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Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers
    #29669576 -

This thread is to discuss the possibility of getting non-active-producing mycelium to start producing active compounds (psilocin and psilocybin)

My intuition tells me that this would take 100s if not thousands of agar transfers to cross an active species (Psilocybe Cubensis) with a non active species (Pleurotus Ostreatus)

I am aware that anything is possible, yet I have not heard of two mushrooms of different genuses being able to cross. No matter, when there is a will there is a way.


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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: langten69]
    #29669614 -

Scientists have been able to produce psilocybin from yeast and, to a lesser extent, from bacteria.

I recall reading how different types of mushrooms evolved psychedelics twice, each using their own unrelated different ways to produce psilocybin.

You may be on to something. Fascinating project.


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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: veggie]
    #29669688 -

That is truly fascinating, do you have a link to the article which says some mushrooms evolved psychadelics twice?

Ive been doing research and apparently there is a third option as well...




Convergent evolution:
Different fungi independently evolved machinery capable of producing psilocybin

Horizontal gene transfer:
One lineage acquired an already-existing psilocybin biosynthetic cluster from another

Gene loss/divergence:
Some lineages subsequently lost or modified parts of the machinery


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Edited by langten69 (09/15/26 07:39 PM)

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: langten69]
    #29669701 -

Link to the article posted in News.

https://www.shroomery.org/forums/showflat.php/Number/29354490#29354490


Massospora cicadina likely the third, See related article, also in News.

https://www.shroomery.org/forums/showflat.php/Number/27308678#27308678

Edited by veggie (09/15/26 07:57 PM)

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: veggie]
    #29669737 -

Amazing, it seems as though fungus infecting insects as a propogation vector is a theme (like cordyceps)

My intuition tells me that the process of one inactive fungus gaining active compounds will be a result of the inactive fungus consuming/copulating with the active fungus (over many iterations) until it takes on the active traits in itself whilst keeping the morphology of itself or a hybrid of the 2. Or vice versa


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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: langten69]
    #29669871 -

Insights from Alan Rockefeller about how Inocybe acquired the genes:

"It's probably horizontal gene transfer.... these genera are not closely related. The psilocybin gene cluster is the same gene cluster. It didn't evolve independently in some way. This gene cluster got into all these other mushrooms."

Here's a longer article that argues the same thing Rockefeller did in that video.

Now, the other article veggie posted suggests fiber caps make the compounds in a different way. That would be evidence of convergent evolution instead. But if it's the exact same gene cluster, it's an evidence of horizontal gene transfer. So it may well be both.

Perhaps experimentally, you could get one mushroom to transfer the gene to another. But how would you make sure it transfers this one gene, how long would it take? This sounds like something of a 20-40 year lifelong research project to figure out how to do that.

Edited by CreonAntigone (09/16/26 12:48 AM)

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: CreonAntigone]
    #29669894 -

In the video, the filmer states that it was potentially the symbiotic bacteria that introduced the Psilocybin producing genes.

I suppose the ways to test it would be to either:

1) do a classic potency test where you eat the potential mushroom you suspect may have inherited the alkaloid producing gene

2) wait until the dna reading technology of the world becomes competent enough to be aware of the specific gene cluster you are testing for in a sample

Option 1 seems the most viable for now. For example, you want to make psychadelic oyster mushrooms. At each iteration of a successful "cross", you would then fruit the mushroom normally and collect spores, then eat a portion of the raw/dehydrated oyster cross to test for potency


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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: langten69]
    #29669905 -

If you watch the whole Rockefeller video he talks about how you can use a PCR test to look for specific genes. The technology to do all that stuff exists right now, you'd just need a particularly good lab.

And rather than to eat and test which is subjective and prone to bias, there are also psilocybin tests to determine the percent alkaloids (also discussed in the video).

There are direct genomic methods that would be easiest (eg many gene splicing methods). Other methods include transfers mediated by a specialized bacteria, Agrobacterium tumefaciens. Also, "bacterium to Saccharomyces cerevisiae conjugation followed by DNA exchange via bacterial conjugative plasmids" (I'm not exactly sure what this means).

But more in line with the thread, I wonder if this method would eventually work: Extensively growing let us say a co-culture of two mushrooms, like growing cubes and buttons in the same substrate over multiple generations. You'd want to keep doing this until you find a button mushroom that shows a hit for any percent of psilocybin on the psilocybin test showing that some genes would be transferred. Then you take button, that has shown promise to have taken the gene, and isolate and stabilize the culture. And then potentially, to do it again and grow it in culture with cubes again until it potentially takes more of the gene, and then isolate and stabilize that culture.

You'd want to have a genetic hodgepodge. Probably spores. Start button mushroom spores on the same dish as psilocybin spores. No genes would be transferred right away. In nature this process probably takes something like hundreds to thousands of years. You'd want to repeatedly iterate. Even then I think it would be complete chance on if it'd happen. It's just that the method would be trying to increase the chances by having the genes grow repeatedly together, thus eventually it may happen.

The studies suggest HGT happens more often if there's some kind of pressure. It's possible perhaps that growing next to psilocybin mushrooms very closely may pressure the other target species to at least learn to tolerate it, possibly increasing the chance of HGT. Here's an example of HGT induced by co-culturing, Aspergillus was able to acquire resistance to anti-fungals when grown next to a resistant strain in this study, under the pressure of applying the antifungal. They use something called plasmids though to achieve this, and someone else will have to explain exactly how that works, as I don't quite get it.

Edited by CreonAntigone (09/16/26 02:13 AM)

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: CreonAntigone]
    #29669922 -

My research tells me that a plasmid is a vehicle for delivering specific gene programs, which makes sense as to how the scientists used it to get the Aspergillus to resist antifungals.

So lets say that we create a genetic hodgpodge of our two candidate cultures, whilst also introducing the pressure necessary to get them to 'tolerate' each other. Essentially we need to get them to play nice with each other, until they feel comfortable to share gentic information between them.

As far as the plasmids go, i need more research on them. I do recall hearing cultivators using snake venom as a plasmid to break down the barriers of two different cultures which resulted in a cross between the two...


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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: langten69]
    #29669943 -

Well it's an interesting question if cooperation could have any role. Could they share genes from cooperation, or might one try to essentially steal the gene to win a competition?

The first goal would to be to find a culture that successfully grows both button (or any other gourmet) and cubes in the same tub without either killing the other. That would be hard, definitely not impossible. People on here have had that accidentally growing one type of contaminant mushroom in their tubs. But usually one or the other is much weaker/looks sick. Although, georgeM  grew Schizophylum Commune and cubes in this same cake and described them as 'living in harmony', whether you agree with that assessent or not.

Or, maybe a tub/cake is the wrong way to go. Maybe they'd share genes more readily on the agar stage, and you can already make the gene transfer happen at the culture stage, then later produce psilocybin-containing buttons, say. I think this is more promising.
Psilocin has antifungal properties. Possibly the other mushroom may eventually have to steal genes containing enzymes that process psilocin in order to survive in their setting - and the gene cluster that makes psilocybin includes enzymes degrading psilocin into the more stable psilocybin and could have survival benefits.

If it's possible by cooperation no extra pressure is needed. I could see three possible selective pressures you could do though.

A. Adding huge amounts of exogenous psilocin to the substrate and/or to petris. In the petri stage this might be faster at inducing change than in substrate stage. Psilocybin mycelium will be fine, but the other mushroom may struggle when it didnt before. This induces pressure to possibly gene transfer. But I'm not sure if it could 'steal' the gene in that circumstance if it even needed to - might need to be some requisite bacteria.

B. Let us say you have a plate or tub of Psilocybe mushrooms that overcame a trichoderma infection. Culture the trichoderma. It might be vulnerable to psilocybe metabolites, including psilocybin. Then this trichoderma is introduced to the petri containing both mushrooms. The non-active may have to acquire the gene to fend off this competitor.

C. Probably what happened in nature, psilocybin can be effective against certain bugs. Some bugs could be introduced, in which only mushrooms that produce a good insecticide are favored, making the other mushroom be pressured to acquire psilocybin genes.

B and C are theoretically interesting but involve using insects and contaminant molds, unpredictable. I think A would be really interesting and more reliable.

But the thing is, even if you introduce the selective pressures for a mushroom to benefit from producing or tolerating psilocybin, would it be able to acquire it from another species? Perhaps the mushrooms would have to become buddy-buddy first and part of the same network for such sharing to be possible. Or perhaps it would only happen if they were enemies. Or perhaps there are rare and hard to acquire bacteria needed in the first place for it to happen and it may not happen despite thousands of trials. Several different possibilities.

Edited by CreonAntigone (09/16/26 03:47 AM)

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: CreonAntigone]
    #29669956 -

My intuition tells me that repeated and prolonged exposure of both cultures of one another in order to buddy up is the most efficient route, like in a really big petri dish.

Take the two cultures and place samples very close to each other and basically let them fight off a common enemy (TBD) in the middle, which would require them to share cultural information between the two, leading to a hybrid culture on the other end.


I suppose that if we could get the two cultures to become familiar with each other over a big enough petri, the two cultures must be interwoven at some point.

Ive read where 2 cultures that interweave are able to produce fruiting bodies of both cultures in one tub as you have stated, but with a 50/50 ratio.

Maybe enough iterations of this process will lead the 2 cultures' spores of becoming compatible with each other enough to hybridize.


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When my pies have nine lives?"

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: CreonAntigone]
    #29669963 -

Yeah I think that's a better idea. It may not even require a competitior at all. Reading more about how fungi interact, they have moments of fusion between hyphae and then there is cell death, and possible sharing of genetic information. Just growing next to each other could be enough.

But the interesting thing is you could keep doing this again and again. After the petri is finished, you could take the part that is button mushroom and then transfer it to a new one, and expose the petri to new cube spores on the other side. Potentially even more information/interaction. Or maybe instead of new spores reculture the proven-friendly cube culture again and just keep developing the same cultures.

After each dish you can test to see if any psilocybin is found in the button mycelium (or whatever species you choose) without needing to grow any mushrooms yet.

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: CreonAntigone] * 1
    #29669967 -

I like this idea, im going to test it out onto pyrex dishes like This


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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: veggie]
    #29670076 -

veggie said:
Scientists have been able to produce psilocybin from yeast and, to a lesser extent, from bacteria.

I recall reading how different types of mushrooms evolved psychedelics twice, each using their own unrelated different ways to produce psilocybin.

You may be on to something. Fascinating project.


I was researching this recently.

Think it's one of the deer mushrooms that create baocistis which isn't uncommon in certain fungi though it produces different enzymes altogether and goes through a different method of creating psilocybin than the average cube

Cool read that had me really inspired to get my hands on the fungi n mass spread it for the cult scene though I've been unable to find a way to secure a genetic sample

And I read something about a psilocybin producing hybrid of a tabacco plant which is very closely related to hops and marijuana if u didn't know so psychedelic beer might be around the corner

Endgame goal use psilocybin producing hops to grow psilocybin producing fungi for insane potencys previously unobtainable? :popcorn:


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An enigma some caerulescens n a mono 😎

Edited by bloodssonnew (09/16/26 07:57 AM)

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Re: Transgenetic Expression : Getting Nonactive Mycelium to Become Active Producers [Re: bloodssonnew]
    #29670104 -

They have a lot of psilocybin-producing stuff via gene editing, including a bacteria strain. If someone got their hands on that bacteria, I mean they could make as much psilocybin as in 50 tubs in a week, with a single big fermenter.

Quote:
"In less than 2 days, we successfully produced approximately 300 mg/L of psilocybin under simple conditions with easily sourced equipment and supplies"


One liter, in 2 days, the equivalent of 30 grams dry if the mushrooms were 1% potency.

Now imagine if you got a couple of carboy fermenters, at 25 liters.

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