DNA Authentication and Chemical Analysis of
            <i>Psilocybe</i>
            Mushrooms Reveal Widespread Misdeterminations in Fungaria and Inconsistencies in Metabolites

Bradshaw, Alexander J; Backman, Talia; Ramírez-Cruz, Virginia; Forrister, Dale L.; Winter, Jaclyn M.; Guzmán-Dávalos, Laura; Furci, Giuliana; Stamets, Paul; Dentinger, Bryn T. M.

doi:10.1128/aem.01498-22

Cited by 11 publications

(16 citation statements)

References 80 publications

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“…In comparison, our data set provided sequence representation for 27 taxa not previously reported. In particular, the inclusion of public data was vital for our work as it allowed us to pinpoint specimens that are likely misidentified at the species level, a common issue in museum fungaria and Psilocybe in particular (Figure 2, Table 1) (Andrew et al 2019; Bradshaw et al 2022).…”

Section: Resultsmentioning

confidence: 99%

“…These results suggest an Asian rather than African origin for the progenitor of P. cubensis . However, sequences labeled as P. natalensis (KwaZulu-Natal, South Africa) deposited in NCBI do show close relationships with P. cubensis (Bradshaw et al 2022), although these sequences do not have voucher information included, rendering them dubious. Nonetheless, these results align with African Psilocybe assigned morphologically to the Cubensae complex by Froese et al (2016).…”

Section: Discussionmentioning

confidence: 99%

“…While taxonomy of Psilocybe has been the subject of a handful of studies spanning the past 60+ years (Singer and Smith 1958;Hofmann et al 1959;Guzmán 1983Guzmán , 1995Guzmán , 2005Borovička et al 2011;Ramírez-Cruz et al 2013b;Cortés-Pérez et al 2020), the ambiguous legal status of Psilocybe specimens and their rampant misidentification has hindered modern taxonomic research on them (Bradshaw et al 2022). The only multi-locus molecular phylogenetic studies included 24 (accounting for found synonyms) species and up to three gene regions, with many branches without strong statistical support and lacking most of the currently recognized species (Borovička et al 2011(Borovička et al , 2015Ramírez-Cruz 2013a).…”

Section: Introductionmentioning

confidence: 99%

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Phylogenomics of the psychoactive mushroom genusPsilocybeand evolution of the psilocybin biosynthetic gene cluster

Bradshaw

Ramírez-Cruz

Awan

et al. 2022

Preprint

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Psychoactive mushrooms in the genus Psilocybe have immense cultural value and have been used for centuries in Mesoamerica. Despite a recent surge in interest in these mushrooms due to emerging evidence that psilocybin, the main psychoactive compound, is a promising therapeutic for a variety of mental illnesses, their phylogeny and taxonomy remain substantially incomplete. Moreover, the recent elucidation of the psilocybin biosynthetic gene cluster is known for only five species of Psilocybe, four of which belong to only one of two major clades. We set out to improve the phylogeny for Psilocybe using shotgun sequencing of 71 fungarium specimens, including 23 types, and conducting phylogenomic analysis using 2,983 single-copy gene families to generate a fully supported phylogeny. Molecular clock analysis suggests the stem lineage arose ~66 mya and diversified ~53 mya. We also show that psilocybin biosynthesis first arose in Psilocybe, with 4-5 possible horizontal transfers to other mushrooms between 40 and 22 mya. Moreover, predicted orthologs of the psilocybin biosynthetic genes revealed two distinct gene orders within the cluster that corresponds to a deep split within the genus, possibly consistent with the independent acquisition of the cluster. This novel insight may predict differences in chemistry between the two major clades of the genus, providing further resources for the development of novel therapeutics.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Phylogenomics of the psychoactive mushroom genusPsilocybeand evolution of the psilocybin biosynthetic gene cluster

Bradshaw

Ramírez-Cruz

Awan

et al. 2022

Preprint

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“…60 The ITS tree was constructed under the TPM2u+F+G4 substitution model and the rpb1 tree under the TNe+G4 substitution model, determined by the included model test in IQ-TREE (Ver 1.6.11). 61,62 Support for topologies was assessed by 1000 ultrafast bootstrap replicates.…”

Section: Methodsmentioning

confidence: 99%

Cultivation, chemistry, and genome ofPsilocybe zapotecorum

Miller,

Jacobs,

Rockefeller

et al. 2023

Preprint

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Psilocybe zapotecorumis a strongly blue-bruising psilocybin mushroom used by indigenous groups in southeastern Mexico and beyond. While this species has a rich history of ceremonial use, research into its chemistry and genetics have been limited. Herein, we detail mushroom morphology and report on cultivation parameters, chemical profile, and the full genome sequence ofP. zapotecorum. First, growth and cloning methods are detailed that are simple, and reproducible. In combination with high resolution microscopic analysis, the strain was barcoded, confirming species-level identification. Full genome sequencing reveals the architecture of the psilocybin gene cluster inP. zapotecorum,and can serve as a reference genome for Psilocybe Clade I. Characterization of the tryptamine profile revealed a psilocybin concentration of 17.9±1.7 mg/g, with a range of 10.6-25.7 mg/g (n=7), and similar tryptamines (psilocin, baeocystin, norbaeocystin, norpsilocin, aeruginascin, 4-HO-tryptamine, and tryptamine) in lesser concentrations for a combined tryptamine concentration of 22.5±3.2 mg/g. These results showP. zapotecorumto be a potent – and variable –Psilocybemushroom. Chemical profiling, genetic analysis, and cultivation assist in demystifying these mushrooms. As clinical studies with psilocybin gain traction, understanding the diversity of psilocybin mushrooms will assure that psilocybin therapy does not become synonymous with psilocybin mushrooms.

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“…Much of the research on the therapeutic effects of psychedelics has focused on psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine), which is found in various genera of mushrooms, most notably the genus Psilocybe (Bradshaw et al, 2022; Dodd et al, 2022; Gotvaldova et al, 2022). In the past decade, a number of early phase trials have suggested that psilocybin-assisted psychotherapy may have rapid-acting and long-lasting therapeutic effects in depression, anxiety, substance use, smoking cessation, migraine, and other disorders (Carhart-Harris et al, 2016; Griffiths et al, 2016; Ross et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Sub-acute effects of psilocybin on EEG correlates of neural plasticity in major depression: Relationship to symptoms

et al. 2023

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Background: Evidence suggests that serotonergic psychedelics (e.g. psilocybin), have rapid-acting and long-lasting antidepressant effects after a single dose. However, the mechanism underlying these effects remain unclear. One proposed mechanism is that these drugs promote neuroplasticity. However, this has not been conclusively demonstrated in humans. Aims: We hypothesized that relative to placebo, psilocybin would: (1) increase electroencephalographic (EEG) correlates of neuroplasticity, (2) reduce depression symptoms, and (3) changes in EEG would correlate with improvements in depression. Methods: In this double-blind, placebo-controlled, within-subject study, individuals with major depressive disorder (MDD; n = 19) were administered placebo followed by psilocybin (0.3 mg/kg) in a fixed order (placebo, followed by psilocybin 4 weeks later). EEG indices of neuroplasticity (tetanus-induced long-term potentiation) as assessed via auditory evoked theta (4–8 Hz) power and measures of depression (GRID Hamilton Rating Scale for Depression-17 (GRID-HAM-D-17)) were measured at several time-points after placebo and psilocybin (24 h and 2 weeks after each session). Results: EEG theta power doubled in amplitude 2 weeks after a single psychedelic dose of psilocybin but not after placebo. Further, improvements in depression symptoms 2 weeks after psilocybin were correlated with increases in theta power. Conclusions: The increased theta power observed represents evidence of sustained changes in the brain following psilocybin. Given the correlation with enhancement in depressive symptoms, changes in theta may represent an EEG biomarker of the sustained effects of psilocybin, and may shed light on potential mechanisms of psilocybin’s antidepressant effect. Taken together, these results complement the emerging notion that psilocybin, and perhaps other psychedelics, can produce long-term alterations in neuroplasticity.

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DNA Authentication and Chemical Analysis of Psilocybe Mushrooms Reveal Widespread Misdeterminations in Fungaria and Inconsistencies in Metabolites

Cited by 11 publications

References 80 publications

Phylogenomics of the psychoactive mushroom genusPsilocybeand evolution of the psilocybin biosynthetic gene cluster

Phylogenomics of the psychoactive mushroom genusPsilocybeand evolution of the psilocybin biosynthetic gene cluster

Cultivation, chemistry, and genome ofPsilocybe zapotecorum

Sub-acute effects of psilocybin on EEG correlates of neural plasticity in major depression: Relationship to symptoms

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