A draft reference assembly of the Psilocybe cubensis genome.

We describe the use of high-fidelity single molecule sequencing to assemble the genome of the psychoactive Psilocybe cubensis mushroom. The genome is 46.6Mb, 46% GC, and in 32 contigs with an N50 of 3.3Mb. The BUSCO completeness scores are 97.6% with 1.2% duplicates. The Psilocybin synthesis cluster exists in a single 3.2Mb contig. The dataset is available from NCBI BioProject with accessions PRJNA687911 and PRJNA700437.

Ptr ToxA requires multiple motifs for complete activity.

Ptr ToxA was the first proteinaceous necrosis-inducing toxin identified and cloned from the wheat pathogen, Pyrenophora tritici-repentis. How this protein causes necrosis in sensitive wheat cultivars is not known. In an effort to understand the structural features of Ptr ToxA required for induction of necrosis, we employed a combination of site-directed mutagenesis and peptide inhibition studies. Mutagenesis was carried out on conserved motifs within the active domain of Ptr ToxA. Proteins with mutations of potential casein kinase 2 phosphorylation sites but not protein kinase C phosphorylation sites have significantly reduced activity. Additionally, mutations in a region with high homology to amino acids surrounding and including the RGD cell attachment motif of vitronectin result in proteins with significantly less activity than Ptr ToxA. The importance of the vitronectin-like motif was confirmed by a decrease of Ptr ToxA-induced activity when coinfiltrated with peptides corresponding to amino acids within this motif. Reduction in Ptr ToxA activity by competition with mutant proteins demonstrates the necessity of multiple motifs for Ptr ToxA activity.