Protocol for generating protoplasts of Trichoderma koningiopsis and transformation of plasmid DNA
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Updated: May 23, 2025

A Robotic Platform for High-throughput Protoplast Isolation and Transformation
Published on: September 27, 2016
Sophie Jin1, Fabrizio Alberti1
1School of Life Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.
Abstract:
Trichoderma koningiopsis is a filamentous fungus that produces numerous bioactive molecules and has the potential to be used as a biological control and plant growth-promoting agent. Here, we present a protocol for generating protoplasts of T. koningiopsis. We then describe four techniques for the stable transformation of plasmid DNA: polyethylene glycol (PEG)-mediated transformation of protoplasts, Agrobacterium-mediated transformation, electroporation of spores, and lipofection of mycelia. We then detail procedures for confirming the expression of a spCas9 gene using fluorescence microscopy.

