Initially, I wasn’t even thinking about mushroom breeding. While working with the PF Albino and growing a few pure white mushrooms, I decided to check out the gills under the microscope. I wanted to see if these mushrooms were spore-less, like everyone assumed. Surprisingly, spores were observed, and like the rest of the fruit body, the spores were colorless. With some effort, a light partial spore print, that was barely visible on foil, was produced from a very mature cap. The spores appeared white in deposit but there weren’t many and they were difficult to see. To check for spore viability, I placed some of the white spores on malt agar in petri dishes.

The spores germinated in only a few days. I noticed that there were very few colonies growing and they were spaced some distance apart. I assume this was due to the low number of spores and maybe low viability. The isolated germinations made me think that each colony might be from a single spore (monokaryotic). If this was true, I might be able to use these monokaryotic mycelia for breeding experiments. Maybe, even make an Albino Penis Envy! To prevent fusing with other mycelia from nearby spores and generating a dikaryotic mycelium, I quickly transferred the tiniest samples of tissue to new agar plates.

At this time, I should go over the very basics of mushroom sex for those that aren’t familiar with mushroom genetics. At its simplest, two compatible spores are needed to produce a mushroom producing dikaryotic mycelium. A mycelium generated from a single spore (monokaryotic) won’t produce mushrooms (usually) but is very useful for breeding new mushroom varieties. Crossing different monokaryotic mycelia is by simple mixing or allowing them grow together across a medium such as an agar plate.

To confirm that the mycelium was monokaryotic, a small sample was placed under the microscope to look for clamp connections. A clamp connection looks like a little bump or bulge between hyphal cells and indicates nuclear movement within a dikaryotic mycelium. A mycelium with no clamp connections should be monokaryotic, which is what I wanted to see. I want to keep this simple, so that is all I’m going to cover on clamp connections. More information and pictures can be easily found online.

At this same time, I was reading my newly acquired copy of “Genetics and Breeding of Edible Mushrooms” by A.C. Chang. I noted this passage on nuclear migration on page 47.

In many Basidiomycetes a phenomenon occurs following hyphal fusion which permits the establishment of a heterokaryotic condition in growing cells. This phenomenon is nuclear migration by which is meant the migration of nuclei through the established mycelium of a confronting strain. Nuclear migration was first described by Buller from observations of the formation of clamp connections, indicative of dikaryotic hyphae, at a distance from the site of fusion of compatible strains greater than could be explained by hyphal growth. Nuclear migration rates of different species may vary widely from 0.5mm/h in Coprinus lagopus (Buller, 1931) to 40mm/h in Coprinus congregatus (Ross, 1976). The advantage to the organism of nuclear migration is that the heterokaryotic condition is not limited to those heterokaryotic cells that have been formed by plasmogamy, but can be established rapidly in large numbers of cells and be perpetuated in the growing cells as is the case in the clamp forming dikaryotic hyphae of Basidiomycetes

I am pretty lazy and I didn't want to spend time isolating more single spores from other cubes and then do several controlled crosses and hope for the best. I was now aware of a mechanism where a monokaryotic mycelium can become a dikaryotic mycelium by nuclear migration, and not just at the new growth at the point of mycelial contact. Essentially the entire monokaryotic culture becomes dikaryotic by replicating and moving nuclei in a sort of a chain reaction through the already existing mycelial network.

Sooooooo.......... here is what I did.

I put the microscopically confirmed monokaryoitic PF Albino culture into a jar of sterile grain (grass seed in this case) and let it colonize almost completely. Every grain was covered with mycelium, but it would need a few more days to get really dense. Then I made a fresh solution (adding just dry spores also worked) of normally pigmented Penis Envy (PE) cubensis spores and injected that into the nearly colonized grain. I shook the jar to mix. The idea is to deny any uncolonized substrate to the injected PE spores for colonization if they combined (mated) with other PE spores. In theory, the PE spores will germinate and some will combine with other nearby germinated PE spores, but some will combine with the PF Albino mycelium. Nuclear migration should then transform the entire jar of monokaryotic PF Albino mycelium into several strains of dikaryotic PF Albino X Penis Envy mycelium. There shouldn't be any substrate left for any significant amount of pure Penis Envy dikaryotic mycelium to grow and later form mushrooms.

The colonized grain needs to be fruited directly and not spawned to a bulk substrate. Surprisingly, this method worked on the first attempt and produced some exceptionally normal looking Psilocybe cubensis mushrooms, with no hints of mutations or abnormalities. Since both varieties are supposedly descendants of early collections from the Amazon, this may be a glimpse into what the original mushrooms looked like when first collected.

PF Albino x Penis Envy (F1) mushrooms grown from cased grass seed. (above)

Now that we have varietal hybrid mushrooms, the hard part is over. I can just take spore prints of this generation and use them for new multi spore cultures. The first generation is genetically 50% of each parent and all the mushrooms look the same. But the later generations are where the action is, and we should expect to see all sorts of crazy combinations of traits. And that is what I observed.

A few examples of the F2 generation showing various forms. (above)

After selecting spores from promising specimens from the F2 generation, I finally got an early version of what I was looking for. An albino mushroom with a penis shaped cap in the F3 generation and another interesting form that I called the PE Uncut.

The very first Albino Penis Envy (APE) 2006

After a few more generations for stabilization and we have the APE we have today. As with the original Penis Envy, the reduced caps don’t produce or release many spores. But spores are present and can be swabbed from the gills in adequate amounts for propagation. I also developed an in-house method of separating and concentrating spores with a centrifuge for syringes.

I created the APE purely with aesthetics in mind and didn’t intend to produce a super potent variety of Psilocybe cubensis. Anecdotal reports always rated the APE highly potent but real results finally came to light in 2022 with the advent of laboratory testing in the Psilocybin Cup by Hyphae Labs, where APE ranked at or near the top in potency.

https://www.oaklandhyphae510.com/post/oakland-hyphae-hyphae-labs-present-the-spring-psilocybin-cup-2022-final-results

And again in 2023 with Colorado’s first Psychedelic Cup.

https://www.5280.com/what-we-learned-at-colorados-first-psychedelic-cup/

John Workman

The Penis Envy: A Classic Psilocybe cubensis Variety Selected for High Potency.

The Penis Envy variety of Psilocybe cubensis is extremely popular due to its high potency and distinct phallic appearance. Originally only available from the Homestead Book Company and later the more obscure MycoTech, both of which no longer exist.

The origin story of the Penis Envy (PE) variety of cubensis was clouded in myth and mystery until Hamilton Morris’s 2021 podcast where he interviews Richard Guitierrez (Rich Gee), the developer of the PE. The podcast is now available on Youtube (https://www.youtube.com/watch?v=QRArM5Ev4EE) Originally called “Amazons” The name “Penis Envy” was coined by Gee around 1987 based on its appearance and comments by some exotic dancers. The first convincing documented photograph of the PE is on the cover of a 1995 self-published and extremely rare book titled “Cubensis Aquarium Gardening” by Rich Gee. Although the Penis Envy name isn’t used in the book, the distinctive shaped caps are unmistakable.

Cover of “Cubensis Aquarium Gardening” showing an early version of Penis Envy

As the story goes, around 1975, Gee extracted spores from dried Psilocybe cubensis Amazonian mushrooms, purchased in California. The mushrooms were claimed to be grown from the direct descendants of Terrence McKenna’s Amazonian spore collection. Gee was specifically selecting for large mushrooms with a strong bluing reaction when damaged. Blue bruising is a reasonably good indicator of potency. By 1978 or so, the original Amazon spores were incrementally improved in potency and size and acquired the reduced penis cap. Gee did not directly select for this penis shaped cap. The phallic shape appeared as a result of a random genetic mutation that was associated with high potency. Around 2000, the PE variety was becoming very well known on magic mushroom forums like the Shroomery. Since spore production was low, it was in high demand with limited supply. Sporeworks first started selling PE in 2002 after working out a method of spore extraction and concentration.

Cultivation of Psilocybe cubensis Penis Envy variety (2002)

Early attempt at microscopy of Penis Envy spores.

So now we have two mutant varieties of P. cubensis, both distinct in appearance and of high potency. Since they both looked very different from each other, it should be visually obvious if a hybridization is successful. I also thought an Albino Penis Envy (APE) would look cool. I had never done any mushroom breeding before, but I had some ideas on how to go about it. The final part of this 3-part series will cover the hybridization and selection method for the APE.

John Workman

Stay tuned for Part Three!

Part One

Over the years, since it was first released in 2006, many websites have generated their own narratives on the origin of the APE. Either through ignorance or intentional misinformation for their own purposes, the true history has been obscured by time. To set the record straight, I will document in excruciating detail the real story here. Much of this work was originally documented on the Shroomery.org forums, where much of it still remains.

The short version (TLDR) is that APE is an in-house varietal hybrid of the PF Albino and Penis Envy strains of Psilocybe cubensis generated by Sporeworks.com.

First a little ground background on the parentage of the APE.

The PF Albino: An Ultraviolet Induced Mutation by a Pioneer in Home Psilocybe cubensis Cultivation

The Psylocybe Fanaticus (PF) TEK magic mushroom revolution began in September 1991, with its introduction in High Times magazine by Robert 'Billy' Mcpherson aka Psylocybe Fanaticus. The PF TEK was an early simplified method of growing Psilocybe cubensis mushrooms using a hypodermic syringe containing mushroom spores and water (novel at the time) and small ½ pint cakes of steamed brown rice flour and vermiculite. Not requiring a pressure cooker or messy composts like earlier methods, the PF TEK opened magic mushroom cultivation to more casual hobbyists. The PF TEK became very popular as the early internet accelerated the spread of information. Unfortunately, Psylocybe Fanaticus was raided and shut down in February 2003. Further details of the raid is on this archive of the original website (https://www.seanet.com/~rwmcpherson/pfcase.htm). Sadly, Robert 'Billy' Mcpherson died in 2011 at the age of 65.

The PF Albino was a chance mutation of the standard PF Classic strain (aka Matias Romero and Amazonian) of Psilocybe cubensis sold by Psylocybe Fanaticus. The PF Classic was selected specifically to do well using the PF TEK method. Around the year 2000, for unstated reasons, PF started using fluorescent UV lights during his PF Classic production runs. The UV exposure unintentionally induced albinism and eventually caused enough genetic damage that fruiting was impossible. Prints of UV exposed pigmented mushrooms, that were growing on the same PF cakes as the albinos, were distributed on a limited basis. It was hoped that the albinism trait was present in these normal looking spores and would be revealed in later multispore cultures. This was indeed true, and PF albino mushrooms began to circulate in the magic mushroom community. I was gifted a UV exposed PF print and started working with it in 2005. PF assumed that the PF Albino was sporeless, but I was able to find colorless spores on the gills. Albino spores were essential to what was to come.

Microscopy of the gill of PF Albino showing 4-spored basidia with unpigmented spores

PF Classic grown from UV exposed spores 

Clone of Albino form

Stay tuned for Part Two...

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