Transformation of Fungi Using Shock Waves
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Genetic transformation of fungi faces major challenges. Most standard methods have a low frequency of transformation, problems of reproducibility, and are not suitable for high throughput experimentation. Additionally, there are many species of fungi that have proved recalcitrant to transformation by biolistics, electroporation, or agrobacterium transformation. We have developed a new method, based on the use of underwater shock waves that is highly efficient, simple, widely applicable, and not dependent on one particular type of tissue. Intact spores, conidia, or mycelia can be used. The method consists of the application of a few hundred shock waves to cell suspensions. Acoustic cavitation, i.e., the growth and violent collapse of microbubbles contained in the suspension, is responsible for a transient cell permeabilization. Several fungal species including, Aspergillus niger, Fusarium oxysporum, Phanerochaete chrysosporium, Trichoderma reesei, and Mycosphaerella fijiensis have been transformed successfully by this method. As an example, the methodology to transform Aspergillus niger is described in this chapter. Information on the basics of shock wave-mediated transformation of fungi is also included. Our main conclusion is that shock wave-mediated transformation is an attractive alternative for efficient fungal transformation.
KeywordsFilamentous fungi Genetic transformation Shock waves Biotechnology Shock wave-mediated transformation
The authors would like to thank Nancy Coconi, Claudia León, Elizabeth Ortiz, René Preza, Ángel Luis Rodríguez, and Guillermo Vázquez for technical assistance. This work was supported by DGAPA, UNAM Grant Number IN108410, and CONACYT Grant Number 22655.
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