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Theory about a massive downside of MS grows


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#1 Moby

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Posted 17 February 2021 - 09:00 AM

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


Edited by Moby, 17 February 2021 - 09:04 AM.

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#2 sandman

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Posted 17 February 2021 - 09:21 AM

 

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

 

 

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

 

Where exactly is your source on this? I do not believe this to be anywhere remotely true.

 

ALl that I see tends to say that the "quality" improves with domestication such as size of fruiting bodies and quantity.

 

I am unaware of any gas chromatography showing wild cubes as stronger if that what you mean. Please do enlighten me if you know of any sources.

 

The only way I could se your theory holding any water would be if by improving the size and appearance through selection we are decreasing the psilocin et al but I don't see that happening, likely the reverse.

 

Most BIG growers throw out weak strains and focus on growing the best isolations that are strong and look good. They probably don't want people coming back asking for refunds all the times

 

And print from like this flood the market because they are the ones supplying the spores to the websites also.


Edited by sandman, 17 February 2021 - 09:34 AM.

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#3 jrh

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Posted 17 February 2021 - 09:43 AM

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

 

In the wild, "fittest" means best able to reproduce and pass on genetic information to the next generation. All wild mushroom grows use MS so MS can't inherently be bad, or we wouldn't have wild mushrooms any longer.

 

I'm not sure what "quality" means or whether it has anything to do with genetics vs growing conditions.

 

In the home, we grow things we find interesting, whether because it's a funny color or shape, or has more actives, or can survive crappy growing conditions. The popularity of PE suggests that we're not trying to cultivate mushrooms based on their ability to reproduce in the wild.

 

So I guess I disagree both with your basic premise and the assumption that we should change domestic cultivation practices if it were true.


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#4 Microbe

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Posted 17 February 2021 - 10:21 AM

That's some good info. When two compatible hyphae join and clamp (not fuse). They each receive a copy of the others chromosomes so it is possible that weaker genetics are mixed in. However stronger genetics will dominate as far speed, fruiting capability, primary and secondary metabolite production and etc. Although the hyphae at this point have 2 sets of chromosomes, the nuclei have not fused therefore the dikaryotic phase is technically not diploid, since the nuclei remain unfused until shortly before spore production. So in simple terms the strongest genetics are in control at this point.

Right before spore production, the nuclei fuse becoming double haploid. This is where the weak genetics get their chance. During spore production a process occurs called genetic recombination. Imagine throwing all these genetics in a bag and shaking it up them dumping them out, these are the new genetics. They could have digressed, stayed the same, or improved. I do want to note that the genome sequence does not change or it would be altogether another species......don't quote me on that as in known to be silly at times.

Imagine if you will genomes are keyholes and each keyhole unlocks or locks a specific trait or phenotype. During genetic recombination, keys are inserted in keyholes at random and locked or unlocked. Now is this really random? Do the dominant genomes still have the upper hand during this process? I can't really answer either of those questions but my money is on the dominant genetics holding the master key ring.

I can tell you, i never had a sub par MS grow. Maybe its because i never shot spores in to a jar and ran with it. I always dilute spores by flaking spores into a jar of sterilized water then mixing up before drawing into a syringe. I then place a single drop from a 21 gauge or smaller needle. I have had plates not germinate because they didn't get any spores. I also believe that natural selection takes place in home grows starting on agar plates, or grain, and etc. The weaker genetics are weeded out by dominant genetics even if at the end, they fused. A MS culture will isolate itself if given enough area.

If we do get some bad genetics when germinating a spore, as cultivators we need to run quality control and eliminate weak genetics. But then its like who are we to determine who lives and dies and i don't care for that either. Toss the print out side and give it a chance because it matters lol.

I absolutely disagree with wild mushrooms being picked fresh are better then domesticated varieties being homegrown. Im telling you, and with the most confidence, any cube i have grown, is far better then any wild cube i have ever consumed. If i currently had some, i would find a way to get some to you....that's how confident i am with that statement.

Good info you posted. I have said this a few times already but im just now brushing up on all the mycology terminology and processes (biological) so as i continue through some reads, im certain i will visit this thread because i like talking about this stuff as long as it doesn't turn in to a argument. Nothing wrong with bouncing info back and forth and an occasional healthy debate. Im finally back in a good place and i will be damned if i give anyone at topia the power to take that from me. If it gets out of control anywhere, i will simply act like a airport and announce my departure and leave lol


Note: im not saying any of us will argue, i just want to make it clear that i will not. We have so much we can learn from each other.

Edited by Microbe, 17 February 2021 - 10:43 AM.

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#5 rockyfungus

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Posted 17 February 2021 - 10:56 AM

Just to throw some other processes into the mix...

Not sure if mushroom genetics work the same way, hard to find studies. Epigenetics may play a role as well.

What are you doing to the genetics to change what pieces of the genome are activated. Stress, diet, age, viruses, bacteria, can activate regions of the genome that are normally not activated.
Let alone viruses/bacteria can inject new cellular information into a genome.


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#6 Microbe

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Posted 17 February 2021 - 11:08 AM

Just to throw some other processes into the mix...

Not sure if mushroom genetics work the same way, hard to find studies. Epigenetics may play a role as well.

What are you doing to the genetics to change what pieces of the genome are activated. Stress, diet, age, viruses, bacteria, can activate regions of the genome that are normally not activated.
Let alone viruses/bacteria can inject new cellular information into a genome.

Absolutely correct! All that you mentioned can have influence on which genome is turned on or off. Rosecomb is a perfect example Epigenetics. You can observe bacteria living inside hyphae and that could also have some influence. What if mushrooms are so smart, they can actually copy the dna of the symbiotic bacteria, and recreate it on its own? Mushrooms are the most advanced and intelligent life form (biologically) when you really get to know them. They can do anything they chose to do......i i hope the mushrooms don't decide to say, "fuck it, lets start eating people" uh oh lol

Edited by Microbe, 17 February 2021 - 11:10 AM.

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#7 groo

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Posted 17 February 2021 - 11:16 AM

I have been growing mushrooms for 8 years and I EVEN me have trouble separating what is LIE or TRUTH. Pissed there is legit writings on mushrooms jumbled with all the shitty old information. an you theorize how to get that information and books and posts marked as inaccurate and known lists of books with the dishonest concepts exposed so when someone posts that they put their pf tek jars in the complete dark, because they read Paul amethysts first book.


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#8 coAsTal

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Posted 17 February 2021 - 11:17 AM

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.

 


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#9 Microbe

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Posted 17 February 2021 - 11:32 AM

I have been growing mushrooms for 8 years and I EVEN me have trouble separating what is LIE or TRUTH. Pissed there is legit writings on mushrooms jumbled with all the shitty old information. an you theorize how to get that information and books and posts marked as inaccurate and known lists of books with the dishonest concepts exposed so when someone posts that they put their pf tek jars in the complete dark, because they read Paul amethysts first book.

There is a lot of info out there for sure and even a lot of dated info. Keep in mind with all this information we have, most of it is still anecdotal but with peer review. The science of mushrooms is ever evolving as we learn more about them. They are very complex organisms and who's to say that mushrooms are not evolving themselves forcing us to continue to study and learn their very hidden teachings.

Perhaps 30 years ago, mushrooms did need to be kept in the dark during colonization, but through our practices, they evolved to not require this. Im simply using this as an example you mentioned. Some dated info now, may not have been wrong at that time based on the current evolution of mushrooms. Evolution in mushrooms happens pretty damn quick.....probably faster then most other organisms other then viruses or bacteria.

You are right, and the only way to determine the inaccurate verse accurate info, is to follow the science. If you understand the basic functioning of mushrooms, you will be able to determine what is clearly BS info.

#10 Moby

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Posted 17 February 2021 - 12:07 PM

Hhm.

 

The "wild varieties are superior" is something i picked up over multiple sources, which all were not study based.

In Vendors that sell freshly collected variety as more agressive and/or far more potent and in shroomery somewhere.

 

I understand that vendors try to sell their stuff and polish the infos like crazy but together with the one time I've read it on shroomery where someone said with confidence they are more potent, its something that stayed in my mind until now.

 

I want to remind you that the headline here is "Theory about a massive downside of MS grows"

 

and not "wild cubes are superior".

 

The wild cube part is something I added because it made sense in the whole picture of my theory BUT that doesnt mean the whole theory is based on the wild cubes part.

I mentioned that the idea came into my mind because different pieces of "info" together made sense and painted the picture of this theory in my head.

I'm not trying to sell anything as facts and was careful with my wording.

 

If you leave that out and look at the rest of what my mind put together (based on what I've read over the time) then I think its not crazy stupid to assume that MS grows could actually have a negative effect on the produced spores. With "negative effect" I mean constant lost potential with every MS print generation" due to fusing of weak with strong strains which then spread their mixed genetic through the spores.

__

 

Gonna answer and read in steps, didn't expect so much text to read in such a short time.

Grateful for every bit of information I can get to understand better and build a more valuable opinion


Edited by Moby, 17 February 2021 - 12:11 PM.

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#11 TVCasualty

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Posted 17 February 2021 - 12:07 PM

I can tell you, i never had a sub par MS grow.

 

Me neither.

 

 

 

 

One of the reasons why I haven't participated in very many threads about cloning or making isolates is that I quit doing that years ago. I didn't mention it much since it went against the common assertion that isolates are superior and I didn't have anything but anecdotal observations to offer as a counter-argument. But those observations really began to add up over time.

 

I noticed that after printing my best clusters (I select for cluster size and density over individual specimen size) and growing the next crop from that print and then printing the best cluster from that grow and so on I eventually got results that were extremely consistent and no less reliable than growing isolates. So I ditched isolates to save lots of time and energy. Potency remained consistent insofar as a teaspoon of my honey mix didn't noticeably vary between grows. I assumed that "hybrid vigor" was playing a role in that even if the strain was also inbred, technically-speaking. It became apparent over time that inbreeding doesn't work in higher fungi like it does in plants and animals.

 

I didn't know that stuff about heterozygosity at the time. But it makes sense based on what I've seen. My favorite strain for making lots of mushrooms has been grown out more generations than I can recall over the past 12 years or so (when I began focusing on it for this multispore breeding program). I also have a PF Classic that was sent with the instructions to the original PF TEK when I bought my first syringe from the ad in High Times. I've grown that strain repeatedly from MS since 1998, so it's probably pretty stable in its inherent weirdness now (not nearly as many generations as my go-to strain though).


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#12 Microbe

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Posted 17 February 2021 - 12:08 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.

I like how he mentioned incompatibility zone, i always refered to it as the neutral zone. It is there where you can chemically force non compatible hyphae to mate or cross. I would disagree with him where he states that when compatible mycelium meet that they fuse.

Mushrooms spend most of their life cycle as haploid or having only a single set of chromosomes. Instead they clamp.....they do not fuse nor do the genes combine at this point, but instead are exchanged in the form of a nuclei with a pair of chromosomes. This is why they are considered to be dykaryotic at this point.....the 2 nuclei. IMO this is kind of misleading because it still only functioning off its own genetics at this point and is only transporting the other nuclei or egg/sperm if you will, so therefore this to me would still be monokaryotic at this point but it is what it is because it does technically have two nuclei.

Fusion and gene combining does not take place up until right before spore production. Fusion takes place near or in the Basidia, and now after fusion, the diploid cells (because the two nuclei have fused and genetics are now combined from each parent) undergoes meiosis to produce haploid spores. The beloved MS spores that we talking about.

I skimmed through that and will read it again but very good info from what i read thus far.


Where is workman at? Imma going to go have a talk with him

Edited by Microbe, 17 February 2021 - 12:41 PM.

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#13 sandman

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Posted 17 February 2021 - 12:17 PM

nerds  :wub:


Edited by sandman, 17 February 2021 - 12:18 PM.

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#14 Microbe

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Posted 17 February 2021 - 01:47 PM

Hhm.

The "wild varieties are superior" is something i picked up over multiple sources, which all were not study based.
In Vendors that sell freshly collected variety as more agressive and/or far more potent and in shroomery somewhere.

I understand that vendors try to sell their stuff and polish the infos like crazy but together with the one time I've read it on shroomery where someone said with confidence they are more potent, its something that stayed in my mind until now.

I want to remind you that the headline here is "Theory about a massive downside of MS grows"

I think its not crazy stupid to assume that MS grows could actually have a negative effect on the produced spores. With "negative effect" I mean constant lost potential with every MS print generation" due to fusing of weak with strong strains which then spread their mixed genetic through the spores.

I agree with you mostly on this. However i believe that the genetic combination of a stabilized strain of any given variety, the base genetics even after recombination, will still consist of those dominant traits no matter where they land and are more likely to show. Now who is to say that the dominant genes are the desired one. What if a strain had superior genetics such as, good flavor, color, potency, vitamin production, anti-oxidant, and so on, but was inadequate fruiter......then yeah i can totally see your point here. At the end of the day, and in a commercial setting, yield is the heavy hitter, followed by quality ie flavor, aroma, color, shape and etc. I don't know how we can determine this though until we fruit them. I feel MS grows work well because its isolating itself in front of our eyes rather running across agar, grain, or bulk subs.

What we need to learn to do, is manipulate and control each individual genome. How cool would that be? I said this earlier about mushrooms, but if we did this, they probably decide to add us to their diet. Not to get off topic but there is nifty gadget called a crisper (i think) where you can change the dna sequence of bacteria in your home. It comes with a strain of E. colli and you follow the directions to make a anti-biotic resistant strain. Now by changing the DNA sequence, its probably another species but my point here, is this shit is possible. As crazy as it sounds, and im a little off topic, but eventually being able to alter genomes of mushrooms, may not be that far fetched.

Edit: oh and anytime posts like this pop up, expect a lot of text to read when you return. Posts like this, for me anyway, i swallow the hook, line, and sinker

Edited by Microbe, 17 February 2021 - 04:01 PM.

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#15 rockyfungus

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Posted 17 February 2021 - 03:01 PM

Fresh cubes in a field do hit differently, but I think it's the dopamine of the hunt lol



#16 TVCasualty

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Posted 17 February 2021 - 04:17 PM

Fresh cubes in a field do hit differently, but I think it's the dopamine of the hunt lol

 

I also suspect that we're underestimating fresh potency vs. dry. The assumption is that fresh are 90% water so when dry they'll be 10% of the potency of fresh because they're 10% of the weight. But that neglects to account for the presence of psilocin in fresh mushrooms that's oxidized in the drying process.

 

So in my opinion and experience, 30 grams of fresh cubes will be more potent eaten fresh than they would be if they were dried to ~3g first.


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#17 rockyfungus

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Posted 17 February 2021 - 05:17 PM

Yeah 70grams split between two in the field always way more intense then 3.5 dry. It's def. that psilocin>psilocybin, way more visual for me.



#18 jrh

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Posted 17 February 2021 - 06:46 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.

#19 rockyfungus

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Posted 17 February 2021 - 07:09 PM

chop coarsely with knife, add lemon juice (questionable if it does anything, your stomach is HCL...), bring to boil, discard coarse mush, plug nose...

I just chop coarsely, boil, discard, add some ginger tea, maybe honey, chug...


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#20 Moby

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Posted 18 February 2021 - 11:10 AM

Saved the whole page as pdf and will read it in a while.

I don't want to be part of this community anymore and will leave.

 

Has nothing to do with this thread here, dont worry!

 

Best of success everyone






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