{"title":"Gut microbiota in epilepsy: How antibiotics induce dysbiosis and influence seizure susceptibility","authors":"Shangnan Zou , Xiaofeng Yang , Liemin Zhou","doi":"10.1016/j.micres.2025.128225","DOIUrl":"10.1016/j.micres.2025.128225","url":null,"abstract":"<div><div>Epilepsy, a widespread chronic neurological disorder, has recently come under scrutiny for its potential association with the intricate dynamics of gut microbiota. Numerous investigations into the microbiota-gut-brain axis have revealed a close relationship between gut microbiota and epilepsy, suggesting gut microbiota as a potential treatment strategy. In clinical practice, a longstanding correlation has been observed between some kinds of antibiotics and the potential to induce seizures. Consequently, we have conceived a hypothesis that antibiotics might impact seizure activity by modulating the gut microbiota and influencing the physiological processes within the microbiota-gut-brain axis. In this review, our primary objective is to present the existing evidence and theoretical foundations supporting the hypothesis that dysbiosis within the gut microbiota may play a significant role in the pathophysiology of epilepsy. Furthermore, we aim to summarize the possible mechanisms between microbiota-gut-brain axis and epilepsy, offering insights into the selection of appropriate antibiotics for long-term epilepsy management and enhancing therapeutic efficacy through modulation of the gut microbiota. Further research is necessary to fully elucidate the intricate relationship between gut microbiota ecosystem and epilepsy. Exploring these connections holds promise for advancing our understanding of epilepsy pathogenesis and improving patient treatment and care.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128225"},"PeriodicalIF":6.1,"publicationDate":"2025-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144090130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The relationship between tryptophan metabolism and gut microbiota: Interaction mechanism and potential effects in infection treatment","authors":"Tongchao Pei , Wenweiran Li , Ziyang Zhou , Qinyu Zhang , Guohong Yu , Sokun Yin , Hui Chen , Jianguo Tang","doi":"10.1016/j.micres.2025.128211","DOIUrl":"10.1016/j.micres.2025.128211","url":null,"abstract":"<div><div>Human health is influenced by the gut microbiota, particularly in aspects of host immune homeostasis and intestinal immune response. Tryptophan (Trp) not only acts as a nutrient enhancer but also plays a critical role in the balance between host immune tolerance and gut microbiota maintenance. Both endogenous and bacterial metabolites of Trp, exert significant effects on gut microbial composition, microbial metabolism, the host immunity and the host-microbiome interface.</div><div>Trp metabolites regulate host immunity by activating aryl hydrocarbon receptor (AhR), thereby contributing to immune homeostasis. Among Trp metabolites, AhR ligands include endogenous metabolites (such as kynurenine), and bacterial metabolites (such as indole and its derivatives). Here, we present a comprehensive analysis of the relationships between Trp metabolism and 14 key microbiota, encompassing fungi (e.g., <em>Candida albicans</em>, <em>Aspergillus</em>), bacteria (e.g., <em>Ruminococcus gnavus</em>, <em>Bacteroides, Prevotella copri</em>, <em>Clostridium difficile</em>, <em>Escherichia coli</em>, lactobacilli, <em>Mycobacterium tuberculosis</em>, <em>Pseudomonas aeruginosa</em>, <em>Staphylococcus aureus</em>, <em>Helicobacter. Pylori</em>), and viruses (e.g., SARS-CoV-2, influenza virus). This review clarifies how the gut microbiota regulates Trp metabolism and uncovers the underlying mechanisms of these interactions. And increased mechanistic insight into how the microbiota modulate the host immune system through Trp metabolism may allow for the identification of innovative therapies that are specifically designed to target Trp absorption, Trp metabolites, the gut microbiota, or interactions between Trp and gut microbiota to treat both intestinal and extra-intestinal inflammation as well as microbial infections.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128211"},"PeriodicalIF":6.1,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144090446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lin Zhang , Keyan Chen , Wei Liu , Zhuo Tian , Mingshen Yin , Baolin Sun , Qiuhong Niu
{"title":"Molecular mechanism whereby Bacillus nematocida BN16 activates the “Trojan Horse” response against nematodes","authors":"Lin Zhang , Keyan Chen , Wei Liu , Zhuo Tian , Mingshen Yin , Baolin Sun , Qiuhong Niu","doi":"10.1016/j.micres.2025.128212","DOIUrl":"10.1016/j.micres.2025.128212","url":null,"abstract":"<div><div>This study determined the molecular mechanisms by which <em>Bacillus nematocida</em> BN16 responds to nematode predation. The spores generated by BN16 act as a defense against nematodes, employing a “Trojan horse” strategy. The metabolite 6-methyl-2-heptanone triggers the defense response. To mount an effective defense, BN16 employs transcriptional regulators, including SigA, NtdR, and PksA, that modulate the expression of genes such as <em>mtnD</em> and <em>pycA</em> to enhance the synthesis of 2-heptanone. Subsequently, the enzymes YxjG and YdaC convert 2-heptanone to 6-methyl-2-heptanone, stimulating spore formation via Spo0M. Under nematode predation, <em>B. subtilis</em> BS168 activates metabolic pathways associated with environmental adaptation. However, these responses are generally inadequate to prevent the physical damage inflicted by nematode mouthparts. This study has deepened our understanding of species-specific defense strategies within the genus <em>Bacillus</em> and has advanced the development of pesticide-free pest control methods. The results have identified a novel interaction between microbe and their hosts.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128212"},"PeriodicalIF":6.1,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144107087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yinya Pan , Xinlu Zhao , Qiongyun Chen , Tao Zhao , Yichun Ma , Hai Wu , Ying Xiang , Ping Jiang , Wenjun Li , Qiong Yan , Shangtao Mao , Yufei Tao , Lei wang , Yun Zhu , Guifang Xu
{"title":"Faecalibacterium Prausnitzii extracellular vesicles regulating macrophage differentiation via homologous recombination repair in colitis model","authors":"Yinya Pan , Xinlu Zhao , Qiongyun Chen , Tao Zhao , Yichun Ma , Hai Wu , Ying Xiang , Ping Jiang , Wenjun Li , Qiong Yan , Shangtao Mao , Yufei Tao , Lei wang , Yun Zhu , Guifang Xu","doi":"10.1016/j.micres.2025.128217","DOIUrl":"10.1016/j.micres.2025.128217","url":null,"abstract":"<div><div>Inflammatory Bowel Disease (IBD) is characterized by chronic inflammation influenced by the depletion of beneficial gut microbiota, a critical factor in disease onset and progression. This study investigates the therapeutic potential of extracellular vesicles (EVs) derived from <em>Faecalibacterium prausnitzii</em> (<em>F.p</em> EVs), a commensal bacterium whose reduction is linked to IBD. Our research demonstrates that <em>F.p</em> EVs are preferentially taken up by macrophages, where they exert their anti-inflammatory effects through the enhancement of homologous recombination (HR) repair mechanisms. Specifically, <em>F.p</em> EVs upregulate the expression of key proteins involved in HR repair, such as BRCA1 and BRCA2, thereby reducing DNA damage and inhibiting the cGAS-STING pathway, which is central to the inflammatory response. This modulation of macrophage function results in decreased pro-inflammatory cytokine production and enhanced intestinal barrier integrity. By elucidating these mechanisms, our study provides a clear understanding of how <em>F.p</em> EVs can be used to target fundamental aspects of IBD pathology, laying the groundwork for the development of more effective and targeted therapies.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128217"},"PeriodicalIF":6.1,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144090445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mehran Khan , Atul Kumar Srivastava , Mir Muhammad Nizamani , Muhammad Asif , Ali Kamran , Lingfeng Luo , Sanwei Yang , Songshu Chen , Zhiqiang Li , Xin Xie
{"title":"The battle within: Discovering new insights into phytopathogen interactions and effector dynamics","authors":"Mehran Khan , Atul Kumar Srivastava , Mir Muhammad Nizamani , Muhammad Asif , Ali Kamran , Lingfeng Luo , Sanwei Yang , Songshu Chen , Zhiqiang Li , Xin Xie","doi":"10.1016/j.micres.2025.128220","DOIUrl":"10.1016/j.micres.2025.128220","url":null,"abstract":"<div><div>Phytopathogen interactions are complicated and constantly evolving, driven by a never-ending war amongst the host’s immune defenses and the pathogen's virulence strategies. This comprehensive review examines the intricate mechanisms of effector-triggered immunity (ETI) and how pathogen effectors use host cellular progressions to promote infection. This review article investigates the modification of Phytopathogen effectors and plant resistance proteins, highlighting the role of meta-population dynamics and rapid adaptation. Additionally, it highlights the influence of environmental impact and climate change on host-pathogen interactions, describing their significant impact on disease dynamics and pathogen evolution. Effector proteins are crucial in sabotaging plant immunity, with bacterial, fungal, oomycete, and nematode effectors targeting common host protein networks and phytohormone pathways. Additionally, the review discusses advanced approaches for classifying effector targets, such as bioinformatics and single-cell transcriptomics, highlighting their importance in developing effective disease management strategies. Further insights are described into how effectors control phytohormone pathways, shedding light on how pathogens exploit host signaling. This review covers structural studies and protein modeling that have advanced effector prediction and our understanding of their functions and evolution, while providing an overview of phytopathogen interactions and future directions for effector research.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128220"},"PeriodicalIF":6.1,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144090447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lijuan Ding , Lin Cui , Yuheng Wang , Beau Dronsella , Xu Liu , Huiying Luo , Huoqing Huang , Xiaoyun Su , Yingguo Bai , Jie Zhang , Tao Tu , Xing Qin , Yuan Wang , Yaru Wang , Bin Yao , Xianli Xue , Gang Guo , Xiaolu Wang
{"title":"Fructose metabolism in Entner-Doudoroff pathway-deficient Cupriavidus necator H16 depends on the Calvin shunt","authors":"Lijuan Ding , Lin Cui , Yuheng Wang , Beau Dronsella , Xu Liu , Huiying Luo , Huoqing Huang , Xiaoyun Su , Yingguo Bai , Jie Zhang , Tao Tu , Xing Qin , Yuan Wang , Yaru Wang , Bin Yao , Xianli Xue , Gang Guo , Xiaolu Wang","doi":"10.1016/j.micres.2025.128222","DOIUrl":"10.1016/j.micres.2025.128222","url":null,"abstract":"<div><div>As a facultative chemolithoautotrophic bacterium, <em>Cupriavidus necator</em> H16 uses the Entner-Doudoroff (ED) pathway for heterotrophic growth on carbohydrates such as fructose and the Calvin cycle for lithoautotrophic carbon dioxide fixation. In a previous study, we found that an ED pathway-deficient <em>C. necator</em> strain can survive on fructose, but the underlying metabolic pathway remained unclear. This study aimed to elucidate the metabolic mechanism of fructose metabolism in this ED pathway-deficient <em>C. necator</em> strain. First, the metabolic characteristics of fructose catabolism in the deficient strain were examined. Then, the roles of glycolysis/gluconeogenesis, the Calvin shunt, and the non-oxidative pentose phosphate pathway (non-OxPPP) in the metabolism of fructose were identified through comparative transcriptomic analysis combined with <sup>13</sup>C tracer experiments. Further growth experiments using knockout strains of key genes involved in these pathways confirmed that the non-OxPPP compensates for the blocked ED pathway to metabolize fructose and provide a precursor for the Calvin shunt, thereby driving subsequent carbon fluxes. Additionally, phosphoglycolate salvage pathways, particularly the malate cycle, are crucial for recycling glycolate-2-phosphate produced during RuBisCO-catalyzed oxidation. This study revealed a novel fructose metabolism mechanism in <em>C. necator</em> and highlighted its metabolic flexibility, thereby deepening our understanding of its carbon utilization strategies and providing a theoretical basis for further metabolic engineering research.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128222"},"PeriodicalIF":6.1,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144107088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sk Soyal Parvez , Anupam Mondal , Kalpna Sharma , Andrei Stecca Steindorff , Igor V. Grigoriev , Utpal Bakshi , Avishek Banik
{"title":"Trichoderma sp. strain AM6 whole-genome guided untargeted metabolomics: Terpenoid backbone synthesis and modulation of VOCs in tea (Camellia sinensis L.)","authors":"Sk Soyal Parvez , Anupam Mondal , Kalpna Sharma , Andrei Stecca Steindorff , Igor V. Grigoriev , Utpal Bakshi , Avishek Banik","doi":"10.1016/j.micres.2025.128215","DOIUrl":"10.1016/j.micres.2025.128215","url":null,"abstract":"<div><div>Industrial cash crop tea is a cherished drink for its bioactive components like terpenoids and flavonoids shaping its flavor and health benefits. <em>Trichoderma</em> species are potent biocontrol agents and plant growth regulators, with unexplored potential in modulation of <em>C. sinensis</em> terpenoid biosynthesis. Genome sequencing of a tea root-associated <em>Trichoderma</em> sp. strain AM6 revealed a genome size of 39.91 Mbp, comprising 446 contigs organized into 406 scaffolds, with 98.8 % completeness. Single scaffold mitochondrial genome assembly of 34,430 bp in length with a GC content of 28.03 % encodes a total of 49 genes including 27 tRNA, 2 rRNA, and 20 protein-coding genes. Metabolic pathway analysis indicates exclusive reliance on the mevalonate pathway for terpenoid biosynthesis in <em>Trichoderma</em>, unlike <em>C. sinensis</em>, which utilizes both the mevalonate and non-mevalonate (MEP/DOXP) pathways. Untargeted LC-ESI-MS/MS analysis of <em>Trichoderma</em> identified 11,841 secondary metabolites, including 34 monoterpenoids, 72-diterpenoids, and 76-sesquiterpenoids, emphasizing its metabolic diversity. Comparative phylogenomic study positioned it as a phylogenetically distinct species with unique adaptive traits. Untargeted GC-MS shows high volatile abundance from microbial consortia (T5) followed by only <em>Trichoderma</em> treatment (T2) compared to control (T1). Terpenoid transcripts of <em>C. sinensis</em> from the plant-microbe consortium assembly sets illuminate upregulation of genes assigned to 3-hydroxy-3-methylglutaryl-Co-A reductase (HMGCR) and downregulation of genes assigned to 1-Deoxy-D-xylulose-5-phosphate (DXS), indicating metabolic shift towards more mevalonate pathway activity influenced by this novel <em>Trichoderma</em> strain itself and in combination with other tea root-associated microbes.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128215"},"PeriodicalIF":6.1,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144115054","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xuhang Wang , Wanqiu Lin , Linlin Ye , Xinru Chen , Jianluan Ren , Feng Xue , Jianjun Dai , Fang Tang
{"title":"Caspase-8 drove apoptosis of BMECs to promote neutrophil infiltration and DE205B clearance in meningitis","authors":"Xuhang Wang , Wanqiu Lin , Linlin Ye , Xinru Chen , Jianluan Ren , Feng Xue , Jianjun Dai , Fang Tang","doi":"10.1016/j.micres.2025.128223","DOIUrl":"10.1016/j.micres.2025.128223","url":null,"abstract":"<div><div>Avian pathogenic <em>Escherichia coli</em> (APEC), a significant virulence reservoir for human extraintestinal pathogenic <em>E. coli</em> (ExPEC), poses an escalating zoonotic risk through the food chain. Our previous study demonstrated that the poultry-derived strain DE205B shared high genetic similarity with the neonatal meningitis-associated <em>E. coli</em> (NMEC) strain RS218 and induced meningitis in a rat model. Here, we further demonstrated that DE205B crossed the blood-brain barrier (BBB) via a transcellular pathway without compromising barrier integrity. During this process, brain microvascular endothelial cells (BMECs) trigger limited RIPK1-independent apoptosis. Mechanistically, caspase-8 activation in BMECs drove the release of proinflammatory mediators, thereby promoting neutrophil recruitment into the cerebrospinal fluid (CSF). These neutrophils facilitated bacterial clearance through the formation of neutrophil extracellular traps (NETs). <em>In vivo</em> pharmacological inhibition of caspase-8 attenuated the ability of BMECs to recruit neutrophils, exacerbating meningitis progression. These findings suggested that limited apoptosis of BMECs contributed positively to APEC clearance in the brain. Collectively, this study systematically elucidated mechanisms underlying DE205B-mediated BBB invasion and host immune responses, providing insights into its cross-species pathogenic potential and zoonotic implications.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128223"},"PeriodicalIF":6.1,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144115548","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yan Guo , Zhiying Liu , Jianfeng Wang , Xuming Deng , Liuqing He , Yong Zhang , Hongtao Liu , Jiazhang Qiu
{"title":"Equol neutralizes toxin B to combat Clostridioides difficile infection without disrupting the gut microbiota","authors":"Yan Guo , Zhiying Liu , Jianfeng Wang , Xuming Deng , Liuqing He , Yong Zhang , Hongtao Liu , Jiazhang Qiu","doi":"10.1016/j.micres.2025.128219","DOIUrl":"10.1016/j.micres.2025.128219","url":null,"abstract":"<div><div><em>Clostridioides difficile</em> (<em>C. difficile</em>) toxin B (TcdB) is essential for <em>C. difficile pathogenicity</em>. TcdB induces apoptosis in host cells by internalizing and utilizing its glycosyltransferase activity to modify members of the small GTPase protein family through glycosylation. The intestinal environment is critical for the colonization of <em>C. difficile</em>, and the use of broad-spectrum antibiotics disrupts the balance of the gut microbiota, leading to increased susceptibility of the host to <em>C. difficile</em>. At present, the mainstream clinical approach for treating <em>C. difficile</em> infection (CDI) involves antibiotic therapies such as vancomycin, which disrupt the gut microbiota and are associated with a considerable risk of infection recurrence. Therefore, there is an urgent clinical need to develop new strategies to combat CDI. Here, we have identified a natural compound, equol, which inhibits the TcdB-mediated glycosylation of Rac1 through direct interaction, thereby reducing TcdB-induced cell death. Equol functions as an inhibitor of indoleamine 2,3-dioxygenase 1 (IDO1) and tryptophan 2,3-dioxygenase (TDO), effectively suppressing the conversion of tryptophan to kynurenine in the intestinal tract while preserving the integrity of the gut microbiota. Concurrently, equol exhibits robust antioxidant properties, which markedly reduced TcdB-mediated oxidative damage and subsequent cell death. These findings suggest that equol holds therapeutic potential for the treatment of CDI.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128219"},"PeriodicalIF":6.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143948176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Juan Pablo Morán Torres, Jun Lyu, Xiaoyi Chen, Antonia M. Klaas, Peter Jan Vonk, Luis G. Lugones, Hans de Cock, Han A.B. Wösten
{"title":"Single and combinatorial gene inactivation in Aspergillus niger using selected as well as genome-wide gRNA library pools","authors":"Juan Pablo Morán Torres, Jun Lyu, Xiaoyi Chen, Antonia M. Klaas, Peter Jan Vonk, Luis G. Lugones, Hans de Cock, Han A.B. Wösten","doi":"10.1016/j.micres.2025.128204","DOIUrl":"10.1016/j.micres.2025.128204","url":null,"abstract":"<div><div><em>Aspergillus niger</em> is a saprotroph, a pathogen, an endophyte, a food spoiler and an important cell factory. Only a minor fraction of its genes has been experimentally characterized. We here set up a CRISPR/Cas9 mutagenesis screen for functional gene analysis using co-transformation of a pool of gene editing plasmids that are maintained under selection pressure and that each contain a gRNA. First, a pool of gRNA vectors was introduced in <em>A. niger</em> targeting five genes with easy selectable phenotypes. Transformants were obtained with all possible single, double, triple, quadruple and quintuple gene inactivation phenotypes. Their genotypes were confirmed using the gRNA sequences in the transforming vector as barcodes. Next, a gRNA library was introduced in <em>A. niger</em> targeting > 9600 genes. Gene <em>nsdC</em> was identified as a sporulation gene using co-transformation conditions that favored uptake of one or two gRNA construct(s) from the genome-wide vector pool. Together, CRISPR/Cas9 vectors with a (genome-wide) pool of gRNAs can be used for functional analysis of genes in <em>A. niger</em> with phenotypes that are the result of the inactivation of a single or multiple genes.</div></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"298 ","pages":"Article 128204"},"PeriodicalIF":6.1,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143937648","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}