Submitted to: Biology Methods and Protocols
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/4/2019
Publication Date: 2/18/2019
Citation: Nichols, N.N., Hector, R.E., Frazer, S.E. 2019. Genetic transformation of Coniochaeta sp. 2T2.1, key fungal member of a lignocellulose-degrading microbial consortium. Biology Methods and Protocols. 4:1-5. https://doi.org/10.1093/biomethods/bpz001.
DOI: https://doi.org/10.1093/biomethods/bpz001

Interpretive Summary: A fungus named strain 2T2.1 occupies a unique ecological niche as a member of a consortium of microbes that degrades fibrous biomass. The fungus also has potential use as a platform for biotechnological applications such as production of enzymes for biomass degradation. This work identified selectable genetic markers and developed a transformation method for strain 2T2.1. Additionally, a fluorescent version of the microbe was constructed. This feature makes the microbe amenable for understanding how members of the consortium interact and respond to their environment.

Technical Abstract: Coniochaeta sp. strain 2T2.1 was identified as a key member of a microbial consortium that degrades lignocellulosic biomass. Due to its ecological niche and its ability to also grow in pure culture on wheat, protocols for transformation and antibiotic selection of Coniochaeta sp. 2T2.1 were established. Hygromycin was found to be a reliable selectable transformation marker, and the mammalian codon-optimized green fluorescent protein was expressed and used to visualize fluorescence in transformed cells of strain 2T2.1.