The basidiomycete yeast Rhodosporidium toruloides (also known as Rhodotorula toruloides) accumulates high concentrations of lipids and carotenoids from diverse carbon sources. It has great potential as a model for the cellular biology of lipid droplets and for sustainable chemical production. We developed a method for high-throughput genetics (RB-TDNAseq), using sequence-barcoded Agrobacterium tumefaciens T-DNA insertions. We identified 1,337 putative essential genes with low T-DNA insertion rates. We functionally profiled genes required for fatty acid catabolism and lipid accumulation, validating results with 35 targeted deletion strains. We identified a high-confidence set of 150 genes affecting lipid accumulation, including genes with predicted function in signaling cascades, gene expression, protein modification and vesicular trafficking, autophagy, amino acid synthesis and tRNA modification, and genes of unknown function. These results greatly advance our understanding of lipid metabolism in this oleaginous species and demonstrate a general approach for barcoded mutagenesis that should enable functional genomics in diverse fungi.

中文翻译：

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产油酵母Rhodosporidium toruloides脂质代谢的功能基因组学

担子菌酵母 Rodosporidium toruloides（也称为 Rhodotorula toruloides）从不同的碳源中积累高浓度的脂质和类胡萝卜素。它具有作为脂滴细胞生物学模型和可持续化学生产模型的巨大潜力。我们开发了一种高通量遗传学方法 (RB-TDNAseq)，使用序列条形码的根癌农杆菌 T-DNA 插入。我们确定了 1,337 个具有低 T-DNA 插入率的假定必需基因。我们对脂肪酸分解代谢和脂质积累所需的基因进行功能分析，验证了 35 个靶向缺失菌株的结果。我们确定了一组高可信度的 150 个影响脂质积累的基因，包括在信号级联、基因表达、蛋白质修饰和囊泡运输中具有预测功能的基因，自噬、氨基酸合成和 tRNA 修饰，以及功能未知的基因。这些结果极大地促进了我们对这种含油物种中脂质代谢的理解，并展示了一种用于条形码诱变的通用方法，该方法应该能够在不同真菌中实现功能基因组学。