Scientists (Michaela Schmull, et al) followed in the footsteps of ethnobotantists Wade Davis and Jim Yost who, during the late 20th century, traveled through South America to document a myriad of indigenous esoteric plant use. One of the more unique organisms that was utilized was a vibrantly colored lichen, Dictyonema huaorani, documented to have tryptamine-like entehogenic qualities. The scientist were granted access to the Harvard stored Davis & Yost collection that they were able to sample; providing further evidence of possible presence of multiple 5-methoxy-tryptamines.
Article Hypothesis and Author Note
Gaining access to the only collected sample of the lichen Dictyonema huaorani, stored at the Harvard Fungarium (Fig 1, FH holotype), these scientists sought to investigate the presence of entheogenic tryptamines as well as piece together some phylogenetic insights into what branch the symbiotic organisms fruits from on the tree of life. Posited by Davis and Yost, who sampled it and inquired from the Waorani their historical use of the organism, noted for having powerful and arguably negative use connotations of 'bad spirits and curses' within the Waorani culture. The Scientists hypothesize that this species can be found in the forest canopy of the dense Amazon forest only, similar to other species of Dictyonema with the "same morphotype that were collected in a dense humid secondary rainforest in Canto ́n San Lorenzo, Ecuador." They believe this might explain the reason for the limited number of collections of this species.
Further continuing the look into entheogen production, cultural use, symbiotic relationships and how they exist in many different layers and forms, I could not resist the chance to talk about 'psychedelic lichen'. I came accross this discussion piece amongst numerous different circles, internet forums, discussion groups; but only recently stumbled accross the information readily avaiable! While considered to be used in invoking 'evil spirits', the Dictyonema sample that Davis and Yost were lead to has not only been tested before for its entheogenic properties, but multiple times. A good lesson to take from these spiritual practices are that not all entheogens are utilized for health, healing, nor the good of the practitioner. Learning important lessons from indigenous pracititioners about what chemical constituents are harmful to our biology is key in helping us understand what historical plant and fungal preparations are going to be key in the coming entheogen reneissance. Listen, and learn well.
A Rare Pair: Basidiomycetes & Cyanobacteria
Lichen are unique organisms, in that 'it' is actually a 'them' as lichen are a symbiosis between fungi (mycobiont) and another organism capable of photosynthesis, typically algae or cyanobacteria (photobiont). The photobiont is typically layered in upper, light facing, cell layers where hyphae 'nets' cradle them and absorb photo-metabolites; with lower, structural, mycelial mats interwoven in the central cortex anchored by more hyphae called rhizines (Fig 2). Of all the lichen out there, less than 1% exist where the mycobiont is a basidomycete instead of an ascomycete fungus, much less a cyanobacteria instead of algae. [Lawrey 2007] [Bungartz 2009] The former of which is what makes the genus Dictyonema more closely related to mushrooms than it is to most other lichens. Additionally, there is the point that when cyanobacteria are the photobionts, they also aid in nitrogen fixation as well as photosynthesis (Fig 3)! In three different instances in which this relationship has arisen, the mycobiont have lost a crucial gene for energy production and are dependent upon the photobiont for survival. [Pogoda 2018] This being the case, it is interesting to note that the photobiont typically lives, even flourishes when outside the mycobiont, who cannot survive without its photosynthetic partner.
'Nɇnɇndapɇ' and the 'ido's' curse
While entheogen use is widely known in many different cultures world-wide; it is most notably so by the peoples and cultures of South America. Famous ethnobotanists Wade Davis and James A. Yost, at the behest of the Harvard Botanical Museum, traveled across the Amazon river basin and Andes mountain range documenting and sampling various flora and funga. Among the many peoples visited were the Waorani found in the north-eastern forests of Ecuador; specifically the Napo, Orellana, and Pastaza provinces (Fig 4). Davis and Yost claimed the tribe contained around 600 people before the turn of the century.
The Scientists ascertained that the Waorani were thought to be hunters and gatherers pre-contact with some communities who have adopted a more western lifestyle engaging in tourism; while still other communities continue to live in isolation, noted as having impact on wildlife populations mainly through hunting. The Waorani lands originally occupied over 20,000 square-kilometers before first-contact by outsiders in 1958. [Rival 2002] Since then, their remaining lands, assigned to them in the 1990s, cover about 6,800 square-kilometers adjacent to Yasunı ́National Park. [Rival 2002] These lands have recently been threatened particularly by oil exploration (Doughty et al. 2010; Kane 1993, 1994;Keefe 2012); but thanks to both local and international support have won landmark legal battle protecting against the sale of their lands.
Their language, 'huao tetedo', the Scientists note, has historically been documented as being unrelated to any other known language. Also unique to the Waorani is the fact that entheogen use is considered 'to be an aggressive anti-social act; so [any] shaman, or 'ido', who desires to project a curse takes the drug alone...at night in the secrecy of the forest or in an isolated house." Under the influence of hallucinogens the 'ido' can call on the 'wenae' (malevolent spirits) to 'wreck evil'; for the in Waorani culture there are no benevolent spirits, and only the ido who invoked the spirit can ask it to undo its curse. [Davis 1983] Among those entheogens thee Waorani exposed Davis and Yost to 'nɇnɇndapɇ', the sample of D. huaorani they collected and tested three decades later for this article. Not only identified by 'ido' as the one used in the reported 'cursing' and death of another shaman, the 'ido' also reported that it can cause sterility; however, it is unsure if the cause is due to compounds in the lichen or if used as an after-the-fact explanation of barrenness. [Davis 1983]
Decoding Barcoding - Fungal Targets
While not the main topic of discussion, genetics, barcoding, and organism ID is crucial to better understanding the organism and the role it plays within its ecosystem. Genomes for organisms can be widely variable, and we have previously looked into some genetics of other entheogenic fungi for reference. Typically, there are specific genes that we use to classify different species from each other based on differences in the coding sequence of those target genes. For fungi they are centered around the genes that build ribosomes (ITS1, ITS2, & LSU) as well as a the genes that build RNA polymerase (RPB1 & RPB2).
Discussing ribosomal genes in fungi, the internal transcribed spacer (ITS) is about 600 base pairs long in the ribosomal tandem repeat gene cluster of the nuclear genome (Fig 5). The region is flanked by the DNA sequences for the ribosomal 18S subunit or small subunit (SSU) at the 5‘ end, and by the 28S subunit or large subunit (LSU) at the 3‘ end. [Xu 2016] [Wurzbacher 2019] The ITS itself consists of two separate parts, ITS1 and ITS2, which are separated from each other by the 5.8S subunit nestled in-between them. Like the flanking 18S and 28S subunits, the 5.8S subunit contains a highly conserved DNA sequence, as they code for structural parts of the ribosome, which is a key component in intracellular protein synthesis. It is for this highly-conserved region interspersed with a highly variable region that makes the ribosome gene a great target for barcoding fungi. [Schoch 2012]
Less used, but still highly valuable is the nuclear gene coding for RNA polymerase. The study specifically chose the second largest sub-unit due to it having available sequences available in literature to reference. "Other studies also used RPB2, the second-largest subunit of the RNA polymerase II, e.g. for a phylogenetic study shedding light on the relationships among early-diverging lineages in the fungal kingdom" [James 2006]
Conclusion and Caveats
The Scientists performed in-depth genetic analysis as well as rigorous chemical analysis of the three-decade old Davis and Yost sample. The genetic results, based on an assembly of three key barcoding genes, pointed towards D. huaorani as separating out from all other Dictyonema species almost every time (Fig 6). This pilot investigation into genetics is a great start, but full genome sequencing of both the mycobiont and the photobiont will be necessary to 1) determine the location of entheogen production (likely the mycobiont), and 2) allow for better phylogenetic assemblies of the entire plant/lichen/fungal family tree. Of the three chemical analysis performed, the final analysis the Scientists did was on an LC/MS and showed the likely presence of 5-methoxy-N-methyl-tryptamine (5-MeO-NMT), 5-methoxy-dimethyl-tryptamine (5-MeO-DMT), and 5-methoxy tryptamine(5-MeOT) based on database values. Analyses of the >30-year-old specimen was able to show that entheogenic compounds may be produced by D. huaorani. Even in light of this fact, the lack of a fresh sample, better understanding of ecosystem, and even lack of analytical standards depict the vast depth of unknown information that this single organism signals towards as a one representative of the diverse Amazonian apothecary.
Support or Reject Hypothesis?
The Scientists were able to show the possible presence of 5-Methoxy-tryptamines via LC-MS analysis of a dried sample.
The scientists were unable to find any other collections or samples in any other collections.
Reviewing the data and the conclusions put forth, the hypothesis proposed by the Scientists was supported. The reason(s) for acceptance are quoted below (emphasis and parentheticals mine):
"Altogether, the [analytical chemistry] results are suggestive of the presence of tryptamine and psilocybin in Dictyonema huaorani, but comparison of larger amounts of fresh material of this species with authentic standards are necessary to confirm these tentative findings."
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