Abstract
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.
Highlights
Antarctica, one of the most extreme ecosystems in the world, is inhabited by a myriad of microorganisms that have adapted to this harsh environment using several cellular and molecular strategies (Pearce, 2012)
In the case of hygromycin B, the growth of P. verrucosus FAE27 was inhibited at 20 μg/ml (Figure 1)
The most widely methods used for transformation are Agrobacterium tumefaciens-mediated transformation (ATMT), polyethylene glycol (PEG)-mediated protoplast transformation, and electroporation (Li et al, 2017)
Summary
Antarctica, one of the most extreme ecosystems in the world, is inhabited by a myriad of microorganisms that have adapted to this harsh environment using several cellular and molecular strategies (Pearce, 2012). Studies prospecting bioactivities in filamentous fungi isolated from Antarctica have gained much interest during last decades, showing that these fungi produce metabolites with interesting biological activities Several studies have shown that extracts of these fungi produce bioactive metabolites with potential application, including antibacterial, antifungal, tripanocidal, herbicidal, and antitumoral activities (Furbino et al, 2014; Henríquez et al, 2014; Gonçalves et al, 2015; Gomes et al, 2018; Purić et al, 2018; Vieira et al, 2018). Pseudogymnoascus strains of Antarctic origin have shown ability to produce cold-adapted enzymes with potential application in several industrial processes (Loperena et al, 2012; Poveda et al, 2018)
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