Advances, Problems, and Prospects of Genetic Transformation of Fungi

Poyedinok, Natalia L.; Blume, Ya. B.

doi:10.3103/s009545271802007x

Cited by 12 publications

(12 citation statements)

References 202 publications

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“…Additionally, various highly effective methods for studying fungal genomic variability are applied using various systems of genetic transformation. The modification of genes through their transformation to create fungal mutant lines is an essential step in investigations of gene function and relationships (Poyedinok & Blume, 2018). Such strategies will facilitate analyses of fungal genetic classification and improve the biosynthetic activity of many biotechnologically important Pleurotus strains.…”

Section: Discussionmentioning

confidence: 99%

Green potential ofPleurotusspp. in biotechnology

Секан

Myronycheva

Karlsson

et al. 2019

PeerJ

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Background The genus Pleurotus is most exploitable xylotrophic fungi, with valuable biotechnological, medical, and nutritional properties. The relevant features of the representatives of this genus to provide attractive low-cost industrial tools have been reported in numerous studies to resolve the pressure of ecological issues. Additionally, a number of Pleurotus species are highly adaptive, do not require any special conditions for growth, and possess specific resistance to contaminating diseases and pests. The unique properties of Pleurotus species widely used in many environmental technologies, such as organic solid waste recycling, chemical pollutant degradation, and bioethanol production. Methodology The literature study encompasses peer-reviewed journals identified by systematic searches of electronic databases such as Google Scholar, NCBI, Springer, ResearchGate, ScienceDirect, and ISI Web of Knowledge. The search scheme was divided into several steps, as described below. Results In this review, we describe studies examining the biotechnological feasibility of Pleurotus spp. to elucidate the importance of this genus for use in green technology. Here, we review areas of application of the genus Pleurotus as a prospective biotechnological tool. Conclusion The incomplete description of some fungal biochemical pathways emphasises the future research goals for this fungal culture.

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Section: Discussionmentioning

confidence: 99%

Green potential ofPleurotusspp. in biotechnology

Секан

Myronycheva

Karlsson

et al. 2019

PeerJ

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“…In some PEG-mediated transformation protocols, the use of linear plasmids is recommended to increase the frequency of transformation. However, results obtained are variable and depend on the fungal species used (Poyedinok and Blume, 2018). In our case, we observed that the linearization of plasmid, in combination with PEG-mediated transformation, did not yield transformants (Table 1).…”

Section: Discussionmentioning

confidence: 99%

Genetic Transformation of the Filamentous Fungus Pseudogymnoascus verrucosus of Antarctic Origin

et al. 2019

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Cold-adapted fungi isolated from Antarctica, in particular those belonging to the genus Pseudogymnoascus, are producers of secondary metabolites with interesting bioactive properties as well as enzymes with potential biotechnological applications. However, at genetic level, the study of these fungi has been hindered by the lack of suitable genetic tools such as transformation systems. In fungi, the availability of transformation systems is a key to address the functional analysis of genes related with the production of a particular metabolite or enzyme. To the best of our knowledge, the transformation of Pseudogymnoascus strains of Antarctic origin has not been achieved yet. In this work, we describe for the first time the successful transformation of a Pseudogymnoascus verrucosus strain of Antarctic origin, using two methodologies: the polyethylene glycol (PEG)-mediated transformation, and the electroporation of germinated conidia. We achieved transformation efficiencies of 15.87 ± 5.16 transformants per μg of DNA and 2.67 ± 1.15 transformants per μg of DNA for PEG-mediated transformation and electroporation of germinated conidia, respectively. These results indicate that PEG-mediated transformation is a very efficient method for the transformation of this Antarctic fungus. The genetic transformation of Pseudogymnoascus verrucosus described in this work represents the first example of transformation of a filamentous fungus of Antarctic origin.

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“…The characterization of A. fumigatus still extensively depends on the use of dominant selectable markers, particularly the hygromycin B phosphotransferase ( hph ) gene and the pyrithiamine resistance ( ptrA ) gene. hph , which confers resistance to hygromycin, encodes a phosphotransferase from Escherichia coli , and is one of the most commonly utilized dominant selectable markers to genetically modify organisms, from bacteria to mammalian cells [ 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 ]. For use in Aspergilli , the hph gene is usually under control of the Aspergillus nidulans gpdA promoter [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

“…A major drawback of drug resistance markers nowadays is related to the generation of genetically modified organisms, but this problem mostly concerns species that are industrially relevant [ 18 , 19 , 20 ]. In the case of A. fumigatus , which is not used in production processes and is studied for its effect on human health as pathogen, drug resistance markers have the advantages of not needing a specific host strain, allowing the stable integration of constructs with high efficiency and generating mutant strains that are easy to select and purify [ 7 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Inducible Selectable Marker Genes to Improve Aspergillus fumigatus Genetic Manipulation

Baldin

Kühbacher

Merschak

et al. 2021

JoF

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The hygromycin B phosphotransferase gene from Escherichia coli and the pyrithiamine resistance gene from Aspergillus oryzae are two dominant selectable marker genes widely used to genetically manipulate several fungal species. Despite the recent development of CRISPR/Cas9 and marker-free systems, in vitro molecular tools to study Aspergillus fumigatus, which is a saprophytic fungus causing life-threatening diseases in immunocompromised hosts, still rely extensively on the use of dominant selectable markers. The limited number of drug selectable markers is already a critical aspect, but the possibility that their introduction into a microorganism could induce enhanced virulence or undesired effects on metabolic behavior constitutes another problem. In this context, here, we demonstrate that the use of ptrA in A. fumigatus leads to the secretion of a compound that allows the recovery of thiamine auxotrophy. In this study, we developed a simple modification of the two commonly used dominant markers in which the development of resistance can be controlled by the xylose-inducible promoter PxylP from Penicillium chrysogenum. This strategy provides an easy solution to avoid undesired side effects, since the marker expression can be readily silenced when not required.

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Advances, Problems, and Prospects of Genetic Transformation of Fungi

Cited by 12 publications

References 202 publications

Green potential ofPleurotusspp. in biotechnology

Green potential ofPleurotusspp. in biotechnology

Genetic Transformation of the Filamentous Fungus Pseudogymnoascus verrucosus of Antarctic Origin

Inducible Selectable Marker Genes to Improve Aspergillus fumigatus Genetic Manipulation

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