Fluorescent Random Amplified Microsatellites (F-RAMS) analysis of mushrooms as a forensic investigative tool

Kallifatidis, Beatrice; Borovička, Jan; Stránská, Jana; Drábek, Jir̆ı́; Mills, DeEtta K.

doi:10.1016/j.fsigen.2013.10.009

Cited by 9 publications

(6 citation statements)

References 24 publications

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“…Psychoactive mushrooms have been used as entheogens during transcendental ceremonies by Mexican inhabitants, mainly by shamans, for thousands of years. Mushrooms used for religious and spiritual rituals are called “magic mushrooms.” 1–3 There are many mushroom species that contain high levels of psychotropic tryptamines, including the genera Pluteus , Panaeolus , Inocybe , Gymnopilus , Galerina , Pholiotina , and especially Psilocybe 4–7 . In the 1950s, these magic mushrooms were introduced to Europe thanks to Gordon Wasson, who brought them from Mexico from the shaman Maria Sabina of the Mazatec tribe 8,9 .…”

Section: Introductionmentioning

confidence: 99%

“…Gordon Wasson passed these Mexican mushrooms on to Roger Heim, who identified them as Psilocybe mexicana , and then passed them on to Albert Hofmann, who in 1958 isolated psilocybin and psilocin from them 10,11 . Psilocybin and its hydrolyzed metabolite psilocin are well‐known psychoactive tryptamines of mushrooms, which are controlled substances 5,12,13 . Nevertheless, other fungal tryptamine analogs have not been explored in detail.…”

Section: Introductionmentioning

confidence: 99%

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Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis

Gotvaldová

Hájková

Borovička

et al. 2020

Drug Testing and Analysis

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Psilocybin, psilocin, baeocystin, norbaeocystin, and aeruginascin are tryptamines structurally similar to the neurotransmitter serotonin. Psilocybin and its pharmacologically active metabolite psilocin in particular are known for their psychoactive effects. These substances typically occur in most species of the genus Psilocybe (Fungi, Strophariaceae). Even the sclerotia of some of these fungi known as “magic truffles” are of growing interest in microdosing due to them improving cognitive function studies. In addition to microdosing studies, psilocybin has also been applied in clinical studies, but only its pure form has been administrated so far. Moreover, the determination of tryptamine alkaloids is used in forensic analysis. In this study, freshly cultivated fruit bodies of Psilocybe cubensis were used for monitoring stability (including storage and processing conditions of fruiting bodies). Furthermore, mycelium and the individual parts of the fruiting bodies (caps, stipes, and basidiospores) were also examined. The concentration of tryptamines in final extracts was analyzed using ultra‐high‐performance liquid chromatography coupled with mass spectrometry. No tryptamines were detected in the basidiospores, and only psilocin was present at 0.47 wt.% in the mycelium. The stipes contained approximately half the amount of tryptamine alkaloids (0.52 wt.%) than the caps (1.03 wt.%); however, these results were not statistically significant, as the concentration of tryptamines in individual fruiting bodies is highly variable. The storage conditions showed that the highest degradation of tryptamines was seen in fresh mushrooms stored at −80°C, and the lowest decay was seen in dried biomass stored in the dark at room temperature.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis

Gotvaldová

Hájková

Borovička

et al. 2020

Drug Testing and Analysis

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“…Kallifatidis et al ( 52 ) used a method based on the analysis of fluorescent random amplified microsatelites to distinguish between closely related species that cannot be differentiated using sequencing of the ITS region (eg, Psilocybe moravica , P. serbica, and P. bohemica ). Using this method, a high degree of intraspecific variation can also be observed.…”

Section: Discussionmentioning

confidence: 99%

Practical aspects of genetic identification of hallucinogenic and other poisonous mushrooms for clinical and forensic purposes

Kowalczyk

Sekuła²,

Mleczko³

et al. 2015

Croat Med J

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AimTo assess the usefulness of a DNA-based method for identifying mushroom species for application in forensic laboratory practice.MethodsTwo hundred twenty-one samples of clinical forensic material (dried mushrooms, food remains, stomach contents, feces, etc) were analyzed. ITS2 region of nuclear ribosomal DNA (nrDNA) was sequenced and the sequences were compared with reference sequences collected from the National Center for Biotechnology Information gene bank (GenBank). Sporological identification of mushrooms was also performed for 57 samples of clinical material.ResultsOf 221 samples, positive sequencing results were obtained for 152 (69%). The highest percentage of positive results was obtained for samples of dried mushrooms (96%) and food remains (91%). Comparison with GenBank sequences enabled identification of all samples at least at the genus level. Most samples (90%) were identified at the level of species or a group of closely related species. Sporological and molecular identification were consistent at the level of species or genus for 30% of analyzed samples.ConclusionMolecular analysis identified a larger number of species than sporological method. It proved to be suitable for analysis of evidential material (dried hallucinogenic mushrooms) in forensic genetic laboratories as well as to complement classical methods in the analysis of clinical material.

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“…Nuclear DNA was extracted from a small piece of dried or frozen fungal biomass (a piece of basidiome) using the Nucleo-Spin ® Plant II extraction kit (Macherey-Nagel) according to the manufacturer’s instructions. Three distinct molecular markers were used to study phylogeny of selected Psilocybe species: partial sequences of the divergent D1-D2 domain at the 5’ end of the nuclear LSU rRNA gene ( Maruyama et al 2003 , 2006 ), ITS rDNA region containing ITS1, 5.8S, and ITS2 sequences ( Pornpakakul et al 2009 , Kallifatidis et al 2014 ), and part of the elongation factor 1-α (EF1-α) ( Antonín et al 2009 , 2013 , Tomšovský et al 2010 ). Oligonucleotide primers and PCR conditions were used as published previously ( Borovička et al 2011 ).…”

Section: Methodsmentioning

confidence: 99%

Phylogenetic and chemical studies in the potential psychotropic species complex of <I>Psilocybe atrobrunnea</I> with taxonomic and nomenclatural notes

Borovička¹,

Oborník²,

Stríbrný³

et al. 2015

Pers - Int Mycol J

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Five Psilocybe species with unresolved systematic position (P. atrobrunnea, P. laetissima, P. medullosa, P. pelliculosa, and P. silvatica) were investigated using four molecular markers (EF1-α, ITS, LSU, and IGS). Phylogenetic analysis revealed that with the exception of P. laetissima, which is now rightfully classified in the genus Leratiomyces, all investigated species belong to Psilocybe sect. Psilocybe. For the first time, psychotropic compounds psilocin and psilocybin were detected in P. medullosa using gas chromatography-mass spectrometry. On the contrary, neither psilocin, nor psilocybin was detected in P. atrobrunnea and negative results were also obtained from mycelia grown in vitro on tryptamine/tryptophan-amended media. These results strongly suggest that biosynthesis of these alkaloids was lost in P. atrobrunnea. With the exception of minor differences detected in EF1-α marker, all sequences of American and European collections of P. atrobrunnea were identical. On the other hand, a thorough nomenclatural study revealed that the name P. atrobrunnea must be considered dubious; the oldest available candidate name, P. fuscofulva, was therefore adopted. The molecular data suggests that morphologically identical American P. silvatica and European P. medullosa likely represent distinct species; epitypes of both taxa were therefore designated.

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Fluorescent Random Amplified Microsatellites (F-RAMS) analysis of mushrooms as a forensic investigative tool

Cited by 9 publications

References 24 publications

Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis

Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis

Practical aspects of genetic identification of hallucinogenic and other poisonous mushrooms for clinical and forensic purposes

Phylogenetic and chemical studies in the potential psychotropic species complex of <I>Psilocybe atrobrunnea</I> with taxonomic and nomenclatural notes

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