Fungal Biology and Biotechnology最新文献

{"title":"Preservation stress resistance of melanin deficient conidia from Paecilomyces variotii and Penicillium roqueforti mutants generated via CRISPR/Cas9 genome editing.","authors":"Sjoerd J Seekles,&nbsp;Pepijn P P Teunisse,&nbsp;Maarten Punt,&nbsp;Tom van den Brule,&nbsp;Jan Dijksterhuis,&nbsp;Jos Houbraken,&nbsp;Han A B Wösten,&nbsp;Arthur F J Ram","doi":"10.1186/s40694-021-00111-w","DOIUrl":"https://doi.org/10.1186/s40694-021-00111-w","url":null,"abstract":"<p><strong>Background: </strong>The filamentous fungi Paecilomyces variotii and Penicillium roqueforti are prevalent food spoilers and are of interest as potential future cell factories. A functional CRISPR/Cas9 genome editing system would be beneficial for biotechnological advances as well as future (genetic) research in P. variotii and P. roqueforti.</p><p><strong>Results: </strong>Here we describe the successful implementation of an efficient AMA1-based CRISPR/Cas9 genome editing system developed for Aspergillus niger in P. variotii and P. roqueforti in order to create melanin deficient strains. Additionally, kusA^- mutant strains with a disrupted non-homologous end-joining repair mechanism were created to further optimize and facilitate efficient genome editing in these species. The effect of melanin on the resistance of conidia against the food preservation stressors heat and UV-C radiation was assessed by comparing wild-type and melanin deficient mutant conidia.</p><p><strong>Conclusions: </strong>Our findings show the successful use of CRISPR/Cas9 genome editing and its high efficiency in P. variotii and P. roqueforti in both wild-type strains as well as kusA^- mutant background strains. Additionally, we observed that melanin deficient conidia of three food spoiling fungi were not altered in their heat resistance. However, melanin deficient conidia had increased sensitivity towards UV-C radiation.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"8 1","pages":"4"},"PeriodicalIF":0.0,"publicationDate":"2021-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-021-00111-w","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25539748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fungal sensing skin.","authors":"Andrew Adamatzky, Antoni Gandia, Alessandro Chiolerio","doi":"10.1186/s40694-021-00110-x","DOIUrl":"10.1186/s40694-021-00110-x","url":null,"abstract":"<p><strong>Background: </strong>A fungal skin is a thin flexible sheet of a living homogeneous mycelium made by a filamentous fungus. The skin could be used in future living architectures of adaptive buildings and as a sensing living skin for soft self-growing/adaptive robots.</p><p><strong>Results: </strong>In experimental laboratory studies we demonstrate that the fungal skin is capable for recognising mechanical and optical stimulation. The skin reacts differently to loading of a weight, removal of the weight, and switching illumination on and off.</p><p><strong>Conclusion: </strong>These are the first experimental evidences that fungal materials can be used not only as mechanical 'skeletons' in architecture and robotics but also as intelligent skins capable for recognition of external stimuli and sensorial fusion.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"8 1","pages":"3"},"PeriodicalIF":0.0,"publicationDate":"2021-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7972235/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25488172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A fully automated pipeline for the dynamic at-line morphology analysis of microscale Aspergillus cultivation.","authors":"Roman Jansen,&nbsp;Kira Küsters,&nbsp;Holger Morschett,&nbsp;Wolfgang Wiechert,&nbsp;Marco Oldiges","doi":"10.1186/s40694-021-00109-4","DOIUrl":"https://doi.org/10.1186/s40694-021-00109-4","url":null,"abstract":"<p><strong>Background: </strong>Morphology, being one of the key factors influencing productivity of filamentous fungi, is of great interest during bioprocess development. With increasing demand of high-throughput phenotyping technologies for fungi due to the emergence of novel time-efficient genetic engineering technologies, workflows for automated liquid handling combined with high-throughput morphology analysis have to be developed.</p><p><strong>Results: </strong>In this study, a protocol allowing for 48 parallel microbioreactor cultivations of Aspergillus carbonarius with non-invasive online signals of backscatter and dissolved oxygen was established. To handle the increased cultivation throughput, the utilized microbioreactor is integrated into a liquid handling platform. During cultivation of filamentous fungi, cell suspensions result in either viscous broths or form pellets with varying size throughout the process. Therefore, tailor-made liquid handling parameters such as aspiration/dispense height, velocity and mixing steps were optimized and validated. Development and utilization of a novel injection station enabled a workflow, where biomass samples are automatically transferred into a flow through chamber fixed under a light microscope. In combination with an automated image analysis concept, this enabled an automated morphology analysis pipeline. The workflow was tested in a first application study, where the projected biomass area was determined at two different cultivation temperatures and compared to the microbioreactor online signals.</p><p><strong>Conclusions: </strong>A novel and robust workflow starting from microbioreactor cultivation, automated sample harvest and processing via liquid handling robots up to automated morphology analysis was developed. This protocol enables the determination of projected biomass areas for filamentous fungi in an automated and high-throughput manner. This measurement of morphology can be applied to describe overall pellet size distribution and heterogeneity.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"8 1","pages":"2"},"PeriodicalIF":0.0,"publicationDate":"2021-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-021-00109-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25441255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of scleroglucan production by Sclerotium rolfsii by lowering pH during fermentation via oxalate metabolic pathway manipulation using CRISPR/Cas9.","authors":"Tianlong Bai,&nbsp;Teng Wang,&nbsp;Yan Li,&nbsp;Na L Gao,&nbsp;Lixin Zhang,&nbsp;Wei-Hua Chen,&nbsp;Xiushan Yin","doi":"10.1186/s40694-021-00108-5","DOIUrl":"https://doi.org/10.1186/s40694-021-00108-5","url":null,"abstract":"<p><strong>Background: </strong>Sclerotium rolfsii is a potent producer of many secondary metabolites, one of which like scleroglucan is an exopolysaccharide (EPS) appreciated as a multipurpose compound applicable in many industrial fields.</p><p><strong>Results: </strong>Aspartate transaminase (AAT1) catalyzes the interconversion of aspartate and α-ketoglutarate to glutamate and oxaloacetate. We selected AAT1 in the oxalate metabolic pathway as a target of CRISPR/Cas9. Disruption of AAT1 leads to the accumulation of oxalate, rather than its conversion to α-ketoglutarate (AKG). Therefore, AAT1-mutant serves to lower the pH (pH 3-4) so as to increase the production of the pH-sensitive metabolite scleroglucan to 21.03 g L^-1 with a productivity of up to 0.25 g L^-1·h^-1.</p><p><strong>Conclusions: </strong>We established a platform for gene editing that could rapidly generate and select mutants to provide a new beneficial strain of S. rolfsii as a scleroglucan hyper-producer, which is expected to reduce the cost of controlling the optimum pH condition in the fermentation industry.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"8 1","pages":"1"},"PeriodicalIF":0.0,"publicationDate":"2021-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-021-00108-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25386120","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The fungal sexual revolution continues: discovery of sexual development in members of the genus Aspergillus and its consequences.","authors":"Valeria Ellena,&nbsp;Michael Sauer,&nbsp;Matthias G Steiger","doi":"10.1186/s40694-020-00107-y","DOIUrl":"https://doi.org/10.1186/s40694-020-00107-y","url":null,"abstract":"<p><p>Asexuality was considered to be a common feature of a large part of fungi, including those of the genus Aspergillus. However, recent advances and the available genomic and genetic engineering technologies allowed to gather more and more indications of a hidden sexuality in fungi previously considered asexual. In parallel, the acquired knowledge of the most suitable conditions for crossings was shown to be crucial to effectively promote sexual reproduction in the laboratory. These discoveries not only have consequences on our knowledge of the biological processes ongoing in nature, questioning if truly asexual fungal species exist, but they also have important implications on other research areas. For instance, the presence of sexuality in certain fungi can have effects on their pathogenicity or on shaping the ecosystem that they normally colonize. For these reasons, further investigations of the sexual potential of Aspergillus species, such as the industrially important A. niger, will be carried on.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"7 1","pages":"17"},"PeriodicalIF":0.0,"publicationDate":"2020-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-020-00107-y","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38746261","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent advances in the intellectual property landscape of filamentous fungi.","authors":"Silvia Hüttner,&nbsp;Anton Johansson,&nbsp;Paulo Gonçalves Teixeira,&nbsp;Puck Achterberg,&nbsp;Ramkumar B Nair","doi":"10.1186/s40694-020-00106-z","DOIUrl":"https://doi.org/10.1186/s40694-020-00106-z","url":null,"abstract":"<p><p>For centuries, filamentous fungi have been used in the making of food and beverages, and for decades for the production of enzymes and pharmaceuticals. In the last decades, the intellectual property (IP) landscape for fungal technology has seen an ever increasing upward trend, introducing new and promising applications utilising fungi. In this review, we highlight fungi-related patent applications published during the last 5 years (2015-2020), identify the key players in each field, and analyse future trends. New developments in the field of fungal technology include the increased use of filamentous fungi as a food source (mycoprotein), using fungi as biodegradable materials, in wastewater treatment, in integrated biorefineries and as biological pest agents. Biotechnology companies in Europe and the US are currently leading when it comes to the number of patents in these areas, but Asian companies and research institutes, in particular in China, are becoming increasingly important players, for example in pesticide formulation and agricultural practices.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"7 1","pages":"16"},"PeriodicalIF":0.0,"publicationDate":"2020-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-020-00106-z","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38687884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selection markers for transformation of the sequenced reference monokaryon Okayama 7/#130 and homokaryon AmutBmut of <i>Coprinopsis cinerea</i>.","authors":"Bastian Dörnte, Can Peng, Zemin Fang, Aysha Kamran, Cut Yulvizar, Ursula Kües","doi":"10.1186/s40694-020-00105-0","DOIUrl":"10.1186/s40694-020-00105-0","url":null,"abstract":"<p><strong>Background: </strong>Two reference strains have been sequenced from the mushroom <i>Coprinopsis cinerea</i>, monokaryon Okayama 7/#130 (OK130) and the self-compatible homokaryon AmutBmut. An adenine-auxotrophy in OK130 (<i>ade8-1</i>) and a <i>para</i>-aminobenzoic acid (PABA)-auxotrophy in AmutBmut (<i>pab1-1</i>) offer selection markers for transformations. Of these two strains, homokaryon AmutBmut had been transformed before to PABA-prototrophy and with the bacterial hygromycin resistance marker <i>hph</i>, respectively.</p><p><strong>Results: </strong>Gene <i>ade8</i> encodes a bifunctional enzyme with an N-terminal glycinamide ribonucleotide synthase (GARS) and a C-terminal aminoimidazole ribonucleotide synthase (AIRS) domain required for steps 2 and 5 in the de novo biosynthesis of purines, respectively. In OK130, a missense mutation in <i>ade8-1</i> rendered residue N231 for ribose recognition by the A loop of the GARS domain into D231. The new <i>ade8</i> ⁺ vector p<i>Cc</i>Ade8 complements the auxotrophy of OK130 in transformations. Transformation rates with p<i>Cc</i>Ade8 in single-vector and co-transformations with <i>ade8</i> ⁺-selection were similarly high, unlike for <i>trp1</i> ⁺ plasmids which exhibit suicidal feedback-effects in single-vector transformations with complementation of tryptophan synthase defects. As various other plasmids, unselected p<i>Cc</i>Ade8 helped in co-transformations of <i>trp1</i> strains with a <i>trp1</i> ⁺-selection vector to overcome suicidal effects by transferred <i>trp1</i> ⁺. Co-transformation rates of p<i>Cc</i>Ade8 in OK130 under adenine selection with nuclear integration of unselected DNA were as high as 80% of clones. Co-transformation rates of expressed genes reached 26-42% for various laccase genes and up to 67% with <i>lcc9</i> silencing vectors. The bacterial gene <i>hph</i> can also be used as another, albeit less efficient, selection marker for OK130 transformants, but with similarly high co-transformation rates. We further show that the <i>pab1-1</i> defect in AmutBmut is due to a missense mutation which changed the conserved PIKGT motif for chorismate binding in the C-terminal PabB domain to PIEGT in the mutated 4-amino-4-deoxychorismate synthase.</p><p><strong>Conclusions: </strong><i>ade8-1</i> and <i>pab1-1</i> auxotrophic defects in <i>C. cinerea</i> reference strains OK130 and AmutBmut for complementation in transformation are described. p<i>Cc</i>Ade8 is a new transformation vector useful for selection in single and co-transformations of the sequenced monokaryon OK130 which was transformed for the first time. The bacterial gene <i>hph</i> can also be used as an additional selection marker in OK130, making in combination with <i>ade8</i> ⁺ successive rounds of transformation possible.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"7 ","pages":"15"},"PeriodicalIF":0.0,"publicationDate":"2020-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-020-00105-0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38493033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fungal research in Japan: tradition and future.","authors":"Norio Takeshita","doi":"10.1186/s40694-020-00104-1","DOIUrl":"https://doi.org/10.1186/s40694-020-00104-1","url":null,"abstract":"","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"7 ","pages":"14"},"PeriodicalIF":0.0,"publicationDate":"2020-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-020-00104-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38420258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular vesicles from the apoplastic fungal wheat pathogen <i>Zymoseptoria tritici</i>.","authors":"Erin H Hill, Peter S Solomon","doi":"10.1186/s40694-020-00103-2","DOIUrl":"10.1186/s40694-020-00103-2","url":null,"abstract":"<p><strong>Background: </strong>The fungal pathogen <i>Zymoseptoria tritici</i> is a significant constraint to wheat production in temperate cropping regions around the world. Despite its agronomic impacts, the mechanisms allowing the pathogen to asymptomatically invade and grow in the apoplast of wheat leaves before causing extensive host cell death remain elusive. Given recent evidence of extracellular vesicles (EVs)-secreted, membrane-bound nanoparticles containing molecular cargo-being implicated in extracellular communication between plants and fungal pathogen, we have initiated an in vitro investigation of EVs from this apoplastic fungal wheat pathogen. We aimed to isolate EVs from <i>Z. tritici</i> broth cultures and examine their protein composition in relation to the soluble protein in the culture filtrate and to existing fungal EV proteomes.</p><p><strong>Results: </strong><i>Zymoseptoria tritici</i> EVs were isolated from broth culture filtrates using differential ultracentrifugation (DUC) and examined with transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). <i>Z. tritici</i> EVs were observed as a heterogeneous population of particles, with most between 50 and 250 nm. These particles were found in abundance in the culture filtrates of viable <i>Z. tritici</i> cultures, but not heat-killed cultures incubated for an equivalent time and of comparable biomass. Bottom-up proteomic analysis using LC-MS/MS, followed by stringent filtering revealed 240 <i>Z. tritici</i> EV proteins. These proteins were distinct from soluble proteins identified in <i>Z. tritici</i> culture filtrates, but were similar to proteins identified in EVs from other fungi, based on sequence similarity analyses. Notably, a putative marker protein recently identified in <i>Candida albicans</i> EVs was also consistently detected in <i>Z. tritici</i> EVs.</p><p><strong>Conclusion: </strong>We have shown EVs can be isolated from the devastating fungal wheat pathogen <i>Z. tritici</i> and are similar to protein composition to previously characterised fungal EVs. EVs from human pathogenic fungi are implicated in virulence, but the role of EVs in the interaction of phytopathogenic fungi and their hosts is unknown. These in vitro analyses provide a basis for expanding investigations of <i>Z. tritici</i> EVs <i>in planta,</i> to examine their involvement in the infection process of this apoplastic wheat pathogen and more broadly, advance understanding of noncanonical secretion in filamentous plant pathogens.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"7 ","pages":"13"},"PeriodicalIF":0.0,"publicationDate":"2020-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7501697/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38412195","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of the human estrogen receptor within a synthetic transcription factor in <i>Trichoderma reesei</i>.","authors":"Christian Derntl,&nbsp;Robert Mach,&nbsp;Astrid Mach-Aigner","doi":"10.1186/s40694-020-00102-3","DOIUrl":"https://doi.org/10.1186/s40694-020-00102-3","url":null,"abstract":"<p><strong>Background: </strong>Synthetic gene expression systems offer a possibility for controllable and targeted induction of the expression of genes of interest, which is a fundamental technique necessary for basic research and industrial applications. The human estrogen receptor α contains a ligand binding domain that enforces dimerization and nuclear import upon binding of the inducer 17β-estradiol. In this study, we tested the potential of this ligand binding domain to be used in filamentous fungi as an auto-regulatory domain in a synthetic transcription factor.</p><p><strong>Results: </strong>We constructed the synthetic transcription factor SynX by fusing the DNA-binding domain of Xyr1 (Xylanase Regulator 1), the transactivation domain of Ypr1 (Yellow Pigment Regulator 1), and the ligand binding domain of the human estrogen receptor α. SynX is able to strongly induce the gene expression of xylanases and an aldose reductase by addition of 17β-estradiol, but SynX does not induce gene expression of cellulases. Importantly, the induction of xylanase activities is mostly carbon source independent and can be fine-tuned by controlling the concentration of 17β-estradiol.</p><p><strong>Conclusion: </strong>The ability of SynX to induce gene expression of xylanase encoding genes by addition of 17β-estradiol demonstrates that the ligand binding domain of the human estrogen receptor α works in filamentous fungi, and that it can be combined with a transactivation domain other than the commonly used transactivation domain of herpes simplex virion protein VP16.</p>","PeriodicalId":52292,"journal":{"name":"Fungal Biology and Biotechnology","volume":"7 ","pages":"12"},"PeriodicalIF":0.0,"publicationDate":"2020-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40694-020-00102-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38238727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}