Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulans
Abstract Extracellular vesicles (EVs) are increasingly recognized as an important mechanism for cell–cell interactions. Their role in fungi is still poorly understood and they have been isolated from only a handful of species. Here, we isolated and characterized EVs from Aureobasidium pullulans, a b...
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| Format: | Article |
| Language: | English |
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BMC
2022-11-01
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| Series: | Fungal Biology and Biotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40694-022-00146-7 |
| _version_ | 1811328496589340672 |
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| author | Anja Černoša Cene Gostinčar Teja Lavrin Rok Kostanjšek Metka Lenassi Nina Gunde-Cimerman |
| author_facet | Anja Černoša Cene Gostinčar Teja Lavrin Rok Kostanjšek Metka Lenassi Nina Gunde-Cimerman |
| author_sort | Anja Černoša |
| collection | DOAJ |
| description | Abstract Extracellular vesicles (EVs) are increasingly recognized as an important mechanism for cell–cell interactions. Their role in fungi is still poorly understood and they have been isolated from only a handful of species. Here, we isolated and characterized EVs from Aureobasidium pullulans, a biotechnologically important black yeast-like fungus that is increasingly used for biocontrol of phytopathogenic fungi and bacteria. After optimization of the isolation protocol, characterization of EVs from A. pullulans by transmission electron microscopy (TEM) revealed a typical cup-shaped morphology and different subpopulations of EVs. These results were confirmed by nanoparticle tracking analysis (NTA), which revealed that A. pullulans produced 6.1 × 108 nanoparticles per milliliter of culture medium. Proteomic analysis of EVs detected 642 proteins. A small fraction of them had signal peptides for secretion and transmembrane domains. Proteins characteristic of different synthesis pathways were found, suggesting that EVs are synthesized by multiple pathways in A. pullulans. Enrichment analysis using Gene Ontology showed that most of the proteins found in the EVs were associated with primary metabolism. When sequencing the small RNA fraction of A. pullulans EVs, we found two hypothetical novel mil-RNAs. Finally, we tested the biocontrol potential of EVs from A. pullulans. The EVs did not inhibit the germination of spores of three important phytopathogenic fungi—Botrytis cinerea, Colletotrichum acutatum, and Penicillium expansum. However, exposure of grown cultures of C. acutatum and P. expansum to A. pullulans EVs resulted in visible changes in morphology of colonies. These preliminary results suggest that EVs may be part of the antagonistic activity of A. pullulans, which is so far only partially understood. Thus, the first isolation and characterization of EVs from A. pullulans provides a starting point for further studies of EVs in the biotechnologically important traits of the biocontrol black fungus A. pullulans in particular and in the biological role of fungal EVs in general. |
| first_indexed | 2024-04-13T15:26:09Z |
| format | Article |
| id | doaj.art-ea11fc6dbddf4f7a997fb232e5bf7531 |
| institution | Directory Open Access Journal |
| issn | 2054-3085 |
| language | English |
| last_indexed | 2024-04-13T15:26:09Z |
| publishDate | 2022-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Fungal Biology and Biotechnology |
| spelling | doaj.art-ea11fc6dbddf4f7a997fb232e5bf75312022-12-22T02:41:30ZengBMCFungal Biology and Biotechnology2054-30852022-11-019111710.1186/s40694-022-00146-7Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulansAnja Černoša0Cene Gostinčar1Teja Lavrin2Rok Kostanjšek3Metka Lenassi4Nina Gunde-Cimerman5Department of Biology, Biotechnical Faculty, University of LjubljanaDepartment of Biology, Biotechnical Faculty, University of LjubljanaInstitute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of LjubljanaDepartment of Biology, Biotechnical Faculty, University of LjubljanaInstitute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of LjubljanaDepartment of Biology, Biotechnical Faculty, University of LjubljanaAbstract Extracellular vesicles (EVs) are increasingly recognized as an important mechanism for cell–cell interactions. Their role in fungi is still poorly understood and they have been isolated from only a handful of species. Here, we isolated and characterized EVs from Aureobasidium pullulans, a biotechnologically important black yeast-like fungus that is increasingly used for biocontrol of phytopathogenic fungi and bacteria. After optimization of the isolation protocol, characterization of EVs from A. pullulans by transmission electron microscopy (TEM) revealed a typical cup-shaped morphology and different subpopulations of EVs. These results were confirmed by nanoparticle tracking analysis (NTA), which revealed that A. pullulans produced 6.1 × 108 nanoparticles per milliliter of culture medium. Proteomic analysis of EVs detected 642 proteins. A small fraction of them had signal peptides for secretion and transmembrane domains. Proteins characteristic of different synthesis pathways were found, suggesting that EVs are synthesized by multiple pathways in A. pullulans. Enrichment analysis using Gene Ontology showed that most of the proteins found in the EVs were associated with primary metabolism. When sequencing the small RNA fraction of A. pullulans EVs, we found two hypothetical novel mil-RNAs. Finally, we tested the biocontrol potential of EVs from A. pullulans. The EVs did not inhibit the germination of spores of three important phytopathogenic fungi—Botrytis cinerea, Colletotrichum acutatum, and Penicillium expansum. However, exposure of grown cultures of C. acutatum and P. expansum to A. pullulans EVs resulted in visible changes in morphology of colonies. These preliminary results suggest that EVs may be part of the antagonistic activity of A. pullulans, which is so far only partially understood. Thus, the first isolation and characterization of EVs from A. pullulans provides a starting point for further studies of EVs in the biotechnologically important traits of the biocontrol black fungus A. pullulans in particular and in the biological role of fungal EVs in general.https://doi.org/10.1186/s40694-022-00146-7Extracellular vesiclesAureobasidium pullulansBiocontrolProteomicssRNA |
| spellingShingle | Anja Černoša Cene Gostinčar Teja Lavrin Rok Kostanjšek Metka Lenassi Nina Gunde-Cimerman Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulans Fungal Biology and Biotechnology Extracellular vesicles Aureobasidium pullulans Biocontrol Proteomics sRNA |
| title | Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulans |
| title_full | Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulans |
| title_fullStr | Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulans |
| title_full_unstemmed | Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulans |
| title_short | Isolation and characterization of extracellular vesicles from biotechnologically important fungus Aureobasidium pullulans |
| title_sort | isolation and characterization of extracellular vesicles from biotechnologically important fungus aureobasidium pullulans |
| topic | Extracellular vesicles Aureobasidium pullulans Biocontrol Proteomics sRNA |
| url | https://doi.org/10.1186/s40694-022-00146-7 |
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