diverse wildfowl exhibit distinct antifungal susceptibility profiles driven by genetic and non-genetic determinants","authors":"Oscar Romero , Magalie Galarneau , Samantha Pladwig , Boyan Liu , Sherri Cox , Jennifer Geddes-McAlister","doi":"10.1016/j.fgb.2025.104016","DOIUrl":"10.1016/j.fgb.2025.104016","url":null,"abstract":"<div><div>Invasive aspergillosis (IA) is a fungal infection caused by <em>Aspergillus</em> species affecting humans and animals, including birds. Such infections have severe impacts on host health, with the efficacy of current treatment options dwindling against rising rates of antifungal resistance. This scenario represents a critical One Health challenge influenced by climate change at the intersection of animal, human, and environmental health. In this study, we isolated and identified four fungal isolates from infected wildfowl in Southern Ontario, Canada, as <em>Aspergillus fumigatus.</em> Antifungal susceptibility assays against amphotericin B, itraconazole, voriconazole and terbinafine were performed following the Clinical and Laboratory Standards Institute guidelines for filamentous fungi. All strains displayed similar sensitivity to amphotericin B and itraconazole, whereas differences were observed in the response to voriconazole and terbinafine. Next, we performed whole genome sequencing integrated with a comparative genomic analysis to define differences across isolates potentially influencing antifungal susceptibility. As expected, the isolates were phylogenetically similar but demonstrated distinct clustering with <em>A. fumigatus</em> isolate AfB6 mapping closely with the ATCC reference strain compared to the other isolates (i.e., AfB2, AfB8, and AfB7). Notably, single nucleotide polymorphisms (SNPs) were detected across the strains with some correlation between SNPs in antifungal resistance-associated genes and susceptibility profiles; however, antifungal tolerance towards terbinafine was not directly correlated with genetic factors. These data suggest that <em>A. fumigatus</em> isolated from wildfowl with lethal infections from Southern Ontario, Canada, have varying levels of susceptibility to known antifungals and that drivers beyond the anticipated genetic factors influence antifungal response.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"180 ","pages":"Article 104016"},"PeriodicalIF":2.4,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144290517","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}
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