Fungal Genetics and Biology最新文献

{"title":"Putative APSES family transcription factor mbp1 plays an essential role in regulating cell wall synthesis in the agaricomycete Pleurotus ostreatus","authors":"Hayase Kojima ,&nbsp;Moriyuki Kawauchi ,&nbsp;Yuitsu Otsuka ,&nbsp;Kim Schiphof ,&nbsp;Kenya Tsuji ,&nbsp;Akira Yoshimi ,&nbsp;Chihiro Tanaka ,&nbsp;Shigekazu Yano ,&nbsp;Takehito Nakazawa ,&nbsp;Yoichi Honda","doi":"10.1016/j.fgb.2024.103936","DOIUrl":"10.1016/j.fgb.2024.103936","url":null,"abstract":"<div><div>The clade A APSES family transcription factors (Mbp1, Swi4, and Swi6) contribute to cell wall synthesis regulation in fungi. Herein, evolutionary relationships among these proteins were clarified by phylogenetic analysis using various ascomycetes and basidiomycetes, and then the detailed function of Mbp1 in cell wall synthesis regulation was analyzed in <em>Pleurotus ostreatus</em>. Our phylogenetic analysis revealed that Mbp1 and Swi6 are widely conserved among various fungi, whereas Swi4 is a protein specific for Saccharomycotina. In <em>P. ostreatus</em>, two putative clade A APSES family transcription factors, protein ID 83192 and 134090, were found and identified as Mbp1 and Swi6, respectively. The <em>mbp1</em> gene was then disrupted through homologous recombination using <em>P. ostreatus</em> 20b strain (Δ<em>ku80</em>) as a host to obtain <em>mbp1</em> disruption strains (Δ<em>mbp1</em>). Disruption of <em>mbp1</em> significantly decreased the growth rate and shortened aerial hyphae, suggesting that Mbp1 is involved in mycelial growth, especially aerial hyphal growth. Furthermore, thinner cell walls, decreased relative percentage of β-glucan, and downregulation of all β-glucan synthase genes were observed in Δ<em>mbp1</em> strains. Therefore, Mbp1 plays an essential role in β-glucan synthesis regulation in <em>P. ostreatus</em>. Disruption of <em>mbp1</em> also impacted the expression profiles of chitin synthase genes, septum formation, and sensitivity to a chitin synthesis inhibitor, suggesting that Mbp1 also regulates chitin synthesis. In conclusion, Mbp1 is responsible for normal mycelial growth and regulates β-glucan and chitin synthesis in <em>P. ostreatus</em>. To the best of our knowledge, this is the first report on the detailed function of Mbp1 in cell wall synthesis regulation in fungi.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"175 ","pages":"Article 103936"},"PeriodicalIF":2.4,"publicationDate":"2024-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142382525","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}

{"title":"Optimizing fungal extracellular vesicle proteomic profiling through combined analysis of in-solution and in-gel digestion","authors":"Daniel A. Salgado-Bautista ,&nbsp;Eduardo Callegari ,&nbsp;Meritxell Riquelme","doi":"10.1016/j.fgb.2024.103935","DOIUrl":"10.1016/j.fgb.2024.103935","url":null,"abstract":"<div><div>Proteomics offers a powerful tool to identify proteins within diverse microbial organisms, environments, and organelles, including extracellular vesicles (EVs). Fungal EVs are of particular interest due to their roles in cellular development and communication. While several methods exist to isolate EVs from cells, a universally accepted approach for EV protein characterization is lacking. This study investigated in-solution digestion (SD) and in-gel digestion (GD), for characterizing proteins from <em>Neurospora crassa</em> EVs, followed by LC-MS/MS analysis. GD identified three to four-times more proteins than SD while using the same number of unique peptides. Although GD requires a higher amount of starting sample, it offers a more comprehensive protein identification for fungal EVs, potentially preventing the omission of crucial data.</div></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"175 ","pages":"Article 103935"},"PeriodicalIF":2.4,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142309170","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":"Phyllosticta paracitricarpa is synonymous with the EU quarantine fungus P. citricarpa based on phylogenomic analyses","authors":"Valerie A. van Ingen-Buijs ,&nbsp;Anouk C. van Westerhoven ,&nbsp;Petros Skiadas ,&nbsp;Xander C.L. Zuijdgeest ,&nbsp;Sajeet Haridas ,&nbsp;Christopher Daum ,&nbsp;Kecia Duffy ,&nbsp;Jie Guo ,&nbsp;Hope Hundley ,&nbsp;Kurt LaButti ,&nbsp;Anna Lipzen ,&nbsp;Jasmyn Pangilinan ,&nbsp;Robert Riley ,&nbsp;Jie Wang ,&nbsp;Mi Yan ,&nbsp;Francis Martin ,&nbsp;Kerrie Barry ,&nbsp;Igor V. Grigoriev ,&nbsp;Johannes Z. Groenewald ,&nbsp;Pedro W. Crous ,&nbsp;Michael F. Seidl","doi":"10.1016/j.fgb.2024.103925","DOIUrl":"10.1016/j.fgb.2024.103925","url":null,"abstract":"<div><p><em>Phyllosticta citricarpa</em> is an important citrus-pathogen and a quarantine organism in the European Union. Its recently described relative, <em>P. paracitricarpa</em>, is very closely related and not listed as a quarantine organism. <em>P. paracitricarpa</em> is very difficult to distinguish from <em>P. citricarpa</em>, since its morphological features overlap and the barcoding gene sequences that were originally used to delimit them as distinct species have a low number of species-specific polymorphisms that have subsequently been shown to overlap between the two clades. Therefore, we performed extensive genomic analyses to determine whether the genetic variation between <em>P. citricarpa</em> and <em>P. paracitricarpa</em> strains should be considered to represent infraspecific variation within <em>P. citricarpa</em>, or whether it is indicative of distinct species. Using a phylogenomic analysis with 3,000 single copy ortholog genes and whole-genome comparisons, we determined that the variation between <em>P. citricarpa</em> and <em>P. paracitricarpa</em> can be considered as infraspecies variation within <em>P. citricarpa</em>. We also determined the level of variation in mitochondrial assemblies of several <em>Phyllosticta</em> species and concluded there are only minimal differences between the assemblies of <em>P. citricarpa</em> and <em>P. paracitricarpa</em>. Thus, using several orthogonal approaches, we here demonstrate that variation within the nuclear and mitochondrial genomes of other <em>Phyllosticta</em> species is larger than variation between genomes obtained from <em>P. citricarpa</em> and <em>P. paracitricarpa</em> strains. Thus, <em>P. citricarpa</em> and <em>P. paracitricarpa</em> should be considered as conspecific.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"175 ","pages":"Article 103925"},"PeriodicalIF":2.4,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1087184524000628/pdfft?md5=551c0118874a22b2fde3b8d86173256b&pid=1-s2.0-S1087184524000628-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142146921","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}

{"title":"Population genomics of Fusarium graminearum isolates from the Americas","authors":"Upasana Dhakal ,&nbsp;Wei Yue ,&nbsp;John F. Leslie,&nbsp;Christopher Toomajian","doi":"10.1016/j.fgb.2024.103924","DOIUrl":"10.1016/j.fgb.2024.103924","url":null,"abstract":"<div><p>Fusarium head blight (FHB) is a major disease of wheat and barley worldwide and is caused by different species in the genus <em>Fusarium</em>, <em>Fusarium graminearum</em> being the most important. We conducted population genomics analyses using SNPs obtained through genotyping by sequencing of over 500 isolates of <em>F. graminearum</em> from the US Upper Midwest, New York, Louisiana, and Uruguay. PCA and STRUCTURE analyses group our isolates into four previously described populations: NA1, NA2, Southern Louisiana (SLA) and Gulf Coast (GC). Some isolates were not assigned to populations because of mixed ancestry. Population structure was associated with toxin genotype and geographic origin. The NA1, NA2, and SLA populations are differentiated (<em>F_ST</em> 0.385 – 0.551) but the presence of admixed isolates indicates that the populations are not reproductively isolated. Patterns of linkage disequilibrium (LD) decay suggest frequent recombination within populations. <em>Fusarium graminearum</em> populations from the US have great evolutionary potential given the high recombination rate and a large proportion of admixed isolates. The NA1, NA2, and Southern Louisiana (SLA) populations separated from their common ancestral population roughly at the same time in the past and are evolving with moderate levels of subsequent gene flow between them. Genome-wide selection scans in all three populations revealed outlier regions with the strongest signatures of recent positive natural selection. These outlier regions include many genes with unknown function and some genes with known roles in plant–microbe interaction, fungicide/drug resistance, cellular transport and genes that are related to cellular organelles. Only a very small proportion of outlier regions are shared as outliers among the three populations, suggesting unique host-pathogen interactions and environmental adaptation.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"174 ","pages":"Article 103924"},"PeriodicalIF":2.4,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1087184524000616/pdfft?md5=abdc32c0ab09b219944f197979901205&pid=1-s2.0-S1087184524000616-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141879861","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}

{"title":"Differential impacts of furfural and acetic acid on the bioenergetics and fermentation performance of Scheffersomyces stipitis","authors":"José J. Saucedo-Gutierrez ,&nbsp;Monserrat Escamilla-García ,&nbsp;Aldo Amaro-Reyes ,&nbsp;Andrés Carrillo-Garmendia ,&nbsp;Luis A. Madrigal-Pérez ,&nbsp;Carlos Regalado-González ,&nbsp;José Á. Granados-Arvizu","doi":"10.1016/j.fgb.2024.103914","DOIUrl":"10.1016/j.fgb.2024.103914","url":null,"abstract":"<div><p>Lignocellulosic material is a leading carbon source for economically viable biotechnological processes; however, compounds such furfural and acetic acid exhibit toxicity to yeasts. Nonetheless, research about the molecular mechanism of furfural and acetic acid toxicity is still scarce in yeasts like <em>Scheffersomyces stipitis</em>. Thus, this study aims to elucidate the impact of furfural and acetic acid on <em>S. stipitis</em> regarding bioenergetic and fermentation parameters. Here, we provide evidence that furfural and acetic acid induce a delay in cell growth and extend the lag phase. The mitochondrial membrane potential decreased in all treatments with no significant differences between inhibitors or concentrations. Interestingly, reactive oxygen species increased when the inhibitor concentrations were from 0.1 to 0.3 % (v/v). The glycolytic flux was not significantly (<em>p</em> &gt; 0.05) altered by acetic acid, but furfural caused different effects. Ethanol production decreased significantly (4.32 g·L^-1 in furfural and 5.06 g·L^-1 in acetic acid) compared to the control (26.3 g·L^-1). In contrast, biomass levels were not significantly different in most treatments compared to the control. This study enhances our understanding of the effects of furfural and acetic acid at the mitochondrial level in a pentose-fermenting yeast like <em>S. stipitis</em>.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"174 ","pages":"Article 103914"},"PeriodicalIF":2.4,"publicationDate":"2024-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141735735","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":"Final piece to the Fusarium pigmentation puzzle – Unraveling of the phenalenone biosynthetic pathway responsible for perithecial pigmentation in the Fusarium solani species complex","authors":"Mikkel Rank Nielsen ,&nbsp;Trine Sørensen ,&nbsp;Tobias Bruun Pedersen ,&nbsp;Klaus Ringsborg Westphal ,&nbsp;Lorena Díaz Fernández De Quincoces ,&nbsp;Teis Esben Sondergaard ,&nbsp;Reinhard Wimmer ,&nbsp;Daren W. Brown ,&nbsp;Jens Laurids Sørensen","doi":"10.1016/j.fgb.2024.103912","DOIUrl":"10.1016/j.fgb.2024.103912","url":null,"abstract":"<div><p>The <em>Fusarium solani</em> species complex (FSSC) is comprised of important pathogens of plants and humans. A distinctive feature of FSSC species is perithecial pigmentation. While the dark perithecial pigments of other <em>Fusarium</em> species are derived from fusarubins synthesized by polyketide synthase 3 (PKS3), the perithecial pigments of FSSC are derived from an unknown metabolite synthesized by PKS35. Here, we confirm in FSSC species <em>Fusarium vanettenii</em> that PKS35 (<em>fsnI</em>) is required for perithecial pigment synthesis by deletion analysis and that <em>fsnI</em> is closely related to <em>phnA</em> from <em>Penicillium herquei</em>, as well as <em>duxI from Talaromyces stipentatus</em>, which produce prephenalenone as an early intermediate in herqueinone and duclauxin synthesis respectively. The production of prephenalenone by expression of <em>fsnI</em> in <em>Saccharomyces cerevisiae</em> indicates that it is also an early intermediate in perithecial pigment synthesis. We next identified a conserved cluster of 10 genes flanking <em>fsnI</em> in <em>F. vanettenii</em> that when expressed in <em>F. graminearum</em> led to the production of a novel corymbiferan lactone F as a likely end product of the phenalenone biosynthetic pathway in FSSC.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"174 ","pages":"Article 103912"},"PeriodicalIF":2.4,"publicationDate":"2024-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1087184524000495/pdfft?md5=9015af29abe9500f83c7493c0cdf145c&pid=1-s2.0-S1087184524000495-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141617608","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}

{"title":"High phenotypic and genotypic plasticity among strains of the mushroom-forming fungus Schizophyllum commune","authors":"Ioana M. Marian ,&nbsp;Ivan D. Valdes ,&nbsp;Richard D. Hayes ,&nbsp;Kurt LaButti ,&nbsp;Kecia Duffy ,&nbsp;Mansi Chovatia ,&nbsp;Jenifer Johnson ,&nbsp;Vivian Ng ,&nbsp;Luis G. Lugones ,&nbsp;Han A.B. Wösten ,&nbsp;Igor V. Grigoriev ,&nbsp;Robin A. Ohm","doi":"10.1016/j.fgb.2024.103913","DOIUrl":"10.1016/j.fgb.2024.103913","url":null,"abstract":"<div><p><em>Schizophyllum commune</em> is a mushroom-forming fungus notable for its distinctive fruiting bodies with split gills. It is used as a model organism to study mushroom development, lignocellulose degradation and mating type loci. It is a hypervariable species with considerable genetic and phenotypic diversity between the strains. In this study, we systematically phenotyped 16 dikaryotic strains for aspects of mushroom development and 18 monokaryotic strains for lignocellulose degradation. There was considerable heterogeneity among the strains regarding these phenotypes. The majority of the strains developed mushrooms with varying morphologies, although some strains only grew vegetatively under the tested conditions. Growth on various carbon sources showed strain-specific profiles. The genomes of seven monokaryotic strains were sequenced and analyzed together with six previously published genome sequences. Moreover, the related species <em>Schizophyllum fasciatum</em> was sequenced. Although there was considerable genetic variation between the genome assemblies, the genes related to mushroom formation and lignocellulose degradation were well conserved. These sequenced genomes, in combination with the high phenotypic diversity, will provide a solid basis for functional genomics analyses of the strains of <em>S. commune</em>.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"173 ","pages":"Article 103913"},"PeriodicalIF":2.4,"publicationDate":"2024-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1087184524000501/pdfft?md5=4b9314edeb77dde82d99b95832663019&pid=1-s2.0-S1087184524000501-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141617609","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}

{"title":"Characterization of linoleate dioxygenases in basidiomycetes and the functional role of CcLdo1 in regulating fruiting body development in Coprinopsis cinerea","authors":"Jing Chen ,&nbsp;Rong Qu ,&nbsp;Qiurong Chen ,&nbsp;Ziyu Zhang ,&nbsp;Siting Wu ,&nbsp;Mengyu Bao ,&nbsp;Xinyue Wang ,&nbsp;Lei Liu ,&nbsp;Siqi Lyu ,&nbsp;Jialu Tian ,&nbsp;Linna Lyu ,&nbsp;Cigang Yu ,&nbsp;Sheng Yuan ,&nbsp;Zhonghua Liu","doi":"10.1016/j.fgb.2024.103911","DOIUrl":"10.1016/j.fgb.2024.103911","url":null,"abstract":"<div><p><em>Coprinopsis cinerea</em>, a model fungus, is utilized for investigating the developmental mechanisms of basidiomycetes. The development of basidiomycetes is a highly organized process that requires coordination among genetic, environmental, and physiological factors. Oxylipins, a class of widely distributed signaling molecules, play crucial roles in fungal biology. Among oxylipins, the sexual pheromone-inducing factors (psi factors) have been identified as key regulators of the balance between asexual and sexual spore development in Ascomycetes. Linoleate dioxygenases are enzymes involved in the biosynthesis of psi factors, yet their specific physiological functions in basidiomycete development remain unclear. In this study, linoleate dioxygenases in basidiomycetes were identified and characterized. Phylogenetic analysis revealed that linoleate dioxygenases from Basidiomycota formed a distinct clade, with linoleate dioxygenases from Agaricomycetes segregating into three groups and those from Ustilaginomycetes forming a separate group. Both basidiomycete and ascomycete linoleate dioxygenases shared two characteristic domains: the N-terminal of linoleate dioxygenase domain and the C-terminal of cytochrome P450 domain. While the linoleate dioxygenase domains exhibited similarity between basidiomycetes and ascomycetes, the cytochrome P450 domains displayed high diversity in key sites. Furthermore, the gene encoding the linoleate dioxygenase <em>Ccldo1</em> in <em>C. cinerea</em> was knocked out, resulting in a significant increase in fruiting body formation without affecting asexual conidia production. This observation suggests that secondary metabolites synthesized by CcLdo1 negatively regulate the sexual reproduction process in <em>C. cinerea</em> while not influencing the asexual reproductive process. This study represents the first identification of a gene involved in secondary metabolite synthesis that regulates basidiocarp development in a basidiomycete.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"173 ","pages":"Article 103911"},"PeriodicalIF":2.4,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141499690","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":"Dual auxotrophy coupled red labeling strategy for efficient genome editing in Saccharomyces cerevisiae","authors":"Jianhua Li ,&nbsp;Taorui Wu ,&nbsp;Jialong Wang ,&nbsp;Youlong Chen ,&nbsp;Wenxin Zhang ,&nbsp;Lijun Cai ,&nbsp;Shufang Lai ,&nbsp;Kaihui Hu ,&nbsp;Wensong Jin","doi":"10.1016/j.fgb.2024.103910","DOIUrl":"10.1016/j.fgb.2024.103910","url":null,"abstract":"<div><p>The homologous recombination strategy has a long history of editing <em>Saccharomyces cerevisiae</em> target genes. The application of CRISPR/Cas9 strategy to editing target genes in <em>S. cerevisiae</em> has also received a lot of attention in recent years. All findings seem to indicate that editing relevant target genes in <em>S. cerevisiae</em> is an extremely easy event. In this study, we systematically analyzed the advantages and disadvantages of homologous recombination (HR) strategy, CRISPR/Cas9 strategy, and CRISPR/Cas9 combined homology-mediated repair (CRISPR/Case9-HDR) strategy in knocking out BY4742 <em>ade2</em>. Our data showed that when the <em>ade2</em> was knocked out by HR strategy, a large number of clones appeared to be off-target, and 10 %–80 % of the so-called knockout clones obtained were heteroclones. When the CRISPR/Cas9 strategy was applied, 60% of clones were off-target and the rest were all heteroclones. Interestingly, most of the cells were edited successfully, but at least 60 % of the clones were heteroclones, when the CRISPR/Cas9-HDR strategy was employed. Our results clearly showed that the emergence of heteroclone seems inevitable regardless of the strategies used for editing BY4742 <em>ade2</em>. Given the characteristics of BY4742 defective in <em>ade2</em> showing red on the YPD plate, we attempted to build an efficient yeast gene editing strategy, in which the CRISPR/Cas9 combines homology-mediated repair template carrying an <em>ade2</em> expression cassette, BY4742(<em>ade2Δ0</em>) as the start strain. We used this strategy to successfully achieve 100 % knockout efficiency of <em>trp1</em>, indicating that technical challenges of how to easily screen out pure knockout clones without color phenotype have been solved. Our data showed in this study not only establishes an efficient yeast gene knockout strategy with dual auxotrophy coupled red labeling but also provides new ideas and references for the knockout of target genes in the monokaryotic mycelium of macrofungi.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"173 ","pages":"Article 103910"},"PeriodicalIF":2.4,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141428311","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":"Glucose-induced endocytic degradation of the maltose transporter MalP is mediated through ubiquitination by the HECT-ubiquitin ligase HulA and its adaptor CreD in Aspergillus oryzae","authors":"Shoki Fujita ,&nbsp;Hinako Tada ,&nbsp;Yuka Matsuura ,&nbsp;Tetsuya Hiramoto ,&nbsp;Mizuki Tanaka ,&nbsp;Takahiro Shintani ,&nbsp;Katsuya Gomi","doi":"10.1016/j.fgb.2024.103909","DOIUrl":"10.1016/j.fgb.2024.103909","url":null,"abstract":"<div><p>In the filamentous fungus <em>Aspergillus oryzae</em>, large amounts of amylolytic enzymes are inducibly produced by isomaltose, which is converted from maltose incorporated via the maltose transporter MalP. In contrast, the preferred sugar glucose strongly represses the expression of both amylolytic and <em>malP</em> genes through carbon catabolite repression. Simultaneously, the addition of glucose triggers the endocytic degradation of MalP on the plasma membrane. In budding yeast, the signal-dependent ubiquitin modification of plasma membrane transporters leads to selective endocytosis into the vacuole for degradation. In addition, during glucose-induced MalP degradation, the homologous of E6AP C-terminus-type E3 ubiquitin ligase (HulA) is responsible for the ubiquitin modification of MalP, and the arrestin-like protein CreD is required for HulA targeting. Although CreD-mediated MalP internalization occurs in response to glucose, the mechanism by which CreD regulates HulA-dependent MalP ubiquitination remains unclear. In this study, we demonstrated that three (P/L)P<em>x</em>Y motifs present in the CreD protein are essential for functioning as HulA adaptors so that HulA can recognize MalP in response to glucose stimulation, enabling MalP internalization. Furthermore, four lysine residues (three highly conserved among <em>Aspergillus</em> species and yeast and one conserved among <em>Aspergillus</em> species) of CreD were found to be necessary for its ubiquitination, resulting in efficient glucose-induced MalP endocytosis. The results of this study pave the way for elucidating the regulatory mechanism of MalP endocytic degradation through ubiquitination by the HulA–CreD complex at the molecular level.</p></div>","PeriodicalId":55135,"journal":{"name":"Fungal Genetics and Biology","volume":"173 ","pages":"Article 103909"},"PeriodicalIF":2.4,"publicationDate":"2024-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S108718452400046X/pdfft?md5=0ca55b700d218ad72826eac18f3e841e&pid=1-s2.0-S108718452400046X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141405089","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}