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.

本研究在丝状真菌深绿木霉（Trichoderma atroviride）中开展了随机条形码标记T-DNA测序（Random Barcode Tagged T-DNA Sequencing，简称RB-TDNAseq）实验。所构建的突变体库用于高通量适合度筛选，以鉴定参与氨基酸生物合成途径以及特定碳源利用过程的功能基因。通过整合本研究筛选得到的突变体适合度谱与相同生长条件下获取的转录组数据，对全基因组水平的基因功能进行了系统性探究。结合高通量遗传筛选与RNA测序技术，本研究成功鉴定出木糖利用与半胱氨酸生物合成所必需的保守基因。本数据集为丝状真菌领域提供了极具价值的功能基因组学资源，并搭建了可扩展的全基因组基因功能发现平台，既可为真菌生物学的基础研究提供支撑，也可服务于旨在提升生物防治、生物技术与可持续农业水平的应用研究。