Optimized electroporation for efficient evaluation of genetic elements in Dichomitus squalens.

Abstract

Dichomitus squalens, a promising white-rot basidiomycete for industrial enzyme production, necessitates efficient genetic manipulation systems to fully leverage its biotechnological potential. Although established methods such as protoplast-mediated and Agrobacterium tumefaciens-mediated transformations are effective in D. squalens, they are complex and time-consuming. This study introduces the electroporation transformation system for D. squalens, which is simpler and timesaving. By optimizing electroporation parameters, we obtained 77 ± 11 transformants per μg of DNA. Furthermore, we validated the suitability of the Nourseothricin N-acetyl transferase gene as a selectable marker and the NanoLuciferase gene as a bioluminescent reporter in D. squalens using our refined electroporation protocol. This study expands the toolkit for genetic engineering in D. squalens, offering greater flexibility for future molecular investigations. The development of this electroporation system not only enhances the ease of genetic manipulation in D. squalens but also provides a foundation for further exploration of its enzymatic capabilities and potential applications in biotechnology. The streamlined protocol allows for more efficient and rapid genetic engineering, facilitating the study of gene function and the development of improved strains for industrial purposes.