Fig. 4: The distribution and overlap of MSDIN genes across Agaricales spp.

a An UpSet plot depicting the overlap in MSDIN core sequences between species based on our analyses and previously published results (Tables S2 and S4). Set size reflects the total number of unique MSDIN core sequences reported for all genomes of a species; differences in this metric should be interpreted cautiously because species may differ in number of available genomes. Intersection size depicts the MSDINs present in the species marked with a solid black circle(s) underneath each column (e.g., Amanita bisporigera has 26 MSDINs not found in other species; the MSDIN IWGIGCNP (α-amanitin) is found in all species except Clavaria fumosa and Amanita polypyramis). MSDINs identified from published studies were not re-validated. b Phylogenetic relationships of MSDIN-producing species across Agaricales determined from the consensus of 289 maximum-likelihood trees of single-copy orthologs’ protein sequences (i.e., BUSCOs). Terminal branch lengths are not calculated. All nodes separating species have posterior probabilities above 0.93. The presence of the MSDIN associated processing enzyme, POPB, and counts of MSDIN sequences are depicted in the inner and outer rings (respectively). While we searched 249 genomes for MSDINs (Table S3), the tree only includes taxonomic families with species found to have at least one MSDIN and POPB. Genomes found to have both MSDIN and POPB genes are highlighted with a star, where the size of star also reflects the total MSDIN count found in each species. ^† These MSDIN sequences are associated with mature products with characterized bioactivity as toxins. From left to right: phalloidin (AWLATCP), phallacidin (AWLVDCP), β-amanitin (IWGIGCDP), and α-amanitin (IWGIGCNP). * The genome of Amanita brunnescens, a species without MSDINs, encodes a sequence found in a small and early-divergent POPB subclade (Fig. S2). More work is needed to clarify if this gene is functional.