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Phenotypic determination in P. cubensis: does the differentiation phase define everything? * 2
    #29675168 -

I decided to open a separate thread for my question because I think I'm going to ramble a bit.

I've spent days turning over the topic of morphological definition in P. cubensis, because during different tests I ran with different setups — I got results that only raised more questions.

If I understand correctly, phenotypic determination occurs during the mycelium's differentiation phase into hyphal knots. That's when "the genetic program" is being defined by several factors, including environmental ones (CO₂, RH, temperature, etc.). As I understand it — the primordium is really the mushroom already defined with a "predetermined" phenotype — "the genetic program" executing under those conditions.

For example, if we get phenotypes with long stems and small caps (spaghettis) — were it the conditions during the mycelial differentiation phase that determined this phenotype, and not the subsequent conditions?

Over these two months I ran different setups in my individual fruiting chambers for GT PF-Tek (MS) cakes and observed how they favored certain behaviors over others. Greater passive FAE toward the top, more manual intervention (misting + FAE), gave me lower pinning but a few very large specimens. Less passive FAE toward the sides (EZ Dialed), less manual intervention resulted in higher pinning and more medium-sized specimens. PF-Tek to Bulk in a larger volume of space (height) produced enormous, heavy specimens — whereas a smaller volume of space (height) resulted in slightly more mushrooms but somewhat smaller.




Harvested weights were very similar, with an upward trend toward setups with less manual intervention.

Is this because the phenotype established during the differentiation phase expresses itself more easily when subsequent conditions aren't disturbed?

If we change post-pinning conditions, are we actually working against its "genetic program"?

If we disturb conditions from the start — will it determine phenotypes that express themselves best under those conditions of constant disruption?

The same genetics won't express themselves the same way on a PF-Tek cake as in a bulk — if only because of substrate verticality, water management, relationship with gravity, and the way CO₂ is expelled. CO₂, being denser than air, tends to accumulate on horizontal surfaces, creating a microenvironment of elevated CO₂ right where hyphal knots are differentiating. Nor will they behave the same on a cake resting on vermiculite versus on a rack in a watertub that provides more space for CO₂ to fall away.

I observed that my GTs, when they managed to produce a cluster, always did so on the vertical sides of the cakes — never on the flat top or in bulk.



What would happen if instead of making a horizontally flat bulk, we created a slope? A diagonal surface?

I also understand these results are more likely when working with MS vs Clone. Because I deduce that a clone is precisely homogeneous genetics, from which all hyphal knots will respond similarly by starting from the same "genetic program".

This makes me rethink some things and pay much more attention to the differentiation phase — something I perhaps hadn't kept as present. Especially when it comes to selecting specimens.

And going forward — when selecting, I'll definitely split into two lines: cakes and bulks. One doesn't necessarily perform just as optimally in another "medium".

I also have a couple of cakes that encapsulated small areas of Pseudomonas, and I've been digging into research on Agaricus bisporus and its relationship with C8 volatiles in primordium formation. Because of this I'm going to fruit them with a coir roll to give a better chance at further "encapsulation". Just in case — no dunk. But I want to see how this translates to Cubensis. Which genetics will express themselves under these factors.

Honestly, the topic feels overwhelming and I don't know how far off the mark I am — but I'd really appreciate it if anyone knowledgeable could shed some light on my shipwreck.

Hugs :heart:


--------------------
"Little shark in da bathtub"

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: Belintaf] * 1
    #29675184 -

Here is my 2 cents...

This sounds like the age old question Of nature vs nurture, of which I take the position of a pure centrist (50/50)

Quote:
For example, if we get phenotypes with long stems and small caps (spaghettis) — were it the conditions during the mycelial differentiation phase that determined this phenotype, and not the subsequent conditions?


I think its both simply because of the (extreme) example of your powers to alter the growing environment the mushroom resides in (ie. You could drown the mycelium, or take a blow torch to the pins)

I realize no one would ever do this but to me it shows that phenotypical expression has to be completely fluid in order for organisms to adapt to their environment.

Quote:
Is this because the phenotype established during the differentiation phase expresses itself more easily when subsequent conditions aren't disturbed?

If we change post-pinning conditions, are we actually working against its "genetic program"?

If we disturb conditions from the start — will it determine phenotypes that express themselves best under those conditions of constant disruption?


As ive stated before, I take the position that phenotypes are 100% fluid in order to make the most of the environment the specific genetics just so happens to find itself in.

I like to think of mycelium as a civilization of humans as an analogy. I see mushrooms as people and the mycelium as the total knowledge/conscious base that allows humans to functions

If the civilization lives in total anarchy (ie. cultivator fucking with the conditions) the mycelium will have to adapt its fruiting strategy to such chaos, as opposed to the world peace strategy where things always go smoothly and every point of the mycelium has perfect conditions and all the time in the world to make the best fruiting bodies it knows how to

Quote:
What would happen if instead of making a horizontally flat bulk, we created a slope? A diagonal surface?


If you havent already, I recommend reading Pastywhyte's bathtub grow log. I asked him what he learned from it and he stated that one of the things was substrate being level. He stated that due to the tub being uneven, the more shallow portion fruited faster due to there being less sub to colonize





--------------------
"Why try for high fives,
When my pies have nine lives?"
DUBJUG TEK
PLASTIC-EATING MYCELIUM
TRANSGENETIC EXPRESSION 🧪
JOIN CAKE WARS III TODAY
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Edited by langten69 (09/22/26 05:27 AM)

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: langten69] * 1
    #29675198 -

Well, I will hopefully have some useful data for you in 2 to 3 weeks.

My Eungella clone culture is throwing up fatty albinos.



I am going to spawn a pint of millet in a cranberry juice bottle cut in half, with air holes at the top. Why? Because I have seen a disposition for fruits to get tall in those sort of containers where there is a lot of vertical space.

If the same culture outputs taller fruits, that would be an environmental cue favoring or triggering certain phenos to show up. Whether that is a pheno or just an environmental cue is a different question. But if the stem is less spongey, or is hollow, doesn't create the fat bump, and if the caps change, those would be notable differences from the same culture.

Now, what would be very interesting is to see if a standard color comes in instead of the albino (not likely with a clone, but if it was MS, the original mutation tray had both), the idea being because melanin helps mushrooms gets stretchier and taller, which is a reason why albinos tend to be shorter and squattier like Yetis.

Maybe the mycelium acts like its playing a card game. It has a hand drawn with available tactics, and given the situations, it plays those cards according to its intuition of what will work.


--------------------
I call them Huskies cause they Mush!


The Official Wild Cubensis Thread
Instant Pot Agar
Gnocchi Water Agar
Let's Talk Cowtek

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: Huskies] * 1
    #29675214 -

Google "phenotypic plasticity".

I read somewhere that these changes in appearance can also carry through to subsequent generations like kind of a genetic feedback loop where
conditions can have an influence on genetic changes.


--------------------
🍄PANAEOLUS IN MONOTUBS🍄
🍄OFFICIAL SEMILANCEATA THREAD🍄



⭐Baba's Write Ups and Link Collection⭐

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: Baba Yaga]
    #29675218 -

:whathesaid:

Its no different then getting a sun tan, and the more you tan the tanner you get


--------------------
"Why try for high fives,
When my pies have nine lives?"
DUBJUG TEK
PLASTIC-EATING MYCELIUM
TRANSGENETIC EXPRESSION 🧪
JOIN CAKE WARS III TODAY
MODERN FRUITING CONDITIONS (CUBES)
REVITALIZE THE CUBENSIS GENEPOOL
BOOKMARKED THREADS 📖

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: Huskies]
    #29675219 -

Langten69, thanks for mentioning the bathtub case (I was already familiar with it). Though what the bathtub actually demonstrates is the slope of the base, responsible for draining water toward the drain — which determines substrate thickness. Not the substrate itself forming a pronounced slope on its surface.

From what I understand (and I could be wrong), hyphal knots are potential pins, but if environmental conditions during differentiation aren't adequate — the mycelium "reabsorbs"/"recycles" them, and those that did "win the selection" under those same conditions become established and commit as primordia.

I'm not saying what comes after doesn't matter — but if that's the case, it no longer determines morphology. Conditions of humidity, temperature, available space and CO₂ modulate the speed of execution and can cause damage, cracking or slowdown, but they don't rewrite the "morphological program."

And that's precisely what I'm asking about.

Because if phenotypes were 100% fluid as you say, we wouldn't see so many differences between specimens on MS — they would all look like the same phenotype under the same conditions.

Thanks for your response, Huskies. I find your work very interesting and will gladly follow it.

These are exactly the questions I'm asking myself, and the "cards" analogy feels accurate — it's the same thing I'm thinking. That a given genetics has X options for X conditions and responds accordingly.

Which might also explain why many genetics revert to being a simple Cubensis when fruiting in the wild.


--------------------
"Little shark in da bathtub"

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: Belintaf] * 1
    #29675231 -

Just to be clear, when I say fluid, I mean that the environment plays a 50% part in determining morphology and the genetics play the other 50%

Since a MS is a genetic lottery, that is why the phenotypes vary as opposed to a clone

Quote:
From what I understand (and I could be wrong), hyphal knots are potential pins, but if environmental conditions during differentiation aren't adequate — the mycelium "reabsorbs"/"recycles" them, and those that did "win the selection" under those same conditions become established and commit as primordia.

I'm not saying what comes after doesn't matter — but if that's the case, it no longer determines morphology. Conditions of humidity, temperature, available space and CO₂ modulate the speed of execution and can cause damage, cracking or slowdown, but they don't rewrite the "morphological program."


I 100% agree with this. I believe the "morphological program" is like what huskies said in that its basically like a sort of blueprint that can alter based on the environmental cues. Thats what I mean by fluid.

I do truly believe that mycelium plays everything by ear and that there is nothing set in stone when it comes to the final morphology since the environment can (relatively) change at any moment in the wild


--------------------
"Why try for high fives,
When my pies have nine lives?"
DUBJUG TEK
PLASTIC-EATING MYCELIUM
TRANSGENETIC EXPRESSION 🧪
JOIN CAKE WARS III TODAY
MODERN FRUITING CONDITIONS (CUBES)
REVITALIZE THE CUBENSIS GENEPOOL
BOOKMARKED THREADS 📖

Edited by langten69 (09/22/26 06:51 AM)

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: langten69] * 1
    #29675272 -

^ specifically in cases of albinos and Leucistics reverting back to standard cube colorings in the wild or outdoors, which I think has been documented a few times.

Being without melanin could be a major disadvantage in sunlight, so it would show them playing differently cards so to speak.

Makes me wonder what the leucistic mutation might help with. Since they tend to sporulate less, AND that would appear higher contrast against grass for animals with two cones such as cows and horses, my hypothesis is that they are more likely to be eaten, leaving the spores inside of the fruit and thus inside of the ruminant of choice.




--------------------
I call them Huskies cause they Mush!


The Official Wild Cubensis Thread
Instant Pot Agar
Gnocchi Water Agar
Let's Talk Cowtek

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: Huskies] * 2
    #29675295 -

I apologize in advance if I am stepping on anyone’s toes here. I skimmed the responses but this may be redundant. Also a bit dusty on my knowledge here.

Epigenetic factors (environment, nutrition, location etc) can instigate significant phenotypical diversity i.e. twins raised in separate geographic, monetary, environmental conditions and pressures etc diverging in physical appearance. The underlying genetic base, the genotype, is unchanged.

A mushroom “clone” is not analogous to a plant clone. Taking a cutting from a plant will result in a true one to one DNA match. Mushrooms are much different. Any given “clone” may contain many (hundreds or thousands or more) individual genetics working in conjunction with one another.

The exception being growing out and selecting monokaryon cultures, which are true clones. Of course they must be bred to form dikaryons and fruit.

Hope that helps and isn’t too confusing. As always I may be wrong as well.

:mushroom2:

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: phytophthora]
    #29675312 -

Quite the opposite, phythophthora. I genuinely appreciate your opinion, thank you.

I'm not sure if it's exactly the "phenotypic plasticity" BabaYaga mentions, but if we're talking about the epigenetic imprint inherited by the next generation that includes previous conditions — a "new" adaptation — that would explain the different behaviors of a "clone," as you say.

From my ignorance I see it as information transmitted by experience to the next generation, which is then expressed during differentiation.

Another question comes to mind — if one decides to mechanically influence morphology from the outside, will the result be partially inherited in the next generation under the same conditions? Because I don't think the feedback loop is that broad — though I genuinely don't know.

And here I'll admit I had no idea about monokaryon cultures — but I'll take the opportunity and look into it with interest. Thanks for the chance to learn something new.

Huskies, what you're describing reminds me of the behavior of the black truffle — ascocarps full of spores that learned to propagate through animals.


--------------------
"Little shark in da bathtub"

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Re: Phenotypic determination in P. cubensis: does the differentiation phase define everything? [Re: Belintaf] * 1
    #29675318 -

Epigenetics are more or less the genotypes response to environmental factors. They can change throughout the life of the organism.

Epigenetic changes can very rarely alter the DNA being passed to the next generation, but the amount varies by species and the vector of influence respectively.

Mechanical manipulation probably won’t even begin to move the needle in regards to heritability.

Mushroom tissue biopsies that are further cultured are, as I stated previously, not unique genetic individuals. If I take an agar wedge from a plate (originating from multispore or tissue biopsy from a mushroom produced from multispore inoculation) and snag other cultures from different areas of the plate and put them to grain I can end up with genetically sectored and unique gene clusters in each grain spawn container, and they will perform differently, regardless of external epigenetic factors.

Edited by phytophthora (09/22/26 10:17 AM)

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