An efficient shortened genetic transformation strategy for filamentous fungus Trichoderma reesei

J Gen Appl Microbiol. 2020 Jan 31;65(6):301-307. doi: 10.2323/jgam.2019.02.001. Epub 2019 Jun 21.


The filamentous fungus Trichoderma reesei is one of the most important fungi for the production of cellulases and xylanases, which can be used for biofuel production from lignocellulose. We aimed to develop an effective selection marker system for more extensive functional genomic studies in the fungus T. reesei, and to construct better industrial transformants for producing cellulases. Here, we present a novel effective G418 selection marker to use a codon-optimized neomycin phosphotransferase II gene nptII to transform T. reesei. We developed an effective and erasable selection marker, lcNG, and a combined genetic transformation system for gene manipulation in T. reesei using a two-Agrobacterium-mediated transformation method. This transformation strategy combines two steps in the transformation protocol, which saves 15-30-day's time. The system could be a useful tool for the genetic engineering of T. reesei.

Keywords: G418; Trichoderma reesei; optimized codons; two-Agrobacterium-mediated transformation.

MeSH terms

  • Cellulases / metabolism
  • Genetic Engineering / methods
  • Genome, Fungal
  • Kanamycin Kinase / genetics
  • Transformation, Genetic*
  • Trichoderma / enzymology
  • Trichoderma / genetics*


  • Kanamycin Kinase
  • Cellulases