tree frog said:

Small correction, Panaeolus bisporus is psuedo-homothallic, so n+n even from a single spore.  Not all of these will be compatible mating types, so often they will still cross with another strain (the bisporus strain on the plate, not with like cyans).

And this part is my opinion, not proof.  Though it's an informed opinion from reading David Moore's textbook.  But I believe that by the time we spawn the war is over.  The genetics have already sorted themselves out through the parasexual cycle, and the weaker strains died out.  I think this often can happen with pans by t2.  Especially in the two spored species.  And part of the reason I believe this is because of the break and shake.  The substrate acts as a colony, not a bunch of small colonies spread through a large substrate.

So, and again just my opinion, the benefit of cloning isn't in narrowing down the strains.  But in epigenetic markers.  The cloned strain has learned what conditions to expect.  And an isolate from that same plate prior to cloning has not.

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tree frog said:

I think that's a lot bigger than the space I am using.

When I was using a tent I had an HVAC tube with a 4" exhaust fan on it.  So sucking out instead of blowing.

But I use four computer 3" fans and the speed is adjustable. Right now they are on one.

One exhaust on each fruiting chamber.  And two on the climate chamber blowing in.  But the humidifier also pushes quite a bit of air in.

They all run together on a timer.  Not that I really trust the humidity sensor at such high humidity but it's always at 100 basically.  They tolerate high humidity better at cooler temps (72-74 at night and 74-76 day).

My setup looks like this.






And there's like six seed mats. Two on top of the fruiting chambers. And four under the climate chamber.

I know that doesn't really answer your question. But you need to calculate the square footage of your space, and then compare that to the CFM of the fan.

Either that or just get adjustable fan so you can dial it in using the fruit morphology.

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Quote:

Many recent studies have demonstrated that stressful conditions that are experienced by parents can influence the offspring’s vulnerability to many pathological conditions, including psychopathologies—primarily related to a disruption in stress response mechanisms. These effects may even endure for several generations. Nevertheless, the mechanisms of this phenomenon have not been detailed, and only recently have scientists examined epigenetics to answer this question.

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Kunta Kente said:

Ah that's essentially what I have with the Joc tent. (which i guess is the point haha). Do you dial it in from common settings just based on the surface of the substrate? Or could you get from wall condensation / drippage (like that's too much fog)? I'm setting my tent / timers up and literally just need to press on as many many jars are about to be shaken.

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Kunta Kente said:

tree frog said:

Small correction, Panaeolus bisporus is psuedo-homothallic, so n+n even from a single spore.  Not all of these will be compatible mating types, so often they will still cross with another strain (the bisporus strain on the plate, not with like cyans).

And this part is my opinion, not proof.  Though it's an informed opinion from reading David Moore's textbook.  But I believe that by the time we spawn the war is over.  The genetics have already sorted themselves out through the parasexual cycle, and the weaker strains died out.  I think this often can happen with pans by t2.  Especially in the two spored species.  And part of the reason I believe this is because of the break and shake.  The substrate acts as a colony, not a bunch of small colonies spread through a large substrate.

So, and again just my opinion, the benefit of cloning isn't in narrowing down the strains.  But in epigenetic markers.  The cloned strain has learned what conditions to expect.  And an isolate from that same plate prior to cloning has not.

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Ah didn't know that about bisporus, very cool.

I 100% agree on your "isolated early" statement. Just from the shear fact that we are selecting small sections to take to T1 and T2 likely pares it down to 1-2 genome on T1 and 1 genome on T2.


~~Opinion ahead, (not a TC or even veteran grower, just a bio major and interested)~~
From my reading, I would state this about cloning: You do it to either rescue a culture that you lost the agar of, or to isolate genetics if you had a dirty multispore bulk.

My understanding of epigenetics is the changes are dependent on the current state (current media). So mycellium strongly pushes for tiny changes to optimze for agar while there, and then again separately on grain, and then again separately on bulk.

By nature epigenetic changes are not permanent and can easily be overwritten or drift from favor. So, while in bulk, the colony is NOT optimizing for agar or grain. So, we clone (let's say to into colonies B and C, same genome, same epigenetics.)

Now we are back to agar where bulk doesn't matter at all. We (mycelium) care about only about agar environment performance. B and C will perform essentially identical on agar and grain because those are highly homogenous media. But, while on agar, while on grain, they are again reoptimizing for those medias.


Now back to bulk: B and C are presented with similar but not perfectly identical substrates. Localized sectors (pins, air pockets, and especially corners vs middle) are all different, and so optimization must REOCCUR at those places.

So each and every fruit is constantly optimizing alone. You clone from the middle and it's a different optimization pattern than the side wall, because those are entirely different conditions. Then trying to get a whole tub of the best middle guy or the best side guy would likely both result in simply the same base genome still performing well under relatively similar bulk conditions.



Sorry for again the long write up, I like talking about the background science (as in I'm not saying "NO I'M RIGHT" just saying what I think).

TLDR: combining media specific state optimizations (agar | grain | bulk) and differences in the single bulk environment (the middle is more dry and airy than edges), epigenetics are likely to transient to effectively select for, and performance is much more likely related to the original genomic strength (that should be preserved at the original 'master' T2 for future resampling)

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tultedspoon said:

Kunta Kente said:

tree frog said:

Small correction, Panaeolus bisporus is psuedo-homothallic, so n+n even from a single spore.  Not all of these will be compatible mating types, so often they will still cross with another strain (the bisporus strain on the plate, not with like cyans).

And this part is my opinion, not proof.  Though it's an informed opinion from reading David Moore's textbook.  But I believe that by the time we spawn the war is over.  The genetics have already sorted themselves out through the parasexual cycle, and the weaker strains died out.  I think this often can happen with pans by t2.  Especially in the two spored species.  And part of the reason I believe this is because of the break and shake.  The substrate acts as a colony, not a bunch of small colonies spread through a large substrate.

So, and again just my opinion, the benefit of cloning isn't in narrowing down the strains.  But in epigenetic markers.  The cloned strain has learned what conditions to expect.  And an isolate from that same plate prior to cloning has not.

---

Ah didn't know that about bisporus, very cool.

I 100% agree on your "isolated early" statement. Just from the shear fact that we are selecting small sections to take to T1 and T2 likely pares it down to 1-2 genome on T1 and 1 genome on T2.


~~Opinion ahead, (not a TC or even veteran grower, just a bio major and interested)~~
From my reading, I would state this about cloning: You do it to either rescue a culture that you lost the agar of, or to isolate genetics if you had a dirty multispore bulk.

My understanding of epigenetics is the changes are dependent on the current state (current media). So mycellium strongly pushes for tiny changes to optimze for agar while there, and then again separately on grain, and then again separately on bulk.

By nature epigenetic changes are not permanent and can easily be overwritten or drift from favor. So, while in bulk, the colony is NOT optimizing for agar or grain. So, we clone (let's say to into colonies B and C, same genome, same epigenetics.)

Now we are back to agar where bulk doesn't matter at all. We (mycelium) care about only about agar environment performance. B and C will perform essentially identical on agar and grain because those are highly homogenous media. But, while on agar, while on grain, they are again reoptimizing for those medias.


Now back to bulk: B and C are presented with similar but not perfectly identical substrates. Localized sectors (pins, air pockets, and especially corners vs middle) are all different, and so optimization must REOCCUR at those places.

So each and every fruit is constantly optimizing alone. You clone from the middle and it's a different optimization pattern than the side wall, because those are entirely different conditions. Then trying to get a whole tub of the best middle guy or the best side guy would likely both result in simply the same base genome still performing well under relatively similar bulk conditions.



Sorry for again the long write up, I like talking about the background science (as in I'm not saying "NO I'M RIGHT" just saying what I think).

TLDR: combining media specific state optimizations (agar | grain | bulk) and differences in the single bulk environment (the middle is more dry and airy than edges), epigenetics are likely to transient to effectively select for, and performance is much more likely related to the original genomic strength (that should be preserved at the original 'master' T2 for future resampling)

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Disclaimer: I'm not an expert
I'm thinking there can be some small changes, but I think most is decided from the spores that combine into the mycelium network, and once mycelium branches are formed and venture off and those combinations take place, the changes from then on are more subtle if it's even capable of recomining again and again I don't know. I don't think there's substantial changes as it encounters different media (over time if mycelium was running long distances maybe it could, probably moreso mutations by chance, but the reason it fruits is to make spores for changes too, like the mycelium will run, fruit, spores, whatever takes takes, mycelium runs then fruits again, and then spores again to survive changing environments.

This reminds me I've probably been using not the best prints. I was using recent prints, but I should probably be using older prints when I was finding my specimens more frequently under easier conditions to mimic. I've probably accidentally selected for harder specimens to get bigger flushes from.

I think the mushroom memory may work in such a way that when you take a clone, you are cloning mycelium that will fruit. Not all mycelium is fruiting mycelium, I believe RR said that somewhere. So by taking mycelium from a clone, you know you will have mycelium that is capable of putting out a fruit if conditions are similar. The growth styles for how it performs in spawn are already baked into the genome. I don't think the mycelium has to relearn growing through agar (which actually kind of sounds stupid when you think about it, for reasons people use agar to begin with) or spawn again, the whole lifecycle is already mostly (I say mostly because always possibilities of mutations or selections) determined. The mycelium is so advanced (possibly of alien origin), that it is able to separate from primordia into stem myceliums and cap myceliums, from the root mycelium (something like this I'm close but not exact).

Maybe people are cloning from the middle and the sides, but I think more importantly people are cloning phenotypes or fruiting styles (clusters for big flushes). I don't think master growers are expecting their tubs to have vastly different environments from the middle vs. the sides. There's probably more normal environmental variation that occurs in a regular routine misting.

This is not advice.

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tree frog said:

That's true when you clone it's guaranteed to be a fruiting strain. It's possible to accidentally isolate a mono, though not easy.  And some nuclei can be vegetatively compatible but not fruiting compatible.  But again this is rare because of the way the nuclei are favored throughout the mating life cycle.

Epigenetics is a whole different type of learning, you can think of it as a modifier that's applied to a gene. Telling it to activate or not.  And there is evidence that this type of learning also passes into the future generations.  I don't know how much evidence we have of that in fungi, but it is evident in animals.

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will use as i remmber the indrotuction of a CRISPR conference i folow few years ago...

if a give to each of you the exact same bucket of lego block with a book of blue print.

and i ask you to build something using the book.

majority will just take a random print and build it (we got a shroom)

some are less bright and will fuck up with the plan (we got somme albinos)

some are funny guy and will just treat the book as a block itself( we got enigma)

and we have somme realy dumm ass that will just put everything in a stack and glue it all together ( we got blob)

when we proceed to clooning we restrict the acces to somme page of the book. but even without acces to this page the actual configuration relevant to this page is still a possibility for the funny guy and the less bright one, for the dummy let said that i wont bet on him......

But with CRISPR we dont care about the book. we just remove the block that we dont want from the bucket....


ok the actualy exemple wasnt shroom... but it serve the same purpose

the  "book" it self is stock in the nucleide  in the cellular core. you cant touch it by clonning. what we do is playing on the probability, and by succesive transfert and selection is almost like trying scratch the funny guy and the less bright one, but we can do nothing about some one who will choose to change is mindset and we got revert.

fahtster said:

3rd ish flush ziplock.  This tray had 25% worm castings and the rest plain CV

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