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Theory about a massive downside of MS grows


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#21 Baphom3t

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Posted 18 February 2021 - 12:20 PM

I've always found fresh seem to onset faster and have a higher probability to be more intense.
I second TV's assessment of Fresh vs. Dry no matter if they were wild or grown indoors. There is also talk about closed cap vs. open cap, stem vs. cap, and blah, blah, blah. This sort of debate has been going on since I can remember.
Is it not possible the atmospheric/meteorological conditions in the grow area affect potency as each grow that is done, is done under slightly different conditions. I.E, the difference between outside being 85 with 80% humidity compared to 72 with 70% humidity should in essence relate to potency.
This is an interesting topic. I look to see where this goes.


Edited by Baphom3t, 18 February 2021 - 12:21 PM.


#22 Moby

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Posted 18 February 2021 - 12:39 PM

I think the thought of fresh = more potent compared to dry is not only logic, its reality, isn't it?

 

Here something from a study about alkaloid contend in the caps vs stems:

 

"The fruiting bodies were separated into caps and stipes to determine the tryptamine concentrations in these mushroom parts. These experiments were performed on dried fungal fruiting bodies in six replicates. On average, the caps contained 0.01 wt.% of aeruginascin, 0.07 wt.% of baeocystin, 0.88 wt.% of psilocybin, 0.01 wt.% of norbaeocystin, and 0.06 wt.% of psilocin. In stipes, we found <0.01 wt.% aeruginascin, 0.03 wt.% baeocystin, 0.47 wt.% psilocybin, <0.01 wt.% norbaeocystin, and 0.01 wt.% psilocin. There was approximately 50 % less baeocystin, psilocybin, and norbaeocystin in the stipes than in the caps. The stipes contained 32 % less aeruginascin and 85 % less psilocin than the caps. The total content of tryptamine alkaloids in the stipes was approximately 50 % less than in the caps. These results are slightly different from an older study, which states that the psilocin content is higher in the stipes than in the caps in P. cubensis, but a similar distribution of psilocybin (higher levels in the caps than in the stipes) was observed in Psilocybe samuiensis.52 Our results correspond with the published work.26 The parametric two-sample unpaired t-test with Welch correction in the R program found that the results were not statistically significant, as the p-value was 0.3756. This may be due to the large difference between the alkaloid concentrations in the individual fruiting bodies. This means that although the average content of tryptamines in caps is higher than in stipes, due to the SD, where there is high variability between individual fruiting bodies, it cannot be said that this statement applies to all fruiting bodies"

 

The result is that generally it was noticeable that the caps have roughly twice the amount of psilocybin than the stems but since strains are different and its hard to say if all strains produce fruits with this property, it yet can't be called as generally proven


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#23 FunG

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Posted 18 February 2021 - 03:30 PM

Moby, you are a asset to this forum and community!

I've read similar research studies that also concluded the same findings so what your posting must be accurate as it can be.

I like a good technical read from time to time, I'm just a dumbed down grower, this is where peoples education comes out to shine.
Clearly some of you are either attending or have graduated university or college grade bio chemistry courses cause the info I've been reading is obviously advanced studies.
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#24 Microbe

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Posted 18 February 2021 - 06:41 PM

Moby, you are a asset to this forum and community!

I've read similar research studies that also concluded the same findings so what your posting must be accurate as it can be.

I like a good technical read from time to time, I'm just a dumbed down grower, this is where peoples education comes out to shine.
Clearly some of you are either attending or have graduated university or college grade bio chemistry courses cause the info I've been reading is obviously advanced studies.

There are some people here with degrees but the bulk of us are autodidact in our own research and studies. There some really advanced shit that can be applied to really make things interesting and fun. For example one my primary focus was on bacteria floras and symbiotic relationships inside the culture and in substrates to increase fruiting potential. Don't ask, its buried in my bipolar thread "Microbe Randoms" and i was going through some serious shit at the time, and even though a mod cleaned it up a little for me, it still gets a little wonky or WTF just happened moments lol. There some good topics in there and most of my dialogue pertaining to it, is anecdotal.

You read about something and then you come across something else you need to read to be able to connect the dots and its a vicious cycle and one can easily find themselves deep in the rabbit hole.

If you find yourself in that thread and want to discuss something, tag me in a quote as i don't always get alerts on taptalk or they are delayed. Or you can always DM me.

Anyway keep doing your thing. I can see that you have a good skill set.
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#25 FLASHINGROOSTER

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Posted 18 February 2021 - 11:39 PM

Is there a lemon tek for freshies? I tried eating 30 g fresh the other day and my stomach pushed them all the way through my gut in 10 mins. I ended up with teeth chattering chills, but none of the good stuff you'd want.

 

Do people make shakes with freshies? Wonder if it would help with the digestion part of it



#26 FunG

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Posted 19 February 2021 - 01:07 AM

You can hot water extract blue juice from fresh mushrooms (the water actually pigments blue) but only if they're fresh, dried and it's a brown color. But anyways.

I use to poor it into a empty 2L pop bottle and cap it tight to preserve the blue color. Itll stay blue along as the cap is placed on tight after each use but anyways. I'd use the extract to make everything from coffee to hot chocolates, jellow, smoothies, cool aid etc

I find the extract easier on the stomach then eating dried tissue, fresh tissue also doesnt produce that heavy feeling in the gut.
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#27 rockyfungus

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Posted 19 February 2021 - 10:13 AM

 

Is there a lemon tek for freshies? I tried eating 30 g fresh the other day and my stomach pushed them all the way through my gut in 10 mins. I ended up with teeth chattering chills, but none of the good stuff you'd want.

 

Do people make shakes with freshies? Wonder if it would help with the digestion part of it

 

I've always made a tea with them. I bet you could chew em up real good, swallow the juice, and spit the fungi out (grosssssssss)


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#28 bezevo

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Posted 19 February 2021 - 11:53 AM

LOOK UP Paul Stamets paper on ICE COLD WATER Extracting of freshies  .Paul called his idea he called it Blue Juice .. it'sa interesting paper.

...

here is a thread about Paul Stamets  Ice extraction of freshies .. seems Paul Stamets Disagrees with FunG  on how to do it .. hummm  ...  opinions vary heh

 

WHATCH THE VID  coorsMikey posted

 

https://mycotopia.ne...y-paul-stamets/

 

heres a good versionn

 

<iframe width="560" height="315" sandbox="allow-same-origin allow-scripts allow-popups" src="https://hyperreal.tu...63-dccfd31315cb" frameborder="0" allowfullscreen></iframe>


Edited by bezevo, 19 February 2021 - 05:15 PM.

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#29 hyphaenation

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Posted 19 February 2021 - 01:53 PM

I remember years ago discovering the freshies-turn-water-blue by trial & error accident using a blender...
 

  • post-104353-0-53889100-1549731628_thumb.

     
  • post-104353-0-68375300-1549731655_thumb.

https://mycotopia.ne...-4#entry1395997

Also it's interesting to actually juice shrooms... In a wheat-grass style hand-cranked juicer you can easily separate the juice and the fiber.
 

https://mycotopia.ne...-3#entry1395868
 

post-104353-0-05114100-1477531025_thumb.

 

 

post-104353-0-77135400-1477531010_thumb.

 

post-104353-0-37477000-1477535800_thumb.

There is a movie of the first juicing!

Avi attachment:

https://mycotopia.ne...tach_id=1131432

In this experiment I paid no mind to keeping air/light away from the juice... I just wanted to see the process. This juicing was done with P.cyanescens. I'll try some with Redboy cube when I have some on hand. This was awhile ago and I can't quite remember if I tried freezing the juice or not. I believe I tried and it would not freeze and stayed "slushy" but I will have to repeat experiment.










 


Edited by hyphaenation, 19 February 2021 - 02:40 PM.

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#30 FLASHINGROOSTER

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Posted 19 February 2021 - 06:38 PM

 

 

Is there a lemon tek for freshies? I tried eating 30 g fresh the other day and my stomach pushed them all the way through my gut in 10 mins. I ended up with teeth chattering chills, but none of the good stuff you'd want.

 

Do people make shakes with freshies? Wonder if it would help with the digestion part of it

 

I've always made a tea with them. I bet you could chew em up real good, swallow the juice, and spit the fungi out (grosssssssss)

 

Agreed, tea is my only method of ingestion these days. One can't help but wonder if your missing out on something by not digesting all of the mushroom though. That being said it sure didn't stop me from blasting off to planet 9 when I took and 8g tea dose a month ago. That seems to be my upper limit of effectiveness. The last few times I usually wake up on the floor and after a few minutes of trying to grasp reality I wonder what happened. It's sort of like waking up from a black out drunk, except in this case I feel fucking fantastic and have trippy visuals to look at. Anyone else have issues with time loss like that? I suppose I should mention I do smoke weed most of the time with mush but not sure what effect that would have at the higher doses



#31 rockyfungus

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Posted 19 February 2021 - 06:45 PM

I've blacked out once when I was a young man. I'm a tiny tiny dude and it took 3-4 guys over 6 ft to hold me down while blacked out. They were ready to knock my ass out.

I had to put together every memory, sensation, synapse, sinew, fiber of my being to come back. I was stuck in an endless loop of me fractaling into my existence and remembering reality. Scared me away from Mush for years and I swore by acid only. Now I wont touch any blotters/liquid, you never know what RC it could be.



#32 ElPirana

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Posted 22 February 2021 - 10:56 PM

I think somewhere in this thread there were comments related to MS grows being consistently good quality.  I've had some grows over the past year that have been poor in potency, but thinking back these have been a lot of grows from clones.  So maybe by bad luck I was picking a mushroom to clone that happened to have genetics with a weak potency.  It's just that cloning makes it sooo easy to keep grows going.... oh well. 

 

I just noc'd up some agar jars a few days ago w/ Alacab spores and have good growth already.  I'm going to use these to purely move forward as MS grows and not make any clones, maybe just make prints from choice mushrooms instead like TV mentioned.  I'm always willing to change things up.


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#33 Sharonlovesu2

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Posted 25 February 2021 - 01:07 PM

The title says it, its a theory that I developed over time by putting different pieces together.

Feel free to throw your thoughts in here, to discuss or even to change my mind, I want to talk about it.

In MS grows (multispore) we have many strains present, agressive ones, trashy ones, extremely potent ones, strains that are not even able to produce fruits etc.

The organism is a complex and complicated system.
Anastomosis:

"In mycology, anastomosis is the fusion between branches of the same or different hyphae. Hence the bifurcating fungal hyphae can form true reticulating networks. By sharing materials in the form of dissolved ions, hormones, and nucleotides, the fungus maintains bidirectional communication with itself. The fungal network might begin from several origins; several spores, several points of penetration, each a spreading circumference of absorption and assimilation. Once encountering the tip of another expanding, exploring self, the tips press against each other in pheromonal recognition, fusing to form a genetic singular that can cover hectares called a genet.

For fungi, anastomosis is also sex. In some fungi, two different haploid mating types - if compatible - merge. Somatically, they form a morphologically similar mycelial wave front that continues to grow and explore. The significant difference, is that in each septated unit is binucleate, containing two unfused nuclei, i.e. one from each parent that do not undergo karyogamy."


'Better' strains fuse with 'lower quality' strains when the expanding tips meet each other and find compatible strains.

This not only happens when for example plates are inoculated with spores, it also happens when grain colonizes and when the colonized grain is broken up and mixed with whatever kind of bulk substrate.

The recolonizing parts (the whole thing) has mycelium colonizing into all directions and at parts that appear to be fully colonized there again the tips of the colonizing mycelium from every single grain meet the tips from the neigbor grain where possibly Anastomosis can happen.


To safe a genetic its common practice to take spore prints.

The present genetic in every spore print is directly influenced by the choice of the fruits we decide to print.


Here are the first pieces put together to get an idea of what I try to say.

If the genetic of our prints is influenced by the quality of the fruiting strains, then does growing MS grows, where constantly good with not so good strains fuse, not 'degenerate' our home grown genetics ?


I did read that freshly collected wild growing Cubensis are of much better quality than domesticated ones.

I could imagine that outside in alot rougher conditions only the strongest strains are able to survive and produce fruits.

They then drop spores and again only the toughest strains are able to survive and produce fruits (natural selection).

In our home lab setting we always give our best to provide the best possible conditions for spores on agar plates to germinate, for grain to colonize and to have optimal fruiting conditions.

Compared to the "natural growing Cubensis" in nature there is a huge difference because here in our home cultivation we also give weaker strains the opportunity to 'compete' and mix their genetic into 'the pot'.

By bulking we provide a second opportunity to mix weaker strains with 'better' ones.



For the case the theory is accurate, I'd like to throw a hypothesis into the room.

"Not only selecting strong mycelium from agar and sorting out weak strains prior to any fruiting, does work against the degeneration of all of the home grown varieties; also inoculating germination plates with heavily diluted spore solutions or just a tiny amount of spores (a needle tip) would also make sure that we have less potential to heavily mix up good with not so good genes."

___________


Feel free to chime in and tell your thoughts or to change my mind, everyone is welcome

Brilliant topic, thank you.
I think this resonates with me as I'm extremely new to this craft and recently got my hands on tampanensis spores from an awesome seller ,as a gift I was given an additional spore print of a wild Florida cubensis strain so yay soo much more exciting than before. can I ask how you'd use it (the wild print?)
What would you suggest fora start? Always to agar? Thank you again for your topic.
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#34 BirdsArentReal

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Posted 03 April 2022 - 07:30 PM

 

Love how this thread is developing-- even though the OP started from an incorrect premise, it's the pursuit of knowledge that's valuable to everyone!

 

I'm very interested in breeding excellent examples of wild(er) strains, and if it's OK, I'd like to post something shared with me by a friend that was written by others elsewhere that might help add useful information to the discussion:

 

i have a culture, named corumba, got it from a guy in brazil.
it would always give me great results from MS, on various substrate.

i have another two named TLY and Z-strain, both would give the same results from generation to generation, MS of course.

print grow then print grow then print grow, they would always give good big massive flushes

i just leave this here

Workman said:
Mushrooom genetics are a little strange since a single mushroom produces spores that can then act as both parents for a new mycelium.  Essentially, you are selfing or inbreeding each time you do a multispore grow.

Now consider a wild collection of Psilocybe cubensis with a high heterozygosity.  This basically means that most or all of each pair of genes in the mushroom are different from each other.  Its the same gene location with the same basic function, but different versions.  For example, if there is a single gene for height, you might have a version that gives short mushrooms and a version that gives tall mushrooms.  If heterozygosity is high, you have one of each which may result in medium mushrooms unless one of the height genes is dominant.

Now, when you do multispore from a single mushroom you randomly get a mix of all the genes.  Sticking to our height gene example, you could get two short copies, two tall copies or one of each.  Obviously the strains with two short copies will be short and the ones with two tall copies will be tall.

Lets say we liked the short mushrooms so we saved that one and took a spore print for later.  In this example the tall version of the height gene is lost to later generations.  There is a net loss of heterozygosity.  Over the entire genome the loss is about 50% per generation.

So mathematically we can figure out how many sequential multispore generations we need until the heterozygosity is reduced to an insignificant level and the strain is stable even from multispore.

Starting with a presumably high (~100%) heterozygosity from a wild collection.  In reality, the heterozygosity is probably lower than 100%, but its an easy number to start with.

100% wild print
50% 1st generation from wild print
25% 2nd generation from 1st generation print
12.5% 3rd generation.....
6.25% 4th generation.....
3.12% 5th generation.....
1.56% 6th generation.....
0.78% 7th generation.....

You can see that the heterozygosity drops off quickly in the first few generations and is less than 1% after the 6th generation.  This highlights the importance of choosing the best traits early on when there are more to choose from.  Attempting to isolate traits in well established strains results in only minimal improvements unless spontaneous mutations increase the heterozygosity in a positive way (rare).

In summary:

Popular classic strains in circulation have all been grown well beyond 6 generations and are relatively stable from multispore with little need for isolation.

New strains, from wild material or cross breeding between different strains of the same species, can be stabilized fairly quickly with 6 or 7 generations of sequential multispore grows.

Selection is most important early in the process and if good genes are bred out, they are gone forever.  Archiving original or early generation prints is recommended for preserving heterozygosity for later selective breeding.  Continuous isolation of a bad strain with hopes of significant improvement is futile.

Does that help?
If I may jump in here on the breeding true from spores aspect.

You lose on average 50% of heterozygosity with each strain generated from multispore (analogous to selfing in plants). What this means is, the more sequential generations you do via multispore, the less variability you will see in the later generations until the variability is nearly undetectable. There will come a point that the only variability will be from new random mutations.

OK, so what is this point, or how many generations until you can be confident you have a true breeding strain? With a wild strain you would assume 100% heterozygosity. Previously domesticated strains will obviously have less heterozygosity but since that is unknown it doesn't hurt to be conservative.

Mathematically you can see the reduction in variability with each generation. It drops off quickly and then around generation 5 or 6 the gains in stability drop off dramatically. After generation 7, with heterozygosity less than 1%, the rate of random mutations will outpace any small amount of remaining variabilty loss in later generations.

Wild 100%
F1 50%
F2 25%
F3 12.5%
F4 6.25%
F5 3.125%
F6 1.5625%
F7 0.78125%

Generate several strains from the spore print and you should expect to see huge differences between these new strains if the heterozygosity is high. Choose the strain(s) with traits you want (in this case, cap color and size) and take prints. Repeat with these new prints. You can probably feel pretty confident that after doing this 5 or 6 times that the strain is stable, especially if
you don't see any new variants in the later generations.

Good luck and I hope this is understandable.
one of my favs
breeding


Yes it is possible and mushrooms do have sex. Of course you have to stick to the same species so technically crossing one strain of cubensis with another strain of cubensis isn't a hybrid. But it still can be used to generate novel strains.

Spores, like eggs and sperm, have only half the chromosomes of somatic cells. A singe spore germinates and grows a thin monokaryotic mycelium until it comes in contact with another strand of mycelium from a different spore. At the point of contact the two myceliums fuse and genes recombine into a dikaryotic mycelium that grows thicker and faster and is "hopefully" capable of fruiting.

The difficult part in breeding is isolating individual spores and growing out the monokaryotic mycelium. "The Mushroom Cultivator" (Stamets and Chilton) covers the spore diltution technique on pages 340-341.

There is another method of crossing strains that isn't as well understood or well known called anastomosis. This is mentioned on page 8 of "The Mushroom Cultivator". Anastomosis is where two dikaryotic myceliums fuse, exchange genetic material and form a new strain. This can sometimes be seen in casings containing two different strains where a few mushrooms seem to be intermediate between the two parent strains. Anastomosis can be done easily on agar where the two different fruiting strains are allowed to grow together in a single petri dish. Typically, a zone of incompatibility forms where the two strains meet. Even though it seems that the two strains are completely rejecting each other, genetic exchange is usually taking place. If a small wedge is taken from the incompatibility zone and culture out to fruiting, new strains often result mixed in with the parent strains. For some reason the crosses appear more abundantly in later flushes. It is suggested that strains very different in appearance are chosen for crossing by this method so that they are easily recognized when they occur.
inbreeding
Inbreeding?  Well, it isn't generally good long term, but it is a useful tool for enhancing and isolating certain traits.

Each multispore culture is selfing (breeding with one's self).  This doesn't seem to be a problem initially, but sequential multispore cultures reduces heterozygosity.  What I mean by sequential multispore is using prints from your current cultivation to start the next cultivation and so on.  A better idea is to save prints from your earliest cultivation and use those to preserve your "strain" as long as possible.  Cubensis spores should last at least 10+ years if kept cool and dry, but viability drops yearly, so you may need to use large amounts of spores to revive a culture from very old spores.

The loss of heterozygosity means that the spores will produce fruits that show less and less variability from multispore, which sounds fine.  You get a nice uniform crop with little variability, so its similar in behavior to a clone.  But this also makes your culture vulnerable to random mutations.  Random mutations that can be recessive and invisible to the grower.  The PE mutation is a good example.

The PE mutation is recessive, so lets say you made a multispore culture from a print, maybe you selected out a single strain on agar or just injected tons of spores into a substrate.  You got a crop of nice looking mushrooms and printed some.  You gave away all the prints you made and just have one print remaining.  Unknown to you, the mushroom you saved the print from was generated by a pair of spores, where one had been hit by a cosmic ray that damaged a key gene necessary for proper cap development.  The mushroom looks fine because each of it's cells also contains the nuclei from the other spore with an undamaged gene.

You only have the one print, you want to grow it out again and expect the same results as before.  But something is wrong, several mushrooms look like PE, with weird malformed caps.  The selfing, or inbreeding has resulted in pairs of the damaged gene appearing in 25% of the generated strains.  Many of the mushrooms in the crop look fine, but most of them will also contain the damaged gene.  And this is just one trait.  More mutations can sneak in over time, almost all of them are bad, some can make the mycelium unable to even fruit.

So save your earliest prints, save slants of your best cultures.  Combining the spores of different varieties can increase heterozygosity which can then be selectively selfed to produce a new custom made variety.

I know it's confusing and I probably didn't explain it very well, but maybe I helped.
more on inbreeding
I should point out that common sense dictates that inbreeding is bad since it can expose recessive genetic defects or undesirable traits.  Ideally you want high heterozygosity to give a particular mushroom strain a wide range of available genes, which in turn makes it better adapted to unpredictable environmental conditions.  This could be achieved by cloning a wild vigorous specimen or by crossing two very different strains.  In my experience, the mushrooms resulting from such a cross are very vigorous and productive (hybrid vigor).

The problem is that to keep this strain going you have to keep the mycelium going.  This isn't a problem with edible cultures but we need stable spores.  The spores from a high heterozygosity strain will be genetically recombined and won't produce the original strain  (although with aggressive isolation you could get something close).  Multispore grows will be terrible, not because the newly generated strains are bad, but because they are so genetically different from each other.  These different strains don't get along nicely in the same tray which reduces productivity.  If we can reduce the differences between the strains generated by multispore to some minimal level, they tend to cooperate better with each other and act more like a single strain.

The point is that we are dealing with spores not clones.  Many (most) Psilocybe growers use syringes to generate multispore grows with no isolation. In this context, stabilized strains are desirable.  If everyone could trade/sell mycelium this wouldn't be an issue.

 

This is very interesting info that pertains to another post I have made recently about senescence being reached by repeatedly using MS. Im stealing it!


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