Abstract
In 2004, 59 cases of food poisoning linked to the consumption of the mushroom Pleurocybella porrigens were reported in Japan, resulting in 17 fatalities due to acute encephalopathy. In 2023, we found three components of this mushroom-a lectin (PPL), pleurocybelline (PC), and pleurocybellaziridine (PA)-which are collectively implicated in its toxicity in mice. Although we reported the genomic data for P. porrigens in 2013, detailed annotation was not provided at that time. In addition, the biosynthetic pathway of PA, the compound responsible for acute encephalopathy, has not previously been reported in any Basidiomycetes species, including P. porrigens. In this study, we obtained the genome sequence of P. porrigens using the PacBio Revio System. De novo assembly with hifiasm resulted in a higher N50 value compared to previous assemblies, and gene annotation was conducted through BLAST search, GO analysis, KEGG pathway analysis, and carbohydrate-active enzyme identification. In addition, differential gene expression analysis revealed a significant up-regulation of 1427 genes in the fruiting bodies and 1436 genes in the mycelia. Although the gene encoding TqaL, a PA biosynthetic enzyme reported in the genus Penicillium, was not found in P. porrigens, a detailed examination of the BLASTx search identified genes with Fe/αKG oxygenase activity similar to TqaL. Three-dimensional structural modeling using AlphaFold3 revealed that the active site cavities of TqaL and the identified protein were in close proximity. Furthermore, the validity of the differential gene expression analysis between the fruiting bodies and the mycelia was confirmed by quantitative reverse-transcription PCR.