Self-fusion and fusion cell isolation of transformants derived from white rot fungus Phanerochaete sordida YK-624 by simple visual method

• Published in: Journal of bioscience and bioengineering 2020-02, Vol.129 (2), p.146-149
• Main Authors: Mori, Toshio, Kondo, Ojiro, Sumiya, Tomoki, Kawagishi, Hirokazu, Hirai, Hirofumi
• Format: Article
• Language: English
• Subjects: Cell Separation - methods; Chlamydospore; Fungal Proteins - genetics; Fungal Proteins - metabolism; Phanerochaete - cytology; Phanerochaete - genetics; Phanerochaete - isolation & purification; Phanerochaete - metabolism; Phanerochaete sordida YK-624; Recombinant Proteins - metabolism; Self-compatibility; Self-fusion
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1016/j.jbiosc.2019.08.011

In order to develop a simple method for crossing two transformants, we first attempted to elucidate the fusion type (self-compatibility or -incompatible) of Phanerochaete sordida YK-624. Two transformants expressing green or red fluorescent protein derived from an auxotrophic mutant were constructed. Each recombinant protein fluoresced by expression as a fused protein with glyceraldehyde-3-phosphate dehydrogenase. On co-culture of both transformants, a number of sequential hyphal cells emitting dual fluorescence were formed at the contact areas of both hyphae. Some of the single cells isolated as protoplasts and chlamydospore from the co-cultures also expressed these fluorescent proteins. These results suggest that P. sordida YK-624 possesses a self-compatible fusion system. In addition, transformant strains with different fluorescence derived from this fungus can readily undergo self-fusion and nuclear interchange events by confrontational and mixed cultivation, and we developed a simple method that allows fused cells to be isolated as chlamydospores.