Whole-genome functional characterization using mutants from a randomly barcoded insertional library in Trichoderma atroviride
We performed RB-TDNAseq (Random Barcode Tagged T-DNA Sequencing) experiments in the filamentous fungus Trichoderma atroviride. This mutant library was used for high-throughput fitness screening to identify genes involved in amino acid biosynthetic pathways and in the utilization of specific carbon sources. Genome-wide gene function was interrogated by integrating mutant fitness profiles from our screens with transcriptional data obtained under the same growth conditions.Through the combination of high-throughput genetic screens and RNA sequencing, we identified conserved genes required for xylose utilization and cysteine biosynthesis. This dataset provides a valuable functional genomics resource and establishes a scalable platform for genome-wide gene function discovery in filamentous fungi, enabling both fundamental studies of fungal biology and applied research aimed at improving biocontrol, biotechnology, and sustainable agriculture.